Faculty Bibliography

To examine the biological plausibility of the adverse health effects of ambient paniculate matter (PM), we have studied the pulmonary and cardiovascular effects of ambient PM in an animal model of pulmonary hypertension. A centrifugal aerosol concentrator was used to concentrate ambient paniculate matter up to 10-fold. Normal and monocrotaline-treated rats were exposed nose-only for 3 hrs to filtered air or concentrated ambient PM ranging from 160 to 900 ?g/m3. At 3 hrs post-exposure, the percentage of neutrophils in peripheral blood was significantly elevated in PM-exposed animals while the percentage of lymphocytes was decreased. This demargination of neutrophils occurred in both normal and monocrotaline-treated animals. The hematological changes returned to control levels by 24 hrs after exposure. Evidence that these changes were a result of exposure to the particulate and not the gaseous components of urban air was demonstrated by a lack of hematological changes in animals exposed to HEPA-filtered air from the centrifugal concentrator. Heart rate and core temperature were monitored in unrestrained normal and monocrotaline-treated rats for 1 hr prior to, during, and 24 hrs after, single or multiple exposures to PM. No consistent changes in heart rate or core temperature were observed after exposure to concentrated ambient PM. Pulmonary injury, as evidenced by increased protein levels in lavage fluid, occurred in monocrotaline-treated but not in normal animals exposed to greater than 360 ?g/m3 PM. The observed pattern of hematological changes suggests an activation of the sympathetic stress response which appeared to manifest pulmonary injury only in animals with compromised health

Humans exhibit an acute inflammatory response in the lungs after controlled laboratory exposure to ozone. The present study was designed to test whether biomarkers of inflammation are detectable in humans exposed to ozone and associated copollutants under natural conditions outdoors. Bronchoscopy with bronchoalveolar lavage (BAL) was carried out on 19 normal volunteer joggers from Governors Island, NY, who exercised in the afternoon during the 1992 summer (S1) season. Fifteen subjects were retested during the following, low ozone, winter season (W). The BAL protocol involved an initial instillation of 20 ml saline followed by four sequential 50-ml saline washes carried out in both the right middle lobe and the lingula. The eight 50-ml samples were pooled as the 'alveolar' sample. Analyses performed on the alveolar lavage samples included cell differentials, release of IL-8, TNF-alpha, and reactive oxygen species (ROS) by pooled cells, and levels of IL-8, protein, LDH, fibronectin, alpha1-antitrypsin (alpha1-AT), complement fragment 3a (C3a), and prostaglandin E2 (PGE2) in lavage fluids. Release of ROS by stimulated BAL cells was lower in S1 than in W (p = 0.03). In contrast, LDH levels in BAL fluids were 2-fold higher in S1 than in W (p = 0.02), as were IL-8 (p = 0.12) and PGE2 (p = 0.06). These results suggest a possible ongoing inflammatory response in the lungs of recreational joggers exposed to ozone and associated copollutants during the summer months

This study examined the production of stored mucosubtances in rats after repeated exposure to aerosolized endotoxin, a common contaminant of bioaerosols. Male Fischer 344 rats were exposed to aerosolized saline (sham control) or endotoxin (target concentrations of 0.05, 0.5, and 5.0 micrograms/m3) for 3 h/day, 5 days/week for 4 weeks. Following the final exposure, the left lung of each animal was lavaged and the right lung and nasal cavity were fixed with buffered formalin. Morphometric examination of Alcian blue/Periodic acid Schiffs-stained (AB/PAS) lung sections demonstrated dose-dependent increases in stored intraepithelial mucosubstances in the intrapulmonary airways of endotoxin-exposed rats. Threefold and eightfold increases in stored mucosubstances were observed in generation 5 airways of animals exposed to 0.5 or 5.0 microgram/m3 endotoxin, respectively (p < .05). This mucous cell metaplasia in the intrapulmonary airways was not accompanied by evidence of lung inflammation or increased AB/PAS-staining high molecular weight material in lavage fluid. Furthermore, despite significant deposition of endotoxin aerosols (mass median aerodynamic diameter of 1.9 microns) in the nasal cavity, no significant changes in stored mucosubstances were observed in the nasal septum. In animals repeatedly exposed to 5.0 micrograms/m3 endotoxin and allowed to recover for 1 month, stored mucosubstances in the intrapulmonary airway were still more than fivefold greater than control values. Thus, in rats, repeated exposure to inhaled endotoxin produced a persistent mucous cell metaplasia only in the intrapulmonary airways

