Effect of cigarette smoking on nitric oxide, structural, and mechanical properties of mouse arteries.

Cigarette smoking (CS) is a major risk factor for vascular disease. The aim of this study was to quantitatively assess the influence of CS on mouse arteries. We studied the effect of short-term (6 wk) and long-term (16 wk) CS exposure on structural and mechanical properties of coronary arteries compared with that of control mice. We also examined the reversibility of the deleterious effects of CS on structural [e.g., wall thickness (WT)], mechanical (e.g., stiffness), and biochemical [e.g., nitric oxide (NO) by-products] properties with the cessation of CS. The left and right coronary arteries were cannulated in situ and mechanically distended. The stress, strain, elastic modulus, and WT of coronary arteries were determined. Western blot analysis was used to analyze endothelial NO synthase (eNOS) in the femoral and carotid arteries of the same mice, and NO by-products were determined by measuring the levels of nitrite. Our results show that the mean arterial pressure was increased by CS. Furthermore, CS significantly increased the elastic modulus, decreased stress and strain, and increased the WT and WT-to-radius ratio compared with those of control mice. The reduction of eNOS protein expression was found only after long-term CS exposure. Moreover, the NO metabolite was markedly decreased in CS mice after short- and long-term exposure of CS. These findings suggest that 16 wk of CS exposure can cause an irreversible deterioration of structural and elastic properties of mouse coronary arteries. The decrease in endothelium-derived NO in CS mice was seen to significantly correlate with the remodeling of arterial wall.

Methods for modeling particle deposition as a function of age.

The purpose of this paper is to review the application of mathematical models of inhaled particle deposition to people of various ages. The basic considerations of aerosol physics, biological characteristics and model structure are presented along with limitations inherent in modern modeling techniques. Application of the models to children and senescent adults has been largely based on extrapolating anatomical and physiological data from young adults to match the changes observed during growth and aging. Sample results are included for total particle deposition and deposition in the bronchial and pulmonary regions. The models proposed provide particle deposition predictions that are consistent with the scant measurements available. The models discussed appear to be on firm theoretical grounds, but they are largely limited in application to simple aerosols and average individuals. Also, additional validation of the computational predictions is needed.

Respiratory tract responses to repeated inhalation of an oxidant and acid gas-particle air pollutant mixture.

The purpose of this study was to examine a broad range of toxicologic responses in rats exposed to a multi-component pollutant atmosphere. Cumulative and adaptive respiratory tract responses to 3 concentrations of an inhaled particle-oxidant mixture were examined in Fisher 344 N rats exposed 4 h/day, 3 days/week for 4 weeks. The mixtures contained O3, NO2, NH4HSO4, carbon particles, and HNO3 vapor. Irritant-induced, rapid-shallow breathing responses were present during the first 4-h exposure to medium and high concentrations. Successive exposures showed diminished responses in medium concentrations and exacerbated responses in high concentrations. At the end of 4 weeks, rats exposed to high concentrations exhibited lung lesions. Lavaged pulmonary macrophages showed dose-dependent depressions of Fc-receptor binding and phagocytosis. Lung tissue macrophages showed dose-dependent increases in acid phosphatase staining density and carbon particles. Respiratory tract clearance of tracer particles was not significantly affected by the exposures. Broncho-alveolar epithelial permeability was increased by the high concentration. Epithelial cell-proliferation labeling showed a dose-dependent increase at all levels of the respiratory tract. Progressively exacerbated breathing-pattern responses at high concentrations were associated with lung lesions and high cell-proliferation labeling in the nose transitional epithelium and terminal bronchioles. Attenuating or adaptive breathing-pattern responses occurred in the presence of smaller, but in many cases still significant, compromise of respiratory functions. Either attenuating or exacerbated breathing-pattern responses can occur in the presence of a significant dose-dependent compromise of other respiratory functions and lung tissue injury.

Uncertainties relating to the health effects of particulate air pollution: the US EPA's particle standard.

