Paul E. Morrow

Possible Mechanisms to Explain Dust Overloading of the Lungs

This paper briefly reviews the available evidence on dust overloading of the lungs, a condition which has come to the forefront in many recently reported chronic inhalation studies. A general hypothesis is developed that dust overloading, which is typified by a progressive reduction of particle clearance from the deep lung, reflects a breakdown in alveolar macrophage (AM)-mediated dust removal due to the loss of AM mobility. The inability of the dust-laden AMs to translocate to the mucociliary escalator is correlated to an average composite particle volume per alveolar macrophage in the lung. When this particulate volume exceeds approximately 60 micron3/AM, on the basis of a uniform distribution of particles over the AM pool size (approximately 2.5 X 10(7) cells) in the Fischer 344 rat, the overload effect appears to be initiated. When the distributed particulate volume exceeds approximately 600 micron3 per cell, the evidence suggests that AM-mediated particle clearance virtually ceases and agglomerated particle-laden macrophages remain in the alveolar region. This paper considers possible mechanisms why these particle-laden cells are immobilized, viz., one is based on excessive particle-cell, cell-cell chemotactic interactions, and migratory inhibition factors; the other considers the volumetric increase by phagocytized particles, per se, as leading to an inability of the AM to spread and migrate probably through a competitive requirement for surface membrane and cytoskeleton in both endocytotic and migratory functions.

Ultrafine Particle Deposition in Humans During Rest and Exercise

Ultrafine particles (diameter < 100 nm) may be important in the health effects of air pollution, in part because of their predicted high respiratory deposition. However, there are few measurements of ultrafine particle deposition during spontaneous breathing. The fractional deposition for the total respiratory tract of ultrafine carbon particles (count median diameter = 26 nm, geometric standard deviation = 1.6) was measured in 12 healthy subjects (6 female, 6 male) at rest (minute ventilation 9.0 ± 1.3 L/min) using a mouthpiece exposure system. The mean ± SD fractional deposition was 0.66 ± 0.11 by particle number and 0.58 ± 0.13 by particle mass concentration, similar to model predictions. The number deposition fraction increased as particle size decreased, reaching 0.80 ± 0.09 for the smallest particles (midpoint count median diameter = 8.7 nm). No gender differences were observed. In an additional 7 subjects (2 female, 5 male) alternating rest with moderate exercise (minute ventilation 38.1 ± 9.5 L/min), the deposition fraction during exercise increased to 0.83 ± 0.04 and 0.76 ± 0.06 by particle number and mass concentration, respectively, and reached 0.94 ± 0.02 for the smallest particles. Experimental deposition data exceeded model predictions during exercise. The total number of deposited particles was more than 4.5-fold higher during exercise than at rest because of the combined increase in deposition fraction and minute ventilation. Fractional deposition of ultrafine particles during mouth breathing is high in healthy subjects, and increases further with exercise.

Dust overloading of the lungs: Update and appraisal

This article reviews recent studies which involve, or impact on, the condition of dust overloading in the lungs of several species, especially the Fischer 344 rat. Its main purpose is to provide an update of the overload concept and new information of possible mechanistic relevance. At present, the most likely general explanation for the suppression of particle transport by the alveolar macrophage (AM) and the development of concurrent events, e.g., increased interstitial dust uptake and prolonged inflammatory response, is the persistent, possibly excessive, elaboration of chemotactic and chemokinetic factors by the AM. The induction of these interrelated events is hypothesized as related to the volume of dust phagocytized by the AM pool. The review concludes, inter alia, that information is badly needed on dust overload in nonrodent species and on the normal role of the AM in dust removal from the human lungs.

Toxicological data on NOx: an overview.

This overview is based on experimental and epidemiological studies of NOx toxicity during the past decade. Approximately 130 published studies are cited and about one-fourth of these are discussed briefly under one of the following headings: acute and subacute studies, chronic low-level studies, human studies, and special studies. The latter section examines a selection of comparatively unique investigations, including several devoted to the pulmonary uptake and retention of NO2, and several examining the potential tumorigenicity of NO2. For each major section of the overview, a critical evaluation is attempted in terms of the impact of the appropriate studies on the extant NOx toxicological data base and on the current and planned air quality standards for NOx.

