Ozone absorption in the human nose during unidirectional airflow.

This study addresses the effect of gas flow rate and ozone (O(3)) concentration on the uptake of this air pollutant in the nose. A nasal exposure system was developed in which a constant flow of humidified air (V) containing a constant concentration of O(3) (C(inlet)) entered one nostril and then exited the other nostril while a subject closed the velopharyngeal aperture. Experiments were conducted on 10 healthy nonsmokers for whom O(3) concentration was measured at the inlet nostril and the outlet nostril to determine the fraction of inhaled O(3) that was absorbed into the nasal mucosa (Lambda(nose)). Lambda(nose) decreased from 0.80 +/- 0.02 to 0.33 +/- 0.02 (SE) when V was increased from 3 to 15 l/min and C(inlet) was fixed at 0.4 ppm. Analysis of these data with a mathematical model indicated that O(3) uptake was limited by diffusion reaction through mucus, rather than by convective diffusion through the respired gas. A small decrease in Lambda(nose) from 0.36 +/- 0.02 to 0.32 +/- 0.01 was also observed when C(inlet) was increased from 0.1 to 0.4 ppm at a fixed V of 15 l/min. This may have been due to nonlinear reaction kinetics between O(3) and reactive substrates in mucus or an active response by a physiological process such as mucus secretion or transepithelial water influx.

Longitudinal distribution of ozone and chlorine in the human respiratory tract: simulation of nasal and oral breathing with the single-path diffusion model.

In the single-path model of the respiratory system, gas transport occurs within a conduit of progressively increasing cross-sectional and surface areas by a combination of flow, longitudinal dispersion, and lateral absorption. The purpose of this study was to use bolus inhalation data previously obtained for chlorine (Cl(2)) and for ozone (O(3)) to test the predictive capability of the single-path model and to adjust input parameters for applying the model to other exposure conditions. The data, consisting of uptake fraction as a function of bolus penetration volume, were recorded on 10 healthy nonsmokers breathing orally as well as nasally at alternative air flows of 150, 250, and 1000 ml/s. By employing published data for airway anatomy, gas-phase dispersion coefficients, and gas-phase mass transfer coefficients while neglecting diffusion limitations in the mucus phase, the single-path model was capable of predicting the uptake distribution for O(3) but not the steeper distribution that was observed for Cl(2). To simultaneously explain the data for these two gases, it was necessary to increase gas-phase mass transfer coefficients and to include a finite diffusion resistance of O(3) within the mucous layer. The O(3) reaction rate constants that accounted for this diffusion resistance, 2 x 10(6) s(-1) in the mouth and 8 x 10(6) s(-1) in the nose and lower airways, were much greater than previously reported reactivities of individual substrates found in mucus.

A hybrid computational fluid dynamics and physiologically based pharmacokinetic model for comparison of predicted tissue concentrations of acrylic acid and other vapors in the rat and human nasal cavities following inhalation exposure.

To assist in interspecies dosimetry comparisons for risk assessment of the nasal effects of organic acids, a hybrid computational fluid dynamics (CFD) and physiologically based pharmacokinetic (PBPK) dosimetry model was constructed to estimate the regional tissue dose of inhaled vapors in the rat and human nasal cavity. Application to a specific vapor would involve the incorporation of the chemical-specific reactivity, metabolism, partition coefficients, and diffusivity (in both air and tissue phases) of the vapor. This report describes the structure of the CFD-PBPK model and its application to a representative acidic vapor, acrylic acid, for interspecies tissue concentration comparisons to assist in risk assessment. By using the results from a series of short-term in vivo studies combined with computer modeling, regional nasal tissue dose estimates were developed and comparisons of tissue doses between species were conducted. To make these comparisons, the assumption was made that the susceptibilities of human and rat olfactory epithelium to the cytotoxic effects of organic acids were similar, based on similar histological structure and common mode of action considerations. Interspecies differences in response were therefore assumed to be driven primarily by differences in nasal tissue concentrations that result from regional differences in nasal air flow patterns relative to the species-specific distribution of olfactory epithelium in the nasal cavity. The results of simulations with the seven-compartment CFD-PBPK model suggested that the olfactory epithelium of the human nasal cavity would be exposed to tissue concentrations of acrylic acid similar to that of the rat nasal cavity when the exposure conditions are the same. Similar analysis of CFD data and CFD-PBPK model simulations with a simpler one-compartment model of the whole nasal cavities of rats and humans provides comparable results to averaging over the compartments of the seven-compartment model. These results indicate that the general structure of the hybrid CFD-PBPK model applied in this assessment would be useful for target tissue dosimetry and interspecies dose comparisons for a wide variety of vapors. Because of its flexibility, this CFD-PBPK model is envisioned to be a platform for the construction of case-specific inhalation dosimetry models to simulate in vivo exposures that do not involve significant histopathological damage to the nasal cavity.

