Synergistic interaction of ozone and respirable aerosols on rat lungs. III. Ozone and sulfuric acid aerosol

Previously we have demonstrated that a synergistic interaction, as evaluated by several biochemical, toxicological, and morphological responses of the lung, results from exposure of rats to ozone (O3) in conjunction with moderate concentrations of acidic, but not neutral, aerosols. To extend these studies, groups of rats were continuously exposed to either O3 or sulfuric acid aerosol alone, or to combinations of these pollutants. Pulmonary responses from these rats were measured by assay, after exposures for 6 hr to 7 days, of total lavageable protein content, total lung tissue protein content after 5, 7, or 9 days of exposure, or apparent collagen synthesis rates from lung tissue after 7 days of exposure. While the lavageable protein content from rats exposed for 3 days to 0.1 or 1.0 mg/m3 of sulfuric acid aerosol alone was not different from control values, significant elevations from control values were observed from groups exposed to 0.12, 0.20, or 0.64 ppm of O3. Synergy was demonstrated by this assay upon exposure of rats to 0.20 ppm of O3 in conjunction with 0.1, 0.5, or 1.0 mg/m3 of sulfuric acid aerosol. Similarly, the tissue protein content from rats exposed to 0.1 or 1.0 mg/m3 of sulfuric acid aerosol alone was indistinguishable from control values. Significant elevations from control values were observed by this assay from groups of rats exposed to 0.64 or 0.20 ppm of O3, and a synergistic interaction was demonstrated between 0.64 ppm of O3 and 1.0 mg/m3 of sulfuric acid aerosol. Furthermore, synergy was observed by quantification of increased total lung protein between 0.20 ppm of O3 + 40 micrograms/m3 and higher concentrations of sulfuric acid aerosol. Values of the lung collagen synthesis rate from rats exposed to 0.1, but not 1.0, 0.5, or 0.04 mg/m3 of sulfuric acid aerosol were significantly higher than values from lungs of control animals. Significant elevations from control values were also observed by this assay from groups of rats exposed to 0.64 or 0.20 ppm of O3. A synergistic interaction was demonstrated by the collagen synthesis rate assay between groups of rats exposed to 0.64 ppm of O3 + 0.20 mg/m3 and higher concentrations of sulfuric acid aerosol or between groups exposed to 0.20 ppm of O3 + 40 micrograms/m3 and higher concentrations of sulfuric acid aerosol. These results demonstrate synergy between O3 and sulfuric acid aerosol upon exposure to concentrations of each pollutant at or near peak hourly ambient levels in polluted urban atmospheres.     

Carcinogenicity of sodium dichromate and chromium (VI/III) oxide aerosols inhaled by male Wistar rats

In inhalation chambers, male Wistar rats of the strain TNO-W74 were continuously exposed to submicron aerosols of sodium dichromate and to a pyrolyzed Cr(VI)/Cr(III) (3:2) oxide mixture. The sodium dichromate (Na2Cr2O7) aerosol had the chromium concentrations of 25, 50 and 100 micrograms/m3, the chromium oxide mixture (Cr5O12) had the chromium concentration of 100 micrograms/m3. After 18 months of inhalation the rats were held under conventional conditions for a further year. The experimental groups consisted of 20 rats and the control group of 40 rats. More than 90% of the rats in each group reached 2 years. At the end of the study the mortality rates amounted to 35%, 45% and 25% in the 3 sodium dichromate aerosol groups, respectively, and 50% in the chromium oxide mixture aerosol group, which was not significantly different from that of the controls (42.5%), living under the same conditions in filtered fresh air. In all sodium dichromate exposed groups significant effects were neither found clinically nor from hematology and clinical chemistry compared to the controls. In the chromium oxide mixture group, however, there was a number of significant findings. Elevated white and red blood cell counts and serum cholesterol as well as decreased serum total immunoglobulin levels at different stages of the study were observed together with few local lung effects determined histopathologically in this group. We assume that these effects are mainly due to the increased chromium lung burden of the rats. At the end of the study the lung chromium retention was about 10 times higher for the rats exposed to chromium oxide versus sodium dichromate at an aerosol Cr-concentration of 100 micrograms/m3, while the kidney chromium retention was measured to be nearly equal in both groups. Three primary lung tumors (2 adenomas and 1 adenocarcinoma) and 1 malign tumor of the pharynx were found at the highest Cr-concentration (100 micrograms/m3) of the sodium dichromate aerosol, 1 primary adenoma of the lung was in the chromium oxide mixture group exposed also to a Cr-concentration of 100 micrograms/m3. No primary lung tumors were observed in the other experimental and control groups. These results indicate a weak carcinogenicity at 100 micrograms/m3 for the rats continuously exposed to submicron Na2Cr2O7 and Cr5O12 aerosols. Thus, there may be a small carcinogenic risk from occupational relevant chromium air levels. However, results have to be confirmed with larger animal populations.     

