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Metal oxides containing either Ni alone (NiO's) or both Ni and Cu (Ni-CuO's) are encountered during Ni refining. Six NiO compounds calcined at temperatures ranging from less than 650 to 1045 degrees and four Ni-CuO's containing from 6.9 to 28% Cu and 44 to 69% Ni were screened for their in vitro cytotoxicity to alveolar macrophages (AM). NiO's were less toxic to rat AM than were the Ni-CuO compounds. The toxicity of the Ni-CuO compounds increased with increasing Cu content and decreasing Ni content of the molecules, indicating that the toxicity was due to the Cu content of the molecules. AM obtained from beagle dogs, F344/N rats, and B6C3F1 mice displayed the following species sensitivities: dog greater than rat = mouse, with dog AM being most sensitive. The observed differences in species sensitivities correlated with differences in the phagocytic abilities of dog, rat, and mouse AM, with the ranking of phagocytic abilities of the AM in decreasing order of ability being dog greater than rat greater than mouse.

To study the changes in collagen metabolism that occur in the pathogenesis of pulmonary fibrosis, female rats were exposed to 0, 0.57, and 1.1 ppm ozone for 19 hr/day for 11 days and sacrificed 12 or 60 days after initiation of exposure. The lungs of rats sacrificed at 12 days after initiation of exposure to 1.1 ppm had interstitial pneumonia characterized by a mixed inflammatory cell infiltrate, type II cell hyperplasia, and fibroplasia, a proliferation of the collagen-producing cells; increased cathepsin D and macrophage elastase activity, indicating macrophage-induced proteinolysis; a reduced percentage of the increased collagen production that was ultrafilterable, indicating a decreased rate of intracellular degradation of newly produced collagen prior to its secretion; and increased lavage fluid hydroxyproline, indicating turnover of extracellular collagenous matrix. Reduced intracellular collagen degradation correlated directly with both increased net collagen production and fibroplasia in rats exposed to 1.1 ppm ozone for 11 days. These changes preceded an increased total lung collagen and the development of modest fibroplasia and fibrosis in the alveolar duct regions by 60 days after the 1.1 ppm ozone exposure was initiated.

The quinoline dye 2-(2'-quinolyl)-1,3-indandione or Solvent Yellow 33 (SY) and the anthraquinone dye 1,4-di-p-toluidinoanthraquinone or Solvent Green 3 (SG) are used in many manufactured products including military smoke grenades. During manufacturing, SY or a combination of both SY and SG can be released into the air, exposing factory workers by inhalation to these dye compounds. The potential inhalation toxicity of these compounds was tested by exposing F344/N rats to different concentrations of SY or SY/SG dye mixture (30:70 w/w) for 6 hr/day, 5 days/week for 4 or 13 weeks. In the 4-week studies, rats were exposed to SY aerosols at average concentrations of 10 +/- 5, 51 +/- 10, or 230 +/- 30 mg/m3 (means +/- SD) or SY/SG aerosols at average concentrations of 11 +/- 5, 49 +/- 11, or 210 +/- 50 mg/m3 (means +/- SD). Rats exposed to the highest concentration of SY or SY/SG had body weights that were approximately 8% or 7% less, respectively, than their controls after exposure. Rats exposed to the highest concentration of SY/SG for 4 weeks also had reduced pulmonary gas exchange efficiency, airflow obstruction, mild pulmonary inflammation, slight Type II pulmonary epithelial cell hyperplasia, and proliferation of vacuolated alveolar macrophages. In the 13-week studies, rats were exposed to SY aerosols at average concentrations of 1.0 +/- 0.2, 10.8 +/- 1.8, or 100 +/- 17 mg/m3 (means +/- SD) or SY/SG aerosols at average concentrations of 1.1 +/- 0.5, 10.2 +/- 3.1, or 101 +/- 23 mg/m3 (means +/- SD). Animals exposed to the highest concentration of SY or SY/SG for 13 weeks had body weights that were approximately 5 or 9% less, respectively, than their controls after exposure and had accumulation of vacuolated alveolar macrophages in lungs. Rats exposed to the highest concentration of SY/SG dye mixture for 13 weeks also had indications of mild pulmonary inflammation and slight Type II pulmonary epithelial cell hyperplasia. Very little SY was found in lungs after any exposures, indicating its clearance from lungs was at a rapid rate. However, significant amounts of the SG component of the SY/SG mixture were detected in lungs after each exposure. Lung clearance half-times of SG from the 13-week exposure were estimated to be approximately 280 days. In summary, neither test material appeared to be highly toxic following inhalation. However, the slightly higher toxicity observed for SY/SG over SY alone is probably related to the longer lung retention of the SG component of the dye mixture.

Daily washing in vivo of the lung with 0.15 M saline did not deplete the Beagle dog lung of surfactant lipids, but rather increased the quantity of surfactant lipid in the tissue. Replacement time for the lung lipids removed by the lavage was approximately 5 hours. This rate is one indication of the time required for movement of surfactant lipid from storage areas to the surface of the alveoli. The increase in tissue surfactant lipid following multiple lavage suggests that the rate of surfactant lipid synthesis is controlled in part by the level of surfactant lipid in the alveoli.

Two reactions that chemically alter primary aromatic amines (PAA) were used to assess the contribution of these compounds to the indirect bacterial mutagenicity of tar from an experimental low Btu gasifier. The first reaction, nitrosation, effectively eliminated the mutagenicity of several PAA standards and a coal oil when run in a low pH media (1.2). When applied to gasifier tar, extensive direct (not requiring metabolic activity) mutagenicity was generated. This direct mutagenicity limited the interpretation of results. When the pH of the reaction media was raised to 2.5, the mutagenicity of PAA standards and the coal oil were still greater than 90% eliminated, however, no direct mutagenicity was observed for the gasifier tar. Furthermore, only 61% of the indirect (requiring metabolic activation) mutagenicity was eliminated. Acetylation reduced the indirect activity of most primary amine standards by greater than 79%. Acetylation of the tar likewise eliminated part, but not all, of the activity, whereas most of the activity of the coal oil was eliminated. These results indicated that a much lower percentage of the mutagenic activity of low Btu coal tar samples was due to primary aromatic amines than was the case for coal oil.

The process gas stream of an experimental pressurized McDowell-Wellman stirred-bed low-Btu coal gasifier, and combustion products of the clean gas were characterized as to their mutagenic properties and chemical composition. Samples of aerosol droplets condensed from the gas were obtained at selected positions along the process stream using a condenser train. Mutagenicity was assessed using the Ames Salmonella mammalian microsome mutagenicity assay (TA98, with and without rat liver S9). All materials required metabolic activation to be mutagenic. Droplets condensed from gas had a specific mutagenicity of 6.7 revertants/microgram (50,000 revertants/liter of raw gas). Methylnaphthalene, phenanthrene, chrysene, and nitrogen-containing compounds were positively identified in a highly mutagenic fraction of raw gas condensate. While gas cleanup by the humidifier-tar trap system and Venturi scrubber led to only a small reduction in specific mutagenicity of the cooled process stream material (4.1 revertants/microgram), a significant overall reduction in mutagenicity was achieved (to 2200 revertants/liter) due to a substantial reduction in the concentration of material in the gas. By the end of gas cleanup, gas condensates had no detectable mutagenic activity. Condensates of combustion product gas, which contained several polycyclic aromatic compounds, had a specific mutagenicity of 1.1 revertants/microgram (4.0 revertants/liter). Results indicate that the process stream material is potentially toxic and that care should be taken to limit exposure of workers to the condensed tars during gasifier maintenance and repair and to the aerosolized tars emitted in fugitive emissions. Health risks to the general population resulting from exposure to gas combustion products are expected to be minimal.