Although the epidemiologic associations between urban particulate air pollution and human mortality and morbidity have been accumulating for several years, the causal agents (a specific chemical component, a specific particle size range, one or more pollutant combinations, etc.), and the physiological mechanisms behind the associations have yet to be identified. Significant questions regarding confounding effects due to weather, indoor air pollutant exposures and co-pollutants (that accompany particulate matter) stubbornly remain. The events in the United States began with recent epidemiological associations, followed by a lawsuit forcing the US EPA to accelerate the standard-setting process, and finally controversy over the scientific basis of the new standard. In contrast to the potential risks posed by particulate air pollution, many of the sources of such particles are positive contributors to human health; control measures to meet the proposed standard may therefore produce offsetting enhanced mortality and morbidity. In order to establish the information required for well-informed public health policies, a substantial research program is needed because of uncertainties relating to, the affected individuals, the potential causal agents, and the consequences of particle-control activities. Not only are the remaining scientific questions significant, but the particle exposure/health effects associations also call into question some of the current scientific assumptions relating to the nature of effects of population exposures to low concentrations of pollutants.

Effects of breathing parameters on sidestream cigarette smoke deposition in a hollow tracheobronchial model.

The effects of variations in cyclic breathing parameters (i.e., tidal volume and breath frequency) have been the subject of few studies devoted to the deposition of submicrometer aerosols in the human respiratory tract. Therefore, a series of experiments was performed to investigate whether the deposition efficiency (DE) of sidestream cigarette smoke is altered by varying tidal volume and breath frequency in a child-size hollow tracheobronchial (TB) model while maintaining a fixed minute ventilation rate of 5 L/min. Under cyclic flow conditions with tidal volumes of 100 mL (50 breaths/min), 250 mL (20 breaths/min), 500 mL (10 breaths/min) and 750 mL (6.7 breaths/min), sidestream cigarette smoke was passed through replicas of an idealized hollow TB model. The smoke deposits were extracted and then quantitated spectrophotometrically. The experiments revealed a significant difference in DE between the 100-mL tidal volume (DE = 6.0%) and the 750-mL tidal volume (DE = 11.1%). Under equivalent steady flow conditions, the mean DE was 21.5%. A trend was evident in the data--DE increased as tidal volume increased (and breathing frequency decreased)--suggesting that the influence of diffusion and secondary flows on DE becomes greater as the air residence time increases and the degree of air turbulence decreases. The results provide evidence of the importance of breathing parameters when attempting to model in vivo deposition of environmental tobacco smoke and other similar-size respirable aerosols.

Deposition of monodisperse particles in hollow models representing adult and child-size tracheobronchial airways.

A series of experiments was performed to determine deposition efficiencies of four sizes of radiolabeled monodisperse particles in custom-made hollow tracheobronchial models. The particles had geometric diameters of about 1, 5, 10, and 15 microm. The tracheobronchial models, consisting of a trachea and two or more additional generations, had dimensions representative of a typical adult, a 7-y-old child, and a 4-y-old child; the child-size models were appropriately scaled-down replicas of the adult-size model. Each deposition experiment was conducted using a steady inspiratory airflow representative of low physical activity for the appropriate age of individual: 20 L min(-1) for the adult; 9 L min(-1) for the 7-y-old; 5.5 L min(-1) for the 4-y-old. The results indicate that deposition efficiency of the particles increased substantially (up to 35 times) in all three models as particle diameter increased from 1-15 microm, undoubtedly as a result of particle impaction and sedimentation-related phenomena. An analysis of variance demonstrated the occurrence of statistically-significant (p < 0.05) main effects of hollow model size and particle size; the interaction between the two parameters was also significant. In general, deposition efficiencies of the various sizes of particles were greater in the child-size models than in the adult-size model; this effect may have risk assessment implications. In addition, the results obtained experimentally agreed more closely with those predicted using a radiation-protection mathematical particle deposition formulation as the particle size increased for each of the sizes of models.

Physical characterization of incense aerosols.

Experiments were performed to study the physical characteristics of smoke aerosols generated by burning three types of stick incense in a 4 m3 clean room. Sidestream cigarette smoke was also examined under the same conditions to provide a comparison. Among the parameters measured were (a) masses of aerosol, carbon monoxide and nitrogen oxides generated by burning the incense or cigarettes, (b) rates of decay of the particles from the air, and (c) estimates of count median particle size during a 7 h period post-burning. There was variability among the types of incense studied with respect to many of the parameters. Also, as a general trend, the greater the initial particulate mass concentration, the more rapid the rate of decay of the smoke. In relation to the quantity of particulate generated, cigarette smoke was found to produce proportionally larger quantities of carbon monoxide and nitrogen oxides than did incense. Due to the fact that burning incense was found to generate large quantities of particulate (an average of greater than 45 mg/g burned, as opposed to about 10 mg/g burned for the cigarettes), it is likely, in cases in which incense is habitually burned in indoor settings, that such a practice would produce substantial airborne particulate concentrations.

