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.     

Endocrine and behavioral traits in low-avoidance Sprague-Dawley rats

In the present series of experiments, we have examined the endocrine profile of two stable colonies of Sprague-Dawley rats, here labeled Stock A, and Stock B, differing markedly in their ability to acquire a conditioned avoidance response. On separate occasions, the animals were subjected to five daily sessions (approximately 20 trials per 15 min session) of conditioned avoidance training, measurements of startle reactivity to an auditory stimulation and open-field spontaneous locomotor activity observations. The experiments were concluded by taking blood samples for later analysis of plasma glucose and plasma levels of the following hormones: insulin, gastrin, CCK, glucagon, somatostatin, oxytocin and corticosterone. The low-performing Stock B animals were characterized by [1] being more reactive to sensory stimulation: higher startle amplitude and shorter startle latency; [2] having higher plasma insulin and corticosterone levels, whereas plasma gastrin and oxytocin were significantly lowered and a strong tendency for a decrease also in plasma CCK. There were no differences in spontaneous locomotor activity between the two substrains. Taking total variability in avoidance performance into account, there was a statistically significant positive correlation between plasma oxytocin, as well as gastrin, levels and avoidance performance. The evidence obtained here, and in other laboratories, suggests that the Stock B animals display hormonal changes indicative of a submissive-defensive reaction pattern. Thus, the avoidance acquisition deficits displayed by the present Sprague-Dawley stocks A and B, are in all probability caused by emotional reactions when challenged with external stimuli requiring active responding.     

Effects of microwave exposure on the hamster immune system. I. Natural killer cell activity

Hamsters were exposed to repeated or single doses of microwave energy and monitored for changes in core body temperature, circulating leukocyte profiles, serum corticosteroid levels, and natural killer (NK) cell activity in various tissues. NK cytotoxicity was measured in a 51Cr-release assay employing baby hamster kidney (BHK) targets or BHK infected with herpes simplex virus. Repeated exposure of hamsters at 15 mW/cm2 for 60 min/day had no significant effect on natural levels of spleen-cell NK activity against BHK targets. Similarly, repeated exposure at 15 mW/cm2 over a 5-day period had no demonstrable effect on the induction of spleen NK activity by vaccinia virus immunization, that is, comparable levels of NK were induced in untreated and microwave-treated animals. In contrast, treatment of hamsters with a single 60-min microwave exposure at 25 mW/cm2 caused a significant suppression in induced spleen NK activity. A similar but less marked decrease in spleen NK activity was observed in sham-exposed animals. Moreover, the sham effects on NK activity were not predictable and appeared to represent large individual animal variations in the response to stress factors. Depressed spleen NK activity was evident as early as 4 h postmicrowave treatment and returned to normal levels by 8 h. Hamsters exposed at 25 mW/cm2 showed an elevated temperature of 3.0-3.5 degrees C that returned to normal within 60 min after termination of microwave exposure. These animals also showed a marked lymphopenia and neutrophilia by 1 h posttreatment that returned to normal by 8-10 h. Serum glucocorticosteroids were elevated between 1 aNd 8 h after microwave treatment. Sham-exposed animals did not demonstrate significant changes in core body temperature, peripheral blood leukocyte (PBL) profile, or glucocorticosteroid levels as compared to minimum-handling controls.     

Prefrontal cortex aspiration in pups and juvenile rats: behavioural changes and recovery of function

Male Wistar rats sustaining prefrontal cortex aspiration or sham operation at 6 days or 30 days of age were submitted to the following behavioural tests: open-field, acquisition and retention of two-way active as well as passive avoidance tasks. In the open-field the locomotor activity proved enhanced in all the aspirated animals and this enhancement lasted for 30 days. In the two-day active avoidance task, an acquisition deficit was observed in both aspirated groups; but when retrained one month later, they were able to acquire the avoidance task like sham-operated rats and no difference appeared between the groups aspirated at 6 or at 30 days of age. Concerning the passive avoidance task, no difference could be detected between aspirated and sham-operated animals of both groups except that the rats aspirated at an early age (6 days) seemed to display a better avoidance ability in the retention test. These behavioural alterations (hyperactivity and impairment of the acquisition of the 2-way active avoidance) resulted from the prefrontal cortex aspiration, at whatever age this aspiration was performed (6 days or 30 days). They disappeared after a postoperative recovery period of about one month, as evidenced by this longitudinal study.     

