Abstract  This criteria document focuses on a review and assessment of the effects on human health and welfare of the nitrogen oxides, nitric oxide (NO) and nitrogen dioxide (NO2), and the related compounds, nitrites, nitrates, nitrogenous acids, and nitrosamines. Although the emphasis is on presentation of health and welfare effects data, other scientific data are presented in order to provide a better understanding of these pollutants in the environment. To this end, separate chapters are included which discuss the nitrogen cycle, sources and emissions, atmospheric chemical processes which transform emissions of nitrogen oxides into related airborne compounds, transport and removal processes, measurement methods, and atmospheric concentrations of nitrogenous pollutants.

Abstract  Many epidemiological studies have shown positive short-term associations between health and current levels of outdoor air pollution. The aim of this study was to investigate the association between air pollution and the number of visits to accident and emergency (A&E) departments in London for respiratory complaints. A&E visits include the less severe cases of acute respiratory disease and are unrestricted by bed availability. Daily counts of visits to 12 London A&E departments for asthma, other respiratory complaints, and both combined for a number of age groups were constructed from manual registers of visits for the period 1992-1994. A Poisson regression allowing for seasonal patterns, meteorological conditions and influenza epidemics was used to assess the associations between the number of visits and six pollutants: nitrogen dioxide, ozone, sulphur dioxide, carbon monoxide, and particles measured as black smoke (BS) and particles with a median aerodynamic diameter of <10 Ám (PM10). After making an allowance for the multiplicity of tests, there remained strong associations between visits for all respiratory complaints and increases in SO2: a 2.8% (95% confidence interval (CI) 0.7-4.9) increase in the number of visits for a 18 Ág x (-3) increase (10th-90th percentile range) and a 3.0% (95% CI 0.8-5.2) increase for a 31 Ág x m(-3) increase in PM10. There were also significant associations between visits for asthma and SO2, NO2 and PM10. No significant associations between O3 and any of the respiratory complaints investigated were found. Because of the strong correlation between pollutants, it was difficult to identify a single pollutant responsible for the associations found in the analyses. This study suggests that the levels of air pollution currently experienced in London are linked to short-term increases in the number of people visiting accident and emergency departments with respiratory complaints.

Abstract  A preliminary testing of eight guinea pigs for D-2,3-diphosphoglycerate (2, 3DPG) content of red blood cells, half of which had been exposed to continuous 0.36 ppm nitrogen dioxide for one week, showed a significant increase (P < .05)for the exposed animals.

Abstract  The expanded Second Edition of Dr. Rothman's acclaimed Modern Epidemiology reflects the remarkable conceptual development of this evolving science and the engagement of epidemiologists with an increasing range of current public health concerns. This landmark work is the most comprehensive and cohesive text on the principles and methods of contemporary epidemiologic research.Coauthored by two leading epidemiologists, with 15 additional contributors, the Second Edition presents a much broader range of concepts and methods than Dr. Rothman's single-authored original edition. Coverage of basic measures and study types is more thorough and includes a new chapter on field methods. New chapters on advanced topics in data analysis, such as hierarchical regression, are also included. A new section covers specific areas of research such as infectious disease epidemiology, ecologic studies, disease surveillance, analysis of vital statistics, screening, clinical epidemiology, environmental and occupational epidemiology, reproductive and perinatal epidemiology, genetic epidemiology, and nutritional epidemiology.

Abstract  With the use of an isolated rat lung model, we investigated pulmonary air space absorption kinetics of the reactive gas NO2 in an effort to determine the contributory role of chemical reaction(s) vs. physical solubility. Unperfused lungs were employed, because vascular perfusion had no effect on acute (0- to 60-min) NO2 absorption rates. We additionally found the following: 1) Uptake was proportional to exposure rates (2-14 micrograms NO2/min; 10-63 ppm; 37 degrees C) but saturated with exposures greater than or equal to 14 micrograms NO2/min. 2) Uptake was temperature (22-48 degrees C) dependent but, regardless of temperature, attained apparent saturation at 10.6 micrograms NO2/min. 3) Lung surface area (SA) was altered by increasing functional residual capacity (FRC). Expanded SA (8 ml FRC) and temperature (48 degrees C) both raised fractional uptakes (greater than or equal to 0.81) relative to 4 ml FRC, 37 degrees C (0.67). Uptake rates normalized per unit estimated SA revealed no independent effect of FRC on fractional uptake. However, temperature produced a profound effect (48 degrees C = 0.93; 4 and 8 ml FRC = 0.54). 4) Arrhenius plots (ln k' vs. 1/T), which utilized derived reactive uptake coefficients (k'), showed linearity (r2 = 0.94) and yielded an activation energy of 7,536 kcal.g-1.mol-1 and Q10 of 1.43, all consistent with a reaction-mediated process. These findings, particularly the effects of temperature, suggest that acute NO2 uptake in pulmonary air spaces is, in part, rate limited by chemical reaction of NO2 with epithelial surface constituents rather than by direct physical solubility.