Ozone uptake in the intact human respiratory tract: relationship between inhaled dose and actual dose.

Inhaled concentration (C), minute volume (MV), and exposure duration (T) are factors that may affect the uptake of ozone (O(3)) within the respiratory tract. Ten healthy adult nonsmokers participated in four sessions, inhaling 0.2 or 0.4 ppm O(3) through an oral mask while exercising continuously to elicit a MV of 20 l/min for 60 min or 40 l/min for 30 min. In each session, fractional absorption (FA) was determined on a breath-by-breath basis as the ratio of O(3) uptake to the inhaled O(3) dose. The mean +/- SD value of FA for all breaths was 0.86 +/- 0.06. Although C, MV, and T all had statistically significant effects on FA (P < 0.0001, P = 0.004, and P = 0.026, respectively), the magnitudes of these effects were small compared with intersubject variability. For an average subject, a 0. 05 change in FA would require that C change by 1.3 ppm, MV change by 46 l/min, or T change by 1.7 h. It is concluded that inhaled dose is a reasonable surrogate for the actual dose delivered to a particular subject during O(3) exposures of <2 h, but it is not a reasonable surrogate when comparisons are made between individuals.

Longitudinal distribution of chlorine absorption in human airways: a comparison to ozone absorption.

The bolus inhalation method was used to measure the fraction of inhaled chlorine (Cl(2)) and ozone (O(3)) absorbed during a single breath as a function of longitudinal position in the respiratory system of 10 healthy nonsmokers during oral and nasal breathing at respired flows of 150, 250, and 1,000 ml/s. At all experimental conditions, <5% of inspired Cl(2) penetrated beyond the upper airways and none reached the respiratory air spaces. On the other hand, larger penetrations of O(3) beyond the upper airways occurred as flow increased and during nasal than during oral breathing. In the extreme case of oral breathing at 1,000 ml/s, 35% of inhaled O(3) penetrated beyond the upper airways and approximately 10% reached the respiratory air spaces. Mass transfer theory indicated that the diffusion resistance of the tissue phase was negligible for Cl(2) but important for O(3). The gas phase resistances were the same for Cl(2) and O(3) and were directly correlated with the volume of the nose and mouth during nasal and oral breathing, respectively.

Longitudinal distribution of chlorine absorption in human airways: comparison of nasal and oral quiet breathing.

The fraction of an inspired chlorine (Cl2) bolus absorbed during a single breath (Lambda) was measured as a function of bolus penetration (VP) into the respiratory system of five male and five female nonsmokers during both nasal and oral breathing at a quiet respiratory flow of 250 ml/s. The correspondence between VP and specific anatomic landmarks was found for each subject by a combination of acoustic reflection and nitrogen washout measurements. For both nasal and oral breathing, Lambda reached approximately 0. 95 at the distal end of the upper airways and reached 1.00 within the lower conducting airways. The values of a regional mass transfer parameter computed from the Lambda-VP data indicated that the resistance to Cl2 diffusion in the airway mucosa was negligible compared with the diffusion resistance in the respired gas. Changing the peak inhaled Cl2 concentration from 0.5 to 3.0 parts/million did not significantly affect the distribution of Cl2 absorption, suggesting that the underlying mass transport and chemical reaction processes were linear with respect to Cl2 concentration.

Enhancement of O2 and CO2 transfer through microporous hollow fibers by pressure cycling.