The surfactant layer covering the gas-exchange region of the lung serves as the initial site of interaction with inhaled oxidant gases. Among the endogenous compounds potentially vulnerable to oxidative injury are surfactant proteins. This study focused on the effect of ozone on surfactant protein A (SP-A) function, content, and gene expression. To determine the time course of response to ozone, guinea pigs were exposed to 0.2-0.8 parts/million (ppm) ozone for 6 h and were killed up to 120 h postexposure. To determine the effect of repeated exposure, animals were exposed to 0.8 ppm ozone for 6 h/day and were killed on days 3 and 5. A significant increase in surfactant's ability to modulate the respiratory burst induced by phorbol 12-myristate 13-acetate in naive macrophages was observed at 24 h after a single 0.8 ppm ozone exposure. Because neutralizing antibodies to SP-A blunted this stimulatory effect, we hypothesized that ozone enhanced the modulatory role of SP-A in macrophage function. This alteration in function was accompanied by an influx of inflammatory cells and only marginal changes in SP-A levels as determined by an enzyme-linked immunosorbent assay. No significant changes in steady-state levels of SP-A mRNA were observed after single or repeated exposure to ozone. Thus the inflammation that accompanies in vivo ozone exposure may result in a change in the structure and thus functional role of SP-A in modulating macrophage activity

Occupational exposure to microbial-contaminated machining fluids is associated with a variety of adverse pulmonary effects including chronic bronchitis and increased sputum production. We have previously demonstrated in F344 rats that inhaled endotoxin can increase the amount of stored intraepithelial mucosubstances (Vs) in the respiratory tract. The purpose of the present study was to examine the effect of endotoxin-contaminated machining fluid aerosols on mucous production. Rats were exposed to aerosols of pyrogen-free water, 1 or 10 mg/m3 used machining fluid, or 10 mg/m3 unused machining fluid for 3 hr/day for 3 days. Twenty-four hours after the final exposure, right lung lobes were lavaged and the nasal cavity and left lung were fixed in formalin. The amount of Alcian blue/periodic acid-Schiff-stained mucosubstances was determined by morphometry. Exposure to 10 mg/m3 used machining fluid (equivalent to 0.8 micrograms/m3 endotoxin) produced a significant increase in Vs in the epithelial lining of both the nasal septum and intrapulmonary airways. These changes in Vs were accompanied by a significant increase in total cells and neutrophils in the lavage fluid. No changes in stored mucosubstances or lavage parameters were found in animals exposed to 1 mg/m3 used machining fluid aerosols. A significant increase in Vs was observed in the nasal septum but not in the intrapulmonary airways of animals exposed to 10 mg/m3 unused machining fluids (no measurable endotoxin). These results suggest that in addition to endotoxin, nonendotoxin components of machining fluids may contribute to the increase in sputum and chronic bronchitis reported for workers exposed to machining fluid aerosols

Occupational exposure to endotoxin-contaminated organic dusts is associated with a variety of adverse pulmonary effects, including chronic bronchitis and sputum production. We have previously demonstrated in F344 rats that inhaled endotoxin rapidly induces an increase in the volume of stored intraepithelial mucosubstances (Vs) in the respiratory tract. The present study examined whether endotoxin-contaminated cotton dust can produce a similar increase in Vs in this animal model. Rats were exposed to air or 1.5 to 15.0 mg/m3 cotton dust for 2 h/d for 3 d. Twenty-four hours after the final exposure, the nasal cavity and lungs were fixed in formalin and the presence of Alcian blue/periodic acid-Schiff-staining mucosubstances determined by morphometry. Exposure to cotton dust produced concentration-dependent changes in Vs in the nasal septum and intrapulmonary airways. Statistically significant increases in Vs were observed in the epithelial lining of the nasal septum of animals exposed to 5.3 and 14.5 mg/m3 cotton dust (equivalent to 2.8 and 8.9 micrograms/m3 endotoxin). Vs in the intrapulmonary airways was also significantly increased at these concentrations. No significant changes were observed in the nasal septum or intrapulmonary airways after exposure to 1.8 mg/m3 cotton dust. These results are consistent with the hypothesis that endotoxin may contribute to the increase in human cases of chronic bronchitis reported in occupational settings in which endotoxin-contaminated dusts are encountered

Components of the complement system are thought to play a role in the respiratory effects of inhaled organic dusts encountered in occupational settings such as grain storage sites and cotton mills. This hypothesis is based upon observations that grain dusts and endotoxin can activate complement in vitro and in vivo and that the complement system appears to contribute to the endotoxin-induced lung injury of septic shock. The present study examined whether the inflammatory effects of inhaled endotoxin and cotton dust were (1) blocked by decomplementing animals with cobra venom factor (CVF) or (2) present in animals genetically deficient in specific components of the complement system. In untreated and sham-treated guinea pigs, inhaled endotoxin produced significant increases in biochemical and cellular indices of pulmonary injury in bronchoalveolar lavage fluid. Treatment of guinea pigs with 200 units CVF/kg at 24 h prior to exposure did not block the biochemical and cellular changes produced by inhaled endotoxin or cotton dust. Guinea pigs deficient in the fourth component of complement (C4) also demonstrated evidence of pulmonary injury roughly equivalent to that of normal guinea pigs exposed to inhaled endotoxin or cotton dust. Finally, mice genetically deficient in the fifth component of complement (C5) did not exhibit a decreased pulmonary response to inhaled endotoxin compared to C5-sufficient mice. These studies suggest that despite in vitro and in vivo evidence of complement activation by endotoxin and extracts of organic dusts, the complement system may not play a major role in the acute respiratory effects of inhaled endotoxin and cotton dust

