Low dose rotenone treatment causes selective transcriptional activation of cell death related pathways in dopaminergic neurons in vivo.

Mitochondrial complex I inhibition has been implicated in the degeneration of midbrain dopaminergic (DA) neurons in Parkinson's disease. However, the mechanisms and pathways that determine the cellular fate of DA neurons downstream of the mitochondrial dysfunction have not been fully identified. We conducted cell-type specific gene array experiments with nigral DA neurons from rats treated with the complex I inhibitor, rotenone, at a dose that does not induce cell death. The genome wide screen identified transcriptional changes in multiple cell death related pathways that are indicative of a simultaneous activation of both degenerative and protective mechanisms. Quantitative PCR analyses of a subset of these genes in different neuronal populations of the basal ganglia revealed that some of the changes are specific for DA neurons, suggesting that these neurons are highly sensitive to rotenone. Our data provide insight into potentially defensive strategies of DA neurons against disease relevant insults.

Effects of acute systemic 3-nitropropionic acid administration on rat activity and acoustic startle.

Spontaneous activity, acoustic startle, and prepulse inhibition (PPI) of acoustic startle were measured in male Sprague-Dawley rats 3-5 h after 0, 10, 15, or 20 mg/kg i.p. 3-nitropropionic acid (3-NP), a mitochondrial toxin. Mean activity was significantly influenced by the 3-NP dose due to decreased activity for 20 mg/kg. Mean startle amplitude was not significantly affected by the 3-NP dose. Means of PPI for prepulses 6 and 12 dBA above background were smaller than means for respective 0 mg/kg doses, but the main effect of 3-NP dose did not reach statistical significance in ANOVA. The changes in measured exploratory-type activity and, possibly, in startle PPI parallel the occurrence of clinical signs exhibited at 3-5 h after 3-NP injection. Neural processing involved in these quantitative behavioral endpoints seems to be affected as energy stores are depleted and degenerative processes are beginning.

Hyperactivity caused by a nitric oxide synthase inhibitor is countered by ultra-wideband pulses.

Potential action of ultra-wideband (UWB) electromagnetic field pulses on effects of N(G)-nitro- L-arginine methyl ester (L-NAME), an inhibitor of nitric oxide synthase (NOS), on nociception and locomotor activity was investigated in CF-1 mice. Animals were injected IP with saline or 50 mg/kg L-NAME and exposed for 30 min to no pulses (sham exposure) or UWB pulses with electric field parameters of 102+/-1 kV/m peak amplitude, 0.90+/-0.05 ns duration, and 160+/-5 ps rise time (mean+/-S.D.) at 600/s. Animals were tested for thermal nociceptive responses on a 50 degrees C surface and for spontaneous locomotor activity for 5 min. L-NAME by itself increased mean first-response (paw lift, shake, or lick; jump) and back-paw-lick response latencies and mean locomotor activity. Exposure to UWB pulses reduced the L-NAME-induced increase in back-paw-lick latency by 22%, but this change was not statistically significant. The L-NAME-induced hyperactivity was not present after UWB exposure. Reduction and cancellation of effects of L-NAME suggest activation of opposing mechanism(s) by the UWB pulses, possibly including increase of nitric oxide production by NOS. The action, or actions, of UWB pulses appears to be more effective on locomotor activity than on thermal nociception in CF-1 mice.

Pulsed microwave induced light, sound, and electrical discharge enhanced by a biopolymer.

Intense flashes of light were observed in sodium bicarbonate and hydrogen peroxide solutions when they were exposed to pulsed microwave radiation, and the response was greatly enhanced by a microwave-absorbing, biosynthesized polymer, diazoluminomelanin. A FPS-7B radar transmitter, operating at 1.25 GHz provided pulses of 5.73 +/- 0.09 micros in duration at 10.00 +/- 0.03 pulses/s with 2.07 +/- 0.08 MW forward power (mean +/- standard deviation), induced the effect but only when the appropriate chemical interaction was present. This phenomenon involves acoustic wave generation, bubble formation, pulsed luminescence, ionized gas ejection, and electrical discharge. The use of pulsed microwave radiation to generate highly focused energy deposition opens up the possibility of a variety of biomedical applications, including targeting killing of microbes or eukaryotic cells. The full range of microwave intensities and frequencies that induce these effects has yet to be explored and, therefore, the health and safety implications of generating the phenomena in living tissues remain an open question.