The effects of DMSO are thought to result from the formation of hydrogen bonds with proton-donor groups on biopolymers, which are stronger than those formed with water. Since DMSO contains methyl groups, however, effects on hydrophobic bonding in proteins could be expected at higher DMSO levels. Our studies of the effects of DMSO on model subunit proteins can be interpreted in the above terms. At a concentration of 20% or less, DMSO changed glutamate dehydrogenase into the inactive monomer and the effects were fully reversible with the activator (ADP). Higher DMSO levels resulted in irreversible inactivation. The predominant effect noted on beta-glucuronidase was irreversible inactivation by 20% or more DMSO at 37 degrees C. Purified beta-glucuronidase exhibited an activation in 20% DMSO at high substrate levels; this resulted from an apparent substrate inhibition in the absence of DMSO. DMSO inhibited the clotting of fibrinogen by purified thrombin, but the major effect appeared to be due to competition between thrombin and DMSO for binding sites on fibrinogen. These effects appear to be largely due to interactions between DMSO and hydrophobic bonding in fibrinogen, although DMSO also appears to interfere with the aggregation of fibrin monomers through its effects on hydrophilic groups. These results suggest that reversible alterations in protein structure are the major effect of exposure of subunit proteins to low DMSO levels at low temperatues, while irreversible denaturation of subunit proteins may be an appreciable effect a higher temperatures and higher DMSO concentrations.

Isoprene (2-methyl-1,3-butadiene), a volatile monomer occurring in the natural environment and used in the manufacture of elastomers, is a close chemical relative of the animal carcinogen 1,3-butadiene. To obtain toxicokinetic data for inhaled isoprene, male F344 rats were exposed in groups of 30 to 14C-labeled isoprene vapor at four concentrations from 8 to 8200 ppm. The percentage of the inhaled isoprene that was metabolized decreased with increasing exposure concentration. The percentage of the total metabolites (that is, non-isoprene-retained 14C) excreted in urine and feces or expired was determined as a function of vapor concentration. About 75% of the total metabolites was excreted in urine. This was independent of inhaled isoprene concentration. After exposure to 8200 ppm, a larger percentage of the metabolites was excreted in feces than after exposure to lower concentrations. Using vacuum line techniques, blood metabolite concentrations were determined as functions of both vapor concentration and exposure duration. At one exposure concentration (1480 ppm) metabolites were measured in the nose, lungs, liver, kidney, and fat, as well as in blood. A mutagenic metabolite, isoprene diepoxide, was tentatively identified in all tissues examined. Between 0.0018 and 0.031% of the inhaled 14C label was tentatively identified as this metabolite in blood. The relative amount of the metabolites present in blood was highest for low concentrations of inhaled isoprene and for shorter exposure durations. Body fat appeared to be a reservoir for both isoprene metabolites and isoprene itself. The appearance of metabolites in the respiratory tract after short exposure durations together with low blood concentrations of isoprene indicated that substantial metabolism of inhaled isoprene in the respiratory tract may occur.

An epithelial cell strain (LEC, lung epithelial cells) was established from the lungs of an adult Fischer-344 rat. The cells had properties of normal, untransformed cells in culture which include contact inhibition of growth, inability to grow in soft agar, and inability to form tumors when injected subcutaneously into syngeneic hosts. Low passage LEC were diploid, while high passage cells were aneuploid with an extra chromosome. LEC resembled type II alveolar cell in vivo in that they had lamellar inclusions in their cytoplasm, microvilli on cell surface, and that they could incorporate [14C] choline into disaturated phosphatidyl choline, a surfactant lipid. The ability of LEC to activate procarcinogens was demonstrated by co-cultivation of LEC with Chinese hamster ovary (CHO) cells in the presence of procarcinogens benzo[a]pyrene and 3-methyl cholanthrene. While these procarcinogens were non-mutagenic to CHO cells which lacked the metabolic activation capacity, they were mutagenic when LEC were co-cultured with the CHO cells. By maintaining LEC cells on collagen gel without overlying medium, the cells could be directly exposed to noxious airborne agents. Using this technique, the cytotoxicity of NO2 was detected. Because of the lung epithelial origin, the ability to metabolize xenobiotics, and the exposure system to airborne agents, the LEC strain can be a useful and relevant in vitro system for the toxicological studies of inhalable materials.

Sialic acid (14.6 mug/mg protein) was quantitated in the non-cellular material removed from the lung of Beagle dogs by lavage. Sialic acid did not affect the dynamic surface tension properties of either the total alveolar lipid removed by lavage or of its major lipids, dipalmitoyl lecithin (DPL) and dipalmitoyl phosphatidyl glycerol (DPG). The presence of divalent cation (Ca2+, Mg2+, Mn2+ and Zn2+) or a lowered pH in the subphase medium lowered the surface tension during the expansion phase of the total alveolar lipid film when it was compressed and expanded on a Wilhelmy trough. Films of DPL behaved similarly, but no pH effect was observed with DPG monolayers. The cation effect manifested itself in the same direction as the value of the individual stability constants (Zn2+ greater than Mn2+ greater than Mg2+ greater than Ca2+) which suggests ionic binding of the cations to the phosphate group of the phospholipids. A physiological advantage of such an effect may lie in the conservation of the energetically favorable low surface tension state achieved during film compression with a minimum of surfactant lipid.

The glucuronide and sulfate conjugates of benzene metabolites as well as muconic acid and pre-phenyl- and phenylmercapturic acids were separated by ion-pairing HPLC. The HPLC method developed was suitable for automated analysis of a large number of tissue or excreta samples. p-Nitrophenyl [14C]glucuronide was used as an internal standard for quantitation of these water-soluble metabolites. Quantitation was verified by spiking liver tissue with various amounts of phenylsulfate or glucuronides of phenol, catechol, or hydroquinone and analyzing by HPLC. Values determined by HPLC analysis were within 10% of the actual amount with which the liver was spiked. The amount of metabolite present in urine following exposure to [3H]benzene was determined using p-nitrophenyl [14C]glucuronide as an internal standard. Phenylsulfate was the major water-soluble metabolite in the urine of F344 rats exposed to 50 ppm [3H]benzene for 6 h. Muconic acid and an unknown metabolite which decomposed in acidic media to phenylmercapturic acid were also present. Liver, however, contained a different metabolic profile. Phenylsulfate, muconic acid, and pre-phenylmercapturic acids as well as an unknown with a HPLC retention time of 7 min were the major metabolites in the liver. This indicates that urinary metabolite profiles may not be a true reflection of what is seen in individual tissues.

This report describes 25 patients with reflux-induced cricopharyngeal dysphagia ultimately requiring surgical management. Eighteen patients underwent cricopharyngeal myotomy alone and seven patients required cricopharyngeal myotomy after an antireflux operation failed to correct this symptom. Cricopharyngeal incoordination was demonstrated at manometry in over 90% of patients. Treatment included cricopharyngeal myotomy, which was extended proximally to the pharynx and distally to the intrathoracic esophagus. Results were excellent to satisfactory in 24 of 25 patients. Pathologic examination of the cricopharyngeal muscle demonstrated a wide variety of myopathic degenerative changes. We stress that cricopharyngeal myotomy may be performed even in the presence of reflux without fear of subsequent aspiration.