Acute toxicity of polyethylene glycol p-isooctylphenol ether in Syrian hamsters exposed by inhalation or bronchopulmonary lavage

Dose-response studies were conducted with Syrian hamsters exposed to polyethylene glycol p-isooctylphenyl ether (Triton X-100) via inhalation or bronchopulmonary lavage. Syrian hamsters were exposed to an aerosol of Triton X-100 with a mass median aerodynamic diameter of 1.5 μm and a concentration of 3.0 mg/liter. Estimated initial lung burdens of Triton X-100 ranged from 800 to 3100 μg. Hamsters were lavaged with concentrations of Triton X-100 ranging from 0.01 to 0.10% in isotonic saline resulting in initial lung burdens of Triton X-100 that ranged from 300 to 3200 μg. The $\text{LD}\text{50}\text{7}$ values were 1700 μg (1300–2100 μg, 95% confidence limits) for the inhalation study and 2100 (1900–2700) μg for the lavage study. The difference between the $\text{LD}\text{50}\text{7}$ values for the two methods of exposure was not significant. However histopathological examination revealed differences in the nature and distribution of pathologic changes observed in animals exposed by the two routes of administration. Animals exposed by inhalation died as a result of ulcerative laryngitis and laryngeal edema with only minimal pulmonary pathologic alterations. Animals exposed by lavage, where the larynx was not exposed to Triton X-100, died from pulmonary edema and acute exudative pneumonia, these results demonstrate the need for careful selection of exposure methods to meet the specific objectives of a toxicology study.     

Inhalation exposure of rats to metal aerosol. I. Effects on pulmonary surfactant and ascorbic acid.

Female albino Wistar rats were exposed to less than 5 microns particles separated from nickel refinery waste. The generated aerosol of 50 mg m-3 mainly consisted of metal oxides, the most toxic being NiO and Cr2O3. The exposure of 5 h per day, 5 days per week, lasted for 4 weeks or 4 months. At the end of the exposure period the amounts of pulmonary surfactant and ascorbic acid were estimated in both exposed and control rats. The amount of pulmonary surfactant was elevated after both exposure times, while ascorbic acid increased significantly (P less than 0.02) only after 4 weeks of exposure.     

Respiratory responses of humans exposed to an aerosol-gas pollutant mixture: multivariate contrast of a complex atmosphere to clean air and sodium chloride aerosol controls

Data from a group of 20 subjects with normal baseline pulmonary function, who were exposed for 2 h to a test atmosphere containing a complex mixture of pollutants, have been contrasted with data from two other groups exposed to presumably non-toxic control atmospheres. Group 1 was exposed to clean air, group 2 was exposed to clean air containing sodium chloride aerosol at 270 micrograms m-3, and group 3 was exposed to the complex atmosphere containing sodium chloride (332 micrograms m-3) and zinc ammonium sulfate (23 micrograms m-3) aerosols plus nitrogen dioxide (0.5 ppm) and sulfur dioxide (0.5 ppm). These atmospheres (ranked according to the presumed relative toxicities of the components; clean air = 0, sodium chloride = 1, complex mixture = 2) were contrasted using multiple regression and partial correlation analyses. The effects of exposure to the complex gas-aerosol mixture on forced expiratory performance were not significantly different from those observed in subjects exposed to clean air or to sodium chloride aerosol.     

Endotoxin or cytokines attenuate ozone-induced DNA synthesis in rat nasal transitional epithelium.