Frequency of micronuclei in the blood and bone marrow cells of mice exposed to ultra-wideband electromagnetic radiation.

PURPOSE: To investigate the extent of genetic damage in the peripheral blood and bone marrow cells of mice exposed to ultra-wideband electromagnetic radiation (UWBR). MATERIALS AND METHODS: CF-1 male mice were exposed to UWBR for 15 min at an estimated whole-body average specific absorption rate of 37 mW x kg(-1). Groups of untreated control and positive control mice injected with mitomycin C were also included in the study. After various treatments, half of the mice were killed at 18 h, and the other half at 24 h. Peripheral blood and bone marrow smears were examined to determine the extent of genotoxicity, as assessed by the presence of micronuclei (MN) in polychromatic erythrocytes (PCE). RESULTS: The percentages of PCE and the incidence of MN per 2000 PCE in both tissues in mice killed at 18 h were similar to the frequencies observed in mice terminated at 24 h. There were no significant differences in the percentage of PCE between control and the mice with or without UWBR exposure; the group mean values (+/- standard deviation) were in the range of 3.1+/-0.14 to 3.2+/-0.23 in peripheral blood, and 49.0+/-3.56 to 52.3+/-4.02 in bone marrow. The mean incidence of MN per 2000 PCE in control and in mice with or without UWBR exposure ranged from 7.7+/-2.00 to 9.7+/-2.54 in peripheral blood and 7.4+/-2.32 to 10.0+/-3.27 in bone marrow. Pairwise comparison of the data did not reveal statistically significant differences between the control and mice with or without UWBR exposure groups (excluding positive controls). CONCLUSION: Under the experimental conditions tested, there was no evidence for excess genotoxicity in peripheral blood or bone marrow cells of mice exposed to UWBR.

Ultra-wideband electromagnetic pulses and morphine-induced changes in nociception and activity in mice.

Mice were exposed to ultra-wideband (UWB) electromagnetic pulses averaging 99-105 kV/m peak amplitude, 0.97-1.03 ns duration, and 155-174 ps rise time, after intraperitoneal administration of saline or morphine sulfate. They were then tested for thermal nociception on a 50 degrees C surface and for spontaneous locomotor activity and its time profile over 5 min. Analysis of results showed no effect of UWB exposure on nociception and activity measures in CF-1 mice after 15-, 30-, or 45-min exposure to pulses at 600/s or after 30-min exposure to UWB pulses at 60/s. Similarly, no effect was seen in C57BL/6 mice after 30-min exposure to pulses at 60/s or 600/s. Although trends in morphine-modified measures seen with UWB pulse repetition frequency could be expected because of increased levels of low-frequency energy, no significant change was seen in normal or morphine-modified nociception or activity after UWB exposure. This indicated lack of effect of the UWB pulses used in these experiments on nervous system components, including endogenous opioids, involved in these behaviors.

Ultra-wideband electromagnetic pulses: lack of effects on heart rate and blood pressure during two-minute exposures of rats.

Exposure to fast-rise-time ultra-wideband (UWB) electromagnetic pulses has been postulated to result in effects on biological tissue (including the cardiovascular system). In the current study, 10 anesthetized Sprague-Dawley rats were exposed to pulses produced by a Sandia UWB pulse generator (average values of exposures over three different pulse repetition rates: rise time, 174-218 ps; peak E field, 87-104 kV/m; pulse duration, 0.97-0.99 ns). Exposures to 50, 500 and 1000 pulses/s resulted in no significant changes in heart rate or mean arterial blood pressure measured every 30 s during 2 min of exposure and for 2 min after the exposure. The results suggest that acute UWB whole-body exposure under these conditions does not have an immediate detrimental effect on these cardiovascular system variables in anesthetized rats.

Sensing platform for acoustic startle responses from rat forelimbs and hindlimbs.