An intravascular gas exchange device for the treatment of respiratory failure consisted of a multitude of blind-ended hollow fibers glued in a pine-needle arrangement to a central gas supply catheter. It has previously been shown that gas desorption rates can be significantly enhanced by cycling gas pressure between a hypobaric level of 130 and an ambient level of 775 Torr. In this study, influences of the cycling frequency (f) and the cycle fraction during which hypobaric pressure is applied (theta) were investigated. Rates of O2 desorption from O2-saturated water and CO2 desorption from CO2-saturated water into a manifold containing 198 fibers, 380 microm in diameter, were measured over a range of f from 0.2 to 1.0 Hz. theta from 0.1 to 0.8, and fiber lengths from 4 to 16 cm. Relative to operation at ambient pressure, pressure cycling increased O2 transfer 3-4 times and CO2 transfer 4-6 times when the water flowed over the fiber manifold at 2.3 l/min. Transfer rates were relatively insensitive tof and theta with 80%-90% of maximum enhancement obtained when theta was as low as 0.2. Transfer rates increased continuously with fiber length, implying that pressure cycling reduced the intra-fiber resistance to gas diffusion. A mathematical diffusion model which utilized only two adjustable parameters, a mass transfer coefficient for O2 and for CO2, simulated the trends exhibited by desorption data.

Application of a hybrid computational fluid dynamics and physiologically based inhalation model for interspecies dosimetry extrapolation of acidic vapors in the upper airways.

This study provides a scientific basis for interspecies extrapolation of nasal olfactory irritants from rodents to humans. By using a series of short-term in vivo studies, in vitro studies with nasal explants, and computer modeling, regional nasal tissue dose estimates were made and comparisons of tissue doses between species were conducted. To make these comparisons, this study assumes that human and rodent olfactory epithelium have similar susceptibility to the cytotoxic effects of organic acids based on similar histological structure and common mode of action considerations. Interspecies differences in susceptibility to the toxic effects of acidic vapors are therefore assumed to be driven primarily by differences in nasal tissue concentrations that result from regional differences in nasal air flow patterns relative to the species-specific distribution of olfactory epithelium in the nasal cavity. The acute, subchronic, and in vitro studies have demonstrated that the nasal olfactory epithelium is the most sensitive tissue to the effects of inhalation exposure to organic acids and that the sustentacular cells are the most sensitive cell type of this epithelium. A hybrid computational fluid dynamics (CFD) and physiologically based pharmacokinetic (PBPK) dosimetry model was constructed to estimate the regional tissue dose of organic acids in the rodent and human nasal cavity. The CFD-PBPK model simulations indicate that the olfactory epithelium of the human nasal cavity is exposed to two- to threefold lower tissue concentrations of a representative inhaled organic acid vapor, acrylic acid, than the olfactory epithelium of the rodent nasal cavity when the exposure conditions are the same. The magnitude of this difference varies somewhat with the specific exposure scenario that is simulated. The increased olfactory tissue dose in rats relative to humans may be attributed to the large rodent olfactory surface area (greater than 50% of the nasal cavity) and its highly susceptible location (particularly, a projection of olfactory epithelium extending anteriorly in the dorsal meatus region). In contrast, human olfactory epithelium occupies a much smaller surface area (less than 5% of the nasal cavity), and it is in a much less accessible dorsal posterior location. In addition, CFD simulations indicate that human olfactory epithelium is poorly ventilated relative to rodent olfactory epithelium. These studies suggest that the human olfactory epithelium is protected from irritating acidic vapors significantly better than rat olfactory epithelium due to substantive differences in nasal anatomy and nasal air flow. Furthermore, the general structure of the hybrid CFD-PBPK model used for this study appears to be useful for target tissue dosimetry and interspecies dose comparisons for a wide range of inhaled vapors.

A CFD-PBPK hybrid model for simulating gas and vapor uptake in the rat nose.

