Acute low-intensity microwave exposure increases DNA single-strand breaks in rat brain cells

Levels of DNA single-strand break were assayed in brain cells from rats acutely exposed to low-intensity 2450 MHz microwaves using an alkaline microgel electrophoresis method. Immediately after 2 h of exposure to pulsed (2 μs width, 500 pulses/s) microwaves, no significant effect was observed, whereas a dose rate-dependent [0.6 and 1.2 W/kg whole body specific absorption rate (SAR)] increase in DNA single-strand breaks was found in brain cells of rats at 4 h postexposure. Furthermore, in rats exposed for 2 h to continuous-wave 2450 MHz microwaves (SAR 1.2 W/kg), increases in brain cell DNA single-strand breaks were observed immediately as well as at 4 h postexposure. © 1995 Wiley-Liss, Inc.     

In vitro study of microwave effects on calcium efflux in rat brain tissue

In this study we investigated the prospect of microwave-induced alteration of ⁴⁵Ca²⁺ efflux from rat neural tissue at low pulse repetition frequencies and low power densities under in vitro conditions. Rat cerebral tissue, preloaded with ⁴⁵Ca²⁺, was exposed to pulsed-microwave radiation (1-GHz carrier frequency) according to one of several PRF-power density exposure schemes: 16 Hz at 0.5, 1.0, 2.0, or 15 mW/cm², or 32 Hz at 1.0 or 2.0 mW/cm² average power density. Measurements of radioactivity in the efflux medium and in the tissue sample were used to calculate an efflux value for each sample. The results indicate that the radiation conditions used did not alter calcium efflux in rat brain tissue.     

Influence of pre- and postnatal exposure of rats to 2.45-GHz microwave radiation on neurobehavioral function

Rats exposed to microwaves prenatally (2,450 MHz, 10 mW/cm2, 3 h/day, days 5-20 of gestation) or perinatally (same as above plus days 2-20 postnatally) were examined by a neurobehavioral test battery on postnatal days 30 and 100. Body mass, locomotor activity, startle to acoustic and air-puff stimuli, fore- and hindlimb grip strength, negative geotaxis, reaction to thermal stimulation, and swimming endurance were assessed. The prenatally and the perinatally exposed rats (male and female) weighted more than sham-exposed rats at 30, but not at 100, days of age. In addition, the perinatally exposed animals had less swimming endurance at 30, but not at 100, days of age relative to sham-exposed rats. For the other measures, only the air-puff startle response was altered and was limited to the prenatally exposed female pups; ie, at postnatal day 30, the startle response was increased in magnitude, and at postnatal day 100, the response was decreased. No other reliable effects were observed. In a second experiment, rats treated as described above were examined for alterations in body mass, locomotor activity, reaction to air-puff stimuli, reaction to thermal stimulation, and swimming endurance at postnatal days 30-36. Again, perinatally exposed rats were larger in body mass and had less swimming endurance compared with sham-exposed rats. The latency to the air-puff startle response was longer in female pups exposed prenatally. These data indicate that altered endurance and gross motor activity result from perinatal exposure to microwave irradiation.     

In vitro effects of microwave radiation on rat liver mitochondria

Liver mitochondria were exposed in vitro at 30°C to microwave radiation (2.45 GHz) during the following states of respiraton: resting, state 1; substrate dependent, state 2; ADP stimulated, state 3; and ADP depleted, state 4. At 10 or 100 mW/g, with succinate as substrate, no effect of exposure was observed on states 1–4 or the respiratory control index (state 3/state 4) of either tightly or loosely coupled mitochondria. When glutamate was used as substrate, no effects were observed at 10 mW/g. However, in the loosely coupled mitochondria the 100 mW/g exposure produced an increase in states 2 and 4 and a decrease in the respiratory control index. The results suggest that the function of loosely coupled mitochondria can be affected at high power levels of microwave radiation.     

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.     

Attempts to alter 45Ca2+ binding to brain tissue with pulse-modulated microwave energy

Rat brain tissue, loaded with ⁴⁵Ca²⁺ by intraventricular injection was exposed in vitro to pulsemodulated 1-GHz (SAR of 0.29 or 2.9 W/kg) or 2.45-GHz radiation (SAR = 0.3 W/kg), and in vivo to 2.06-GHz radiation (SAR of 0.12 to 2.4 W/kg). There were no significant differences in efflux of ⁴⁵Ca²⁺ between the microwave- and sham-irradiated groups.     