The distribution, retention, and fate of 2[3H]aminoanthracene (2-AA) were determined in male Fischer-344 rats after inhalation exposure (70 micrograms/liter; activity mass median diameter = 2.1 micron) for 30 min. Radioactivity was found in the turbinates, trachea, lungs, liver, kidneys, and gastrointestinal tract 20 min after exposure. Lower quantities of radioactivity were found in fat, brain, testes, and muscle. Inhaled 2-AA was excreted predominantly in feces (80%). The remaining 2-AA was excreted in urine. Organic soluble radioactivity was released from urinary conjugates after treatment of urine with acid and with beta-glucuronidase. This result suggests that glucuronides of both ring- and N-hydroxy-2-AA were excreted. The remaining radioactivity was water soluble and accounted for approximately one-half of the total urinary radioactivity. The organic soluble radioactivity found after hydrolysis indicated that inhaled 2-AA is extensively metabolized by rats after inhalation and excreted as conjugated metabolites. Eighty-three percent of orally administered material was absorbed into the body from the GI tract; thus, material cleared by mucociliary action after inhalation would contribute to total body burden. 2-AA, a nitrogen substituted polycyclic aromatic hydrocarbon (PAH), was very similar to other PAHs in its rapid distribution throughout the body and in its route of excretion after inhalation.

The purpose of this study was to determine how prior exposure of rats to graded concentrations of diesel exhaust would affect respiratory tract tissue metabolism of 1-nitropyrene (NP), a known constituent of diesel exhaust. Rats were exposed (whole body) 7 hr/day, 5 days/week for 4 weeks to clean air (controls) or to diluted diesel exhaust containing concentrations of 350, 3300, and 7400 micrograms particles/m3. After exposures, nasal tissue and lungs from rats were tested for their ability to metabolize NP. Rat nasal tissue was incubated for 10 min with 20 microM 1-[14C]NP. Isolated perfused rat lungs were perfused for 90 min with 25 microM 1-[14C]NP. NP metabolites formed in rat nasal tissue and the isolated perfused rat lung were separated by high-pressure liquid chromatography. Exposure of rats to 7400 micrograms particles/m3 for 4 weeks resulted in significant increases (twofold) in rates of NP metabolism in both nasal tissue (440 pmol/mg protein/min) and the isolated perfused rat lung (112 nmol/g lung). Exposure of rats to lower concentrations of diesel exhaust did not increase the rates of NP metabolism in either rat nasal tissue or perfused rat lungs. In all cases, the major metabolites of NP formed in nasal tissue and perfused lungs were 3-, 6-, and 8-hydroxy-1-nitropyrene and 4,5-dihydro-4,5-dihydroxy-1-nitropyrene. A fourfold increase was observed in the amounts of 14C covalently bound in lungs from rats exposed to 7400 micrograms particles/m3. The results from this study indicate that exposure to diesel exhaust particles significantly alters metabolism and subsequent covalent binding of NP.

The present study was designed to characterize the quantity of mucosubstances in surface epithelia of the rat nasal cavity and nasopharynx after short-term ozone exposure. Rats were exposure. Nasal cavities were processed for morphometric analysis of intraepithelial mucosubstances. Compared to controls, rats exposed to 0.12 ppm ozone had increased amounts of stored mucosubstances within epithelium lining the medial aspect of the nasal turbinate, but no change within the epithelium of the nasopharynx. Rats exposed to 0.8 ppm ozone had increased quantities of stored mucosubstances within the transitional and respiratory epithelia lining turbinates and lateral walls of the anterior nasal airway, and significant decreases in stored mucosubstances within the epithelium of the nasal septum at the end of exposure. Seven days after the end of exposure, the amounts of intraepithelial mucosubstances returned to control levels along the septum, but remained greater than those of controls along the turbinates and nasopharynx. We conclude that exposures to ambient levels of ozone induce significant changes in the stored secretory product of nasal epithelium in the rat, and that these changes persist for at least 7 days after cessation of exposure.

Nitroaromatics in general, and 1-nitropyrene in particular, are potent bacterial mutagens and animal carcinogens. Their importance as possible human carcinogens is difficult to assess because they are usually found in the environment as the products of combustion processes, and so they usually exist with many other compounds associated with airborne particles. The experiments reported here were carried out to determine if the inhalation of particle-associated 1-nitropyrene, or the concomitant exposure to an irritant gas, would alter the tissue distribution of 1-nitropyrene or its metabolites, compared to their distribution after inhalation of pure 1-nitropyrene. These experiments were intended to yield insights into the mechanisms involved in the potential carcinogenicity of particle-associated nitroaromatics as inhaled in the environment from automotive emissions and other sources. Groups of Fischer 344 rats inhaled pure 14C-1-nitropyrene aerosols, with and without coexposure to 5 parts per million sulfur dioxide, or 14C-1-nitropyrene adsorbed onto gallium oxide particles, with and without coexposure to sulfur dioxide, for four weeks. Lung retention of 14C-1-nitropyrene was not prolonged by its association with gallium oxide particles or by coexposure to sulfur dioxide. There was a marked inflammatory and fibrogenic response to the gallium oxide particles. Another set of experiments was carried out in which rats were exposed to 14C-1-nitropyrene either as pure aerosol or adsorbed onto carbon black particles. The amount of 14C in the lung that was bound to carbon black particles steadily decreased with time after exposure, compared to total lung 14C, indicating removal of 14C from the particles. Thirty minutes after exposure, the amount of 14C covalently bound to lung macromolecules, expressed as a percentage of calculated deposited radioactivity, was twofold greater for 1-nitropyrene adsorbed onto carbon black than for 1-nitropyrene alone. The amount of covalently bound 14C increased with time after exposure to 14C-1-nitropyrene adsorbed onto carbon black, reaching a level of approximately 1 percent of deposited radioactivity, 10-fold greater than that seen with pure 14C-1-nitropyrene seven to 30 days after exposure. The level of covalently bound 14C declined steadily after exposure to pure 14C-1-nitropyrene. Carbon black particles associated with adsorbed 1-nitropyrene offer the potential of studying DNA adduct formation in the lung, because DNA modification might be greater after inhalation of 1-nitropyrene adsorbed onto carbon black than after inhalation of pure 1-nitropyrene or 1-nitropyrene associated with metal oxides, such as gallium oxide.(ABSTRACT TRUNCATED AT 400 WORDS)

Genetic toxicology studies were conducted on organic dyes and mixtures used in colored smoke munitions. The dyes studied included Solvent Red 1; two different batches (Lot 1 and Lot 2) of Disperse Red 11; terephthalic acid; and a mixture of 25 parts Solvent Red 1, 5 parts Disperse Red 11, and 16 parts terephthalic acid. The dyes were evaluated for their ability to produce mutations in Salmonella bacterial strains and in Chinese hamster ovary (CHO) cells. The dyes were also tested in CHO cells to determine cytotoxicity and the induction of sister chromatid exchanges and chromosome aberration. None of the dyes were genotoxic in the standard Ames assay using bacterial strain TA1535 or TA100 with or without the addition of S-9 or in TA98 and TA1538 without S-9. With S-9, Disperse Red 11 (Lot 2) showed significant mutagenic activity in TA98 and TA1538 which increased as a function of S-9 concentration. However, the maximum level of mutagenic activity detected was low (3.8 revertants/micrograms). The azo dye Solvent Red 1 was also negative in a pre-incubation assay designed to reduce azo compounds to free amines. Solvent Red 1 was cytotoxic to mammalian cells, caused a significant increase in SCE, but was not mutagenic or clastogenic. Disperse Red 11 (Lot 1 and Lot 2) were not cytotoxic or clastogenic but produced an increase in cell cycle time and SCE frequency. Only Disperse Red 11 (Lot 2) increased mutations in the CHO/hypoxanthine-guanine phosphoribosyltransferase (HGPRT) assay. The mutagenic activity of the dye mixture was not significant, suggesting no synergistic interaction between the dyes. These studies demonstrated that none of the dyes was clastogenic and that a contaminant in Disperse Red 11 (Lot 2) may be responsible for the weak mutagenic activity in both mammalian and bacterial cell systems.