Surfactant dysfunction after inhalation of nitric oxide.

To study whether nitric oxide (NO) affects surfactant function, 36 young rats inhaled one of the following humidified environments for 24 h: 1) air; 2) 95% O2; 3) air and 100 parts/million (ppm) NO; and 4) 95% O2 and 100 ppm NO. The treatments did not change the recovery of phospholipid from bronchoalveolar lavage (BAL). Exposure to NO of animals that breathed either air or 95% O2 increased the minimum surface tension of surfactant from BAL at low (1.5 mumol/ml), but not at high (4 mumol/ml), phosphatidylcholine concentration. After inhaled NO, the nonsedimentable protein of BAL decreased the surface activity of surfactant (1 mumol phosphatidylcholine/ml) more than the protein from the controls. NO treatment of animals that breathed either air or 95% O2 affected neither the quantity nor the molecular weight distribution of nonsedimentable protein. Hyperoxia increased the amount of the nonsedimentable protein, whereas NO increased the iron saturation of transferrin. The surfactant fraction and the nonsedimentable protein from BAL were separately exposed to 80 ppm NO in vitro. NO exposure had no effect on the surface activity of surfactant fraction. NO exposure of nonsedimentable protein from the control animals (no NO) increased the inhibition of the surface activity and changed the adsorption spectrum of the protein, suggesting conversion of hemoglobin to methemoglobin. Nonsedimentable protein from NO-exposed animals contained methemoglobin. We propose that surfactant dysfunction caused by inhaled NO is in part due to alteration of protein(s) in epithelial lining fluid that in turn inactivates surfactant.

Effects of nitrogen dioxide on respiratory tract clearance in the ferret.

During growth and development, young children are periodically exposed to relatively high concentrations of various air contaminants, including tobacco smoke and environmental pollutants generated by fossil fuel use. The effects of these exposures on respiratory function and lung development are difficult to determine because of interindividual variation and lack of accurate dosimetry. To provide information on the effects of chronic exposure to a common indoor and outdoor pollutant during lung development, a study was performed to assess the effects of exposure to two concentrations of nitrogen dioxide (NO2; 0.5 or 10 ppm) on tracer particle clearance from the airways of ferrets exposed during postnatal respiratory tract development. Separate groups of ferrets were exposed nose-only to the test atmospheres or clean air 4 h/d, 5 d/wk, for either 8 or 15 wk. Those animals exposed for 8 wk were subsequently housed in a filtered air environment until the particle clearance measurements commenced at 3 wk prior to the end of the 15-wk exposure protocol. Radiolabeled (51Cr) tracer particles were deposited in the respiratory tract of all animals by inhalation, and the clearance rates from the head and thoracic regions were separately monitored for 18 d. No significant effects of the NO2 exposure on head airways clearance were seen. In contrast, the rates of particle clearance from the thorax of both the 8- and 15-wk groups exposed to 10 ppm NO2 were significantly reduced, and did not differ from each other. Thoracic clearance was also reduced in animals exposed to 0.5 ppm, but the rate was not significantly different from that of the clean air exposed controls. These results show that NO2 at moderate concentrations caused highly significant changes in the deep lung of the juvenile ferret, and suggest that impairment of the clearance function may be only slowly recovered after chronic exposure.

Low-cost wind tunnel for aerosol inhalation studies.

A low-cost wind tunnel for aerosol studies has been designed, constructed, and evaluated for aerosol uniformity with 2- and 0.46-micron particles. A commercial nebulizer was used to produce the suspended test particles, and a custom-made, four-hole injector was used to introduce the aerosol into the wind tunnel. A commercially available optical particle counter measured the particle concentration. Performance tests of the velocity profile and particle concentration distribution at two flow rates showed that the system performs well for small particles.

The effect of dead space on inhaled particle deposition.

