Roy J. Rando

Passive cigarette smoke—challenge studies: Increase in bronchial hyperreactivity

Degree and duration of bronchial hyperreactivity (BHR) after environmental tobacco smoke (ETS) inhalation was assessed in 31 smoke-sensitive subjects with asthma who exhibited lower airway symptoms on ETS exposure (group I) and 39 smoke-sensitive subjects without asthma who manifested only upper airway symptoms on cigarette-smoke exposure (group II). Subjects were challenged with ETS for 4 hours in a static-test chamber. The atmosphere was continuously monitored for airborne particulate levels (800 cpm), total suspended particulates (1266 +/- 283 micrograms/m3), and airborne nicotine levels (226 +/- 49 micrograms/m2). Methacholine challenges were performed before and serially after cigarette-smoke exposure, and the provocative dose causing a 20% fall in FEV1 was determined. Five of the 31 smoke-sensitive subjects with asthma and none of the smoke-sensitive subjects without asthma reacted to cigarette-smoke challenge (greater than or equal to 20% fall from baseline FEV1). Thirty-two percent (10/31) of the subjects with asthma demonstrated increased BHR at 6 hours, 29% (9/31) at 24 hours, and 13% (4/31) up to day 14 after ETS challenge. Of the subjects without asthma, 18% (7/39) demonstrated increased BHR at 6 hours, 10% (4/39) at 24 hours, and 8% (3/39) at 3 weeks. These studies demonstrated an increase in BHR after cigarette-smoke challenge in a number of study subjects (although they were clinically asymptomatic) and suggest that prolonged subclinical airway inflammation can occur in the absence of demonstrable change in airway caliber on exposure to ETS.

Identification of triptolide, a natural diterpenoid compound, as an inhibitor of lung inflammation.

Inflammation is associated with various pulmonary diseases and contributes to the pathogenesis of acute lung injury. We previously identified a proinflammatory signaling pathway triggered by G protein-coupled receptors (GPCRs) in which stimulation of G(q)-coupled GPCRs results in activation of the transcription factor NF-kappaB. Because damage to the lung causes the release of multiple mediators acting through G(q)-coupled GPCRs, this signaling pathway is likely to contribute to inflammatory processes in the injured lung. In an effort to identify novel inhibitors of lung inflammation, the National Institutes of Health Clinical Collection, a library of 446 compounds, was screened for inhibitory activity toward production of IL-8 induced by stimulation of the G(q)-coupled tachykinin 1 receptor with substance P in A549 cells. Twenty-eight compounds that significantly inhibited substance P-induced IL-8 production were identified. The most potent inhibitor was triptolide, a diterpenoid compound from Tripterygium wilfordii Hook F, a vine used in traditional Chinese medicine for the treatment of autoimmune diseases. Triptolide inhibited IL-8 production induced by substance P with an IC(50) of 2.3 x 10(-8) M and inhibited NF-kappaB activation in response to an agonist of the protease-activated receptor 2 with an IC(50) of 1.4 x 10(-8) M. Anti-inflammatory effects of triptolide were assessed in vivo using a chlorine gas lung injury model in mice. Triptolide inhibited neutrophilic inflammation and the production of KC (Cxcl1) in the lungs of chlorine-exposed mice. The results demonstrate that triptolide exhibits anti-inflammatory activity in cultured lung cells and in an in vivo model of acute lung injury.

Acute Lung Injury Induced by Chlorine Inhalation in C57BL/6 and FVB/N Mice

Humans may be exposed to chlorine gas via accidental or intentional release, and effective countermeasures for the resulting lung injury are lacking. To develop a model in which therapeutic measures could be evaluated, lung injury induced by chlorine inhalation in two inbred mouse strains was examined. C57BL/6 and FVB/N mice were exposed for 1.1 h to varying doses of chlorine (197–289 ppm-h) and were evaluated for indices of lung injury at different times after exposure (6–48 h). Chlorine induced increases in lung weight that were more evident in FVB/N mice than in C57BL/6 mice. Both strains exhibited sloughing of airway epithelium observed within 6 h after exposure. As judged by Ly-6G immunostaining, chlorine exposure caused widespread neutrophil influx into the lung parenchyma at 6 h followed by a clustering of neutrophils around damaged airways by 24 h. High levels of cellular proliferation revealed by Ki-67 staining were observed in airway epithelium 48 h after exposure. Lavage fluid parameters showed consistent trends in both strains. Lavage fluid protein content was elevated throughout the times examined. Lavage fluid neutrophils were significantly increased beginning 12 h after exposure and were highest at 48 h. The concentration of the neutrophil chemoattractant KC peaked 6 h after exposure and was near baseline by 48 h. In summary, chlorine inhalation resulted in lung injury characterized by edema, epithelial cell death, and neutrophilic inflammation in C57BL/6 and FVB/N mice. Characterization of such responses in these mice will allow testing of therapeutic agents to treat chlorine-induced lung injury.