Compressed air-powered jet mills are used in the chemical and food industries for grinding and classifying powders. We adapted one type of these fluid energy mills as a powder generator for inhalation experiments. The generating system included a jet mill and a screw feeder, the jet mill consisting of an elongated channel, a feeding jet to deliver the material into the channel, and two high-speed air jets. High speed air circulating in the channel created turbulence and centrifugal forces to disperse powder. The jet mill used can be operated from 25 to 100 psig at flow rates of 300 to 900 L/min. Two test materials--a solvent yellow dye and a dye mixture of solvent green and solvent yellow--were used. Both dyes were soft and sticky and could not be dispersed with several other powder generators tested at the concentrations required for toxicity studies. Aerosol concentrations ranging from 10 to 1500 mg/m3 at a flow rate of 400 L/min were obtained by adjusting the feed rate to the jet mill. Stability of aerosol concentration during six-hour continuous generation was 15 to 20%. Comparisons of several generators for producing sticky organic powders are also discussed.

In life-span studies in CD-1 mice and F344/Crl rats, inhaled diluted diesel exhaust was highly fibrogenic in rats but not in mice. This was the case despite the higher lung burden, in mg soot/g lung, achieved in mice compared to rats. We tested the hypothesis that the greater fibrogenicity of the soot in rats was due in part to greater release of mediators of inflammation from alveolar cells in rats compared to mice. Female F344/rats and B6C3F1 mice were exposed for up to 17 days to diluted diesel exhaust containing 3.5 mg/m3 of soot. The lungs of control and soot-exposed animals were lavaged after 2, 12 or 17 days of exposure. The presence of leukotriene (LT)B4, LTC4, prostaglandin (PG)E2, PGF2 alpha and thromboxane (TX) B2 in the lavage fluids and LTB4 and PGF2 alpha in cultured lavage cell supernatants was determined. The total amount of each lavage fluid constituent was normalized to lung weight for species comparisons. Control rats had higher levels of TXB2 (16-fold), and LTB4 (6-fold) and PGE2 (2-fold) than control mice, but control mice had higher amounts of LTC4 (4-fold). Control rats and mice had approximately the same amounts of PGF2 alpha/g lung in bronchoalveolar lavage fluid (BALF). Rats exposed to diesel exhaust had increases in BALF PGF2 alpha and LTB4 that were highest after 2 days of exposure and decreased thereafter. Mice had lesser increases in both parameters. Rat cells recovered from lavage fluid released larger amounts of LTB4 into culture supernatants than mouse cells. The data were consistent with the hypothesis that soot-laden rat alveolar cells release greater quantities of mediators of inflammation than do the alveolar cells in mice.

The responses of guinea pigs and rats to inhaled sulfuric acid aerosols were compared to define species differences and to determine the small-animal model most relevant to human exposures. Rats were exposed for 6 h to 1, 10, and 100 mg H2SO4/m3. Guinea pigs were exposed for 6 h to 1, 10, and 27 mg H2SO4/m3. Tracheal mucous clearance of guinea pigs was slowed 1 d after exposures to 1 mg H2SO4/m3. A tendency toward faster clearance was observed at high concentrations of H2SO4 for both guinea pigs and rats (statistically significant only for the rats). The speeding of mucous clearance was correlated with increases in airway sialic acid and also with the appearance of excess tracheal secretions, detected using scanning electron microscopy in both rats and guinea pigs. The responses of guinea pigs to sulfuric acid exposures were more similar to those reported for humans than were those of rats.

Nickel subsulfide (Ni3S2), nickel sulfate (NiSO4), and nickel oxide (NiO) are encountered occupationally in the nickel refining and electroplating industries, with inhalation being a common route of exposure. The purposes of this study were to evaluate the biochemical responses of lungs of rats and mice exposed for 13 weeks to occupationally relevant aerosol concentrations of Ni3S2, NiSO4, and NiO, to correlate biochemical responses with histopathologic changes, and to rank the compounds by toxicity. Biochemical responses were measured in bronchoalveolar lavage fluid (BALF) recovered from lungs of exposed animals. Parameters evaluated in BALF were lactate dehydrogenase (LDH), beta-glucuronidase (BG), and total protein (TP). Total and differential cell counts were performed on cells recovered in BALF. All compounds produced an increase in LDH, BG, TP, and total nucleated cells, and an influx of neutrophils, indicating the presence of a cytotoxic and inflammatory response in the lungs of exposed rats and mice. Increases in BG were greater than increases in LDH and TP for both rats and mice. Chronic active inflammation, macrophage hyperplasia, and interstitial phagocytic cell infiltrates were observed histologically in rats and mice exposed to all compounds. Statistically significant increases in BG, TP, neutrophils, and macrophages correlated well with the degree of chronic active inflammation. Results indicated a toxicity ranking of NiSO4 greater than Ni3S2 greater than NiO, based on toxicities of the compounds at equivalent mg Ni/m3 exposure concentrations.

The toxicities of 1-nitropyrene (NP) and benzo[a]pyrene (BaP), inhaled alone and in combination with particles and an irritant gas, were examined to evaluate synergisms among the organic compounds, particles, and gas. Groups of F344 rats were exposed 2 h/d, 5 d/wk for 4 wk to atmospheres of pure NP aerosol (7.5 mg/m3), and to these same compounds adsorbed to Ga2O3 particles (27 mg/m3) both with and without coexposure to 5 ppm SO2. Rats were also exposed to Ga2O3 and SO2 alone. Measurements were made of lung burdens of Ga2O3 particles and retention of radiolabeled tracer particles after the cessation of exposure to evaluate effects on particle clearance. Bronchoalveolar lavage fluid was analyzed to assess inflammation and cytotoxicity. Histopathology was examined to assess the nature and extent of lung injury. Particle clearance was significantly impaired (p less than .05) in all groups whose exposure atmosphere included Ga2O3, but was not significantly changed in the other exposure groups.

The analysis of bronchoalveolar lavage fluid has been used as a probe to detect lung injury in toxicological studies and to diagnose the disease state of the lung in humans. To determine how variable the content of lavage fluid from different species is, bronchoalveolar lavage fluids from normal individuals of four species (hamster, rat, guinea pig, and rabbit) were compared for enzymatic and cellular content as well as total protein and sialic acid. In addition, lavage fluid from young adult rats and hamsters was compared to that from older animals. Finally, the effect of the method of lavage on lavage fluid content was evaluated by comparing lavage fluid obtained from an excised lung with that from a lavage performed in vivo. In general, lavage fluids from the four species were similar. However, lavage fluid from guinea pigs had higher numbers of granulocytes and higher mean beta-glucuronidase activities than fluids from other species. Rats had higher mean alkaline phosphatase activities, reflecting higher serum values of this enzyme. Older hamsters had more protein in their lavage fluid than younger animals, and older rats had lower elastase inhibitory activity than young rats. Performing lavage in vivo, as compared to in vitro, did not greatly alter the lavage fluid except for a trend toward a higher level of sialic acid in fluid taken from the living animal.

