Dennis W. Lynch

Sister-chromatid exchanges and chromosome aberrations in lymphocytes from monkeys exposed to ethylene oxide and propylene oxide by inhalation

The ability of long-term exposures to inhaled ethylene oxide (EO) and propylene oxide (PO) to induce sister-chromatid exchanges (SCEs) and chromosome aberrations in peripheral lymphocytes of monkeys was investigated. Five groups of adult male cynomolgus monkeys were exposed at 0 (shared control), 50, or 100 ppm EO, and at 100 or 300 ppm PO (7 hr/day, 5 days/week) for 2 years. EO exposures at 50 and 100 ppm resulted in statistically significant increases in sister-chromatid exchange rates and in the incidence of chromosome aberrations in monkey lymphocytes. Both EO-exposed groups had increased numbers of SCEs/metaphase compared to controls, with the SCEs/metaphase of the EO 100 ppm group also significantly elevated versus the EO 50 ppm group. Variability of SCEs/metaphase within each monkey increased even more than the increase in total SCEs/metaphase group with increasing EO exposure. Chromatid-type aberrations were also significantly increased for both EO 50 and EO 100 ppm groups compared to controls. Statistically significant increases in the number of chromosome-type aberrations (excluding gaps) were found only in the EO 100 ppm group. Combined chromatid- and chromosome-type aberrations were increased in both EO 50 and EO 100 ppm groups. No group differences in the number of gaps were found. In lymphocytes from monkeys exposed at 100 and 300 ppm PO, there were no group differences compared to controls for any variable-chromatid or chromosome-type aberrations, gaps, or SCEs/metaphase. These results indicate that EO is a more potent clastogen than PO and demonstrate, for the first time, statistically significant effects of EO on both SCEs and chromosome aberrations in lymphocytes of nonhuman primates.

A Chronic Inhalation Toxicity Study of Diesel Engine Emissions and Coal Dust, Alone and Combined:

To evaluate the potential health hazards of diesel engine emissions in underground coal mines, inhalation studies were performed using three species of animals. A wide range of toxicological responses was measured. Exhaust was provided by a 425 in.3 displacement four-cycle, water-cooled, naturally aspirated diesel engine (Caterpillar Model 3304) equipped with a water scrubber. Exposures were 7 h/day, 5 days/week, for periods up to 24 months. Micronized coal dust was generated using a Wright dust feeder. Four exposures were evaluated: (1) filtered ambient air, (2) 2 mg/m3 diesel particulate, (3) 2 mg/m3 respirable coal dust, and (4) 1 mg/m3 each of 2 and 3. Gaseous and vapor concentrations were similar in both exposures employing diesel exhaust. Male cynomolgus monkeys, Fischer-344 male and female rats, and female CD-1 mice were the experimental subjects. Monkeys were sacrificed at 24 months, rats at 3, 6, 12, and 24 months, and mice at 1, 3, and 6 months. Gross morphology and histopathology demonstrated that both diesel and coal dust particles are deposited in the lungs and retained in alveolar tissue. Alveolar type II cell hyperplasia and pulmonary lipidosis occurred in rats, being most evident in rats exposed to diesel exhaust alone. There was, however, no evidence of emphysema or chronic bronchitis, and only minimal fibrosis was seen in association with the retained particulate. Both particulates affected the defense mechanisms of the lung. Exposure to coal dust activated responses associated with phagocytosis, whereas exposure to diesel exhaust depressed them. Severity of influenza challenge increased concomitantly with decreased interferon production in diesel-exposed mice. Exposure to diesel emissions did not result in genotoxic effects as measured by increases in sister chromatid exchange, chromosomal aberrations, micronucleus testing, and urine genotoxic assays. Pulmonary function studies in monkeys showed mild obstructive airway disease in coal dust, diesel exhaust, and the combined exposed animals. This effect was most pronounced in monkeys exposed to diesel exhaust. Evidence of restrictive lung disease was not seen in any group. Clearance of F3O4 particles appeared to be stimulated by exposure to diesel exhaust in the first 3 months, but long-term clearance of diesel particulate appeared to be inhibited. No evidence was found for increases in tumorogenicity (rats) or induction of xenobiotic metabolizing enzymes in the lung or liver (rats). Humoral and cellular immunities were not significantly affected by exposure (rats). No adverse seminal effects were observed in monkeys exposed for 2 years. There was no frank evidence of chronic toxicity as demonstrated by changes in mortality, body weight gains, organ-body weight ratios, or clinical parameters in rats or monkeys. Synergistic effects between diesel exhaust and coal dust were not demonstrated.