Increased sputum production and chronic bronchitis are associated with occupational exposure to endotoxin-contaminated organic ducts. The present study examined whether repeated exposure to occupationally relevant concentrations of airborne endotoxin in the F344 rat can alter the volume density of stored intraepithelial mucosubstances (Vs) in the respiratory tract. Rats were exposed to saline or endotoxin aerosols for 3 h/day for 3 days and were killed 24 h after the last exposure. Quantitative histochemistry of Vs in airway epithelium was examined at three distinct levels of the respiratory tract (nose, trachea, and lung). Exposure to endotoxin produced a dose-dependent increase in Vs in the intrapulmonary airways. The quantity of Vs in the intrapulmonary airways was significantly increased in animals exposed to as little as 0.3 micrograms/m3 endotoxin. Significant increases in Vs were observed in the trachea only after exposure to > or = 3.1 micrograms/m3 endotoxin, whereas no significant changes were observed in the nasal airways even at concentrations as high as 52.4 micrograms/m3. These results are consistent with earlier findings in which repeated instillation of endotoxin produced significant increases in Vs in the epithelial lining of the pulmonary airways and demonstrate that inhaled endotoxin may play a role in the increase in sputum and chronic bronchitis reported for workers exposed to organic dusts

In vivo exposure to sulfuric acid aerosols produces profound effects on pulmonary macrophage (PM phi) phagocytic function and cytokine release and perturbs intracellular pH (pHi) homeostasis. Because pHi influences a multitude of cellular processes, we sought to investigate the mechanism by which acid aerosol exposure affects its regulation. Guinea pigs underwent a single or 5 repeated 3-hr exposures to sulfuric acid aerosol (969 and 974 micrograms/m3 for single and repeated exposures, respectively). PM phi harvested immediately after exposure were incubated in HCO3-free media and their pHi recovery from an intracellular acid load was examined. The overall pHi recovery was depressed after single and multiple exposures to sulfuric acid aerosol. delta pHi (the difference between initial pHi and the one measured at 150 sec) decreased by 15.6 and 23.3% (p < 0.05) for single and repeated exposures, respectively. Initial dpHi/dt (maximum pHi recovery rate) after cytoplasmic acidification diminished by 20.3 and 32.2%, which were not statistically significant (p = 0.08 for repeated exposure). To determine whether the activity of the H(+)-ATPase pump the Na(+)-H+ exchanger was specifically altered by the acid exposures, PM phi were first incubated in Na+ and HCO3-free media with NBD-Cl (7-chloro-4-nitrobenz-2-oxa-1,3-diazol, blocking H(+)-ATPase and leaving only the Na(+)-H+ exchanger in effect) and then challenged with 30 mM NaCl. The pHi recovery of PM phi after Na challenge was significantly reduced in acid aerosol exposed guinea pigs (p < 0.05) compared to controls (for delta pHi, 18.2% lower in single exposure and 22.7% in multiple exposure groups; for initial dpHi/dt, 26.9% lower in single exposure and 22.4% in multiple exposure groups). In contrast, the H(+)-ATPase pump was inconsistently affected as indicated by delta pHi and initial dpHi/dt measured in the presence of MIA (amiloride-5-N-methylisobutyl, inhibiting the Na(+)-H+ exchanger and leaving only the H(+)-ATPase pump in effect). These results suggest that in vivo exposure to sulfuric acid aerosols induces alterations in pHi regulation in guinea pig PM phi attributable to changes in Na(+)-H+ exchanger activity

Metal fume fever is an acute self-limited illness induced most commonly by inhalation of zinc oxide fumes. The affected individual characteristically experiences the rapid onset of intense shaking chills, fever, and body aches a few hours after exposure, and symptoms dissipate spontaneously. While the occurrence of metal fume fever appears to be widespread and the current TLV/PEL of 5 mg/m3 and STEL of 10 mg/m3 may not be fully protective, no chronic health sequelae have been documented to date. Nonetheless, as any worker who has experienced a full-blown case will likely testify, metal fume fever remains one of the more noxious short-term illnesses contracted in the workplace, and its prevention deserves serious attention