Occupational exposures to 1,3-butadiene or isoprene occur through their use in the manufacture of rubber and other related polymer products. The purpose of this study was to determine if butadiene or isoprene administration would result in the formation of adducts with blood hemoglobin (Hb), and if such adducts can be used as a measure of previous exposure(s). Male B6C3F1 mice and male Sprague-Dawley rats were injected intraperitoneally with 1, 10, 100, or 1000 mumol [14C]butadiene or 0.3, 3.0, 300, 1000, or 3000 mumol [14C]isoprene per kilogram body weight. Animals were killed 24 hr later. Globin was isolated from blood samples and was analyzed for 14C by liquid scintillation spectroscopy. Hb adduct formation was linearly related to administered doses up to 100 mumol [14C]butadiene or 500 mumol [14C]isoprene per kilogram body weight for mice and rats, respectively. For [14C]butadiene, the efficiency of Hb adduct formation in mice and rats within the linear response range was 0.177 +/- 0.003 and 0.407 +/- 0.019 (pmol of 14C-adducts/mg globin)/(mumol of retained [14C]butadiene/kg body wt), respectively (mean +/- SE; n = 18). For [14C]isoprene, these values for mice and rats were 0.158 +/- 0.035 and 0.079 +/- 0.016 (pmol of 14C-adducts/mg globin)/(mumol of retained [14C]isoprene/kg body wt), respectively (mean +/- SE; n = 12). Hb adducts also accumulated linearly after repeated daily administration of 100 mumol [14C]butadiene or 500 mumol [14C]isoprene per kilogram body wt to mice and rats, respectively, for 3 days. [14C]Butadiene-derived Hb adducts in blood showed lifetimes of approximately 24 and approximately 65 days for mice and rats, respectively, which correlate with the reported lifetimes for red blood cells in these rodent species. Thus, levels of butadiene- or isoprene-derived adducts on Hb in circulating blood may be a useful measure of prior repeated exposures to these compounds.

Male and female F344 rats were exposed 7 hr/day, 5 day/week for up to 30 months to automotive diesel engine exhaust at soot concentrations of 0.35, 3.5, or 7.0 mg/m3 or were sham-exposed to clean air. Rats were terminated at 6-month intervals to measure lung burdens of diesel soot and for histopathology. Other rats either died or were terminated after 30 months of exposure. Lungs were fixed, sectioned into 3-mm slices, and examined by a dissecting microscope to detect tumors. Lesions were stained and examined by light microscopy. Survival and body weight were unaffected by exposure. Focal fibrotic and proliferative lung disease accompanied a progressive accumulation of soot in the lung. The prevalence of lung tumors was significantly increased at the high (13%) and medium (4%) dose levels above the control prevalence (1%). Four tumor types, all of epithelial origin, were observed: adenoma, adenocarcinoma, squamous cyst, and squamous cell carcinoma. Logistic regression modeling demonstrated a significant relationship between tumor prevalence and both exposure concentration and soot lung burden. These results demonstrate that diesel exhaust, inhaled chronically at a high concentration, is a pulmonary carcinogen in the rat.

Studies were completed in F344/N rats and B6C3F1 mice to determine the effect of dose, dose rate, route of administration, and rodent species on formation of total and individual benzene metabolites. Oral doses of 50 mg/kg or higher saturated the capacity for benzene metabolism in both rats and mice, resulting in an increased proportion of the administered dose being exhaled as benzene. The saturating air concentration for benzene metabolism during 6-hr exposures was between 130 and 900 ppm. At the highest exposure concentration, rats exhaled approximately half of the internal dose retained at the end of the 6-hr exposure as benzene; mice exhaled only 15% as benzene. Mice were able to convert more of the inhaled benzene to metabolites than were rats. In addition, mice metabolized more of the benzene by pathways leading to the putative toxic metabolites, benzoquinone and muconaldehyde, than did rats. In both rats and mice, the effect of increasing dose, administered orally or by inhalation, was to increase the proportion of the total metabolites that were the products of detoxification pathways relative to the products of pathways leading to putative toxic metabolites. This indicates low-affinity, high-capacity pathways for detoxification and high-affinity, low-capacity pathways leading to putative toxic metabolites. If the results of rodent studies performed at high doses were used to assess the health risk at low-dose exposures to benzene, the toxicity of benzene would be underestimated.

Studies conducted by the National Toxicology Program on the chronic toxicity of benzene indicated that B6C3F1 mice are more sensitive to the toxic effects of benzene than are F344 rats. A physiological model was developed to describe the uptake and metabolism of benzene in rats and mice and to determine if the observed differences in toxic effects could be explained by differences in the pathways for metabolism of benzene or by differences in uptake of benzene. Major pathways for elimination of benzene included metabolism to hydroquinone glucuronide or hydroquinone sulfate, phenyl glucuronide or phenyl sulfate, muconic acid, and prephenyl mercapturic acid or phenyl mercapturic acid. Model simulations for total benzene metabolized and for profiles of benzene metabolites were conducted for oral or inhalation exposure and compared to data for urinary excretion of benzene metabolites after exposure of rats and mice to [14C]- or [3H]-benzene by inhalation or gavage. Results for total amount of benzene metabolized, expressed per kilogram body weight, indicated that for inhalation exposure concentrations up to 1000 ppm, mice metabolized at least two to three times as much benzene as did rats. Simulations of oral exposure to benzene resulted in more benzene metabolized per kilogram body weight by rats at oral exposures of greater than 50 mg/kg. Patterns of metabolites formed after either route of exposure were very different for F344/N rats and B6C3F1 mice. Rats primarily formed the detoxification metabolite, phenyl sulfate. Mice formed hydroquinone glucuronide and muconic acid in addition to phenyl sulfate. Hydroquinone and muconic acid are associated with pathways leading to the formation of the putative toxic metabolites of benzene. Metabolic rate parameters, Vmax and Km, were very different for hydroquinone conjugate and muconic acid formation compared to formation of phenyl conjugates and phenyl mercapturic acids. Putative toxication pathways could be characterized as high affinity, low capacity whereas detoxification pathways were low affinity, high capacity. Model simulations suggested that for both rats and mice at lower exposure concentrations hydroquinone and muconic acid represented a larger fraction of the total benzene metabolized than at higher exposure concentrations where detoxification metabolites were predominant. Preferential production of a putative toxic metabolite at low exposure concentrations may have important implications in risk assessment for benzene.