Pretreatment of rats with endotoxin (E), a potent inducer of tumor necrosis factor alpha (TNF), and interleukin 1 beta (IL 1), or a combination of TNF and IL1, has been shown to increase levels of lung antioxidant enzymes and protect against pulmonary toxicity associated with hyperoxia. Inhalation of ozone (O3) induces cell injury, followed by increased DNA synthesis, cell proliferation, and secretory cell metaplasia in rat nasal transitional epithelium (NTE). This study was designed to test the effects of E, TNF, and IL1 pretreatment on acute O3-induced NTE cell injury as measured by changes in NTE cell DNA synthesis. Rats were exposed to either 0.8 ppm O3 or air for 6 hr in whole-body inhalation chambers. Immediately before exposure, rats in each group were injected intraperitoneally (ip) with either saline alone or saline containing E (1 microgram/g body wt), TNF (10 micrograms), IL1 (10 micrograms), or both TNF and IL1 (TNF/IL1; 10 micrograms each). Eighteen hours postexposure, rats were injected ip with bromodeoxyuridine (BrdU; 50 micrograms/g body wt) to label cells undergoing DNA synthesis and were euthanized 2 hr later. NTE was processed for light microscopy and immunochemically stained to identify cells that had incorporated BrdU into nuclear DNA. The number of BrdU-labeled NTE nuclei per millimeter of basal lamina was quantitated. There were no significant differences in the number of BrdU-labeled NTE nuclei in air-exposed rats that were injected with E, TNF, IL1, or TNF/IL1 compared with those in saline-injected, air-exposed controls. Rats that were injected with saline and exposed to O3 had approximately 10 times the number of BrdU-labeled NTE nuclei than saline-injected, air-exposed control rats. O3 exposure also induced a significant increase in labeled nuclei in rats that were pretreated with TNF alone. In contrast, pretreatment with E, IL1, or TNF/IL1 attenuated the O3-induced increase in NTE DNA synthesis. These results indicate that both E and the cytokines TNF and IL1 have physiologic effects that can attenuate O3-induced injury or modify the response to NTE cells to O3 exposure.     

The effects of intratracheally administered coarse mode particles on respiratory tract infection in mice

Because coarse mode particles are rarely studied in their existing size ranges (greatest mass about 5-7 microns, aerodynamic diameter), we investigated the effects of four such particles, quartz, ferric oxide, calcium carbonate, and sodium feldspar, on host defenses against bacterial pulmonary infection. Mice which received intratracheal instillations of 10, 33, and 100 micrograms/mouse were exposed within an hour to aerosols of viable Streptococcus, and pneumonia-induced mortality was measured. At 33 and 100 micrograms/mouse, all particles significantly increased mortality. At the lower dose, only Fe2O3 caused a significant increase in mortality. To evaluate potential delayed effects, mice were challenged with the bacteria 24 h after exposure to 100 micrograms particles/mouse. Delaying the challenge did not significantly alter the response, except for the sodium feldspar group for which a partial recovery was observed. When mice exposed to 100 micrograms particles/mouse received aerosols of Klebsiella pneumoniae 24 h later, there was no significant effect on pulmonary bactericidal activity. For the model system used, it appears that Fe2O3, CaCO3, and sodium feldspar have effects roughly equivalent to quartz.     

Lactate dehydrogenase isoenzymes in hamster lung lavage fluid after lung injury

Lactate dehydrogenase (LD) levels and isoenzyme patterns were determined in the cell-free supernatant fractions of lung lavage fluid from hamsters exposed to alpha-quartz, iron oxide, Triton X-100, 100% O₂, or 200 ppm SO₂. The isoenzyme patterns were compared to those derived from hamster lung homogenates, serum, polymorphonuclear neutrophils (PMNs), pulmonary macrophages, and red blood cells. The isoenzyme patterns from alpha-quartz- and iron oxide-exposed animals resembled each other and were similar to that of PMNs. In contrast, the pattern seen after Triton X-100 exposure was similar to those of whole lung homogenates and of red blood cells. A 96-hr exposure to 100% O₂ yielded an LD isoenzyme pattern in lung lavage fluid similar to that of serum. Exposure to SO₂ did not alter LD levels, showing that upper airways damage is not reflected by changes in LD in lung lavage fluid. We conclude that LD isoenzyme patterns of lung lavage fluid can be used to differentiate among types of pulmonary injury and may help identify the sites of injury.     