Cerebrovascular permeability to 86Rb in the rat after exposure to pulsed microwaves

Microwaves (pulsed, 2,450 MHz) at an average power density of 3 W/cm² were applied directly to the head for 5, 10, or 20 min, producing a peak specific absorption rate of 240 W/kg in the brain, which, after a 10-min exposure, resulted in brain temperatures in excess of 43°C. A bolus of ⁸⁶Rb in isotonic saline was injected intravenously and an arterial sample was collected for 20 s to determine cardiac output. Compared with unexposed controls, uptake of ⁸⁶Rb increased most in those regions directly in the path of the irradiation, namely, the occipital and parietal cortex, as well as the dorsal hippocampus, midbrain, and basal ganglia. In a separate group of animals, regional brain-vascular spaces were found to increase with brain temperature. These results support previous observations indicating that reliably demonstrable increases of blood-brain barrier permeability are associated with intense, microwave-induced hyperthermia, and that the observed changes are not due to field-specific interaction.     

Blood-brain barrier permeation in the rat during exposure to low-power 1.7-GHz microwave radiation

The permeability of the blood-brain barrier to high-and low-molecular-weight compounds has been measured as a function of continuous-wave (CW) and pulsed-microwave radiation. Adult rats, anesthetized with pentobarbital and injected intravenously with a mixture of [14C] sucrose and [3H] inulin, were exposed for 30 min at a specific absorption rate of 0.1 W/kg to 1.7-GHz CW and pulsed (0.5-microseconds pulse width, 1,000 pps) microwaves. After exposure, the brain was perfused and sectioned into nine regions, and the radioactivity in each region was counted. During identical exposure conditions, temperatures of rats were measured in eight of the brain regions by a thermistor probe that did not perturb the field. No change in uptake of either tracer was found in any of the eight regions as compared with those of sham-exposed animals.     

Humoral and cell-mediated immune function in adult japanese quail following exposure to 2.45-GHz microwave radiation during embryogeny

Japanese quail, Coturnix coturnix japonica, eggs were subjected to 2.45-GHz CW microwave radiation at 5 mW/cm² (SAR = 4.03 mW/g) during the first 12 days of embryogeny. Following hatching the exposed embryos, as well as nonexposed controls, were reared to 22 weeks of age. Humoral immune potential, as indicated by comparable anti-CRBC antibody, IgM and IgG, levels at 0, 4, and 7 days postimmunization in both exposed and control quail was not affected significantly. However, cell-mediated immune potential, measured by the reaction to intradermal injection of phytohemagglutinin-P in the wing web, was reduced in the exposed females, but not in the exposed males. Additionally, total leukocyte numbers and absolute circulating numbers of lymphocytes, monocytes, and heterophils were increased significantly only in the exposed females. These data show that exposure of Japanese quail during embryogenesis reduced cell-mediated immune potential and induced a general leukocytosis in females.     

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.     

A model of aluminum exposure associated with lipid peroxidation in rat brain

We have developed a rat model to investigate the relationship between aluminum exposure and aluminum accumulation, and with oxidative damage in brain tissues. Intraperitoneal injections of aluminum lactate for 7 wk (the total aluminum dosage per rat was approx 100 mg) significantly increased aluminum levels in the brain. The concentration of lipid peroxidation products (thiobarbituric acid-reactive substances [TBARS]) also increased in the brain following aluminum lactate injections. No significant correlations between the concentrations of aluminum and of TBARS were found in the whole brain. Subcellular analysis revealed that aluminum lactate injections led to a significant increase in the concentration of aluminum in the mitochondrial fraction but had no significant effect on the concentration of peroxides in any subcellular fraction. These results suggest that aluminum accumulation induced by the aluminum lactate administration associates with the acceleration of lipid peroxidation in rat brain. Furthermore, these data indicate that the pro-oxidant effect of aluminum may be indirect and concentration independent. The experimental conditions used here provide an animal model of aluminum accumulation in the brain that should prove useful for further investigations of the mechanisms of aluminum neurotoxicity.     

Single vs. repeated microwave exposure: effects on benzodiazepine receptors in the brain of the rat.