Occupational Exposure of Nonsmoking Nightclub Musicians to Environmental Tobacco Smoke

This study assessed environmental tobacco smoke (ETS) exposures of nonsmoking musicians in nightclub environments using total suspended particulate (TSP), the ultraviolet absorbing fraction of TSP (UVPM), gaseous nicotine, saliva nicotine, saliva cotinine, and perceived smokiness as exposure/dose indicators. Measured exposures were as high or higher than those of other occupational groups studied. TSP ranged from 110 to 1714 micrograms/m3 (mean 502, SD 390 micrograms/m3). UVPM (mean 221, SD 95 micrograms/m3) was associated with gaseous and saliva nicotine concentrations. Paired-sample variation was much higher for TSP than for UVPM. Correlation of TSP with UVPM, gaseous nicotine, and saliva nicotine was poor. Paired-sample gaseous nicotine results were similar, with exposures of 28.0 to 50.0 micrograms/m3 (mean 37.1, SD 6.9 micrograms/m3), and were high compared with previous studies. These results suggested that nightclub musicians may be exposed to higher concentrations of ETS than some other occupational groups. Saliva nicotine results were consistent with those previously reported with regard to the range of values, large variation observed, and increase in saliva nicotine levels observable after only a few hours of exposure. Saliva nicotine results could not be correlated with other measures of exposure and did not appear to be a reliable biological indicator of absorbed dose. Saliva cotinine levels were comparable to other occupational groups studied, but were lower than previous findings for bartenders and waitresses. Levels ranged from 1.7 to 5.0 ng/mL (mean 3.4, SD 0.9 ng/mL), and increased with number of exposures during the workweek, but did not correlate with other ETS indicators.

Inhalation of benzene leads to an increase in the mutant frequencies of a lacI transgene in lung and spleen tissues of mice

The goal of this study was to determine if inhalation of benzene leads to an increase in the mutant frequencies in the tissues of male C57BL/6 mice. Mutant frequencies were measured using a previously described assay in which bacteriophage lambda lacI transgenes are rescued from mouse genomic DNA as infectious phage and scored for their LacI phenotype. Eight experimental mice were exposed to a target concentration of 300 ppm of benzene for 6 h/day x 5 days/week x 12 weeks, and eight control mice were treated similarly except that they were not exposed to benzene. Mutant frequencies were calculated as the ratio of LacI-/total phage recovered from organs of interest. The mean mutant frequency measured in lung tissues of mice exposed to benzene was (10.6 +/- 1.4) x 10(-5), which is about 1.7-fold higher than that of the unexposed controls. In spleen tissues from benzene-exposed mice the mean mutation frequency was (12.6 +/- 4.1) x 10(-5), which is about 1.5-fold higher than that of spleen tissues from unexposed controls. The differences in mean mutant frequencies between benzene-exposed and unexposed lung and spleen tissues are statistically significant. In liver tissues, however, the mean mutant frequencies of benzene-exposed mice and unexposed mice are not significantly different. These results demonstrate that inhaled benzene results in a statistically significant increase in the mutant frequencies in lung and spleen, but not in liver tissues of mice.

Isomeric Composition of Airborne TDI in the Polyurethane Foam Industry

The isomeric composition of airborne toluene diisocyanate has been determined in two plants producing slab stock flexible polyurethane foam. The high performance liquid chromatographic technique used for analysis of the collected samples was optimized for the quantitative and qualitative determination of 2,4 and 2,6-TDI. The data indicates that there is a very large increase in the amount of airborne 2,6-TDI relative to the 2,4 isomer, as compared to the starting material used in the process. The magnitude of the increase was dependent on the stage of production. These results are consistent with the hypothesis of increased offgassing of 2,6-TDI, due to its lower reactivity.

Deviations from Haber's Law for Multiple Measures of Acute Lung Injury in Chlorine-Exposed Mice

Chlorine gas is considered a chemical threat agent that can cause acute lung injury. Studies in the early 20th century on war gases led Haber to postulate that the dose of an inhaled chemical expressed as the product of gas concentration and exposure time leads to a constant toxicological effect (Habers Law). In the present work, mice were exposed to a constant dose of chlorine (100 ppm-h) delivered using different combinations of concentration and time (800 ppm/7.5 min, 400 ppm/15 min, 200 ppm/30 min, and 100 ppm/60 min). Significant effects of exposure protocol on survival evaluated 6 h after exposure were observed, ranging from 0% for the 7.5-min exposure to 100% for the 30- and 60-min exposures. Multiple parameters indicative of lung injury were examined to determine if any aspects of lung injury were differentially affected by the exposure protocols. Most parameters (pulmonary edema, neutrophil influx, and levels of protein, immunoglobulin M, and the chemokine KC [Cxcl1] in lavage fluid) indicated that lung injury was most pronounced for the 15-min exposure and least for the 60-min exposure. In contrast, changes in pulmonary function at baseline and in response to inhaled methacholine were similar following the three exposure regimens. The results indicate that the extent of lung injury following chlorine inhalation depends not only on total dose but also on the specifics of exposure concentration and time, and they suggest that evaluation of countermeasures against chlorine-induced lung injury should be performed using multiple types of exposure scenarios.