Synergistic interaction of ozone and respirable aerosols on rat lungs. II. Synergy between ammonium sulfate aerosol and various concentrations of ozone

Pulmonary responses after continuous exposure of rats to concentrations of ozone (O3) ranging from 0.12 to 0.64 ppm were quantified by measuring tissue collagen synthesis rate, tissue protein and DNA content, and various constituents of bronchoalveolar lavage fluid. After 7 days of exposure to 0.64 ppm of O3, lung collagen synthesis rate and tissue content of protein and DNA were elevated. After shorter durations of exposure to 0.64 ppm of O3, significant elevations were observed in the protein content and the activities of lactate dehydrogenase, acid phosphatase, and N-acetyl-beta-D-glucosaminidase from lavage fluid. After exposure of rats to 0.20 ppm of O3 for 7 days, changes could be detected in both lung collagen synthesis rate and tissue protein content. Total lavagable protein content, a sensitive indicator for O3-induced effects upon the lung, was significantly elevated in lungs of rats exposed to 0.12 or 0.20 ppm of O3. To examine whether a synergistic interaction occurred between 0.20 or 0.64 ppm of O3 and acid aerosols, rats were continuously exposed to O3 with and without concurrent exposure to 5 mg/m3 of ammonium sulfate. A synergistic interaction between 0.20 ppm of O3 and ammonium sulfate aerosol was observed by measurement of total lavagable protein and of lung collagen synthesis rate. These results demonstrate that ammonium sulfate aerosol interacts synergistically with O3 at concentrations of O3 that approach ambient levels.     

Polyhexamethylene guanidine phosphate aerosol particles induce pulmonary inflammatory and fibrotic responses

Polyhexamethylene guanidine (PHMG) phosphate was used as a disinfectant for the prevention of microorganism growth in humidifiers, without recognizing that a change of exposure route might cause significant health effects. Epidemiological studies reported that the use of humidifier disinfectant containing PHMG-phosphate can provoke pulmonary fibrosis. However, the pulmonary toxicity of PHMG-phosphate aerosol particles is unknown yet. This study aimed to elucidate the toxicological relationship between PHMG-phosphate aerosol particles and pulmonary fibrosis. An in vivo nose-only exposure system and an in vitro air–liquid interface (ALI) co-culture model were applied to confirm whether PHMG-phosphate induces inflammatory and fibrotic responses in the respiratory tract. Seven-week-old male Sprague–Dawley rats were exposed to PHMG-phosphate aerosol particles for 3 weeks and recovered for 3 weeks in a nose-only exposure chamber. In addition, three human lung cells (Calu-3, differentiated THP-1 and HMC-1 cells) were cultured at ALI condition for 12 days and were treated with PHMG-phosphate at set concentrations and times. The reactive oxygen species (ROS) generation, airway barrier injuries and inflammatory and fibrotic responses were evaluated in vivo and in vitro. The rats exposed to PHMG-phosphate aerosol particles in nanometer size showed pulmonary inflammation and fibrosis including inflammatory cytokines and fibronectin mRNA increase, as well as histopathological changes. In addition, PHMG-phosphate triggered the ROS generation, airway barrier injuries and inflammatory responses in a bronchial ALI co-culture model. Those results demonstrated that PHMG-phosphate aerosol particles cause pulmonary inflammatory and fibrotic responses. All features of fibrogenesis by PHMG-phosphate aerosol particles closely resembled the pathology of fibrosis that was reported in epidemiological studies. Finally, we expected that PHMG-phosphate infiltrated into the lungs in the form of aerosol particles would induce an airway barrier injury via ROS, release fibrotic inflammatory cytokines, and trigger a wound-healing response, leading to pulmonary fibrosis. A simultaneous state of tissue destruction and inflammation caused by PHMG-phosphate had whipped up a “perfect storm” in the respiratory tract.     

Pulmonary hyperreactivity in cynomolgus monkeys (Macaca fascicularis) from nose-only inhalation exposure to disodium hexachloroplatinate, Na2PtCl6