We studied the effects of single (45 min) and repeated (ten daily 45-min sessions) microwave exposures (2450-MHz, 1 mW/cm2, average whole-body SAR of 0.6 W/kg, pulsed at 500 pps with pulse width of 2 microseconds) on the concentration and affinity of benzodiazepine receptors in the cerebral cortex, hippocampus, and cerebellum of the rat. We used a receptor-binding assay with 3H-flunitrazepam as ligand. Immediately after a single exposure, an increase in the concentration of receptor was observed in the cerebral cortex, but no significant effect was observed in the hippocampus or cerebellum. No significant change in binding affinity of the receptors was observed in any of the brain-regions studied. In rats subjected to repeated exposures, no significant change in receptor concentration was found in the cerebral cortex immediately after the last exposure, which may indicate an adaptation to repeated exposures. Our data also show that handling and exposure procedures in our experiments did not significantly affect benzodiazepine receptors in the brain. Because benzodiazepine receptors in the brain are responsive to anxiety and stress, our data support the hypothesis that low-intensity microwave irradiation can be a source of stress.     

In utero hypoxic ischemia decreases the cholinergic agonist-stimulated poly-phosphoinositide turnover in the developing rat brain

Perinatal hypoxic-ischemic (HI) insult is known to cause cellular and molecular disturbances leading to functional and behavioral abnormalities during brain development. In this study, we examined the effects of an in utero HI insult on poly-phosphoinositide turnover in vivo in the cerebrum and cerebellum as well as cholinergic-stimulated turnover in cortical slices from developing rat brain. In utero HI treatment was carried out by clamping the uterine blood vessels of near-term fetuses for 5, 10 and 15 min followed by resuscitation of the newborn pups. The in vivo protocol for examining poly-PI signaling activity in 2 week-old pup brain involved intracerebral injection of [³H]inositol for 16 hr and subsequent intraperitoneal injection with lithium (8 meq/kg) for 4 hr prior to decapitation. In the control pups, lithium elicited a 2.6 fold increase in labeled inositol phosphate (IP) in the cerebrum as compared to a 1.3 fold increase in the cerebellum. In utero HI insult (5 to 15 min) resulted in a small increase in labeled IP in the cerebrum but not in the cerebellum. Carbachol stimulation of poly-PI turnover was examined in brain slices prelabeled with [³H]inositol in vivo. Incubation of the prelabeled slices with carbachol in the presence of LiCl (10 mM) resulted in a time-, dose- and age-dependent increase in labeled IP. Brain slices from 2 week-old pups that experienced in utero HI-treatment for 10 and 15 min (but not 5 min) showed a significant decrease in carbachol-stimulation of labeled IP as compared with control pups. These results indicate the effects of in utero HI on the choninergic-stimulated poly-PI signaling pathway and its implication on related functional deficits in the developing brain.Abbreviations HI hypoxic-ischemia - poly-PI poly-phosphoinositides - IP inositol monophosphate, lithium     

Cardiovascular, hematologic, and biochemical effects of acute ventral exposure of conscious rats to 2450-MHz (CW) microwave radiation

In this study the influence of acute (6 hr) exposure to 2450 MHz (CW) microwave radiation on certain cardiovascular, biochemical, and hematologic indices was examined in unanesthetized rats. Under methoxyflurane anesthesia, a catheter was inserted into the right femoral artery, which was used for monitoring blood pressure, heart rate, and blood sampling. Colonic temperature was monitored via a VITEK thermistor probe inserted rectally to a depth of 5 cm. The rat was subsequently placed into a ventilated restraining cage which was located inside an anechoic chamber. The temperature and humidity in the chamber were maintained at 22 +/- 0.5 degrees C and 60 +/- 5% (means +/- S.E.), respectively, during the experimental period. Rats (60) were exposed to either 0 (sham) or 10 mW/cm2 (exposed) for 6 hr. During exposure rats were oriented perpendicular to the E-field, and the measured specific absorption rate (SAR) was 3.7 mW/g. In the sham and exposed rats, the preexposure (time 0) mean +/- S.E. arterial blood pressure (MABP), heart rate, and colonic temperature were approximately 120 +/- 5 mmHg, 450 +/- 10 beats/min, and 37.0 +/- 0.2 degrees C, respectively. In the sham-exposed rats these values remained stable throughout the 6-hr exposure period. In the exposed rats, no effects were noted on MABP or colonic temperature; however after 1 hr of exposure, a significant reduction in heart rate was noted (450 versus 400 beats/min). This decrease in heart rate persisted throughout the remainder of the exposure period. None of the hematologic or biochemical parameters examined were affected by the microwave exposure. Although other mechanisms may be responsible, this decrease in heart rate may have been due to subtle cardiovascular adjustments because of microwave-induced heating with a resultant reduction in resting metabolic rate.     

