Acute and subchronic neurotoxicological evaluation of tetrahydrofuran by inhalation in rats.

Groups of adult male and female rats received exposure to tetrahydrofuran (THF) vapor by inhalation in acute or subchronic exposure scenarios. Acute exposure concentrations were 0, 500, 2500, or 5000 ppm for 6 hr. Evaluations conducted immediately after exposure included clinical observations, motor activity assessments (MA), and a battery of functional tests (FOB) designed to reveal nervous system dysfunction. During exposure to 2500 and 5000 ppm, rats had a diminished or absent startle response to a punctate auditory alerting stimulus. Following exposure to 5000 ppm, male and female rats were lethargic, exhibited abnormal gait or mobility, and splayed rear feet. Lethargy and splayed rear feet were also observed in females exposed to 2500 ppm. During the subsequent FOB, males exposed to 5000 ppm had a lower incidence of palpebral closure, higher incidences of slow or absent righting reflex, and a biphasic pattern of reduced motor activity followed by increased motor activity. Females exposed to 5000 ppm had increased incidences of palpebral closure in the open field, increased incidences of slow or absent righting reflex, and decreased motor activity. During the 14-week subchronic exposure series, daily THF exposure concentrations were 0, 500, 1500, or 3000 ppm, and neurobehavioral evaluations occurred on non-exposure days at approximately monthly intervals. Diminished startle responses to an auditory alerting stimulus were observed during exposure to 1500 or 3000 ppm; however, repeated exposures did not cause additional neurobehavioral or pathological effects. This pattern of effects is suggestive of transient sedation. Despite daily reinstatement of acute sedative effects during repeated exposure with up to 3000 ppm, THF did not produce any persistent or cumulative effects on nervous system structure or function. The demonstrated no-observed-effect level of THF for both acute and subchronic exposure was 500 ppm.

Repeated exposure inhalation study of pentane in rats.

Pentane (CAS No. 109-66-0) is a chemical being used as a co-solvent in a polymer production facility with potential for inhalation exposure in humans. To assess the toxicity of pentane, groups of 10 male rats each were exposed by inhalation, 6 hr/day, 5 days/week for 2 weeks to either 0 (control), 1,000, 3,000 or 10,000 ppm. Five rats per group were killed following the 10th exposure; the remaining 5/group were killed after a 14-day post-exposure recovery period. Parameters investigated were clinical signs of toxicity, functional behavior, body weights, clinical pathology, and gross and microscopic pathology including organ weights. No unusual clinical observations were seen in the pentane-treated rats, and body weights were not altered. Test rats generally exhibited normal behavioral responses in the functional observational battery. Increases in serum calcium and phosphorus concentrations were seen in rats exposed to either 3,000 or 10,000 ppm. These were reversible during the 2-week recovery period. No other clinical pathology changes were observed and no pentane-related tissue pathology was seen in any of the groups. The no-observed-adverse-effect level was 1,000 ppm with reversible clinical pathology changes produced at 3,000 and 10,000 ppm.

Subchronic toxicity of cyclohexane in rats and mice by inhalation exposure.