The pulmonary and dermal effects of exposure to Na2PtCl6 were investigated in cynomolgus monkeys (Macaca fascicularis) exposed by the nose-only inhalation and percutaneous routes. Separate inhalation exposures were performed in monkeys at 200 micrograms/m3 and 2 mg/m3 (4 hr/day, biweekly for 12 weeks), while another group of monkeys was percutaneously exposed biweekly by an open patch method. After a 2-week refractory period, serial Na2PtCl6 bronchoprovocation challenges and intradermal Na2PtCl6 sensitivity evaluations were performed. Na2PtCl6 bronchoprovocation in naive control monkeys yielded significant impairments in post-challenge pulmonary mechanics and ventilatory function. These results indicate a pharmacologic or irritant-mediated bronchoconstriction mechanism for acute exposure to this compound. When the post-challenge pulmonary function of animals exposed for the 12-week exposure regimen (across treatments) was compared to pulmonary deficits observed in control animals upon challenge, significantly greater pulmonary deficits were seen in animals exposed at the 200 micrograms/m3 concentration. Exposure at this concentration yielded significant changes in post-challenge average pulmonary flow resistance (RL) and forced expiratory volume in 0.5 sec corrected for vital capacity (FEV0.5/FVC) when compared to control monkey responses. Animals exposed by the percutaneous route or at 2 mg/m3 showed no significant post-challenge pulmonary deficits when compared to control animals. Intradermal Na2PtCl6 sensitivity was found not to be exposure related in the conditions of this experiment.     

Carbon disulphide induced activation of liver UDP glucuronosyltransferase in rats pretreated with phenobarbitone.

Carbon disulphide (CS2) exposure has been shown to activate the UDP glucuronosyltransferase of liver microsomes in rats pretreated with phenobarbitone. Now the nature of CS2 induced activation of the enzymes has been studied further. Phenobarbitone pretreated rats were exposed to 0.15% CS2 for 2 hrs on two successive days. The activity of UDP glucuronosyltransferase was measured from the liver microsomes after the enzymes was activated by incubation of the microsomes with various concentrations of the detergents Triton X-100, digitonin and cetylpyridinium chloride. The exposed animals showed an increased enzyme activity at all applied concentrations of the detergents; therefore in addition to membrane destruction by CS2 exposure, some other mechanism must also be involved in the CS2 induced activation of liver microsomal UDP glucuronosyltransferase. The changes in membrane lipid-protein interactions with l-anilino-8-naphthalene sulphonate (ANS) were also probed. The CS2 exposed animals had more high-affinity binding sites for ANS in their liver microsomal membranes, and in addition the quantum yield of ANS fluorescence was enhanced by CS2. The changes differed from those found after carbon tetrachloride exposure and suggest that, even if the two drugs have some common effects on microsomes, e.g. UDP glucuronosyltransferase activation, P-450 destruction and lipid peroxidation induction, the changes they cause in the microsomal micro-environment differ.     

Inhalation of 1-nitropyrene associated with ultrafine insoluble particles or as a pure aerosol: A comparison of deposition and biological fate

A large number of the environmental particulate pollutants in the atmosphere, including diesel engine exhaust, have a complex mixture of organic compounds associated with them. Organic solvent extracts of many of these particulate pollutants have been shown to contain mutagenic activity which does not require metabolic activation in the Ames bioassay. Much of this direct-acting mutagenic activity has been attributed to nitroaromatic compounds present in these extracts. In the studies reported here, the direct-acting mutagen, [³H]nitropyrene (³H-NP), was used as a model nitroaromatic compound. Rats were exposed to this radiolabeled compound by nose-only inhalation either as a coating (～ 6% by mass) on relatively inert, ultrafine ⁶⁷Ga₂O₃ particles or as a homogeneous ultrafine aerosol. The tissue deposition, retention, and biological fate of each aerosol were investigated and compared. Respiratory tract clearance of ³H radioactivity from each exposure was very rapid with no apparent differences seen in the lung retention of this inhaled compound between each exposure over the course of these studies. Higher ³H-radioactivity levels were seen in stomach and large intestines of rats exposed to the ⁶⁷Ga₂O₃-associated ³H-NP than in the same tissues from rats exposed to the pure ³H-NP aerosol. Rats exposed to the ³H-NP-⁶⁷Ga₂O₃ aerosol excreted the majority of the deposited ³H radioactivity in the feces (75 ± 18%), whereas pure ³H-NP exposed animals excreted a major portion of the radiolabel in the urine (76 ± 18%). It appeared that the major portion of the pure ³H-NP aerosol was cleared from the respiratory tract by direct absorption into blood, while the ⁶⁷Ga₂O₃-associated ³H-NP was cleared by both blood absorption and mucociliary clearance followed by ingestion and fecal excretion. These differences in the deposition and biological fate between the particle-associated NP and the pure NP aerosol may have important implications in terms of the metabolic fate of inhaled nitroaromatic compounds and the health risks associated with human exposures to particulate environmental pollutants that contain this class of compounds.     