In laboratory studies of rodents, the inhalation of organic vapors often results in preferential damage to olfactory epithelium. Such focal lesion formation may be due either wholly or in part to a corresponding nonuniformity in the spatial distribution of vapor uptake within the nasal cavities. As a tool for determining this dose distribution, a mathematical model based on a combination of computational fluid dynamics (CFD) and physiologically based pharmacokinetic (PBPK) modeling was developed for simulating toxicant vapor uptake in the rat nose. The nasal airways were subdivided into four distinct meatuses selected such that each contained a major air flow stream. Each meatus was further divided into four serial regions attached to separate tissue stacks containing mucus, epithelial, and subepithelial compartments. Values for the gas-phase mass transfer coefficients and gas flows in the 16 airway regions were determined by a solution of the Navier-Stokes and convection-diffusion equations using commercially available CFD software. These values were then input to a PBPK simulation of toxicant transport through the 16 tissue stacks. The model was validated by using overall uptake data from rodent inhalation studies for three "unreactive" vapors that were either completely inert (i.e., acetone), reversibly ionized in aqueous media (i.e., acrylic acid), or prevented from being metabolized by an enzyme inhibitor (i.e., isoamyl alcohol). A sensitivity analysis revealed that accurate values of the mass transfer coefficient were not necessary to simulate regional concentrations and uptake of unreactive vapors in the rat nose, but reliable estimates of diffusion coefficients in tissue were crucial for accurate simulations.

A respiratory ozone analyzer optimized for high resolution and swift dynamic response during exercise conditions.

The breath-to-breath determination of total respiratory ozone (O3) uptake requires the monitoring of O3 concentration at the airway opening with an instrument that responds rapidly relative to the frequency of respiration. Originally, the authors developed an analyzer that used the homogeneous chemiluminescent reaction of O3 with 2-methyl-2-butene, but it was suitable only for monitoring O3 during quiet breathing and light exercise (Ben-Jebria and Ultman, Rev Sci Instrum 1989; 60:3004-11, and Ben-Jebria et al., Rev Sci Instrum 1990; 61:3435-39). The improvement of performance characteristics of the aforementioned analyzer enabled the authors to use the newly constructed and self-contained instrument, which used ethylene as the reactant gas, for respiratory O3 monitoring during moderate-to-heavy exercise. Operating at a reaction chamber pressure of 350 torr, an ethylene/sample flow ratio of 4:1, and a sampling flow of 0.6 lpm, the authors achieved an optimum analyzer performance (i.e., 10-90% step-response of 70 msec and a minimum resolution of 0.006 ppm O3). Furthermore, the new instrument did not exhibit the nonlinear calibration and the CO2 interference suffered by the original analyzer. To demonstrate the quality of the new O3 analyzer in a respiratory application (i.e., total O3 uptake), the authors measured a series of single breaths on two subjects who breathed 0.11 and 0.43 ppm O3-in-air mixtures for 15 min during rest, and during moderate and heavy exercise.

Kinetics of protein depletion in rat bronchoalveolar lavage fluid following in vitro exposure to nitrogen dioxide.

Upon inhalation, nitrogen dioxide (NO(2)), a strong oxidizing agent, first comes into contact and reacts with the fluids lining the airways of the respiratory tract. These respiratory tract lining fluids (RTLF) form a barrier between the inhaled toxic pollutant and the epithelium which protects the underlying tissue from inflammation. Proteins, mainly albumin, and antioxidants are the major components of the RTLF. Many studies have utilized human blood plasma to study the interaction of an extracellular fluid with ozone. In this study, we used bronchoalveolar lavage fluids (BALF) as a more specific surrogate for rat RTLF, and we utilized the native fluorescence as a marker to investigate the depletion kinetics of naturally-occurring protein following exposure to NO(2) in a controlled flow reactor system. We also studied the depletion kinetics of albumin in a buffered salt solution. The results indicated that: (1) the decay in fluorescence was linearly dependent on the concentration of NO(2), indicating that protein oxidation was first order with respect to NO(2) concentration in both BALF and in buffered albumin solution; (2) the depletion kinetics of protein in BALF was non-linear with respect to substrate concentration; (3) the rate of protein depletion was much slower in BALF than in a buffered solution of albumin, suggesting that the presence of antioxidants in BALF protected proteins from being oxidized by NO(2); and (4) whereas the addition of ascorbic acid to buffered albumin solution significantly attenuated albumin depletion, the addition of glutathione had no effect. This suggested that the reaction rate constant of ascorbic acid was considerably higher than that of glutathione.

Improvement of a respiratory ozone analyzer.