Effect of GABA receptor agonist phenibut on behavior and respiration of rabbits in the negative emotional situation

The influence of systemic injection of GABA-receptor agonist--phenibut (40 mg/kg, s/c) on open field behavior, behavioral reactivity and changes in respiratory parameters after exposure of negative emotional stimuli was studied in three rabbit groups differentiated by locomotor activity in open field (active, passive and medium-active animals). The injection of phenibut results in decrease of rabbits horizontal locomotor activity and some components of research behavior in open field and also decrease of behavioral reactivity on emotional stimuli. At the same time the probability of both an active orienting exploratory or defensive reactions and passive reactions (freezing) were decreased. The influence of phenibut depended on typological features of rabbits: the most potent effect occurred upon behavior of active rabbits, less on passive animals and practically none on medium-active rabbits. The phenibut injection results in increase of duration of inhalation during exposure to emotional stimuli, whereas it decreased in normal.     

Disruption of imprinting by memory inhibitors

Several amnestic drugs were administered intracranially to day-old chicks at selected times around a 10-min exposure to an imprinting stimulus. The drugs used were monosodium glutamate, ouabain, cycloheximide and amino-iso-butyrate. The chicks were tested for 10 min in the same apparatus two days later, and the time spent following the stimulu was recorded., The index of memory retention was the difference between the time spent following on test and the time spent following on the initial exposure. When compared with saline-injected control, glutamate administered 5 min before the beginning of the initial exposure was effective in producing a reduction in following times and hence amnesia. Ouabian was effective when injeced before the beginning and immediately after the end of the initial exposure; while cycloheximide was effective when administered as late as 5 min after the initial exposure. The effective times of administration for the drugs to produce a reduction in following times were similar to that observed for amnesia in passive avoidance memory tasks. The increase in following shown by the control chicks was not a developmental effect due to the increae in age on test. Experiments involving a choice of stimuli on test support the invovement of a memoryrelated phenomenon in these experiments.     

Microwave-induced increase of water and conductivity in submaxillary salivary gland of rats

Hypersalivation is an important mechanism for heat dissipation by animals without sweat glands. The water content and conductivity (at 20 kHz) in sub-maxillary salivary gland (SSG) and in other tissues were investigated in adult male rats exposed to microwaves (2880 MHz, 1.5 μs pulses at 1000 Hz) or to conventional heat at 40 °C. Eighty rats in one series were exposed, one at a time, for 30 min to microwaves producing a specific absorption rate (SAR) of 4.2,6.3,6.8,8.4,10.8 or 12.6 W/kg. Fifty rats were sham-exposed under similar environmental conditions. In the second series, ten rats were sham-exposed, 33 rats were exposed, one at time, for 15, 30 or 60 min to microwaves at a SAR of 9.5 W/kg, and 32 rats were exposed for similar periods to conventional heat at 40 °C. In rats of the first series colonic temperatures were elevated significantly at a SAR of 4.2 W/kg, while SSG water content and conductivity increased significantly at SAR values of 6.3 W/kg and higher. In the second series of experiments increases in colonic temperature and SSG water content were greater after 15 and 30 min of microwave exposure than after exposure to heat. Also, SSG conductivity was significantly depressed by heat and significantly increased by microwaves after exposure for 15 or 30 min. The results support the hypothesis that water content and conductivity of SSG of rats can be used as a sensitive specific test of a microwave induced thermal response.     

Induction of Enhanced Acoustic Startle Response by Noise Exposure: Dependence on Exposure Conditions and Testing Parameters and Possible Relevance to Hyperacusis