Microsomal UDP-glucuronyltransferase in rat liver: oxidative activation

Activation of microsomal UDP-glucuronyltransferase (UDPGT) activity by treatment of hepatic microsomes with either detergents or Fe(3+)/ascorbate pro-oxidant system has been reported; however, definite mechanisms underlying these effects have not been clarified. In this work, we characterize Fe(3+)/ascorbate-induced activation of UDPGT activity prior to solubilization with Triton X-100 and after the oxidation process provoked the solubilization of the enzyme. We observed a time-dependent increase in UDPGT activity up to 20 min. incubation of the microsomes with Fe(3+)/ascorbate (3-times); after 20 min. incubation, however, we observed a time-dependent decrease in this activity to basal levels after 4 hr incubation. Treatment of microsomes with 0.1% Triton X-100 (5 min.) lead to a similar increase in UDPGT activity; higher detergent concentrations produced a dose-dependent decrease in this activity to basal levels with 1% Triton X-100. Interestingly, UDPGT activity was susceptible to activation only when associated to microsomal membranes and the loss of activation correlated with the solubilization of this activity. UDPGT activation by either Fe(3+)/ascorbate or Triton X-100 was correlated with an increase in p-nitrophenol apparent K(m) and V(max) values. This activation was prevented or reversed by the reducing agents glutathione, cysteine or dithiothreitol when it was induced by the Fe(3+)/ascorbate. Furthermore, the latter provoked a significant decrease in microsomal thiol content, effect not observed after treatment with Triton X-100. Our results suggest that the main mechanism responsible for Fe(3+)/ascorbate-induced UDPGT activation is likely to be the promotion of protein sulfhydryl oxidation; this mechanism appears to be different from detergent-induced UDPGT activation.     

Tracheal and bronchoalveolar permeability changes in rats inhaling oxidant atmospheres during rest or exercise

Permeability of tracheal and bronchoalveolar airways of rats was measured and used to examine the effects of inhaled oxidant-containing atmospheres. The atmospheres studied were (a) ozone (O3) at 0.6 ppm (1.2 mg/m3) or 0.8 ppm (1.6 mg/m3); (b) nitrogen dioxide (NO2) at 6 ppm (11.3 mg/m3) or 12 ppm (22.6 mg/m3); (c) O3 + NO2 at 0.6 ppm (1.2 mg/m3) and 2.5 ppm (4.7 mg/m3), respectively; and (d) a 7-component particle and gas mixture (complex atmosphere) representing urban air pollution in a photochemical environment. The rats were exposed for 2 h. The effects of exercise during exposure were evaluated by exposing additional groups in an enclosed treadmill. Exposure of resting rats to 0.8 ppm O3 increased tracheal permeability to DTPA and bronchoalveolar permeability to diethylenetriamine pentaacetate (DTPA) and bovine serum albumin (BSA) at 1 h after the exposure. Bronchoalveolar, but not tracheal, permeability remained elevated at 24 h after the exposure. Exercise during exposure to O3 increased permeability to both tracers in the tracheal and the bronchoalveolar zones, and prolonged the duration of increased permeability in the tracheal zone from 1 h to 24 h, and in the bronchoalveolar zone from 24 h to 48 h. Permeability in the tracheal and bronchoalveolar zones of rats exposed at rest to 6 or 12 ppm NO2 did not differ from controls. However, rats exposed during exercise to 12 ppm NO2 for 2 h developed a significant increase in tracheal and bronchoalveolar permeability to DTPA and BSA at 1 h, but not at 24 or 48 h, after exposure. Exposure at rest to 0.6 ppm O3 plus 2.5 ppm NO2 significantly increased bronchoalveolar permeability at 1 and 24 h after exposure, although exposure at rest to 0.6 ppm O3 alone increased bronchoalveolar permeability only at 1 h after exposure. Exposure to O3 + NO2 during exercise led to significantly greater permeability to DTPA than did exercising exposure to O3 alone. Resting rats exposed to a complex gas/aerosol atmosphere composed of the above O3 and NO2 concentrations, plus 5 ppm (13.1 mg/m3) sulfur dioxide (SO2) and an aerosol of insoluble colloidal Fe2O3 with an aerosol of manganese, ferric, and ammonium salts, demonstrated increased permeability at 1 and 24 h after exposure. Nitric acid vapor was formed in both the O3 + NO2 atmosphere and the complex gas/aerosol atmosphere.(ABSTRACT TRUNCATED AT 400 WORDS)     