Mathematical models which have been developed to predict the deposition of particles in the conducting airways of the lung require simplified anatomical models of the dimensions and geometry of the bronchial airways. In order to produce valid deposition predictions, the computed volumes of the conducting airways must be realistic in comparison to anatomical dead space. This requirement must be met even as the developing lung grows to maturity and then undergoes aging. The effect of these age-related changes on predicted particle deposition efficiencies has not been well studied. Numerous authors have suggested that differences in lung volumes (total lung capacity, functional residual capacity, dead space and tidal volume) may account for significant variations between predicted or observed particle deposition but no general age-specific relationship has been proposed. New models are proposed to describe changes in dead space as functions of age and body size, and methods to adjust existing anatomical models to various dead space predictions are given. Also, the effect of these modifications to anatomical models on particle deposition efficiencies are simulated for a variety of breathing patterns for models scaled to represent young children, adults, and aged persons.

Inflated, dried whole lung specimens.

A method is described for preparing fully-inflated whole lung specimens that are suitable for instruction or research purposes. Undamaged lungs are removed from the body and then tracheally cannulated and lavaged with tap water more than 250 times. The treatment also includes rinsing blood from vessels with water. A final filling of the lung with alcohol is optional. The multiply rinsed lung is drained and inflated to 30 cm of H2O pressure with dehumidified air and held at that pressure until the tissue is completely dry. The resulting specimens are light in color and appear to be permanent if stored properly.

Growth of the ferret tracheobronchial tree.

Because the ferret is being used increasingly in inhalation toxicology and lung physiology studies, it is necessary to better understand the airway structure of its tracheobronchial tree. Previously published information does not include dimensions of bronchi and bronchioles in either adult or growing ferrets. The airway structure of interest for calculating inhaled particle deposition patterns includes airway lengths, diameters and branching angles in each generation. Measurements of these dimensions were obtained for several selected airway paths on replica casts. Casts were made in-situ in four male litter mates age 14 hours, 9.5 days, 16.5 days and 56 days. These data demonstrate, that as with human lung growth, body length at a given age is a good predictor of airway lengths and diameters. Airway branch angles do not appear to change significantly during growth. Sufficient measurements were made to provide dimensions of a typical tracheobronchial pathway for this species. This pathway begins with the trachea and ends at the terminal bronchiole. The morphometric data were not sufficient to determine whether or not the number of tracheobronchial generations increase or decrease postnatally.

Health effects of acid aerosols formed by atmospheric mixtures.

Under ambient conditions, sulfur and nitrogen oxides can react with photochemical products and airborne particles to form acidic vapors and aerosols. Inhalation toxicological studies were conducted, exposing laboratory animals, at rest and during exercise, to multicomponent atmospheric mixtures under conditions favorable to the formation of acidic reaction products. Effects of acid and ozone mixtures on early and late clearance of insoluble radioactive particles in the lungs of rats appeared to be dominated by the oxidant component (i.e., the mixture did cause effects that were significantly different from those of ozone alone). Histopathological evaluations showed that sulfuric acid particles alone did not cause inflammatory responses in centriacinar units of rat lung parenchyma (expressed in terms of percent lesion area) but did cause significant damage (cell killing followed by a wave of cell replication) in nasal respiratory epithelium, as measured by uptake of tritiated thymidine in the DNA of replicating cells. Mixtures of ozone and nitrogen dioxide, which form nitric acid, caused significant inflammatory responses in lung parenchyma (in excess of effects seen in rats exposed to ozone alone), but did not damage nasal epithelium. Mixtures containing acidic sulfate particles, ozone, and nitrogen dioxide damaged both lung parenchyma and nasal epithelia. In rats exposed at rest, the response of the lung appeared to be dominated by the oxidant gas-phase components, while responses in the nose were dominated by the acidic particles. In rats exposed at exercise, however, mixtures of ozone and sulfuric acid particles significantly (2.5-fold) elevated the degree of lung lesion formation over that seen in rats exposed to ozone alone under an identical exercise protocol.

Aerosol deposition in the nose as a function of body size.

The effect of body size on nasal doses from inhaled aerosols has not been measured directly in people. Two basic types of computational models are used to calculate inhaled particle deposition in adults. One type uses an impaction parameter that incorporates particle aerodynamic diameter and the average airflow rate. The second type uses the nasal pressure drop and particle aerodynamic diameter. Although both types of models have been adjusted to give reasonably accurate deposition efficiencies for adults, they predict very different deposition efficiencies when they are applied to young children. This is not surprising because the airflow-type model has no body-size-dependent parameters, unlike the pressure-drop-type model. The objective of our studies was to test these two types of computational models using idealized hollow nasal models of two sizes, representing the adult and young child. The results indicate that a pressure-drop relationship fits the aerosol deposition data very well. When the properly scaled physiological air flows and minute ventilations are used in a nasal dose calculation, the young child is seen to have potentially larger nasal doses than those of an adult.