A sensing platform with two piezoelectric transducers was designed and fabricated to measure acoustic startle responses from forelimbs and hindlimbs in the rat. Testing with a vibrator showed that separate forces were measured from 5 to 25 Hz with mean sensitivities of 2.395 and 2.022 V/N and mean linearity errors of 3.23 and 2.98% FS for the forelimb and hindlimb sensors, respectively. Forelimb and hindlimb response waveforms of male Sprague-Dawley rats had shapes similar to the commonly recorded wholebody response but were smaller in amplitude.

Modification of acoustic and tactile startle by single microwave pulses.

Single microwave pulses at 1.25 GHz were delivered to the head and neck of male Long-Evans rats as a prestimulus to acoustic and tactile startle. For acoustic startle, pulses averaging 0.96 microsecond in duration were tested with two specific absorption rate (specific absorption) ranges, 15.0-30.0 kW/kg (16.0-44.2 mJ/kg) and 35.5-86.0 kW/kg (66.6-141.8 mJ/kg), delivered 201, 101, 51, 3, and 1 ms before and 1 ms after onset of a startling noise. The low-intensity pulse did not affect peak amplitude, integral, or latency of the whole-body startle response. The high-intensity pulse at 101 and 51 ms inhibited the startle response by decreasing peak amplitude and integral; at 201 and 51 ms latency was increased. The high-intensity pulse at 1 ms enhanced the startle response by increasing peak amplitude and at 3 ms by increasing integral. For tactile startle, either microwave pulses averaging 7.82 microseconds in duration and 55.9-113.3 kW/kg (525.0-1055.7 mJ/kg) or 94 dB SPL clicks were delivered 157, 107, 57, and 7 ms before and 43 ms after onset of a startling air burst. The microwave pulse at 57 ms inhibited the startle response by decreasing peak amplitude; at 157, 107, 57, and 7 ms it increased latency. The microwave pulse at 43 ms after onset enhanced the startle response by increasing peak amplitude. The acoustic click at 157 and 57 ms inhibited the startle response by decreasing peak amplitude; at 157,2 107, and 57 ms it increased latency.(ABSTRACT TRUNCATED AT 250 WORDS)

Inter-beat intervals of cardiac-cell aggregates during exposure to 2.45 GHz CW, pulsed, and square-wave-modulated microwaves.

Inter-beat intervals of aggregated cardiac cells from chicken embryos were studied during 190 s exposures to 2.45 GHz microwaves in an open-ended coaxial device. Averaged specific-absorption rates (SARs) and modulation conditions were 1.2-86.9 W/kg continuous-wave (CW), 1.2-12.2 W/kg pulse modulation (PW, duty cycle approximately 11%), and 12.0-43.5 W/kg square-wave modulation (duty cycle = 50%). The inter-beat interval decreased during microwave exposures at 42.0 W/kg and higher when CW or square-wave modulation was used, which is consistent with established effects of elevated temperatures. However, increases in the inter-beat interval during CW exposures at 1.2-12.2 W/kg, and decreases in the inter-beat interval after PW exposures at 8.4-12.2 W/kg, are not consistent with simple thermal effects. Analysis of variance indicated that SAR, modulation, and the modulation-SAR interaction were all significant factors in altering the inter-beat interval. The latter two factors indicated that the cardiac cells were affected by athermal as well as thermal effects of microwave exposure.

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.

Method to record evoked potentials from the frog eighth nerve.

A method for recording evoked potentials from the eighth nerve of frogs is described. A prominent bipolar wave with latency of 3-6 ms recorded in response to auditory stimuli in Rana catesbeiana is attributable to eighth-nerve activity. The evoked potential provides an integrated response for study of inner ear and peripheral neural activity which complements responses obtained by other recording methods.

Thresholds of cat cochlear nucleus neurons to microwave pulses.

Action potentials of neurons in cat dorsal and posteroventral cochlear nuclei were recorded extracellularly with glass microelectrodes while the head of the cat was exposed to microwave pulses at 915 MHz using a diathermy applicator. Response thresholds to acoustic tones, acoustic clicks, and microwave pulses were determined for auditory units with characteristic frequencies (CFs) from 278 Hz to 39.2 kHz. Tests with pulsatile stimuli were performed for durations of 20-700 mus, principally 20, 70, and 200 mus. Brainstem midline specific absorption rate (SAR) threshold was as small as 11.1 mW/g per pulse, and specific absorption (SA) threshold was a small as 0.6 muJ/g per pulse. Microwave thresholds were generally lower for CF less than 9 kHz, as were most acoustic thresholds. However, microwave threshold was only weakly related to click threshold and CF-tone threshold of each unit.