Developmental changes and regional distribution of phospholipase D and base exchange enzyme activities in rat brain

Phospholipase D (PL-D) activity per mg protein of whole homogenate increased 5.1 fold between Embryonic (E) day 17 and Postpartum (P) day 14 and slightly decreased by P 30 days. This was due to the decrease of PL-D activity of the P₂ fraction. The PL-D activity of P₂ and P₃ fractions increased 11.2 and 6.1 fold respectively between E 17 and P 14. The 3 base exchange enzyme (BEE) activities per mg protein of whole homogenate increased up to P 14 or P 21 and then decreased. This decrease was greater in the P₂ fraction and the P₃ fraction increased after P14. Brains from 1 day to 25 month old rats were dissected into 7 separate regions and both PL-D and BEE activities were measured. In adult rats, the hippocampus and hypothalamus had the highest PL-D activities while medulla+pons and cerebellum had the lowest PL-D activities. The developmental patterns of 5 regions except for hippocampus and hypothalamus were similar. PL-D activity in the hippocampus was maximum at P 7 followed by a steep decrease till P30 suggesting that the PL-D activity of the hypothalamus develops later and that of the hippocampus develops earlier than any other region. The distributions of BEE activities were quite different from those of PL-D activities. In adult rats, the cerebellum had the highest activity while the striatum and medulla+pons had the lowest. The BEE activities of cerebellum were lowest at P 1 and showed steep increase during the next 2 weeks.To whom to address reprint request are to be sent.     

Lack of effect of 2.45-GHz microwave radiation on the development of preimplantation embryos of mice

The development of preimplantation embryos after exposure to microwave radiation was studied. Female CD-1 mice were induced to superovulate, mated, and exposed to 2.45-GHz microwave or sham radiation for 3 h at power densities of 9 mW/cm² and 19 mW/cm² on either day 2 or 3 of pregnancy (plug day was considered day 1). Another group of mice was exposed to heat stress by placing the dams in an environmental room at an ambient temperature of 38 °C and relative humidity at 62% for 3 h on day 2 of pregnancy. All groups were euthanized on day 4 of pregnancy and embryos were recovered by flushing excised uterine horns. Embryos were examined for abnormalities and classified by the developmental stages. They were then treated with hypotonic solution and dissociated for counting blastomeres. Heat stress caused stunted development of embryos, but no remarkable effect of microwave radiation could be found on the development of preimplantation embryos.     

Rhesus monkey behavior during exposure to high-peak-power 5.62-GHz microwave pulses

Limits on the exposure to high-peak-power, short-duration microwave pulses have only recently been adopted. Additional data, however, are needed to understand the effects that may be produced by exposure to high-peak-power pulsed microwaves. Four male rhesus monkeys (Macaca mulatta) were trained on an operant task for food pellet reward to investigate the behavioral effects of very high-peak-power 5.62 GHz microwaves. The operant task required monkeys to pull one plastic lever on a variable interval schedule (VI-25 s) and then respond to color signals and pull a second lever to obtain food. The monkeys were conditioned to perform a color discrimination task using one of three colors displayed by a fiber-optic cable. A red signal was the discriminative stimulus for responding on the first lever. A response on the second lever when a green signal was presented (1 s duration) delivered a food pellet. If a response on the second lever was made in the presence of a white signal, a 30-s timeout occurred. While performing the behavioral task, the monkeys were exposed to microwave pulses produced by either a military radar (FPS-26A) operating at 5.62 GHz or the same radar coupled to a Stanford linear energy doubler (SLED) pulse-forming device (ITT-2972) that enhanced peak power by a factor of nine by adding a high power pulse to the radar pulse. The effects of both types of pulses were compared to sham exposure. Peak field power densities tested were 518, 1270, and 2520 W/cm² for SLED pulses and 56, 128, and 277 W/cm² for the radar pulses. The microwave pulses (radar or SLED) were delivered at 100 pps (2.8 μs radar pulse duration, ≈ 50 ns SLED pulse duration) for 20 min and produced averaged whole-body SARs of 2,4, or 6 W/kg. Compared to sham exposures, significant alterations of lever responding, reaction time, and earned food pellets occurred during microwave exposure at 4 and 6 W/kg but not 2 W/kg. There were no differences between radar or SLED pulses in producing behavioral effects. ©1994 Wiley-Liss, Inc.