Inhalation studies were conducted to determine the potential toxicity and/or potential neurotoxicity of cyclohexane. Groups of rats and mice were exposed to 0, 500, 2000, or 7000 ppm concentrations of cyclohexane vapor 6 hr/day, 5 days/week for 14 weeks. Subgroups of rats and mice were further observed during a 1-month recovery period. Functional observational battery (FOB) and motor activity (MA) behavioral tests were conducted on rats. These tests were conducted prior to the exposure series and during weeks 4, 8, and 13 on non-exposure days. Clinical pathology evaluations were conducted after approximately 7, 13, and 18 weeks. Approximately 14 and 18 weeks after study initiation, tissues from rats and mice were histologically processed and evaluated by light microscopy. During exposure to 2000 or 7000 ppm, rats and mice had a diminished response or an absent response to delivery of a punctate auditory alerting stimulus. Immediately following removal of rats from the inhalation chambers, 7000 ppm males and females and 2000 ppm females displayed a compound-related increase in the incidence of wet and/or stained fur (which occurred in the areas of the mouth, chin, and/or perineum). These signs were transient, were not observed during exposure or prior to exposure the following day, and were not associated with any behavioral or morphological changes. During exposure sessions, mice exposed to 7000 ppm exhibited clinical signs of toxicity which included hyperactivity, circling, jumping/hopping, excessive grooming, kicking of rear legs, standing on front legs, and occasional flipping behavior. Clinical signs of toxicity observed in 7000 ppm mice immediately after exposure included hyperactivity, hyperreactivity, ruffled fur (females only), gait abnormalities, spasms in both rear legs, and excessive grooming (males only). The clinical signs observed in mice during and immediately after exposure were transient, and were not present prior to the subsequent exposure. A few mice exposed to 2000 ppm appeared hyperactive during exposure in the latter portion of the study. There were no compound-related changes in mean body weights, body weight gains, food consumption, food efficiency, or mortality; and there were no ophthalmological abnormalities in rats or mice. In addition, there were no compound-related effects on 37 different behavioral parameters assessed during the FOB or during motor activity tests in rats. Male and female mice exposed to 7000 ppm had slight increases in measures of circulating erythrocyte mass (red blood cells, hemoglobin, hematocrit) and plasma protein concentration (males only). Male rats and male and female mice exposed to 7000 ppm had significantly increased relative liver weights, and 7000 ppm male mice also had significantly increased absolute liver weights at the end of the exposure period. At the end of the 1-month recovery period, absolute and relative liver weights of male and female mice were similar to control. However, relative liver weights of 7000 ppm male rats continued to be significantly higher at the end of the recovery period. Male and female rats exposed to 7000 ppm had a significantly increased incidence of hepatic centrilobular hypertrophy at the end of the exposure period, which was not observed at the conclusion of the 1-month recovery period. No microscopic changes were observed in mice. In rats, the no-observed-effect level (NOEL) for acute, transient effects was 500 ppm based on a diminished/absent response to an auditory alerting stimulus at 2000 ppm and above. The NOEL for subchronic toxicity in rats was 7000 ppm based on the lack of adverse effects on body weight, clinical chemistry, tissue morphology, and neurobehavioral parameters. In mice, the NOEL for acute, transient effects was 500 ppm based on behavioral changes during exposure at 2000 ppm and above. The NOEL for subchronic toxicity in mice is 2000 ppm based on hematological changes at 7000 ppm.

Evaluation of the sensory irritation potential of volatile organic chemicals from carpets--alone and in combination.

Some individuals have reported burning or painful sensations in the eyes or upper respiratory tract when they enter certain indoor environments. Recently, carpets have been suggested as a potential source of organic chemicals that could contribute to this irritation. The sensations are generally termed 'sensory irritation' or 'pungency', and result from stimulation of trigeminal nerve endings. Indoor air quality is typically evaluated based on the concentrations of individual airborne materials, and rarely do concentrations of potentially offending materials exceed levels expected to cause adverse human health effects. In particular, volatile organic chemicals are measured at low levels (ppb) in the indoor environment. Sources of these organics vary considerably, but concentrations of the individual chemicals do not ordinarily exceed irritating levels. It is possible that the chemical mixture may cause effects not predicted by the present data available on individual chemicals. A list of approximately 50 chemicals identified in carpet emissions was developed for the study. Chemicals were selected from over 200, based on the highest frequency of occurrence and/or highest rate of emission. A mouse model of sensory irritation was used to examine the individual chemicals. In this model, mice are exposed to airborne chemicals by inhalation and evaluated for changes in respiratory function parameters. The model relies on chemical stimulation of the trigeminal nerve endings to elicit a response identified by a decrease in respiratory frequency and an alteration in the breathing pattern. Chemicals are compared quantitatively by measuring the airborne concentration required to elicit a 50% depression in respiratory rate (RD50). Some of the chemicals were previously evaluated by this method, while others were recently examined. The emission chemicals tested individually to date have had RD50 values that generally range from 100 ppm to more than 1000 ppm, indicating that human respiratory irritation would not be expected from the individual chemicals at levels measured in the indoor environment. Differences observed in timing of response from one chemical to another and experimental variability will be important considerations for mixtures testing. The potential for combinations of these emission chemicals to cause sensory irritation at low concentrations, resulting in additivity, synergy, or antagonism of the response, will be addressed. These results should have general application for assessing the risk of causing respiratory irritation in humans exposed to combinations of organic chemicals.