The breath-to-breath measurement of total respiratory ozone (O3) uptake requires monitoring O3 concentration at the airway opening with an instrument that responds rapidly relative to the breathing frequency. Our original chemiluminescent analyzer, using 2-methyl-2-butene as the reactant gas, had a 10% to 90% step-response time of 110 msec and a minimal detectable concentration of 0.018 parts per million (ppm) O3 (Ben-Jebria et al. 1990). This instrument was suitable for respiratory O3 monitoring during quiet breathing and light exercise. For this study, we constructed a more self-contained analyzer with a faster response time using ethylene as the reactant gas. When the analyzer was operated at a reaction chamber pressure of 350 torr, an ethylene-to-sample flow ratio of 4:1, and a sampling flow of 0.6 liters per minute (Lpm), it had a 10% to 90% step-response time of 70 msec and a minimal detectable concentration of 0.006 ppm. These specifications make respiratory O3 monitoring possible during moderate-to-heavy exercise. In addition, the nonlinear calibration and the carbon dioxide (CO2) interference exhibited by the original analyzer were eliminated. In breath-to-breath measurements in two healthy men, the fractional uptake of O3 during one minute of quiet breathing was comparable to the results obtained by using a slowly responding commercial analyzer with a quasi-steady material balance method (Wiester et al. 1996). In fact, fractional uptake was about 0.8 regardless of O3 exposure concentration (0.11 to 0.43 ppm) or ventilation rate (4 to 41 Lpm/m2).

Longitudinal distribution of ozone absorption in the lung: effects of nitrogen dioxide, sulfur dioxide, and ozone exposures.

Investigators used an ozone bolus inhalation method to study the effects of continuous exposure to ozone, nitrogen dioxide, and sulfur dioxide on ozone absorption in the conducting airways of human lungs. Healthy, young nonsmokers (6 males, 6 females) were exposed on separate days for 2 h to air containing 0.36 ppm nitrogen dioxide, 0.75 ppm nitrogen dioxide, 0.36 ppm sulfur dioxide, or 0.36 ppm ozone. Every 30 min, the subject interrupted exposure for approximately 5 min, during which he or she orally inhaled five ozone boluses-each in a separate breath. Investigators targeted penetration of the boluses distal to the lips in the 70-130-ml range, which corresponded to the lower conducting airways. The authors computed the change in absorption resulting from exposure (delta lambda) by comparing the amount of each ozone bolus that was absorbed with a corresponding value obtained prior to exposure. Results indicated that ozone exposure caused delta lambda to decrease relative to air exposure (p < .01), whereas both nitrogen dioxide and sulfur dioxide exposures caused an increase in delta lambda that was not significantly different from air exposure. This resulted, at least in part, to an artifact caused by preexposure to ozone boluses. The authors concluded that exposure of the lower conducting airways to nitrogen dioxide or sulfur dioxide increased their capacity to absorb ozone because more of the biochemical substrates that are normally oxidized by ozone were made available. During continuous ozone exposure, this excess of substrate is depleted and the absorption of ozone boluses decreases.

Longitudinal distribution of ozone absorption in the lung: effect of continuous inhalation exposure.

The effect of continuous exposure to ozone on the absorption of ozone in the conducting airways of human lungs was investigated with a bolus-response method. Eleven healthy nonsmoking college students (8 males, 3 females) were exposed at rest for 2 h on 3 separate days to air containing 0 ppm, 0.12 ppm, and 0.36 ppm ozone. A personal inhalation chamber equipped with a head-only clear plastic dome was used for exposure. Every 30 min a subject removed the dome and orally inhaled a series of five ozone-air boluses, each in a separate breath. Penetration of the boluses distal to the lips was targeted in the range of 70-120 ml (corresponding to the central conducting airways). By integrating the inhaled and exhaled-ozone concentration curves, we obtained the absorbed fraction (lambda) and the dispersion (sigma2) of the ozone bolus for each test breath. In addition, the subtraction of baseline measurements made just before exposure enabled us to determine the changes in absorbed fraction (deltalambda) and in dispersion (deltasigma2) that resulted from exposure alone. Absorbed fraction decreased, but sigma2 increased during O3 exposure, and the differences in deltalambda and in deltasigma2 between breathing air and exposure to either 0.12 ppm or 0.36 ppm O3 were significant. We concluded that exposure of the conducting airways to O3 reduced their capacity to absorb O3, possibly by the depletion of biochemical substrates that are normally oxidized by O3.