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

     

Circulating antibody response of mice exposed to 9-GHz pulsed microwave radiation

Female CD-1 mice immunized against the bacterium Streptococcus pneumoniae type III were exposed to 9-GHz pulsed microwaves (pulse repetition rate 970-1,000, pulse width 1.0 microseconds, peak power 1 W/cm2) at an average incident power density of 1 mW/cm2 (calculated SAR congruent to 0.47 W/kg) for 2 h per day for 5 days. Circulating antibody titers for the microwave-exposed animals were not significantly different from those of the sham-irradiated animals, and there were no differences in any of the hematological parameters analyzed, indicating that 9-GHz pulsed microwaves at 1 mW/cm2 do not alter the immune response of mice immunized against S pneumoniae.     

Neuron-specific aminopeptidase and puromycin-sensitive aminopeptidase in rat brain development

Neuron-specific aminopeptidase (NAP) and the ubiquitous puromycin-sensitive aminopeptidase (PSA) were compared in the rat hippocampus during early development. Hippocampus contains the highest amount of NAP determined by a fast-protein liquid chromatography-aminopeptidase analyzer using Leu -naphthylamide as substrate. Both enzymes were found in the hippocampus in all ages. NAP was lower in immature rat; the 19th embryonic-day fetus contained the least. It increased steeply during the prenatal through the early postnatal period, 9-fold by the first month. The rate of increase diminished subsequently, increasing 20% in the second month and 13% in the third. The age-dependent increase in NAP activity was parallel to its protein expression as determined by Western blot. The specific molecular activity (hydrolytic activity/NAP antigenicity) in newborn, 15-day-old, and 30-day-old rats were 1.00, 0.88, and 1.00, respectively. The PSA developmental profile without linear increase in activity was distinct from NAP. PSA activity was higher than NAP in decreasing order, 100–4 times, during the same development span. Similarly, different growth profiles for NAP and PSA were also found in the primary culture of developing cerebellar granule cells. Puromycin (1–5 M) blocked neurite outgrowth and caused apoptosis by nonantibiotic effects. Our data suggest that the synaptosome-enriched NAP plays a role in neuron growth, differentiation, and information programming.     

Effect of 2,450 MHz microwave radiation on the development of the rat brain

Male Sprague-Dawley rats were exposed to 2,450 MHz microwave radiation at an incident power density of 10 mW/cm2 daily for 3 hours from day 4 of pregnancy (in utero exposure) through day 40 postpartum, except for 2 days at the perinatal period. The animals were killed, and the brains removed, weighed, measured, and histologically examined at 15, 20, 30, and 40 days of age. The histologic parameters examined included the cortical architecture of the cerebral cortex, the decline of the germinal layer along the lateral ventricles, the myelination of the corpus callosum, and the decline of the external germinal layer of the cerebellar cortex. In 40-day-old rats, quantitative measurements of neurons were also made. The spine density of the pyramidal cells in layer III of the somatosensory cortex, and the density of basal dendritic trees of the pyramidal cells in layer V were measured in Golgi-Cox impregnated specimens. In addition, the density of Purkinje cells and the extent of the Purkinje cell layer in each lobule were measured in midsagittal sections of the cerebellum stained with thionin. There were no remarkable differences between microwave-exposed and control (sham-irradiated) groups for any of the histologic or quantitative parameters examined; however, the findings provide important information on quantitative measurements of the brain. The data from this study failed to demonstrate that there is a significant effect on rat brain development due to microwave exposure (10 mW/cm2) during the embryonic, fetal, and postnatal periods.(ABSTRACT TRUNCATED AT 250 WORDS)