Auditory unit responses to single-pulse and twin-pulse microwave stimuli.

Responses of units in the cat cochlear nucleus to single microwave pulses with different durations and to twin microwave pulses with different interpulse delays are used to study microwave hearing. Inferred threshold specific absorption rate is less than 6 mW/g; inferred threshold specific absorption, less than 0.5 microJ/g. The existence of responses from units with characteristic frequencies (CFs) from 931 Hz to 25.5 kHz is not consistent with a primary role for head resonance in microwave hearing. Patterns of response amplitude have a periodicity of 1/CF and are fully explained by frequency content of the pulse stimulus and signal processing of the auditory system. For pulses shorter than about 0.24/CF, it is shown that response amplitude is predictably proportional to pulse energy.

Effects of hypothalamic peptide hormones on the electrical activity of Aplysia neurons.

The effect of luteinizing hormone-releasing hormone (LHRH) and thyrotropin-releasing hormone (TRH) on the electrical activity of neurons in the abdominal ganglion of Aplysia californica was studied. Where tested, TRH had no effect. The neurosecretory white-cell neurons were the most responsive of the neurons tested with LHRH. Bath applications of 1 micro M LHRH increased firing rates in 4 of 5 white cells for extended periods of time. The increased rates persisted in an LHRH-deficient bath. A similar result was obtained with bath applications of the LHRH agonist analog D-Ala6, des-Gly10-LHRH-ethylamide. The iontophoresis of LHRH onto white-cell somata either produced no change in electrical activity or initiated an increase in firing rate and bursting patterns which outlasted the application period. Two types of white cells are suggested by the white-cell responsiveness to LHRH. The white cells responsive to the decapeptide are candidate model neurons for studying the membrane actions of LHRH.

Microwave irradiation and instrumental behavior in rats: unitized irradiation and behavioral evaluation facility.

A facility for the exposure of small animals to pulse-modulated microwave radiation (PM MWR) concurrent with their performance of operant behavioral tasks is described. The computer-managed facility comprises an array of 32 individual waveguide exposure cells, each enclosing instrumental conditioning apparatus within a plastic subhousing. The distribution of the microwave electric field intensity within the waveguide was measured by a nonperturbing probe and the modifications induced by the behavioral apparatus and animal within the waveguide determined. Input and interior voltage standing-wave ratios are presented to characterize the design of the chambers and to demonstrate the suitability of the chambers for whole-body irradiation of rat. The specific absorption rate (SAR) is presented utilizing data derived from incremental thermometric examination of saline loads and of selected sites in rat carcasses. This is compared with the whole-body SAR derived from the input/output energy balance equation for the waveguide. The results of continuous monitoring of the SAR by the latter method, while unrestrained rats were engaged in operant and exploratory behavior within the waveguide, are utilized to derive a relationship between chamber input power and the dose rate for adult rats behaviorally active within the waveguide. From these data, we conclude that the experimental array provides a practical method for exposing a large number of animals to PM MWR for long periods of time and coincident with the establishment and/or performance of complex operant behavior.

Slow and rapid responses to CW and pulsed microwave radiation by individual Aplysia pacemakers.

Specific absorption rates (SARs) of microwave energy that altered firing rates were determined for individual pacemaker neurons in the abdominal ganglion of Aplysia californica. A stripline apparatus provided both for artifact-free recording of transmembrane potentials and for precise determination of the rate of absorption of microwave energy. Exposure for two to three minutes at an SAR of only a few mW/g was capable of changing the firing rate of some pacemakers. Two types of responses were observed. The response that was seen in all neurons developed slowly, reaching a steady state in one to three minutes. The other response was seen in a few neurons and occurred within five seconds from the onset of irradiation. Similar responses were obtained for two microwave frequencies, 1.5 and 2.45 GHz. Pulsed radiation induced rapid changes of firing rate more readily than did CW radiation at the same SAR. A convective heating scheme was used to study the effects of temperature changes on the pacemakers' firing rates. Since all of the responses are not readily explained by general heating of the preparation, alternate mechanisms are suggested for the observed effects.