Functional changes in the lungs of guinea pigs exposed to sodium sulfite aerosols

Guinea pigs were exposed head only for 1 hr to submicrometer sodium sulfite aerosols (mass median aerodynamic diameter = 0.36 micron, sigma g = 2.96) at 474, 669, and 972 micrograms SO3(2-)/m3. Respiratory mechanics were measured in unanaesthetized animals before, during, and after exposure. Dose-related increases in resistance and decreases in compliance were observed. At 972 micrograms SO3(2-)/m3, the sodium sulfite aerosol caused a 50% increase in resistance and a 19% decrease in compliance. These changes were still present 1 hr after the end of exposure. The results were used to assess the irritant potency of sodium sulfite aerosol. Another group of guinea pigs was exposed whole body for 1 hr to the same aerosol at 0, 204, 395, and 1152 micrograms SO3(2-)/m3. Immediately after the exposures, lung volume, diffusion capacity for carbon monoxide (DLCO), and wet lung weight were evaluated in anesthetized, tracheotomized animals. As compared to controls, total lung capacity, vital capacity, functional residual capacity, residual volume, and DLCO were all decreased with increasing concentrations of sodium sulfite. Dose-related increases in wet lung weights were also observed. These results were compared with the irritant responses of animals exposed to zinc oxide and sulfur dioxide mixed under different conditions of temperature and humidity.     

Hepatotoxicity of microcystin-LR in fed and fasted rats

The LD50 (25 hr, i.p.) for microcystin-LR in fed rats (122 micrograms/kg) was significantly higher than that in fasted rats (72 micrograms/kg). At doses of 100, 150 and 200 micrograms of microcystin-LR per kg, the median times to death were 31.9, 18.2 and 11.2 hr for fed rats, and 1.8, 1.7 and 1.5 hr for fasted rats. A sublethal dose of microcystin (50 micrograms/kg) afforded protection to fasted, but not fed, rats against a subsequent lethal dose (200 micrograms/kg) challenge given 72 hr later. Biochemical and ultrastructural changes resulting from microcystin-LR (100 micrograms/kg, i.p.) were compared in fed and fasted rats 1 hr after injection. In both groups, liver weight and serum levels of sorbitol dehydrogenase and glucose significantly increased. Plasma membranes, isolated from livers of fed or fasted rats, exhibited similar toxin-induced changes in associated cytoskeletal elements. Liver mitochondria from toxin-treated, fasted rats exhibited complete inhibition of state 3 respiration, while those from toxin-treated, fed rats had ADP/O ratios and respiratory control indices comparable to control values. The primary event responsible for enhanced microcystin hepatotoxicity in the fasted state has not yet been identified. Depletion of glycogen stores and a decreased respiratory capacity may, however, play significant roles in this degenerative process.     

Isolated perfused lung histamine release, lipid peroxidation, and tissue superoxide dismutase from rats exposed to normobaric hyperoxia

Female Sprague-Dawley inbred rats were exposed to either 1 atm of 100% O2 for 24 h, or 65% O2 for 5 days, with or without pretreatment with disulfiram, an inhibitor of lung CuZn-SOD. After O2 exposure, the rats were killed, the lungs removed, and isolated perfused lungs (IPLs) prepared. The IPLs were perfused with modified Krebs-Henseleit buffer, and perfusate histamine, malondialdehyde (MDA), and lung tissue CuZn-SOD activity examined. Disulfiram administration decreased the LT50 of O2-exposed rats from 65 to 36 h. Histamine and MDA in the perfusate from the IPL prepared from rats exposed to 100% O2 for 24 h were markedly increased. When rats were pretreated with disulfiram and exposed to 100% O2 for 24 h, histamine and MDA were increased an additional 77% and 45%, respectively. In separate experiments, 100% O2 exposure significantly decreased lung CuZn-SOD activity by 40% while IPL histamine and MDA were significantly increased. However, exposure of rats to 65% O2 for 5 days decreased lung CuZn-SOD by 69% but did not affect IPL histamine release or perfusate MDA. These studies suggest that IPL histamine release and/or MDA may be an early biochemical marker for pulmonary O2 toxicity, that lung CuZn-SOD activity may not be the only determinant in O2 toxicity, and other defense mechanisms may play a vital protective role during sublethal O2 exposures.     