Longitudinal distribution of O3 absorption in the lung: gender differences and intersubject variability.

Because the National Ambient Air Quality Standard for ozone (O3) is intended to protect the most sensitive individuals in the general population, it is necessary to identify sources of intersubject variation in the exposure-dose-response cascade. We hypothesize that differences in lung anatomy can modulate exposure-dose relationships between individuals, and this results in differences between their responsiveness to O3 at a fixed exposure condition. During quiet breathing, the conducting airways remove the majority of inhaled O3, so the volume of this region should have an important impact on O3 dose distribution. Employing the bolus inhalation method, we measured the distribution of O3 absorption with respect to penetration volume (Vp), and using the Fowler single-breath N2 washout method, we determined the dead space volume (VD) in the lungs of 10 men and 10 women at a fixed respiratory flow of 250 ml/s. On average, the women absorbed O3 at smaller Vp than the men, and the women had smaller VD than the men. When expressed in terms of Vp/VD, the absorption distribution of the men and women was indistinguishable. Moreover, an interpretation of the O3 distribution in terms of an intrinsic mass transfer parameter (Ka) indicated that differences between the O3 dosimetry in all subjects, whether men or women, could be explained by a unique correlation with anatomic dead space: Ka (in s-1) = 610 VD-105 (in ml). Application of this result to measurements of O3 exposure response indicated that previously reported gender differences may be due to a failure in properly accounting for tissue surface within the conducting airways.

Longitudinal distribution of ozone absorption in the lung: simulation with a single-path model.

A one-dimensional unsteady state diffusion model was used as a basis for simulating the absorption (lambda), breakthrough (V(B)), and dispersion (sigma2) of inhaled ozone boluses as a function of penetration (V(P)) into intact human lungs. The model idealized the respiratory system as a single equivalent tube with cross-sectional and surface areas that varied as a function of longitudinal position. Longitudinal gas transport in the lumen of the equivalent tube occurred by the joint action of bulk flow and a dispersion coefficient, D. Lateral absorption between respired gas and the tube wall was characterized by an overall mass transfer coefficient, K. By inputting published values of anatomic dimensions scaled to a 160-ml conducting airway volume, D values previously reported for inert insoluble gases, and K values equal to gas-phase transfer coefficients determined in physical lung models, a reasonable simulation of the lambda-V(P) distribution measured at a 250 ml/sec respiratory flow was obtained. Simulations of the corresponding V(B)-V(P) and sigma2-V(P) distributions both exhibited the correct shapes but underestimated the actual values. Although the addition of an estimated tissue resistance to K resulted in a poorer simulation of the data, an increase in conducting airway volume from a value of 160 ml estimated by the subjects' CO2 dead space to a value of 200 ml substantially improved the V(B)-V(P) and sigma2-V(P) simulations without sacrificing the quality of the lambda-V(P) simulation. We conclude that the inclusion of a tissue diffusion resistance is not necessary to properly simulate bolus inhalation data during quiet breathing, but a reliable measurement of conducting airway volume is crucial.

Small intrapulmonary artery lung prototypes. Mathematical modeling of gas transfer.

Two diffusion models have been developed to analyze gas transfer data previously measured in an intravascular artificial lung consisting of a central gas supply catheter from which are tethered a large number of blind-ended microporous fibers of equal length. A convective-diffusion model (CD) describes the countercurrent transfer of a binary gas pair when gas is supplied at constant pressure conditions, and a well mixed (WM) cycled pressure model predicts transfer when the gas supply pressure is time cycled between compression and vacuum conditions. Regression of gas to gas and liquid to gas excretion data with the CD model resulted in estimates of the liquid phase mass transfer coefficient kAI. Because these values were intermediate between the kAI expected for flow parallel to a cylinder and for flow normal to a cylinder, gas transfer was influenced by both the tethered region of the fiber that was nearly perpendicular to the axis of the test section and the free end of the fiber that rested along the wall of the test section. With a time cycled gas supply pressure, the enhanced carbon dioxide and oxygen excretion predicted by the WM model was similar to the data, but a loss in transfer efficiency with fiber length was not accounted for by the theory.

A portable inhalation system for personal exposure to ozone.