Abstract  The effects of nitrogen dioxide (NO2) on promotion of lung tumorigenesis induced by N-bis(2-hydroxypropyl) nitrosamine (BHPN) were investigated in male Wistar rats. In a preliminary study, the highest non-effective dose of BHPN was found to be 0.5 g per kg body weight. Rats were given a single intraperitoneal injection of BHPN at a dose of 0.5 g per kg body weight or saline at 6 weeks of age, and then exposed to clean air, 0.04 m, 0.4 ppm or 4 ppm of NO2 for 17 months, respectively. The incidence of pulmonary tumors in rats exposed to BHPN plus 4 ppm of NO2 was 12.5%; the tumors were adenomas and adenocarcinomas. Adenomas were found in 4 out of 40 rats (10%) and adenocarcinomas were found in 1 out of 40 rats (2.5%). The tumor incidence in the lungs of rats kept in BHPN plus clean air and BHPN plus 0.04 ppm of NO2 was 2.5% (1/40). In both groups adenomas were found. There was no significant difference in tumor incidence between animals exposed to BHPN plus clean air and to BHPN plus 4 ppm of NO2. No lung tumors were found in the group of BHPN plus 0.4 ppm NO2 and in animals exposed to NO2 without BHPN treatment. A high incidence of alveolar cell hyperplasia was observed in the lungs of rats injected with BHPN, and the effect of NO2 on development of alveolar cell hyperplasia was slight. On the other hand, marked bronchiolar mucosal hyperplasia was found in 17 out of 40 rats (42.5%) in the group of BHPN plus 4 ppm of NO2, and in 1 out of 40 rats (2.5%) in each of the group exposed to clean air, 0.04 ppm or 0.4 ppm of NO2 with BHPN treatment, respectively. The hyperplasia in lungs of rats exposed to 4 ppm of NO2 without BHPN treatment was slighter than that in lung of rat exposed to 4 ppm of NO2 with BHPN treatment. On the other hand, tumor incidence in the nasal cavity of rats in each of group exposed to clean air and NO2 with BHPN treatment was 97-100%. Incidence of tumors in other organs in the groups exposed to clean air and NO2 with and without BHPN treatment was very low, and NO2 had no effect on tumor development in the nasal cavity and other organs whether animals were treated with BHPN or not. Tumors found in the lungs were not metastases of tumors that had developed in the nasal cavity or in other organs. These results suggest that NO2 can promote the development of lung tumors initiated by BHPN, the NO2 concentration which has promoting activity is 4 ppm or higher.

Abstract  Annular denuder-filter pack sampling systems were used to make indoor and outdoor measurements of aerosol strong H+, SO4(2)-, NH4+, NO3- and NO2-, and the gaseous pollutants SO2, HNO3, HONO and NH3 during summer and winter periods in Boston, Massachusetts. Outdoor levels of SO2, HNO3, H+ and SO4(2-) exceeded their indoor concentrations during both seasons. Winter indoor/outdoor ratios were lower than during the summer, probably due to lower air exchange rates during the winter period. During both monitoring periods, indoor/outdoor ratios of aerosol strong H+ were 40-50 percent of the indoor/outdoor SO4(2-) ratio. Since aerosol strong acidity is typically associated with SO4(2-), this finding is indicative of neutralization of the acidic aerosol by the higher indoor NH3 levels. Geometric mean indoor/outdoor NH3 ratios of 3.5 and 23 respectively were measured for the summer and winter sampling periods. For HONO, NH3, NH4+ and NO2-, indoor concentrations were significantly higher than ambient levels. Indoor levels of NO3- were slightly less than outdoor concentrations.