This study examined age-related differences in the physiological responses of rats to inhaled automotive emissions. Previous reports suggested that lung development of animals exposed to oxidant gases early in life might be impaired, or that developing lungs might be more susceptible than adult lungs to inhaled toxicants. There were no previous comparisons in developing and adult lungs of the effects of atmospheres containing particles. The hypothesis tested in this study was that rats exposed to chronically inhaled nitrogen dioxide (NO2) or diesel exhaust during lung development were more susceptible to lung injury than rats that were exposed to these atmospheres as adults. Rats were exposed either throughout the period of lung development or as adults, and health effects in the two groups were compared at the end of exposure. Rats were exposed seven hours per day, five days per week for six months to NO2 at 9.5 ppm, to whole diesel exhaust diluted to a soot concentration of 3.5 mg/m3, or to filtered air as controls. These concentrations were selected to produce mild effects in adults. The younger group (developing) was conceived in the exposure atmospheres and exposed during gestation and through the age of six months, and the older group (adult) was exposed between six and twelve months of age. Health effects were evaluated at the end of six months' exposure and some measurements were repeated six months after the cessation of exposure. Measurements included respiratory function, pulmonary immune responses, lung clearance of radiolabeled particles, airway fluid enzymes, protein and cytology, lung tissue collagen and proteinases, lung burdens of diesel soot, lung morphometry and histopathology. Nitrogen dioxide slightly reduced body weight and altered airway fluid enzymes of both age groups, with a greater number of statistically significant differences detectable in the enzyme levels of animals exposed as adults. Normal lung development, as reflected in the size and functional efficiency at adulthood, was not affected by NO2 in this study. Diesel exhaust altered the airway fluid constituents as well as lung tissue collagen and proteinases of both age groups. Particularly striking was an almost sixfold increase in the percentage of neutrophils, a class of highly phagocytic leukocytes, in the airway fluids of adults after six months of exposure. Exhaust-exposed adults had increased numbers of cells in pulmonary lymph nodes, delayed clearance of both diesel soot and 134Cs-labeled particles, and increased lung weight. These changes did not occur in the rats exposed during development.(ABSTRACT TRUNCATED AT 400 WORDS)

The purpose of this study was to evaluate the in vivo effects of an acute exposure to low levels of ozone on rat pulmonary alveolar macrophages (PAM). Fisher 344 rats exposed to 0.0, 0.12, 0.8, or 1.5 ppm O3 for 6 h were killed immediately after and 3, 18, 42, or 66 h after ozone exposure and their lungs were lavaged. Compared to sham-exposed (control) rats, exposure to 0.12 ppm O3 had no measurable effect on the total number, labeling index (LI), mitotic index (MI), or morphology of rat alveolar macrophages. The number of neutrophils was significantly (p less than or equal to 0.001) greater than in controls at 3, 18, and 42 h after exposure to 1.5 ppm O3 and 42 h after exposure to 0.8 ppm O3. The number of PAM was approximately twice that of controls 42 and 66 h after exposure to 0.8 and 1.5 ppm O3. There was a significant (p less than or equal to 0.001) increase in PAM MI 42 and 66 h after exposure to 1.5 ppm O3 and 42 h after 0.8 ppm O3. The increase in the number of PAM in mitosis was preceded by an increase in PAM LI. The PAM LI was significantly (p less than or equal to 0.001) greater than controls 18 and 42 h after exposure but returned to near normal levels by 66 h after exposure. There was a transient decrease in the mean nuclear/cytoplasmic ratio of PAM from rats exposed to 1.5 ppm O3 18 and 42 h after exposure due to an increase in the mean PAM cytoplasmic area. Comparison of the PAM population doubling time (Dt) and cell cycle time (Ct) suggest that PAM proliferation played a significant role in the observed increase in PAM following exposure to 0.8 and 1.5 ppm O3. These results highlight the dynamic response of PAM to an acute exposure to ozone and suggest that the proliferative response of pulmonary alveolar macrophages may be a useful indicator of pulmonary damage following inhalation of an irritant oxidant.

Diesel exhaust, inhaled chronically at high concentrations, was previously found to be a pulmonary carcinogen in rats. The exhaust-induced tumors were located exclusively in the peripheral lung, although all of the respiratory tract tissues were exposed to the exhaust. The purpose of this study was to determine whether there were differences in the level of DNA adducts among the regions of the respiratory tract that paralleled the site of tumors. Groups of male F344/N rats were exposed 7 hr/day, 5 day/week for 12 weeks to diesel engine exhaust at a soot concentration of 10 mg/m3 or were sham-exposed to air. The maxilloturbinates, ethmoturbinates, trachea, left mainstem bronchus (airway generation 1), axial airway (airway generations 2-12), and peripheral lung tissue were dissected from the respiratory tract. DNA was isolated from the dissected samples and analyzed for the presence of adducts using the 32P-postlabeling assay. Chromatographic maps of DNA adducts demonstrated unique patterns of DNA adducts for each of the regions. The highest level of total DNA adducts occurred in peripheral lung tissue (approximately 20 adducts per 10(9) bases). The level of DNA adducts detected in the nasal tissues was about one-fourth to one-fifth that detected in peripheral lung. There were less than three adducts per 10(9) bases in each of the regions of the major conducting airways (i.e., trachea, bronchi, axial airway). In control rats, levels of DNA adducts ranged from one adduct per 10(9) bases (mainstem bronchi, axial airway) to about nine adducts per 10(9) bases (parenchyma). The data from this study indicate that higher levels of total DNA adducts and exhaust-induced adducts (i.e., exposed minus control adducts) were present in tissues where exhaust-induced tumors were located. These data suggest that DNA adduct levels in discrete locations of the respiratory tract may be good measures of the "effective dose" of carcinogenic compounds.

Sixteen adult Syrian hamsters were exposed 33 times over a 19-day period to enough halothane to induce a deep surgical plane of anesthesia. The exposures caused no significant change in the total amount of lipid in the exposed lungs; however, the percentage of neutral lipid due to cholesterol increased from 29 +/- 5% to 43 +/- 7% while the percentage of free fatty acid decreased from 33 +/- 3% to 22 +/- 5%. Among the phosphoglycerides, there was a slight shift toward more phosphatidyl choline and less phosphatidyl serine, phosphatidyl glycerol, and phosphatidyl ethanolamine.

Benzene is a known human leukemogen and animal carcinogen. To better assess the risks associated with benzene exposure, it would be helpful to determine whether repeated inhalation exposures would affect the metabolism of benzene. The purpose of these experiments was to determine if exposure of F344 rats and B6C3F1 mice to 600 ppm benzene, 6 h/day, 5 days/week for 3 weeks, would affect the subsequent in vivo metabolism of inhaled [14C]benzene.

The objective of this study was to test the hypothesis that lipophilicity (as measured by the octanol/water partition coefficient, P) and/or molecular weight are determining factors in the rate of clearance of organic compounds from the lung. Previous work in our laboratory has shown that organic-soluble compounds such as pyrene, benzo[a]pyrene, 1-nitropyrene, 2-aminoanthracene, phenanthridone, dibenzo[c,g]carbazole, 1,3-dichloropropene, and methyl bromide, all of which have a log P less than 6.1, clear the lung rapidly (t 1/2 less than 12 hr). In the present study, organic compounds (mainly anthraquinone dyes) having a wider range of log P's (1.95-8.65) were instilled into rat lungs and the percentage of the compound retained in the lungs at 24 hr was determined. A positive correlation between the log of the theoretical P and the percentage of the compound retained in lungs at 24 hr was found. The lipophilicity of the series of compounds studied was highly dependent on the molecular weight, so that there was also a positive correlation between the molecular weight of the compounds and the percentage of the compound retained in the lung at 24 hr. To help understand the relative importance of lipophilicity and molecular weight in determining lung retention, an additional compound with a high molecular weight but containing a polar functional group [1,5-di(2-sulfo-p-toluidino)anthraquinone] was studied. The results indicated that the lipophilicity was the more important factor in whether the material was retained in the lung. On the basis of the results of this study, organic-soluble compounds with molecular weights less than 300 Da can be expected to clear the lungs rapidly. Nonpolar, organic-soluble compounds with a molecular weight greater than 300 Da can be expected to clear the lungs more slowly.