Effects of pollutant atmospheres on surface receptors of pulmonary macrophages.

The effects of two multicomponent pollutant atmospheres on the surface receptors (FcR) and phagocytic activity of rat pulmonary alveolar macrophages have been studied. FcR are crucial for the macrophages to become cytotoxic against target cells. The atmospheres were composed of pollutants that are prevalent in the South Coast Air Basin of southern California. Rats were exposed nose-only to a 7-component oxidant-and sulfate-containing atmosphere for 4 h/d for either 7 or 21 consecutive days. In another experiment rats were exposed 5 h/d for 5 consecutive days to another pollutant combination--acid droplets plus carbon-containing dilute diesel engine exhaust. In both experiments matched rats were exposed nose-only to purified air to be used as controls. Each of the atmospheres studied significantly reduced FcR activity for at least 3 d following the exposure, with the group of rats exposed to the 7-component atmosphere for 21 d exhibiting the most pronounced effect. Macrophages from rats exposed to the diesel exhaust plus acid atmosphere and the 7-component atmosphere for 7 d had significantly reduced phagocytic activity for at least 3 d postexposure, while the macrophages from rats exposed to the latter atmosphere for 21 d had phagocytic activity near control values. The decrease in phagocytosis and inhibition of FcR of macrophages suggests an impairment of macrophage function that probably renders the host vulnerable to bacterial and/or viral infections.

Effects of exercise exposure on toxic interactions between inhaled oxidant and aldehyde air pollutants.

Respiratory tract injury resulting from inhalation of mixtures of ozone (O3) and nitrogen dioxide (NO2) and of O3 and formaldehyde (HCHO) was studied in Sprague-Dawley rats under exposure conditions of rest and exercise. Focal inflammatory injury induced in lung parenchyma by O3 exposure was measured morphometrically and HCHO injury to the nasal respiratory epithelium was measured by cell turnover using tritium-labeled thymidine. Mixtures of O3 (0.35 or 0.6 ppm) with NO2 (respectively 0.6 or 2.5 ppm) doubled the level of lung injury produced by O3 alone in resting exposures to the higher concentrations and in exercising exposures to the lower concentrations. Formaldehyde (10 ppm) mixed with O3 (0.6 ppm) resulted in reduced lung injury compared to O3 alone in resting exposures, but exercise exposure to the mixture did not show an antagonistic interaction. Nasal epithelial injury from HCHO exposure was enhanced when O3 was present in a mixture. Mixtures of O3 and NO2 at high and low concentrations formed respectively 0.73 and 0.02 ppm nitric acid (HNO3) vapor. Chemical interactions among the oxidants, HNO3, and other reaction products (N2O5 and nitrate radical) and lung tissue may be the basis for the O3-NO2 synergism. Increased dose and dose rate associated with exercise exposure may explain the presence of synergistic interaction at lower concentrations than observed in resting exposure. No oxidation products were detected in O3-HCHO mixtures, and the antagonistic interaction observed in lung tissue during resting exposure may result from irritant breathing pattern interactions.

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)

Effect of ozone on mean linear intercept in the lung of young beagles.

Although photochemical air pollutants are believed to be associated with respiratory illness, there is also a need to consider their possible effects on postnatal lung maturation. The purpose of this study was to determine whether the maturation of lungs of young beagle dogs might be altered by an inhalation exposure to ozone that represents a severe 5-d episode of photochemical oxidant air pollution. Exposures were at 6 wk of age to purified air, 1 or 2 ppm ozone for 4 h/d on 5 consecutive days. After holding for 6 wk in clean air, lungs were removed and weighed, and the left lung was fixed both by inflation at 30 cm pressure and immersion using buffered formalin. Histologic sections were used for morphometric measurements. Statistical analysis showed that the mean linear intercept (inversely related to lung surface area) was greater than controls (up to about 5%) in the 1 ppm ozone-exposed group. This effect was not seen at 2 ppm ozone, apparently due to large variations in mean linear intercept. No significant differences were seen in body weight, chest girth, lung weight, or volumes of the fixed, inflated lungs. It is concluded that if anatomic maturation of the lung was retarded by this brief regimen of ozone exposure, the effect was small and not likely to have major health consequences.