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A gas exposure system using rotating vessels was improved for exposure of cultured mammalian cells to gaseous compounds in the chromosomal aberration assay. This system was composed of 12 square culture vessels, a device for preparation of air containing test gas, and positive and negative control gases at target concentrations and for supplying these gases to the culture vessels, and a roller apparatus in an incubator. Chinese hamster lung cells (CHL/IU) were grown on one side of the inner surface of the square culture vessel in the MEM medium. Immediately prior to exposure, the medium was changed to the modified MEM. Air in the culture vessel was replaced with air containing test gas, positive or negative control gas. Then, the culture vessels were rotated at 1.0 rpm. The monolayered culture cells were exposed to test gas during about 3/4 rotation at upper positions and alternatively immersed into the culture medium during about 1/4 rotation at lower positions. This system allowed the chromosomal aberration assay simultaneously at least at three different concentrations of a test gas together with positive and negative control gases with and without metabolic activations, and duplicate culture at each exposure concentration. Seven gaseous compounds, 1,3-butadiene, chlorodifluoromethane, ethyl chloride, methyl bromide, methyl chloride, propyne, and vinyl chloride, none of which has been tested to date, were tested on CHL/IU for the chromosomal aberration assay using this gas exposure system. All the compounds except chlorodifluoromethane showed positive responses of the structural chromosomal aberrations, whereas polyploidy was not induced by any of these gases. This improved gas exposure system proved to be useful for detecting chromosomal aberrations of gaseous compounds.

Rats were chronically implanted with electrodes for EEG, EMG and ECG recordings and exposed to 0.1 ppm, 0.2 ppm ozone (O3) or clean air as control for 5 consecutive days. Amounts of wakefulness (W), slow-wave sleep (SWS) and paradoxical sleep (PS) of the O3-exposed rats were not statistically different from respective control values. Compared with control rats, heart rates of the O3-exposed rats decreased and the number of bradyarrhythmic episodes increased with an increase in O3 levels. The O3-induced bradyarrhythmias occurred more prevalently in the order of W, SWS and PS state. Parasympathetic mechanisms were suggested on those cardiac abnormalities.

Acute effects of a single i.p. injection of toluene on circadian rhythms of sleep-wakefulness were investigated in rats which were chronically implanted with EEG and EMG electrodes for polygraphic recordings. The toluene injection produced an initial increase in wakefulness (W) and a subsequent increase in slow-wave sleep (SWS) during the dark period. In an attempt to clarify mechanisms of these biphasic effects of toluene on sleep-wakefulness rhythms, brain monoamines and their metabolites were determined at the times of the initial increase in W and the increased SWS. The initial increase in W was associated with an increase in cortical NA, MHPG and 5-HT together with a decrease in cortical 5-hydroxyindoleacetic acid (5-HIAA), while the increased SWS during the dark period was associated with an increase in 5-HIAA and a concomitant decrease in 3-methoxy-4-hydroxyphenylglycol (MHPG). The toluene-induced changes in sleep-wakefulness seemed to be manifested at lower blood levels of toluene than the behavioral signs of central nervous system (CNS) depression. These biphasic effects of toluene on circadian sleep-wakefulness rhythms are discussed in terms of the reciprocal interactions between central 5-HT and NA neurons.

Muricidal behavior of the rat was induced by repeated s.c. administration of cadmium (Cd) for 15 weeks. The number of muricidal rats increased with increase in the duration of Cd administration in association with greater accumulation of Cd in the olfactory bulb than in any other region of the brain. The whole brain norepinephrine (NE) level of the rats which had been given Cd for 15 weeks was significantly greater than that of the control rats; neither dopamine (DA) nor serotonin (5-HT) level was changed. The Cd-induced muricide is discussed.

Adult male Wistar rats were almost totally deprived of sleep by handling for 24 hr. 5-Hydroxyindolacetic acid concentrations in the dorsal raphe nucleus area and thalamus increased by 140-180%, immediately after sleep deprivation and when the rats had a 3- or 30-min rebound sleep. The higher levels of 5-hydroxyindolacetic acid were still observed after the rats were awakening from a 4-hr sleep. The concentrations of 5-hydroxytryptamine (serotonin) decreased after sleep deprivation and increased during and after sleep, but the differences were not significant. Tryptophan accumulated in the dorsal raphe area and thalamus after sleep deprivation, and an elevated level did not return to baseline concentrations until the rats were awakening. Tryptophan hydroxylase activity did not change in the dorsal raphe area during and after sleep deprivation. These results suggest that the release and synthesis of 5-hydroxytryptamine in the dorsal raphe area and thalamus increased when the rats had a sleep pressure or a rebound sleep after total sleep deprivation. An increased transport of tryptophan into the brain may be closely involved in sleep-inducing mechanisms.

Effects of methylmercury chloride (MMC) on circadian sleep-waking rhythms were examined in rats which had been chronically implanted with EEG and EMG electrodes. Bihourly distributions of wakefulness (W), slow wave sleep (SWS) and paradoxical sleep (PS) and 12-h amounts of W, SWS and PS during light and dark periods were measured before and after MMC administration for 2 successive days at 3 dose levels. A total dose of 10 mg MMC/kg body wt was found to be the threshold for inducing reversible changes in the sleep-waking patterns. A total dose of 30 mg MMC/kg produced an increase in both dark-phase SWS and PS as well as a decrease in light-phase PS at the expense of an increase in light-phase W and a delayed phase of the circadian PS rhythm. The delayed phase of the PS rhythm tended to persist after the increased SWS during the dark period returned to normal. Brain mercury concentrations were measured in order to find the dose-response relationship and the time dependence of the MMC-induced sleep disorder. The sleep-waking disorder was found to appear at lower levels of brain Hg and shorter latency than behavioral disorders of movement and postural maintenance previously reported [5-8].

In an attempt to examine chronic effects of toluene on sleep, spontaneous locomotor activity and drinking behavior, rats were repeatedly administered toluene i.p. at doses of 100 and 200 mg/kg body weight for 14 consecutive days. The 200-mg/kg injections induced a decrease in total sleep on Day 1, an increase in locomotor activity on Days 1 through 4 and an increase in drinking activity on Days 0 through 6 after discontinuation of the daily injections. Both the reduced sleep and the increased locomotor activity appeared during the light period, whereas the drinking activity increased during the dark period. In order to find neurochemical correlates of the toluene-induced changes in behavior, regional concentrations of brain monoamines and their metabolites were determined. The toluene-induced partial insomnia and hyperactivity were associated with lowered concentrations of serotonin in frontal cortex, hippocampus and midbrain and 5-hydroxyindoleacetic acid in midbrain and hypothalamus. The increased drinking activity was associated with increased concentrations of striatal 3,4-dihydroxy-phenylacetic acid and homovanillic acid and hypothalamic noradrenaline and 3-methoxy-4-hydroxyphenylethyleneglycol. Central monoaminergic mechanisms were implicated in the toluene-induced partial insomnia, hyperactivity and hyperdipsia.