Inhibition of rat heart mitochondrial electron transport in vitro: implications for the cardiotoxic action of allylamine or its primary metabolite, acrolein

Allylamine (3-aminopropene) is a specific cardiac toxicant that causes aortic, valvular and myocardial lesions in many species. Myocardial necrosis can be observed 24 h after a single dose. Acute toxicity is believed to involve metabolism of allylamine to highly reactive acrolein (2-propenal). Allylamine has been shown to bind to mitochondria from aorta and heart, suggesting that the subcellular site of injury is at or near the mitochondrion. The present investigation compared the effect of allylamine and its primary metabolite, acrolein, on electron transport and oxidative phosphorylation in mitochondria isolated from rat heart (RHM). Both compounds weakly inhibited mitochondrial electron transport with either the combination of glutamate, malate, and malonate (GMM, NADH-linked) or succinate as substrate. Comparisons of the slopes of concentration-effect regression (range of concentrations tested, 0.20-2.0 mM) lines showed acrolein to have significantly greater inhibitory effects than allylamine (range of concentrations tested, 0.22-6.4 mM) on GMM oxidation, while no significant difference in the abilities of the compounds to inhibit succinate oxidation were observed, indicating site preferences for inhibitory action. The addition of an uncoupling agent could not reverse inhibition with either substrate system. These results indicate that both the parent compound and its proposed metabolite primarily inhibit electron transport with little direct effect on the coupling mechanism. The State III EC50 (effective concentrations for 50% inhibition of control mitochondrial enzyme activities) for allylamine (2.29 mM with succinate as substrate and 1.22 mM with GMM) and acrolein (0.80 mM with succinate as substrate and 0.39 mM with GMM) are probably too great to invoke the direct action of either the parent compound or its oxidized metabolite on mitochondrial electron transport as a primary mechanism in the cardiotoxic action of allylamine.

Workgroup report: Implementing a national occupational reproductive research agenda - Decade one and beyond

The initial goal of occupational reproductive health research is to effectively study the many toxicants, physical agents, and biomechanical and psychosocial stressors that may constitute reproductive hazards in the workplace. Although the main objective of occupational reproductive researchers and clinicians is to prevent recognized adverse reproductive outcomes, research has expanded to include a broader spectrum of chronic health outcomes potentially affected by reproductive toxicants. To aid in achieving these goals, the National Institute for Occupational Safety and Health, along with its university, federal, industry, and labor colleagues, formed the National Occupational Research Agenda (NORA) in 1996. NORA resulted in 21 research teams, including the Reproductive Health Research Team (RHRT). In this report, we describe progress made in the last decade by the RHRT and by others in this field, including prioritizing reproductive toxicants for further study; facilitating collaboration among epidemiologists, biologists, and toxicologists; promoting quality exposure assessment in field studies and surveillance; and encouraging the design and conduct of priority occupational reproductive studies. We also describe new tools for screening reproductive toxicants and for analyzing mode of action. We recommend considering outcomes such as menopause and latent adverse effects for further study, as well as including exposures such as shift work and nanomaterials. We describe a broad domain of scholarship activities where a cohesive system of organized and aligned work activities integrates 10 years of team efforts and provides guidance for future research.

Subchronic inhalation toxicity of isobutyl nitrite in BALB/c mice. I. Systemic toxicity.

The effects of subchronic inhalation exposure to isobutyl nitrite (IBN) on body weight, selected organ weights, hematology, and gross pathology and histopathology of BALB/c mice were evaluated. Mice of both sexes were exposed at 0, 20, 50, or 300 ppm IBN for 6.5 h/d, 5 d/wk for up to 18 wk. Most changes in measured indices occurred in mice exposed at 300 ppm IBN and included decreased thymus weight (females); decreased liver weight (males); decreased white blood cell counts (males); mild focal hyperplasia and vacuolization of the epithelium lining bronchi and bronchioles of the lungs (males and females). Organ weight and hematologic changes, however, were not accompanied by any observed histologic changes. In addition, elevated methemoglobin concentrations were detected in mice of both sexes exposed at 50 and 300 ppm IBN. Body weights were not adversely affected by exposure. These data suggest that mild tissue injury, restricted to the lung, and methemoglobinemia are the major toxic effects observed following exposures of mice to IBN at concentrations up to 300 ppm for 18 wk. No treatment-related effects were noted in mice exposed at 20 or 50 ppm IBN, except for slight elevations in methemoglobin concentrations in mice exposed at 50 ppm.

Inhalation toxicity of methyl n-amyl ketone (2-heptanone) in rats and monkeys

Abstract A subchronic inhalation study was conducted in which male rats and monkeys were exposed at 0 (controls), 131, or 1025 ppm methyl n -amyl ketone (MAK) for 10 months (6 hr/day, 5 days/week). Comprehensive cardiopulmonary, clinical chemistry, metabolism, and tissue distribution studies were performed. No statistically significant alterations of pulmonary function, electrocardiographic, or clinical chemistry parameters were observed. Methyl n -amyl ketone and methyl n -amyl alcohol were detected in urine and serum from monkeys exposed to MAK for 10 months. Inhaled MAK did not induce rat liver microsomal enzymes. Results of tissue distribution studies of [ 14 C]MAK in rats comparing ip and inhalation routes of exposure were similar, and preexposure to unlabeled MAK did not alter the tissue distribution compared to animals without prior exposure to MAK. Liver tissue had the highest level of radioactivity regardless of the route of administration; however, no liver pathology was observed. Urinary excretion accounted for 25% of the administered dose after 12 hr. In general, a lack of toxicity was noted in both species exposed to MAK.