There has been a recent surge of interest in the development of animal models of hyperacusis, a condition in which tolerance to sounds of moderate and high intensities is diminished. The reasons for this decreased tolerance are likely multifactorial, but some major factors that contribute to hyperacusis are increased loudness perception and heightened sensitivity and/or responsiveness to sound. Increased sound sensitivity is a symptom that sometimes develops in human subjects after acoustic insult and has recently been demonstrated in animals as evidenced by enhancement of the acoustic startle reflex following acoustic over-exposure. However, different laboratories have obtained conflicting results in this regard, with some studies reporting enhanced startle, others reporting weakened startle, and still others reporting little, if any, change in the amplitude of the acoustic startle reflex following noise exposure. In an effort to gain insight into these discrepancies, we conducted measures of acoustic startle responses (ASR) in animals exposed to different levels of sound, and repeated such measures on consecutive days using a range of different startle stimuli. Since many studies combine measures of acoustic startle with measures of gap detection, we also tested ASR in two different acoustic contexts, one in which the startle amplitudes were tested in isolation, the other in which startle amplitudes were measured in the context of the gap detection test. The results reveal that the emergence of chronic hyperacusis-like enhancements of startle following noise exposure is highly reproducible but is dependent on the post-exposure thresholds, the time when the measures are performed and the context in which the ASR measures are obtained. These findings could explain many of the discrepancies that exist across studies and suggest guidelines for inducing in animals enhancements of the startle reflex that may be related to hyperacusis.     

Influence of emotional stimuli on behavior and respiration of rabbits with various locomotor activity in the "open field"

Different behavioral reactivity of rabbit groups differentiated by locomotor activity in the "open field" was revealed during exposure to emotional stimuli (rustle, loud sound, pressuring on back of the neck, vibroacoustic tactile stimulation of an ear). In passive rabbits, the active locomotor reactions were induced harder and freezing was obtained easier than in active animals. During exposure to sound stimuli, passive rabbits increased their locomotion more rarely than active animals, pressing on back of the neck produced longer freezing, a threshold of defensive ear shaking in response to a vibroacoustic stimulus in passive animals was highest. Training to mild immobilization increased the threshold of defensive responses in active rabbits and animals of the intermediate type. Changes in respiratory parameters were correlated with behavioral reactions to emotional stimuli. The duration of exhalation and respiratory cycle increased during freezing and increased during enhanced locomotion. The duration of inhalation decreased in response to emotional stimuli irrespective of a behavioral reaction. The respiratory reactions to emotional stimuli differed in rabbits of different groups. The respiratory rate more frequently changed in passive rabbits than in animals of other groups. Passive animals reacted mainly by exhalation, active rabbits and animals from the intermediate group predominantly responded by inhalation.     

Effects of oxytocin and vasopressin on retrieval of passive avoidance response in melatonin-treated and/or pinealectomized male rats

The role of the pineal gland and its hormone-melatonin-as to the impact of vasopressin (VP) and/or oxytocin (OT) on the regulation of behavior was studied, the passive avoidance task being chosen as an experimental model. The results showed that VP facilitated the avoidance latency during the first retention trial; after pinealectomy, however, VP was ineffective in this regard. Intraperitoneal application of OT was ineffective in modifying the passive avoidance latency when compared with respective saline-treated animals. Melatonin alone, when injected to shamoperated animals 30 min before behavioral experiment, did not affect the passive avoidance response in SA- or OT-treated rats, but blocked the VP-induced lengthening of the passive avoidance latency in the first retention trial. In pinealectomized and OT-treated animals the passive avoidance latency during the second retention trial was severely diminished by melatonin when compared to respective control. We conclude that: a) VP needs a regulated pineal function for developing short-term effects on the passive avoidance response and b) the effect of OT on the avoidance latency in pinealectomized rats develops after melatonin treatment as a long-term effect.     

Effects of microwave exposure on the hamster immune system. II. Peritoneal macrophage function

Acute exposure to hamsters to microwave energy (2.45 GHz; 25 mW/cm2 for 60 min) resulted in activation of peritoneal macrophages that were significantly more viricidal to vaccinia virus as compared to sham-exposed or normal (minimum-handling) controls. Macrophages from microwave-exposed hamsters became activated as early as 6 h after exposure and remained activated for up to 12 days. The activation of macrophages by microwave exposure paralleled the macrophage activation after vaccinia virus immunization. Activated macrophages from vaccinia-immunized hamsters did not differ in their viricidal activity when the hamsters were microwave- or sham-exposed. Exposure for 60 min at 15 mW/cm2 did not activate the macrophages while 40 mW/cm2 exposure was harmful to some hamsters. Average maximum core temperatures in the exposed (25 mW/cm2) and sham groups were 40.5 degrees C (+/- 0.35 SD) and 38.4 degrees C (+/- 0.5 SD), respectively. In vitro heating of macrophages to 40.5 degrees C was not as effective as in vivo microwave exposure in activating macrophages to the viricidal state. Macrophages from normal, sham-exposed, and microwave-exposed hamsters were not morphologically different, and they all phagocytosed India ink particles. Moreover, immune macrophage cytotoxicity for virus-infected or noninfected target cells was not suppressed in the microwave-irradiated group (25 mW/cm2, 1 h) as compared to sham-exposed controls, indicating that peritoneal macrophages were not functionally suppressed or injured by microwave hyperthermia.     