The particulate exhaust of diesel engines consists of 0.1--0.2 micrometers mass median diameter particles composed of a carbonaceous core and adsorbed organic compounds. A technique was needed to determine accumulated lung burdens of particles in animals exposed to diesel exhaust as a determinant of dose. A method was developed for determining lung burdens of diesel soot particles in rats at 1 day, and at 1, 5, 15, 33 and 52 weeks after cessation of a subchronic exposure to diluted diesel exhaust. Lung tissue was dissolved in tetramethylammonium hydroxide and the diesel soot separated by centrifugation. The soot was suspended in water by sonication and the light absorption of samples was compared to standard suspensions of diesel soot. Recovery from lungs spiked with 50-1000 micrograms of soot was 89 +/- 5%. Rats exposed over a period of 18 weeks to diluted diesel exhaust at average net diesel particles concentrations of 150, 940 and 4100 micrograms/m3 had lung burdens of 35, 220 and 1890 micrograms/g lung, respectively, one day after the last exposure. The long-term clearance rates of soot had estimated half-times of 87 +/- 28, 99 +/- 4 days, for the low and medium exposure groups, respectively. The clearance half-time for the high level exposure group of 165 +/- 8 days was significantly longer (P less than 0.0001) than those of the other two groups.

2,3-Dichloropropene (2,3-DCP) is a constituent of some commercially available preplant soil fumigants for the control of plant parasitic nematodes. Human exposure potential exists during manufacture of the chemicals or during bulk handling activities. The purpose of this investigation was to determine the disposition and metabolism of 2,3-[14C]DCP in rats after inhalation. Male Fischer-344 rats were exposed nose-only to a vapor concentration of 250 nmol 2,3-[14C]DCP/liter air (7.5 ppm; 25 degrees C, 620 Torr) for 6 hr. Blood samples were taken during exposure, and urine, feces, expired air, and tissues were collected for up to 65 hr after exposure. Urinary excretion was the major route of elimination of 14C (55% of estimated absorbed 2,3-DCP). Half-time for elimination of 14C in urine was 9.8 +/- 0.05 hr (means +/- SE). Half-time for elimination of 14C feces (17% of absorbed 2,3-DCP) was 12.9 +/- 0.14 hr (means +/- SE). Approximately 1 and 3% of the estimated absorbed 2,3-[14C]DCP were exhaled as either 2,3-[14C]DCP or 14CO2, respectively. Concentrations of 14C in blood increased during 240 min of exposure, after which no further increases in blood concentration of 14C were seen. 14C was widely distributed in tissues analyzed after a 6-hr exposure of rats to 2,3-[14C]DCP. Urinary bladder (150 nmol/g), nasal turbinates (125 nmol/g), kidneys (84 nmol/g), small intestine (61 nmol/g), and liver (35 nmol/g) were tissues with the highest concentrations of 14C immediately after exposure. Over 90% of the 14C in tissues analyzed was 2,3-DCP metabolites. Half-times for elimination of 14C from tissues examined ranged from 3 to 11 hr. The data from this study indicate that after inhalation 2,3-DCP is metabolized in tissues and readily excreted.

Solvent yellow (2-(2'-quinolinyl)-1,3-indandione) and solvent green (1,4-di-p-toluidinoanthraquinone) are components of colored smoke munitions and may become airborne and be inhaled by workers during the manufacture of the munitions. Little is known about the disposition of either dye after inhalation. To obtain this information, we exposed male F344/N rats to 14C-solvent yellow aerosols (160 nmol solvent yellow/liter air) or a mixture of 14C-solvent yellow and unlabeled solvent green (340 nmol solvent yellow and 370 nmol solvent green/liter air) for 60 min. After either exposure, solvent yellow was rapidly cleared from the respiratory tract, with a t1/2 of 2-3 hr. Solvent green was retained in the lungs with a minimum estimated t1/2 for clearance of 22 days. Solvent green was not detected in other tissues during the 70-hr postexposure period. After either exposure, high-pressure liquid chromatography analysis of tissues extracts indicated that 40 to 75% of the 14C in liver and kidney consisted of solvent yellow metabolites. Greater than 90% of the 14C in the lungs was unmetabolized solvent yellow. The major pathway for excretion of solvent yellow and solvent yellow metabolites was the feces (74% of the initial body burden); the t1/2 for excretion was 14 hr. Urinary 14C accounted for 14% of the initial body burden and the t1/2 for excretion was 10 hr. Over 90% of the 14C excreted in the urine was solvent yellow metabolites. Very little solvent yellow (2%) was metabolized to 14CO2. By 72 hr after exposure, only 10% of the initial 14C deposited remained in the body.

To determine the effect of exposure concentration and the route of administration on benzene metabolism, male F344/N rats and B6C3F1 mice were orally exposed to 1, 10, and 200 mg benzene/kg, and by inhalation for 6 hr to 5, 50, and 600 ppm benzene vapor. The effect of different exposure rates on the metabolism of benzene was determined by exposing rodents over different time intervals to the same total amount of benzene [constant concentration X time factor (C X T) = 300 ppm.hr]. Water-soluble metabolites constituted greater than 90% of the metabolite dose to the tissues and were used as a measure of the metabolism of benzene via different pathways. Water-soluble metabolites were measured in the blood, urine, liver, lung, and bone marrow from animals killed following oral exposures and during and following inhalation exposures. The total "dose" to the tissue of individual metabolites was determined by the area under the curve (AUC). The results indicated a shift in metabolism from putative toxification pathways to detoxification pathways as the exposure concentration or oral dose increased. In mice, hydroquinone glucuronide and muconic acid (markers of toxification metabolic pathways) represented a greater percentage of the administered dose at low doses than at high doses. At high doses, phenylglucuronide and prephenylmercapturic acid (detoxification products) increased as a percentage of the administered dose. This same metabolic shift was observed in rats, except that hydroquinone glucuronide was a minor metabolite of benzene at all concentrations. The AUC of phenylsulfate (detoxification pathway) was proportional to the exposure concentration in both species. Within the range of C X T factors studied, the rate of the inhalation exposure to benzene did not affect the AUC of metabolites in tissues of rats; however, a high dose rate (600 ppm 0.5 hr) in mice caused a shift in metabolism to phenyl conjugates. The comparison of oral and 6-hr inhalation exposures indicated that, in terms of metabolite dose to tissues, there is no simple relationship between these two routes of administration. An oral dose and an inhalation exposure concentration which produce an equal dose of one metabolite produce very different doses of another metabolite. These studies demonstrated a species difference in benzene metabolism, as well as a metabolic shift in benzene metabolic pathways as the exposure concentration was increased.(ABSTRACT TRUNCATED AT 400 WORDS)

Chronic inhalation of insoluble particles of low toxicity that produce substantial lung burdens of particles, or inhalation of particles that are highly toxic to the lung, can impair clearance. This report describes model calculations of accumulations in lung of inhaled low-toxicity diesel exhaust soot and high-toxicity Ga2O3 particles. Lung burdens of diesel soot were measured periodically during a 24-mo exposure to inhaled diesel exhaust at soot concentrations of 0, 0.35, 3.5, and 7 mg m-3, 7 h d-1, 5 d wk-1. Lung burdens of Ga2O3 were measured for 1 y after a 4-wk exposure to 23 mg Ga2O3 m-3, 2 h d-1, 5 d wk-1. Lung burdens of Ga2O3 were measured for 1 y both studies using inhaled radiolabeled tracer particles. Simulation models fit the observed lung burdens of diesel soot in rats exposed to the 3.5- and 7-mg m-3 concentrations of soot only if it was assumed that clearance remained normal for several months, then virtually stopped. Impaired clearance from high-toxicity particles occurred early after accumulations of a low burden, but that from low-toxicity particles was evident only after months of exposure, when high burdens had accumulated in lung. The impairment in clearances of Ga2O3 particles and radiolabeled tracers was similar, but the impairment in clearance of diesel soot and radiolabeled tracers differed in magnitude. This might have been related to differences in particle size and composition between the tracers and diesel soot. Particle clearance impairment should be considered both in the design of chronic exposures of laboratory animals to inhaled particles and in extrapolating the results to people.