Subchronic toxicity of 4-vinylcyclohexene in rats and mice by inhalation exposure.

This study was conducted to evaluate the subchronic toxicity of 4-vinylcyclohexene (VCH). Male and female Sprague-Dawley rats and B6C3F1 mice were exposed by inhalation to VCH 6 hr/day, 5 days/week for 13 weeks. Rats were exposed to 0, 250, 1000, or 1500 ppm, and mice were exposed to 0, 50, 250, or 1000 ppm. In addition, another group of rats and mice was exposed to 1000 ppm butadiene so that a comparison could be made between the two compounds. Exposure to 1000 ppm VCH resulted in deaths of all male mice and 5/10 female mice on Test Days 11 or 12. Three additional female mice exposed to 1000 ppm VCH died prior to study completion. The most notable compound-related clinical sign was lethargy observed in the 1500 ppm VCH-exposed rats and 1000 ppm VCH-exposed mice. Male rats exposed to 1500 ppm VCH had significantly lower body weights compared to controls, and male and female rats in the 1500 ppm group had significantly lower body weight gains. None of the VCH-exposed animals or butadiene-exposed rats showed any compound-related hematological effects. However, mice exposed to 1000 ppm butadiene exhibited mild macrocytic anemia. Clinical chemistry evaluation and urinalysis showed no compound-related effects in rats exposed to either VCH or butadiene. Male and female rats exposed to 1000 or 1500 ppm VCH or 1000 ppm butadiene had increased absolute and/or relative liver weights, and male rats in these same exposure groups had increased relative kidney weights. Microscopically, increased accumulation of hyaline droplets was observed in the kidneys of male rats from all VCH exposure groups. Although compound-related, the droplets were not accompanied by cytotoxicity. In mice, the most notable adverse histopathological effect was ovarian atrophy in females exposed to 1000 ppm VCH or 1000 ppm butadiene. The atrophy was slightly more severe in the VCH-exposed females than in the butadiene-exposed females. There were no other compound-related pathological effects in male or female mice exposed to VCH. Additionally, butadiene-exposed male mice had decreased testicular weights, accompanied by slight testicular degeneration and atrophy. For VCH exposure, the no-observed-adverse-effect-level is 1000 ppm for rats based on lethargy and lowered body weights and 250 ppm for mice based on mortality and ovarian atrophy.

Evaluation of a method used to test for potential toxicity of carpet emissions.

After a private testing laboratory had reported mortality, neurotoxicity, and respiratory irritation in mice exposed to emissions from heated carpet in a modified application of the sensory irritation test (ASTM E981-84), the studies reported in this paper were conducted to evaluate the method used in testing for carpet toxicity and to see if the reported findings were reproducible. Mice were exposed head-only to offgasses generated by heating carpet (AT #3) to temperatures that ranged from 37 to 70 degrees C. Control mice were simultaneously exposed to heated air. The animals were evaluated for mortality, clinical signs and respiratory irritation. Neurotoxicity was evaluated using functional observational battery and motor activity monitoring. Pathological evaluations of organs and tissues, including the nervous system, were also conducted. The carpet samples heated to higher temperatures produced greater concentrations of total volatile organic compounds than those heated to 37 degrees C. Both carpet-exposed and control mice displayed some effects, such as body weight loss, mortality and pathological lesions, that were due to the exposure system. There was no mortality or neurotoxicity, nor were there clinical signs or pathological lesions as a result of carpet exposures. Mice exposed to carpet heated to 70 degrees C had slightly decreased respiratory rates and an increased incidence of breathing patterns indicative of sensory irritation. Therefore, none of the results reported by the private testing laboratory could be reproduced when this carpet was heated to temperatures below 70 degrees C, and slight sensory irritation was the only effect observed at the 70 degrees C test conditions. Effects from the exposure system itself made interpretation of results difficult, and concurrent controls were considered essential for interpretation of data.

Chronic toxicity/oncogenicity of dimethylformamide in rats and mice following inhalation exposure.