Pulmonary effects of a polyisocyanate aerosol: hexamethylene diisocyanate trimer (HDIt) or Desmodur-N (DES-N)

Desmodur-N (DES-N) or hexamethylene diisocyanate trimer (HDIt), a biuret structure of hexamethylene diisocyanate, is a viscous liquid used for durable coatings and is applied by brushing or spraying. DES-N aerosol has been shown to be primarily a pulmonary irritant following a single exposure in mice. To explore the pulmonary effects of this agent further, groups of guinea pigs were exposed to concentrations ranging from 8 to 121 mg/m3 of DES-N for 3 hr. Prior to and following exposure, each animal was challenged with 10% CO2 in 20% O2 and 70% N2 to evaluate their pulmonary performance. Following a single exposure, these animals displayed a concentration-dependent increase in respiratory rate and decrease in tidal volume, as well as coughing and apnea. Their ventilatory response to 10% CO2 was abnormal and characteristic of a lung restriction response. Some airflow limitation was seen during expiration but this occurred more often during air breathing than during CO2 challenge. With daily exposures repeated for 11 consecutive days, guinea pigs began to adapt to the exposures as indicated by a return to a normal ventilatory response to CO2. This adaptation occurred within the first 5 days of exposures. From Days 6 to 11, there was a demonstrable effect, but the level of response was much less than that following the first exposure. No cumulative effect could be demonstrated with this polyisocyanate and the effect was found to be different than that for mono- or diisocyanates. Acceptable levels of exposure to this polyisocyanate for industrial workers are suggested.     

Deposition, retention, and biological fate of inhaled benzo(a)pyrene adsorbed onto ultrafine particles and as a pure aerosol

The effect of ultrafine, airborne, carrier particles on the deposition, retention, and biological fate of inhaled polycyclic aromatic hydrocarbons (PAHs) was studied. Using a radiolabeled model PAH, [³H]benzo(a)pyrene ([³H]BaP), Fischer-344 rats were exposed by nose-only inhalation (30 min) to this compound, as a coating (15% by mass) on insoluble ⁶⁷Ga₂O₃ particles or as a pure aerosol. These aerosols were produced by vapor condensation methods in a dynamic aerosol generation system. The concentrations of [³H]BaP in the coated and homogeneous aerosols were 0.6 and 1.0 μg/liter of air, respectively, while the mass median diameter of both these aerosols was approximately 0.1 μm. Pulmonary retention of ³H radioactivity was longer in animals exposed to the [³H]BaP coated on the ⁶⁷Ga₂O₃ particles. The time required to clear 90% of the initial lung burden of ⁶⁷Ga₂O₃-associated ³H radioactivity detected 30 min postexposure was approximately 1 day as compared to 4 hr for animals exposed to the pure [³H]BaP aerosol. Tracheal clearance of 90% of the ⁶⁷Ga₂O₃-associated ³H radioactivity (as a fraction of the amount detected 30 min postexposure) required 1 day, while only 1.5 hr were required to clear the same percentage of ³H radioactivity from the tracheas of rats exposed to the pure [³H]BaP aerosol. The rates of clearance of this ³H material to other tissues suggested that a substantial amount of the [³H]BaP coated on ⁶⁷Ga₂O₃ was cleared from lungs by mucociliary clearance and subsequent ingestion, whereas the majority of the pure [³H]BaP aerosol was cleared by direct absorption into blood. In both cases, the ultimate fate of the majority of the [³H]BaP and its metabolites was excretion in feces. However, clearance of the ⁶⁷Ga₂O₃-associated [³H]BaP by ingestion may have been the cause for the higher levels and longer retention times of ³H radioactivity in stomach, liver, and kidneys when compared to the levels found in these same tissues from animals exposed to the pure [³H]BaP aerosol. Thus, particle association of BaP not only increased the respiratory tract retention of this PAH, but also increased the dose of this compound and its metabolites to stomach, liver, and kidneys.