The characteristics of higher nervous activity and of the brain benzodiazepine system in rats subjected to ethanol exposure in the prenatal period

Exposure to ethanol during pregnancy results in the alternation of 3H-diazepam binding to synaptosomal neocortical membranes from the rat offspring. In male experimental rats, 14 days of age, binding level diminished to 11%. In two-month-old control rats Scatchard plot was biphasic. It has been shown that prenatal exposure to ethanol leads to changes in the nature of binding in two-month-age experimental animals, as compared with the control ones. 3H-diazepam binding changes went along with behavioural deviations. In experimental rats locomotor activity was increased in the "open field" test, passive avoidance conditioned reflex retention was decreased and elaboration parameters of active avoidance conditioned reflex were changed, as compared with the control ones. The data obtained show that higher integrative functions were disturbed by prenatal alcoholization. Correlations between benzodiazepine receptor state and behaviour were studied.     

Vasopressin levels in peripheral blood and in cerebrospinal fluid during passive and active avoidance behavior in rats.

Vasopressin (AVP) levels were measured in plasma and cerebrospinal fluid (CSF) of rats during acquisition and retention of a passive avoidance response. Only 5 min after the onset of the retention session a significantly higher level of AVP was found in plasma of animals which displayed a long latency, as compared with the levels of animals which showed a weak passive avoidance response (short latencies), or no passive avoidance behavior at all (controls). Moreover no changes in plasma AVP levels were found in plasma of rats submitted to acquisition or extinction of an active avoidance response. It is suggested that, although an elevated plasma AVP level is associated with strong retention of a passive avoidance response the peripheral circulation as well as the CSF are of minor importance for the transport of this neuropeptide to its site of behavioral action.     

Amphetamine withdrawal: a behavioral evaluation

The effects of withdrawal from long-term amphetamine treatment of intracranial self-stimulation, forced swim-induced immobility, shuttle escape performance, acoustic startle and locomotor activity were evaluated. Mice implanted with stimulating electrodes in the lateral hypothalamus demonstrated stable and reliable rates of self-stimulation responding. After exposure to a chronic schedule of amphetamine treatment response rates were severely depressed. In addition to modifying intracranial self-stimulation responding, amphetamine withdrawal increased the duration of immobility in a forced-swim situation. Although chronic amphetamine exposure induced pronounced behavioral changes in the intracranial self-stimulation and forced swim tasks, drug withdrawal had little effect on shuttle escape performance, acoustic startle and locomotor activity. Based on these findings it was suggested that the development of post-amphetamine depression in the self-stimulation and forced swim paradigms was not related to variations in motoric or arousal mechanisms resulting from amphetamine withdrawal, but rather involved drug-induced changes in motivational processes.     

Modification of acoustic startle by microwave pulses in the rat: a preliminary report.

Single, 1.25-GHz microwave pulses of 0.8- to 1.0-microseconds duration were presented to each of four rats 100 ms before presentation of a startle-inducing acoustic stimulus. This sequential pairing of microwave pulse and acoustic stimulus was found to modify the startle response. At an energy dose to the head of 22-43 mJ/kg per pulse (peak SAR, 23-48 kW/kg), the mean latency to the startle response was longer and the mean amplitude of the response was smaller with respect to control responses that occurred to acoustic stimuli alone. However, at a higher energy dose per microwave pulse in the range of 59-107 mJ/kg (peak SAR, 63-111 kW/kg), the mean latency and amplitude of the startle response were not statistically different from the respective means of control responses.     