A low-cost portable inhalation system was developed for exposing an individual subject to 60-600 parts per billion of ozone in a 30-l clear-plastic head dome. The inhalation system had the following novel features: a canister vacuum cleaner that supplied room air without the need for precleaning or humidification; a 7% oxygen-in-nitrogen feed to a commercial ultraviolet ozonator that avoided an excess production of ozone; a compact inline mixer that assured homogeneous mixing of the 200-300 liters per minute room air supply with the 0.5-1.0 liters per minute of ozonated gas flow, positioning of gas inlet and exhaust hoses on the head dome that provided fresh gas delivery in the vicinity of the mouth; a quick-disconnect neck seal that allowed rapid donning of the head dome by the subject, and mounting of most system components on a small mobile cart. Temperature, humidity, and ozone and carbon dioxide concentrations were measured inside the dome while a subject exercised on a bicycle ergometer. An air flow of 200 liters per minute between rest and light exercise created a suitable microenvironment in the dome. During moderate and heavy exercise, however, a higher flow of 300 liters per minute should be used to suppress the build-up of carbon dioxide and humidity.

Epithelium-linked smooth muscle hyperresponsiveness in ferret tracheae exposed to acrolein.

The effects of acrolein exposure on tissue uptake and airway responses to substance P (SP) and nitroprusside (NIP) were determined in excised ferret tracheae exposed for 60 min to a constant flow of air or 0.3 and 3.5 ppm acrolein-air mixtures. Histological examination indicated that whereas the epithelium of an air-exposed trachea was intact with no apparent injury, acrolein-induced epithelium damage was more pronounced at 3.5 than at 0.3 ppm vapor concentration. The fractional uptake of acrolein into the tracheal tissue continually decreased during the 1 h of exposure and was found to be significantly concentration dependent at the 60 min measurement point. This suggests that the uptake process of acrolein in the mucosal layer is not linear and is dominated by irreversible reaction. In the absence of the neutral endopeptidase inhibitor, phosphoramidon, acrolein significantly increased the maximal response to SP. Pretreatment with phosphoramidon abolished the differential effect of acrolein on airway smooth muscle response to SP. Nitroprusside relaxed acrolein-exposed tracheal rings precontracted with carbachol to their baseline tone, but it induced relaxation of air-exposed tracheal rings below their initial resting tension, indicating the presence of endogenous as well as induced tone. Pretreatment with NIP also abolished the differential effect of acrolein on airway response to carbachol and modified the potency of this agonist. We conclude that acrolein-induced hyperresponsiveness of the underlying airway smooth muscle is linked to inactivation of neutral endopeptidase synthesis as well as to loss of epithelium-derived relaxation factor.

Acrolein-induced smooth muscle hyperresponsiveness and eicosanoid release in excised ferret tracheae.

Acrolein is a ubiquitous toxic air pollutant that can have adverse lung effects. To understand the mechanism governing airway reactivity in relation to acrolein uptake, in vitro experiments were conducted in which excised tracheae from ferrets were exposed for 1 hr to a unidirectional constant flow (100 ml/min) of an acrolein-in-air mixture at several concentrations (0-12.5 ppm). During exposure, acrolein uptake into the trachea was determined by a chromatographic analysis of gas samples taken at the entrance and at the exit of the trachea. Smooth muscle contractility in response to carbachol (CCh), acetylcholine (ACh), and potassium chloride (KCl) was measured following exposure, and eicosanoids released in the perfusate baths were assayed. The results indicate that the fractional uptake into an excised ferret trachea was strongly dependent on inlet concentration, implying that diffusion and reaction processes of acrolein in airway tissue are not linear. Only the low concentration of acrolein caused an increase of eicosanoid release from the exposed tracheae in the perfusate bath; it is possible that, at higher exposure concentration, the epithelium was sloughed off and most of the eicosanoids were lost. Although acrolein did not alter smooth muscle response to KCl, it did increase the contractile responses to CCh and ACh, suggesting an alteration in the pharmacomechanical but not the electromechanical coupling of ferret tracheal smooth muscle; therefore, it is more likely that this hyperresponsiveness occurs primarily by a mobilization of intracellular Ca2+ stores rather than by an increased influx of extracellular Ca2+ through voltage-dependent channels.