Inhalation of Ultrafine Particles Alters Blood Leukocyte Expression of Adhesion Molecules in Humans

Ultrafine particles (UFPs; aerodynamic diameter < 100 nm) may contribute to the respiratory and cardiovascular morbidity and mortality associated with particulate air pollution. We tested the hypothesis that inhalation of carbon UFPs has vascular effects in healthy and asthmatic subjects, detectable as alterations in blood leukocyte expression of adhesion molecules. Healthy subjects inhaled filtered air and freshly generated elemental carbon particles (count median diameter ~ 25 nm, geometric standard deviation ~ 1.6), for 2 hr, in three separate protocols: 10 μg/m3 at rest, 10 and 25 μg/m3 with exercise, and 50 μg/m3 with exercise. In a fourth protocol, subjects with asthma inhaled air and 10 μg/m3 UFPs with exercise. Peripheral venous blood was obtained before and at intervals after exposure, and leukocyte expression of surface markers was quantitated using multiparameter flow cytometry. In healthy subjects, particle exposure with exercise reduced expression of adhesion molecules CD54 and CD18 on monocytes and CD18 and CD49d on granulocytes. There were also concentration-related reductions in blood monocytes, basophils, and eosinophils and increased lymphocyte expression of the activation marker CD25. In subjects with asthma, exposure with exercise to 10 μg/m3 UFPs reduced expression of CD11b on monocytes and eosinophils and CD54 on granulocytes. Particle exposure also reduced the percentage of CD4+ T cells, basophils, and eosinophils. Inhalation of elemental carbon UFPs alters peripheral blood leukocyte distribution and expression of adhesion molecules, in a pattern consistent with increased retention of leukocytes in the pulmonary vascular bed.

Pulmonary Function, Diffusing Capacity, and Inflammation in Healthy and Asthmatic Subjects Exposed to Ultrafine Particles

Particulate air pollution is associated with asthma exacerbations and increased morbidity and mortality from respiratory causes. Ultrafine particles (particles less than 0.1 μ m in diameter) may contribute to these adverse effects because they have a higher predicted pulmonary deposition, greater potential to induce pulmonary inflammation, larger surface area, and enhanced oxidant capacity when compared with larger particles on a mass basis. We hypothesized that ultrafine particle exposure would induce airway inflammation in susceptible humans. This hypothesis was tested in a series of randomized, double-blind studies by exposing healthy subjects and mild asthmatic subjects to carbon ultrafine particles versus filtered air. Both exposures were delivered via a mouthpiece system during rest and moderate exercise. Healthy subjects were exposed to particle concentrations of 10, 25, and 50 μ g/m3, while asthmatics were exposed to 10 μ g/m3. Lung function and airway inflammation were assessed by symptom scores, pulmonary function tests, and airway nitric oxide parameters. Airway inflammatory cells were measured via induced sputum analysis in several of the protocols. There were no differences in any of these measurements in normal or asthmatic subjects when exposed to ultrafine particles at concentrations of 10 or 25 μ g/m3. However, exposing 16 normal subjects to the higher concentration of 50 μ g/m3 caused a reduction in maximal midexpiratory flow rate (−4.34 ± 1.78% [ultrafine particles] vs. +1.08 ± 1.86% [air], p =. 042) and carbon monoxide diffusing capacity (−1.76 ± 0.66 ml/min/mm Hg [ultrafine particles] vs. −0.18 ± 0.41 ml/min/mm Hg [air], p =. 040) at 21 h after exposure. There were no consistent differences in symptoms, induced sputum, or exhaled nitric oxide parameters in any of these studies. These results suggest that exposure to carbon ultrafine particles results in mild small-airways dysfunction together with impaired alveolar gas exchange in normal subjects. These effects do not appear related to airway inflammation. Additional studies are required to confirm these findings in normal subjects, compare them with additional susceptible patient populations, and determine their pathophysiologic mechanisms.

Volumetric loading of alveolar macrophages (AM): a possible basis for diminished AM-mediated particle clearance.