Abstract  Evolution has resorted to nitric oxide (NO), a tiny, reactive radical gas, to mediate both servoregulatory and cytotoxic functions. This article reviews how different forms of nitric oxide synthase help confer specificity and diversity on the effects of this remarkable signaling molecule.

Abstract  The scientific literature suggests that ambient levels of nitrogen dioxide increase susceptibility to respiratory infections. However, this association has not been conclusively demonstrated. The epidemiologic data regarding this relationship are inconclusive because these studies have used parameters of "acute respiratory illness" that are not necessarily related to infectious episodes. Previous animal studies have used either mortality after bacterial infection with virulent bacteria or decreased rate of intrapulmonary killing of bacteria with low virulence. Studies using appropriate bacterial and viral challenge organisms, with morbidity as an endpoint, provide a better basis for extrapolation to humans. The investigations in animals suggest a relationship between nitrogen dioxide and increased susceptibility to respiratory infection, but studies in which functional parameters of host resistance to such infections have been used are few. The aim of this work was to determine the threshold level of acute nitrogen dioxide exposure that would induce increased susceptibility to, and increased severity of, viral and bacterial infections. Physiologic parameters of host resistance to respiratory infections were used as endpoints. A composite picture was developed of dose-response relationships between nitrogen dioxide and the impairment of a spectrum of defense parameters in the murine respiratory tract against viral and bacterial challenges. The salient findings of this study are as follows: (1) the intrapulmonary killing of Staphylococcus aureus was impaired at 5 ppm of nitrogen dioxide; (2) this effect was found at 2.5 ppm or less when nitrogen dioxide exposure was superimposed on lungs predisposed to lowered resistance through immunosuppression with corticosteroids; (3) the adverse effect of nitrogen dioxide occurred at lower concentrations when exposure followed bacterial challenge; and (4) during the course of murine Sendai virus infection, exposure to nitrogen dioxide for four hours per day did not alter the infection in the lungs, but rather it enhanced lung pathology. The implications of these findings are that the antibacterial defenses of the lungs are susceptible to the inhibiting effects of short acute exposures of lower concentrations of nitrogen dioxide when the lungs are predisposed by bacteria present or, even more so, by immunosuppression. The alveolar macrophage phagocytic system is the defense component of the lungs that is most susceptible to the adverse effects of nitrogen dioxide. The finding that nitrogen dioxide increases virus-associated lung damage suggests that the increased severity of the disease process results from the proliferation of the virus to high titers, rather than from alterations of the infective process.

Abstract  While the pulmonary toxicity of NO2 is clearly established, the mechanism by which it is removed from inspired air is poorly understood. Uptake is most likely dependent on chemical reaction since, despite limited per se gaseous NO2 aqueous solubility, uptake proceeds rapidly without ready saturation. We utilized an isolated perfused rat lung model to characterize the effect of dose rate on uptake and transformation. Dose rate was varied via alterations in inspired concentration, tidal volume, and ventilation frequency. Dose equaled the total amount inhaled, uptake the amount removed from inspired air, and transformation the amount of NO2- that accumulated in the perfusate. We found a linear proportionality between both inspired concentration (4-20 ppm) and minute ventilation (45-130 ml/min) and uptake. Fractional uptakes (65%) were similar for all groups. Regression of combined concentration and minute ventilation data yielded a linear relationship between total inspired dose (25-330 micrograms NO2) and both uptake (r2 = 0.99) and transformation (r2 = 0.98). Testing of the functional descriptions resulted in measured uptakes and transformation that fell within a few percentage points of those predicted. We conclude that in acutely exposed isolated lungs (1) NO2 uptake is dependent on total inhaled dose rather than on the variables which serve to affect dose rate, (2) transformation is related to both total inspired dose and uptake, and (3) uptake is more accurately described using a regression equation rather than by use of fractional uptakes.