Subchronic inhalation toxicity of isobutyl nitrite in BALB/c mice. II. immunotoxicity studies

Initial epidemiologic studies of acquired immunodeficiency syndrome (AIDS) occurring in homosexual men identified the use of the inhalants amyl, butyl, and isobutyl nitrite as possible risk factors contributing to the disease. Because of the lack of immunotoxicological data on these chemicals, we studied the effects of subchronic exposure to isobutyl nitrite (IBN) on the immune system. BALB/c mice were exposed to either 50 or 300 ppm IBN for 6.5 h/d, 5 d/wk for up to 18 wk. After 7, 13, or 18 wk of exposure, mice were killed and the following assays were performed. Antibody producing cells were enumerated by a slide plaque assay on animals immunized with sheep red blood cells while still in exposure chambers. The lymphocyte proliferative response to mitogens (phytohemagglutinin, concanavalin A, pokeweed mitogen, and lipopolysaccharide) was tested using several concentrations of each mitogen. Additional mice were immunized with Freunds complete adjuvant 21 d prior to death and were tested for delayed hypersensitivity response to purified protein derivative by a radiometric skin test. Finally, the relative numbers of T cells and T-cell subsets among splenic lymphocytes from exposed and control animals were determined. At the time periods tested there were no discernable immunotoxic effects observed in the exposed animals in any of the assays performed. These results indicate that IBN, at the dosages tested, had no discernable detrimental effect on the immune system of mice.

Chronic Effects of Inhaled Amorphous Silicas in Animals

Rats, guinea pigs, and monkeys were exposed by inhalation (5.5 to 6 h/day, 5 days/week) for up to 18 months at 15 mg/m 3 concentrations of three types of synthetic amorphous silicas: fume, gel, and precipitated. Autopsieson rats were performed after 3, 6, and 12 months of exposure, and on guinea pigs and monkeys after 10 to 18 months of exposure. The most significant finding was the deposition of large quantities of amorphous silica in macrophages in the lungs and tracheal lymph nodes of exposed monkeys. Relatively few or no macrophages containing particles of amorphous silica were found in the lungs and lymph nodes of the guinea pigs and rats. It is also significant that fume silica induced early nodular fibrosis in the lungs of the monkeys. Correlating these histological findings, lung-function studies indicated statistically significant differences in lung volume and ventilatory mechanics measurements between those monkeys exposed to fume silica and the control group. In addition, monkeys exposed to precipitated silica demonstrated significantly lower lung volumes compared with controls, while monkeys exposed to silica gel had significant changes in ventilatory performance and mechanical properties.

Effects of inhalation exposures and intraperitoneal injections of methyl n-amyl ketone on multiple fixed-ratio, fixed-interval response rates in rats☆

Twelve rats were tested daily for 1 hr on a multiple fixed-ratio, fixed-interval schedule of reinforcement. The rats were given ip injections of methyl n-amyl ketone (MAK; 2-heptanone), an industrial solvent, in concentrations ranging from 18 to 175 mg/kg on one to eight occasions each, and were tested 15 min after the injections. When compared to the preceding control (saline) day, there was little or no fixed interval (FI) response rate change at 18 mg/kg, moderate FI rate decreases at 37 and 74 mg/kg and a near cessation of responding at 175 mg/kg MAK. Changes were statistically significant (p<0.01) at 37, 74, and 175 mg/kg. The quantity of MAK which altered behavior was about 5% of the dose that was subsequently determined to be lethal to two-thirds of the test animals. Time course data indicated that the behavioral changes seen at 15 min after 175 mg/kg injections persisted for at least 1 hr but had vanished by 2.25 hr after the injections. Over the course of the study, there was a consistent reduction in the magnitude of the behavioral changes, suggesting that tolerance developed. Inhalation exposures below 1500 ppm did not cause behavioral changes, but those in excess of 1575 ppm had effects similar to the 175 mg/kg injections when testing began 5–10 min after termination of the inhalation exposure. However, a lack of opportunity for replications made it impossible to achieve statistical significance.

Subchronic Inhalation of Triethylamine Vapor in Fischer-344 Rats: Organ System Toxicitya

Male and female F-344 rats were exposed at 0, 25, or 247 ppm triethylamine (TEA) vapor, 6 hr per day, 5 days per week for up to 28 weeks in order to characterize the subchronic organ system toxicity. Rats were weighed biweekly and scheduled sacrifices were performed following about 30, 60, and 120 days of exposure. No statistically significant treatment-related effects on organ weights, hematology, clinical chemistry, or electrocardiographic indices were observed. Body weight gain was not affected by TEA treatment. No physiologic or pathologic evidence of cardiotoxicity was seen in rats exposed to either TEA concentration for up to 28 weeks. No gross or histopathologic lesions attributable to TEA exposure were noted in any of the organs examined, including the nasal passages. This latter finding is in marked contrast to previously reported findings from this laboratory in which squamous metaplasia, suppurative rhinitis, and lymphoid hyperplasia were found in the respiratory epithelium of F-344 rats exposed to the structurally related chemical, diethylamine, under the same conditions as this study (Lynch et al., 1986).