Intermittent exposure of rats to 2450 MHz microwaves at 2.5 mW cm2: behavioral and physiological effects

Long-Evans male adult rats were intermittently exposed for 14 weeks to continuous wave (CW) 2450-MHz microwaves at an average power density of 2.5 mW/cm2. The mean specific absorption rate was 0.70 W/kg (+/- 0.02 SEM). The rats were exposed 7 h/day, 7 days/week in a radiation chamber with a monopole above ground, while housed in Plexiglas cages. Weekly measures of body mass and food intake did not indicate statistically significant effects of microwave irradiation. Assessments of threshold for electric-footshock detection revealed a significant difference between microwave and sham-exposed animals. Assessments of cholinesterase and sulfhydryl groups in blood and 17-ketosteroids in urine did not distinguish the two groups of rats. Behavioral measures made at the end of the 14-week exposure included an open-field test, shuttlebox avoidance performance, and schedule-controlled lever-pressing for food pellets. Statistically significant differences between microwave- and sham-exposed rats were observed for these measures. Examination of adrenal tissue, plasma electrolytes, and organ masses after 14 weeks of exposure revealed no difference between the two groups of rats.     

Psychotropic effects of glutamic acid diethyl ester in mice

In 1 hour after intraperitoneal injection glutamic acid diethyl ester (GED) in doses 200 and 500 mg/kg decreased locomotor activity and exploratory patterns of mice in "open field" test. GED in doses 100 and 200 mg/kg diminished the immobilization period of animals in forced swimming test, that proves the reversal interaction of glutamatergic and catecholaminergic systems in CNS. Glutamate receptors antagonist--GED in doses 100-200 mg/kg disrupted passive avoidance reaction at 30 min before acquisition and retrieval, therefore glutamate receptors are involved into fixation and retrieval of memory engram.     

Circadian and photoperiodic effects of brief light pulses in male Djungarian hamsters

The effects of brief light pulses (1-60 min in duration) on the circadian rhythm of locomotor activity and/or the neuroendocrine-gonadal axis was investigated in male Djungarian hamsters. Exposure of hamsters free-running in constant darkness to a single 1-h pulse of light induced phase-dependent phase shifts in the rhythm of locomotor activity. The general shape of the "phase-response curve" was similar to that observed in other animals; phase-delays and phase-advances were induced by light pulses delivered in the early and late subjective night, respectively, while light pulses during the subjective day induced little or no phase-shift in the activity rhythm. Animals exposed for 7 days to 1-min of light during the night in animals otherwise exposed to 6L:18D resulted in increased levels of serum FSH and testicular weight. Daily exposure to two 1-h or two 10-min pulses of light (but not two 1-min pulses) for 10 days resulted in stable entrainment of the activity rhythm as well as testicular weight gains and serum FSH increases. When two 10-min pulses of light were presented 8 and 16 h apart, some animals showed a short-day entrainment pattern (i.e., locomotor activity confined to the long period of darkness) while other animals showed a long-day entrainment pattern (i.e., locomotor activity confined to the short period of darkness). Importantly, the stimulatory effects of light on neuroendocrine-gonadal activity were clearly dependent on the phase-relationship between the light pulses and the circadian rhythm of locomotor activity.(ABSTRACT TRUNCATED AT 250 WORDS)     

Hyperreflexive behavior in Brattleboro rats

In two experiments we compared the size of the startle response elicited by shock and acoustic stimuli in animals having a congenital absence of vasopressin (Brattleboro rats) with closely related controls, which do synthesize vasopressin. The rats lacking vasopressin were hyperresponsive to both shock and acoustic stimuli. Inhibition of the acoustic startle by shock prestimuli also was greater in these animals. These results are consistent with reports which indicate that vasopressin attenuates responses to noxious stimuli.     

Brain temperature measurements in rats: a comparison of microwave and ambient temperature exposures

In an effort to understand microwave heating better, regional brain and core temperatures of rats exposed to microwave radiation (2450 MHz) or elevated air temperatures were measured in two studies. In general, we have found no substantial evidence for temperature differentials, or "hot spots," in the brain of these animals. In the first study, after a 30-min exposure, no temperature differences between brain regions either after microwave or ambient air exposure were found. However, a highly significant correlation between brain and core temperatures was found and this correlation was the same for both microwave and ambient air heating. In the second study, time-temperature profiles were measured in rats exposed to either 30 mW/cm2 or 36.2 degrees C. In this study, the 30-min exposure period was divided into seven intervals and the change in temperature during each period was analyzed. Only the cortex showed significantly different heating rates between the air heating and microwave heating; however, this difference disappeared after the initial 5 min of exposure.