Abstract  Morphometric procedures were used to determine the number of cells, cell volume, cell diameter, and surface areas of the airways in human and rat lungs. Nuclear sizes of epithelial cells from human bronchi were significantly larger than other lung cell nuclei. The average volume of human ciliated cell nuclei was 310 ± 30 Ám(3) and 167 ± 12 Ám(3) in bronchi and bronchioles, respectively. The smaller nuclei of human bronchioles were comparable to those of alveolar cells. In the pseudostratified epithelium of human bronchi, basal cells had a large surface area in contact with the basement membrane (51.3 ± 4.6 Ám(2) per cell) when compared with ciliated (1.1 ± 0.1 Ám(2)), goblet (7.6 ± 1.2 Ám(2)), or other secretory cells (12.0 ± 2.1 Ám(2)). In the first four airway generations distal to the trachea, basal cells account for 30% of the cells in human airway epithelium and 2% of the cells in rat airway epithelium. Total airway surface area from trachea to bronchioles was 2,471 ± 320 and 27.2 ± 1.7 cm(2) in human and rat lungs, respectively. These direct measurements of airway surface area are less than half of the estimates based on current lung models. The total number of airway epithelial cells were 10.5 x 10(9) for human and 0.05 X 10(9) for rat lungs. For both species, there were 18 times more alveolar cells than bronchial epithelial cells.

Abstract  As part of the EU-funded SAVIAH project, a regression-based methodology for mapping traffic-related air pollution was developed within a GIS environment. Mapping was carried out for NO2 in Amsterdam, Huddersfield and Prague. In each centre, surveys of NO2, as a marker for traffic-related pollution, were conducted using passive diffusion tubes, exposed for four 2-week periods. A GIS was also established, containing data on monitored air pollution levels, road network, traffic volume, land cover, altitude and other, locally determined, features. Data from 80 of the monitoring sites were then used to construct a regression equation, on the basis of predictor environmental variables, and the resulting equation used to map air pollution across the study area. The accuracy of the map was then assessed by comparing predicted pollution levels with monitored levels at a range of independent reference sites. Results showed that the map produced extremely good predictions of monitored pollution levels, both for individual surveys and for the mean annual concentration, with r2 ~ 0À79-0À87 across 8-10 reference points, though the accuracy of predictions for individual survey periods was more variable. In Huddersfield and Amsterdam, further monitoring also showed that the pollution map provided reliable estimates of NO2 concentrations in the following year (r2~0À59-0À86 for n=20).

Abstract  Mice were exposed to 0.5 ppm nitrogen dioxide (NO2) for 6, 18, and 24 hours daily. The lung alveoli were expanded in all mice exposed to NO2 from three to 12 months, with the highest degree of involvement at 12 months; thus the number of expanded alveoli appeared to increase with exposure time. The general impression was of early bronchiolar inflammation with reduction of distal airway size and a concomitant expansion of alveoli. The overall lesions appeared to be consistent with the development of early focal emphysema. Examination of the heart, liver, kidney, and spleen did not reveal any unique pathology. Mice exposed to NO2 and challenged with Klebsiella pneumoniae exhibited a marked reduction in functional lung tissue.

Abstract  Mice were exposed to 0.5 ppM NO2 and then challenged with Klebsiella pneumoniae and killed after 14 days. Although NO2 exposure was conducted from 7 days to 12 months, significant differences from control group were noted only after 3 months exposure. Groups with less exposure had higher, but not significantly so, mortality than did controls. Exposure to NO2 for 6, 18, or 24 hr/day increased half-life clearance after 12 months. Only 24 hr/day exposure increased half-life clearance after only 6 months exposure. The effect of NO2 on bacterial clearance half-time was more an arithmetical progression than logarithmic.

Abstract  One hundred and twenty female mice fed diets containing various levels of vitamin E were continuously exposed to 0.5 ppM, 1.0 ppM nitrogen dioxide (NO2), and filtered air for 17 months. Blood, lung, and liver tissues were assayed for glutathione peroxidase (GSH-peroxidase) activity. Exposure to 0.5 ppM NO2 did not affect blood and lung GSH-peroxidase activity; 1.0 ppM NO2 exposure however, caused suppression of the enzyme. A combination of vitamin E deficiency and 1.0 ppM NO2 exposure resulted in the lowest GSH-peroxidase activities in the blood and lung. High levels of vitamin E in the diet resulted in elevated GSH-peroxidase in the blood and lung. Liver GSH-peroxidase activity was unaffected by either dietary vitamin E or NO2 exposure.