The potential chronic toxicity and oncogenicity of dimethyl-formamide (DMF) was evaluated by exposing male and female rats and mice to 0, 25, 100, or 400 ppm DMF for 6 hr/day, 5 days/week for 18 months (mice) or 2 years (rats). Clinical pathology was evaluated at 3, 6, 12, 18, and 24 (rats only) months. An interim euthanasia for rats occurred at 12 months and hepatic cell proliferation in rats and mice was examined at 2 weeks, 3 months, and 12 months. No compound-related effects on clinical observations or survival were observed. Body weights of rats exposed to 100 (males only) and 400 ppm were reduced. Conversely, body weights were increased in 400 ppm mice. No hematologic changes were observed in either species. Serum sorbitol dehydrogenase activity was increased in rats exposed to 100 or 400 ppm. There were no compound-related effects on the estrous cycle of rats or mice at any concentration. Compound-related morphological changes were observed only in the liver. In rats, exposure to 100 and 400 ppm produced increased relative liver weights, centrilobular hepatocellular hypertrophy, lipofuscin/hemosiderin accumulation in Kupffer cells, and centrilobular single cell necrosis (400 ppm only). In mice, increased liver weights (100 ppm males, 400 ppm both sexes), centrilobular hepatocellular hypertrophy, accumulation of lipofuscin/hemosiderin in Kupffer cells, and centrilobular single cell necrosis were observed in all exposure groups. These observations occurred in a dose-response fashion and were minimal at 25 ppm. No increase in hepatic cell proliferation was seen in mice or female rats. Slightly higher proliferation was seen in male rats exposed to 400 ppm at 2 weeks and 3 months but not at 12 months. Dimethylformamide was not oncogenic under these experimental conditions in either the rat or mouse.

Lung response to ultrafine Kevlar aramid synthetic fibrils following 2-year inhalation exposure in rats.

Four groups of 100 male and 100 female rats were exposed to ultrafine Kevlar fibrils at concentrations of 0, 2.5, 25, and 100 fibrils/cc for 6 hr/day, 5 days/week for 2 years. One group was exposed to 400 fibrils/cc for 1 year and allowed to recover for 1 year. At 2.5 fibrils/cc, the lungs had normal alveolar architecture with a few dust-laden macrophages (dust cell response) in the alveolar airspaces. At 25 fibrils/cc, the lungs showed a dust cell response, slight Type II pneumocyte hyperplasia, alveolar bronchiolarization, and a negligible amount of collagenized fibrosis in the alveolar duct region. At 100 fibrils/cc, the same pulmonary responses were seen as at 25 fibrils/cc. In addition, cystic keratinizing squamous cell carcinoma (CKSCC) was found in 4 female rats, but not in male rats. Female rats had more prominent foamy alveolar macrophages, cholesterol granulomas, and alveolar bronchiolarization. These pulmonary lesions were related to the development of CKSCC. The lung tumors were derived from metaplastic squamous cells in areas of alveolar bronchiolarization. At 400 fibrils/cc following 1 year of recovery, the lung dust content, average fiber length, and the pulmonary lesions were markedly reduced, but slight centriacinar emphysema and minimal collagenized fibrosis were found in the alveolar duct region. One male and 6 female rats developed CKSCC. The lung tumors were a unique type of experimentally induced tumors in the rats and have not been seen as spontaneous tumors in man or animals. Therefore, the relevance of this type of lung tumor to the human situation is minimal.

Use of dose-response data to compare the skin sensitizing abilities of dicyclohexylmethane-4,4'-diisocyanate and picryl chloride in two animal species.

Groups of English smooth-haired guinea pigs and BALB/cBy mice were exposed to dicyclohexylmethane-4,4'-diisocyanate (HMDI) and picryl chloride (PiCl) by topical exposure. Guinea pigs were challenged 7 days later by patch testing and responses graded at 24 hr. Application of high doses of each chemical resulted in extensive erythema and a large proportion of animals became sensitized. Lower doses produced less sensitivity. In mice, sensitivity was assessed by extent of ear swelling upon challenge 5 days following the sensitization exposure. In mice, as well as in guinea pigs, a dose-response relationship was apparent between the sensitizing dose and both severity of response and number of animals responding. For mice, comparison of the dose of chemical required to sensitize 50% of the animals (SD50) yielded 0.40 mg/kg for PiCl and approximately 0.20 mg/kg for HMDI. Use of the SD50 should provide a method for assessing the ability of various chemicals to cause contact sensitivity.