Azodicarbonamide (ADA) is widely used as a blowing agent in the manufacture of expanded foam plastics, as an aging and bleaching agent in flour, and as a bread dough conditioner. Human exposures have been reported during manufacture as well as during use. Groups of male F344/N rats were administered ADA by gavage, by intratracheal instillation, and by inhalation exposure to determine the disposition and modes of excretion of ADA and its metabolites. At 72 hr following gavage, 30% of the administered ADA was absorbed whereas following intratracheal instillation, absorption was 90%. Comparison between groups of rats exposed by inhalation to ADA to achieve body burdens of 24 or 1230 micrograms showed no significant differences in modes or rates of excretion of [14C]ADA equivalents. ADA was readily converted to biurea under physiological conditions and biurea was the only 14C-labeled compound present in excreta. [14C]ADA equivalents were present in all examined tissues immediately after inhalation exposure, and clearance half-times on the order of 1 day were evident for all tissues investigated. Storage depots for [14C]ADA equivalents were not observed. The rate of buildup of [14C]ADA equivalents in blood was linearly related to the lung content as measured from rats withdrawn at selected times during a 6-hr inhalation exposure at an aerosol concentration of 25 micrograms ADA/liter. In a study extending 102 days after exposure, retention of [14C]ADA equivalents in tissues was described by a two-component negative exponential function. The results from this study indicate that upon inhalation, ADA is rapidly converted to biurea and that biurea is then eliminated rapidly from all tissues with the majority of the elimination via the urine.

2,3-Dichloropropene (DCP) is an intermediate used in the manufacture of carbamate herbicides and there is potential for human exposure during the manufacturing process. DCP is a known mutagen in bacteria systems and some structural analogs of DCP are carcinogenic. Since little is known about the disposition of DCP in animals after inhalation, studies were conducted in male Fischer-344 rats to determine the effect of vapor concentration on absorption and excretion. Uptake and elimination of 14C was studied in rats after nose-only inhalation of 17, 240, or 1650 nmol of [14C]DCP vapor/liter of air (0.4, 6, or 40 ppm, respectively, at 760 mm and 25 degrees C) for 6 hr. The percentage of inhaled DCP absorbed averaged 38% and was not statistically different at any vapor concentration, although minute volume was lower during exposure to 1650 nmol/liter. Urine, feces, and expired air were collected from rats for 65 hr after exposure. Rats were sacrificed and tissues, carcass, excreta, and expired air were analyzed for 14C. Routes of 14C excretion were independent of vapor concentration, with 50% of the 14C excreted in urine, 13% in feces, approximately 7% as CO2, and less than 1% as DCP in expired air. Rates of 14C excretion were also independent of vapor concentration, with the half-times averaging 9.9 hr (urine), 13.6 hr (feces), and 0.9 hr (14CO2). Sixty hours after inhalation, 29% of the initial body burden of 14C remained in the carcass. Most was associated with the pelt, but some 14C was found in all tissues. Respiratory tract, GI tract, liver, and kidney were tissues with the highest 14C contents. The results indicate that DCP metabolism and excretion rates are relatively constant throughout the vapor concentration range studied. This suggests that results from more detailed pharmacokinetic studies (and possibly toxicity studies) at one DCP concentration may be extrapolated to other concentrations within this range.

F344 rats were chronically exposed to diesel exhaust at target soot concentrations of 0 (control, C), 0.35 (low, L), 3.5 (medium, M), and 7.0 (high, H) mg/m3. Accumulated lung burdens of diesel soot were measured after 6, 12, 18, and 24 months of exposure. Parallel measurements of particle deposition and clearance were made to provide insight into the mechanisms of particle accumulation in lungs. The fractional deposition of inhaled 67Ga2O3 particles after 6, 12, 18, and 24 months of exposure and of inhaled 134Cs-fused aluminosilicate particles after 24 months were similar for all groups. Progressive increases in lung burdens of soot particles were observed in M and H exposed rats, reaching levels of 11.5 +/- 0.5 and 20.5 +/- 0.8 mg/lung (mean +/- SE), respectively, after 24 months. Rats in the L group had smaller relative increases in lung burden, reaching levels of 0.60 +/- 0.02 mg/lung after 24 months. Tracheal mucociliary clearance measurements, using 99mTc-macroaggregated albumin deposited in the trachea, showed no changes at anytime. There were statistically significant increases in clearance half-times of inhaled radiolabeled particles of 67Ga2O3 as early as 6 months at the H level and 18 months at the M level; no significant changes were seen at the L level. Rats inhaled fused aluminosilicate particles labeled with 134Cs after 24 months of diesel exhaust exposure to measure long-term components of pulmonary clearance. The long-term clearance half-times were 79 +/- 5, 81 +/- 5, 264 +/- 50, and 240 +/- 50 days (mean +/- SE) for the C, L, M, and H groups, respectively. Differences were significant between the C and both the M and H exposure groups (p less than 0.01). Lung burdens of diesel soot were more than expected at the H and M levels and were also associated with impaired particle clearance while smaller responses were observed in both burdens and clearance at the L level.

Methyl bromide is used as a disinfectant to fumigate soil and a wide range of stored food commodities in warehouses and mills. Human exposure occurs during the manufacture and use of the chemical. The purpose of this investigation was to determine the disposition and metabolism of [14C]methyl bromide in rats after inhalation. Male Fischer-344 rats were exposed nose only to a vapor concentration of 337 nmol [14C]methyl bromide/liter air (9.0 ppm, 25 degrees C, 620 torr) for 6 hr. Urine, feces, expired air, and tissues were collected for up to 65 hr after exposure. Elimination of 14C as 14CO2 was the major route of excretion with about 47% (3900 nmol/rat) of the total [14C]methyl bromide absorbed excreted by this route. CO2 excretion exhibited a biphasic elimination pattern with 85% of the 14CO2 being excreted with a half-time of 3.9 +/- 0.1 hr (means +/- SE) and 15% excreted with a half-time of 11.4 +/- 0.2 hr. Half-times for elimination of 14C in urine and feces were 9.6 +/- 0.1 and 16.1 +/- 0.1 hr, respectively. By 65 hr after exposure, about 75% of the initial radioactivity had been excreted with 25% remaining in the body. Radioactivity was widely distributed in tissues immediately following exposure with lung (250 nmol equivalents/g), adrenal (240 nmol equivalents/g), kidney (180 nmol equivalents/g), liver (130 nmol equivalents/g), and nasal turbinates (110 nmol equivalents/g) containing the highest concentrations of 14C. Radioactivity in livers immediately after exposure accounted for about 17% of the absorbed methyl bromide. Radioactivity in all other tissues examined accounted for about 10% of the absorbed methyl bromide. Elimination half-times of 14C from tissues were on the order of 1.5 to 8 hr. In all tissues examined, over 90% of the 14C in the tissues was methyl bromide metabolites. The data from this study indicate that after inhalation methyl bromide is rapidly metabolized in tissues and readily excreted.