Abstract  Exposures to various mixtures of nitrogen dioxide (NO2) and ozone (O3) reduced the resistance of mice to streptococcal pneumonia as evidenced by increased mortality rates and shortened survival time. Daily 3-h exposures (5 d/wk) for 2--6 mo to an air pollutant mixture consisting of 940 microgram/m3 (0.5 ppm) NO2 and 196 microgram/m3 (0.1 ppm) O3 were most effective in reducing the resistance to infection. The decrease in resistance to the infection occurred sooner than the mice continued to be exposed to the air pollutants instead of clean air for 14 d after the respiratory challenge with Streptococcus pyogenes aerosol. After 3 mo of exposure to the pollutant mixture, there was some decrease in the ability of mice to clear inhalated streptococci from their lungs. At the same time the total cell count in the fluid lavaged from the lungs of mice was markedly reduced, as were the viability and phagocytic activity of the alveolar macrophages. Exposure to the pollutants combined with challenge with Streptococcus aerosol resulted in marked morphological changes in lung tissues as seen by scanning electron microscopy.

Abstract  To investigate the potential for up to a near lifetime exposure to high-ambient levels of nitrogen dioxide (NO2) to induce functional lung damage group of rats were exposed to air or a simulated urban profile of NO2 (0.5 ppm background, 1.5 ppm peak) for 1, 3, 13, 52, or 78 weeks. The dynamic, static, and diffusional characteristics of the lung were evaluated postexposure in anesthetized rats. Furthermore, for the 13-, 52-, and 78-week groups, additional animals were tested after a 6-, 26-, or 17-week period in filtered air, respectively. No significant NO2 differences between exposed and control animals were found for the nitrogen washout, compliance, lung volume, or diffusion capacity of carbon monoxide measurements At 78 weeks, how ever, a reduction in ?FEF25% an estimate of convexity in the later portion of the forced expiratory flow volume curve, was observed. Breathing patterns and mechanics were also assessed postexposure in a parallel group of similarly exposed unanesthetized rats. These rats were examined during a filtered air, 4 and 8%, carbon dioxide (CO2) challenge. In the unanesthetized rat, frequency of breathing was significantly decreased and tidal volume, expiratory resistance, and inspiratory and expiratory times tended to increase. For several of these variables, the largest response also occurred at 78 weeks and seemed to be exacerbated by CO2 challenge. For both unnesthetized and anesthetized test groups, the magnitude of the changes in pulmonary function were small and their significance was borderline, thus indicating that near-lifetime exposure to the rat of a high ambient urban profile of NO2 does not lead to dysfunction suggestive of degenerative lung disease.

Abstract  The major aspects that must be considered in studies of the health effects of environmental pollutants are: the direct damage due to the exposure, the role of pre-existing disease, and effects of the exposure on the response to secondary stresses. In experimental studies at concentrations of air pollutants found in urban environments frank toxicological responses are rarely observed. However, exposure to a secondary stress, i.e. respiratory challenge with infectious bacteria, can exacerbate the response of the experimental host. Changes in the resistance to respiratory infections provide a highly sensitive experimental animal model system, which is increasingly used in studies of health effects of air pollutants. This model indicates the impairment of the basic defense mechanisms of the respiratory system by the combined exposure to low concentrations of pollutants and the superimposed bacterial infection. Changes in the resistance to respiratory infections were studied in various species of laboratory animals. S. pyogenes and K. pneumoniae are the bacteria of choice to induce the pulmonary infection. Included in the studies are short-term single and multiple exposures as well as long-term exposures to gaseous pollutants such as O3 and NO2 and particulate pollutants such as sulfates and nitrates. Changes in the resistance are measured as excess mortalities and reduced survival time as compared to those in infected animals not exposed to the pollutants. Other parameters measured ranged from changes in the immune response to changes in retention rates of bacteria in lungs.

Abstract  Swiss Webster mice were exposed to either 4.8 ppm (9024 microgram/m3) nitrogen dioxide (NO2), 0.45 ppm (882 microgram/m3) ozone (O3), or their combination intermittently (8 hr daily) for 7 days, and the effects were studied in the lung by a series of physical and biochemical parameters, including lung weight, DNA and protein contents, oxygen consumption, sulfhydryl metabolism, and activities of NADPH generating enzymes. The results show that exposure to NO2 caused relatively smaller changes than O3, and that the effect of each gas alone under the conditions of exposure was not significant for most of the parameters tested. However, when the two gases were combined, the exposure caused changes that were greater and significant. Statistical analysis of the data shows that the effects of combined exposure were more than additive, i.e., they might be synergistic. The observations suggest that intermittent exposure to NO2 or O3 alone at the concentration used may not cause significant alterations in lung metabolism, but when the two gases are combined the alterations may become significant.