Inhibition of chlorine-induced lung injury by the type 4 phosphodiesterase inhibitor rolipram

Chlorine is a highly toxic respiratory irritant that when inhaled causes epithelial cell injury, alveolar-capillary barrier disruption, airway hyperreactivity, inflammation, and pulmonary edema. Chlorine is considered a chemical threat agent, and its release through accidental or intentional means has the potential to result in mass casualties from acute lung injury. The type 4 phosphodiesterase inhibitor rolipram was investigated as a rescue treatment for chlorine-induced lung injury. Rolipram inhibits degradation of the intracellular signaling molecule cyclic AMP. Potential beneficial effects of increased cyclic AMP levels include inhibition of pulmonary edema, inflammation, and airway hyperreactivity. Mice were exposed to chlorine (whole body exposure, 228-270 ppm for 1 h) and were treated with rolipram by intraperitoneal, intranasal, or intramuscular (either aqueous or nanoemulsion formulation) delivery starting 1h after exposure. Rolipram administered intraperitoneally or intranasally inhibited chlorine-induced pulmonary edema. Minor or no effects were observed on lavage fluid IgM (indicative of plasma protein leakage), KC (Cxcl1, neutrophil chemoattractant), and neutrophils. All routes of administration inhibited chlorine-induced airway hyperreactivity assessed 1 day after exposure. The results of the study suggest that rolipram may be an effective rescue treatment for chlorine-induced lung injury and that both systemic and targeted administration to the respiratory tract were effective routes of delivery.

Wood Dust Sampling: Field Evaluation of Personal Samplers When Large Particles Are Present

Recent recommendations for wood dust sampling include sampling according to the inhalable convention of International Organization for Standardization (ISO) 7708 (1995) Air quality—particle size fraction definitions for health-related sampling. However, a specific sampling device is not mandated, and while several samplers have laboratory performance approaching theoretical for an ‘inhalable’ sampler, the best choice of sampler for wood dust is not clear. A side-by-side field study was considered the most practical test of samplers as laboratory performance tests consider overall performance based on a wider range of particle sizes than are commonly encountered in the wood products industry. Seven companies in the wood products industry of the Southeast USA (MS, KY, AL, and WV) participated in this study. The products included hardwood flooring, engineered hardwood flooring, door skins, shutter blinds, kitchen cabinets, plywood, and veneer. The samplers selected were 37-mm closed-face cassette with ACCU-CAP™, Button, CIP10-I, GSP, and Institute of Occupational Medicine. Approximately 30 of each possible pairwise combination of samplers were collected as personal sample sets. Paired samplers of the same type were used to calculate environmental variance that was then used to determine the number of pairs of samples necessary to detect any difference at a specified level of confidence. Total valid sample number was 888 (444 valid pairs). The mass concentration of wood dust ranged from 0.02 to 195 mg m−3. Geometric mean (geometric standard deviation) and arithmetic mean (standard deviation) of wood dust were 0.98 mg m−3 (3.06) and 2.12 mg m−3 (7.74), respectively. One percent of the samples exceeded 15 mg m−3, 6% exceeded 5 mg m−3, and 48% exceeded 1 mg m−3. The number of collected pairs is generally appropriate to detect a 35% difference when outliers (negative mass loadings) are removed. Statistical evaluation of the nonsimilar sampler pair results produced a finding of no significant difference between any pairing of sampler type. A practical consideration for sampling in the USA is that the ACCU-CAP™ is similar to the sampler currently used by the Occupational Safety and Health Administration for purposes of demonstrating compliance with its permissible exposure limit for wood dust, which is the same as for Particles Not Otherwise Regulated, also known as inert dust or nuisance dust (Method PV2121).

Critical evaluation of continuous monitors for toluene diisocyanate

This paper describes the experiments and the results of an exhaustive and critical laboratory study which evaluated the performance of continuous area and personal monitors popularly used in measuring the concentrations of TDI in industrial environments. The studies consisted of dynamic calibrations, the effect of humidity and temperature on the measurements, and the interferences due to commonly encountered contaminants in a TDI manufacturing plant. The response and resolution of the personal monitor for short-term fluctuations of TDI concentration in time and space are also reported.