Using intratracheal instillation of radioactively labeled plastic microspheres of 3.3 and 10.3 microns diameter at two dose levels, this 7-month study in male Fischer 344 rats was designed to test a volumetric particulate burden hypothesis that has been proposed as a mechanistic basis for the condition of dust overloading of the lungs with highly insoluble particles of very low toxicity and to explain the prolongation of pulmonary particle retention. The study utilized airway and deep lung lavage techniques, scanning electron and optical microscopy of lavaged cells and lungs of sacrificed animals, particle distribution in alveolar macrophages (AM), fecal recovery of radioactive particles, and lung retention measurements by external counting. Microscopic assessments revealed that essentially all of the 3.3- and 10.3-microns-diameter particles were phagocytized by AM within 24 h postinstillation. One phagocytized 10.3-microns particle is capable of producing the hypothesized 600-microns 3/AM overload criterion for virtual AM immobilization. Neither the number nor the volume of 3.3-microns-diameter particles instilled was large enough to produce volumetric overloading assuming uniform distribution of the particles in the lung. In contrast to the 3.3-microns particles, the 10.3-microns particles were apparently sequestered to a greater extent and capable of greatly prolonging AM-mediated clearance of particles from the pulmonary region. The measured pulmonary retention half-times for the small and large particles were 86-109 days and 870-1020 days, respectively. Fecal recovery data closely complemented pulmonary clearance data for both particle sizes. The two-particle approach was found supportive of the volumetric overload hypothesis.

Nitrogen dioxide exposure in vivo and human alveolar macrophage inactivation of influenza virus in vitro

Epidemiologic studies have reported an increased incidence of respiratory infections and illness in association with elevated indoor levels of nitrogen dioxide (NO2). Animal exposure studies have found that brief exposures to peak levels of NO2 produce greater morbidity than continuous lower level exposure. In order to examine the effect of NO2 inhalation on human alveolar macrophages, normal volunteers were exposed sequentially to air or NO2, by double-blind randomization, in an environmental chamber. Two exposure protocols with comparable concentration x time products were used: (a) continuous 0.60 ppm NO2 (n = 9), and (b) background 0.05 ppm NO2 with three 15-min peaks of 2.0 ppm (n = 15). Inhalation of NO2 caused no significant changes in pulmonary function or airway reactivity in either exposure protocol. Alveolar macrophages obtained by bronchoalveolar lavage 3 1/2 hr after exposure to continuous 0.60 ppm NO2 tended to inactivate influenza virus in vitro less effectively than cells collected after air exposure (1.96 vs 1.25 log10 plaque-forming units on Day 2 of incubation, P less than 0.07). Four of nine subjects accounted for the observed impairment in virus inactivation; cells from these four subjects demonstrated an increase in interleukin-1 (IL-1) production after NO2 vs air, whereas the five remaining subjects decreased IL-1 production after NO2. In contrast, intermittent peak exposure did not alter the rate of viral inactivation or IL-1 production. This methodology has the potential to identify pollutant effects on mechanisms of respiratory defense in humans.

Association of 59Iron Oxide with Alveolar Macrophages during Alveolar Clearance

Long-Evans hooded rats were exposed for 2 h to aerosols of hydrated, radiolabeled iron (59Fe) oxide (MMAD = 1.6 micron; sigma g = 3.0) in order to produce a low mass burden of particles (approximately equal to 30 micrograms) in the lung. The kinetics of particle clearance and the association of the particles with alveolar macrophages (AM) were measured. Two to four hours after exposure, lavaged particles were linearly related to AM numbers harvested, and 60% of the 59Fe activity was physically associated with AM. By 24 h, greater than 90% of the lavaged particles were associated with AM. Such an association was found for at least 75% of the particulate burdens in the lungs. If all the 59Fe is assumed to be AM associated, the 59Fe per AM predicts the total AM population size to be 2.14 X 10(7) cells. This number, in conjunction with the alveolar clearance rate of the particles, suggested the number of AM leaving the lung daily was 2.8 X 10(5) cells.

A Point-to-Plane Electrostatic Precipitator for Particle Size Sampling

Abstract A point-to-plane electrostatic precipitator of simple design is described. It was developed as a particle size sampling instrument for use with the electron microscope. The standard operating conditions as well as special design and technical points are described and illustrated. Tests of the instrument have been made using the thermal precipitator as a standard. These comparisons have indicated no important limitation of the point-to-plane precipitator as a particle size sampler. Other advantages and disadvantages of the instrument based on our experience are given. Most of these points are documented by photographs. None of the negative qualities of the sampler appear general or restrictive. Conversely, there are specific attributes which make the point-to-plane electrostatic precipitator a valuable, if not indispensable, tool in aerosol studies.