Abstract  We review gases that can affect oxidative stress and that themselves may be radicals. We discuss O-2 toxicity, invoking superoxide, hydrogen peroxide, and the hydroxyl radical. We also discuss superoxide dismutase (SOD) and both ground-state, triplet oxygen (O-3(2)), and the more energetic, reactive singlet oxygen (O-1(2)). Nitric oxide ((NO)-N-.) is a free radical with cell signaling functions. Besides its role as a vasorelaxant, (NO)-N-. and related species have other functions. Other endogenously produced gases include carbon monoxide (CO), carbon dioxide (CO2), and hydrogen sulfide (H2S). Like (NO)-N-., these species impact free radical biochemistry. The coordinated regulation of these species suggests that they all are used in cell signaling. Nitric oxide, nitrogen dioxide, and the carbonate radical (CO3.-) react selectively at moderate rates with nonradicals, but react fast with a second radical. These reactions establish "cross talk" between reactive oxygen (ROS) and reactive nitrogen species (RNS). Some of these species can react to produce nitrated proteins and nitrolipids. It has been suggested that ozone is formed in vivo. However, the biomarkers that were used to probe for ozone reactions may be formed by non-ozone-dependent reactions. We discuss this fascinating problem in the section on ozone. Very low levels of ROS or RNS may be mitogenic, but very high levels cause an oxidative stress that can result in growth arrest ( transient or permanent), apoptosis, or necrosis. Between these extremes, many of the gasses discussed in this review will induce transient adaptive responses in gene expression that enable cells and tissues to survive. Such adaptive mechanisms are thought to be of evolutionary importance.

Abstract  BACKGROUND: Relatively few studies have been conducted of the association between air pollution and emergency department (ED) visits, and most of these have been based on a small number of visits, for a limited number of health conditions and pollutants, and only daily measures of exposure and response. METHODS: A time-series analysis was conducted on nearly 400,000 ED visits to 14 hospitals in seven Canadian cities during the 1990 s and early 2000s. Associations were examined between carbon monoxide (CO), nitrogen dioxide (NO2), ozone (O3), sulfur dioxide (SO2), and particulate matter (PM 10 and PM2.5), and visits for angina/myocardial infarction, heart failure, dysrhythmia/conduction disturbance, asthma, chronic obstructive pulmonary disease (COPD), and respiratory infections. Daily and 3-hourly visit counts were modeled as quasi-Poisson and analyses controlled for effects of temporal cycles, weather, day of week and holidays. RESULTS: 24-hour average concentrations of CO and NO2 lag 0 days exhibited the most consistent associations with cardiac conditions (2.1% (95% CI, 0.0-4.2%) and 2.6% (95% CI, 0.2-5.0%) increase in visits for myocardial infarction/angina per 0.7 ppm CO and 18.4 ppb NO2 respectively; 3.8% (95% CI, 0.7-6.9%) and 4.7% (95% CI, 1.2-8.4%) increase in visits for heart failure). Ozone (lag 2 days) was most consistently associated with respiratory visits (3.2% (95% CI, 0.3-6.2%), and 3.7% (95% CI, -0.5-7.9%) increases in asthma and COPD visits respectively per 18.4 ppb). Associations tended to be of greater magnitude during the warm season (April - September). In particular, the associations of PM 10 and PM2.5 with asthma visits were respectively nearly three- and over fourfold larger vs. all year analyses (14.4% increase in visits, 95% CI, 0.2-30.7, per 20.6 microg/m3 PM 10 and 7.6% increase in visits, 95% CI, 5.1-10.1, per 8.2 microg/m3 PM2.5). No consistent associations were observed between three hour average pollutant concentrations and same-day three hour averages of ED visits. CONCLUSION: In this large multicenter analysis, daily average concentrations of CO and NO2 exhibited the most consistent associations with ED visits for cardiac conditions, while ozone exhibited the most consistent associations with visits for respiratory conditions. PM 10 and PM2.5 were strongly associated with asthma visits during the warm season.