Ozone-induced changes in muscarinic bronchial reactivity by different testing methods

We examined the effect of ozone (O3) on muscarinic bronchial reactivity in the guinea pig and compared reactivity determined by two different routes of agonist delivery. Reactivity before and from 4 h to 2 days after O3 exposure (3.0 ppm, 2 h) was determined by measuring specific airway resistance upon administration of intravenous acetylcholine and/or aerosolized methacholine challenge in 34 unanesthetized, spontaneously breathing animals. Before exposure, we observed more gradual and reproducible results to intravenous agonist. After exposure, hyperreactivity to parenteral agonist occurred consistently, but not to inhaled agonist. Hyperreactivity demonstrable by either route was similar in magnitude and time course within 14 h of exposure. Two days later, hyperreactivity to inhaled agonist had remitted; that to intravenous drug persisted. Our results indicate that variability in the occurrence and time course of O3-induced hyperreactivity to inhaled agonist may be a consequence of the technique employed. The consistent occurrence of hyperreactivity after O3 to parenteral agonist suggests mechanisms other than airway mucosal hyperpermeability are responsible for this hyperreactivity.     

Ozone-induced inflammation assessed in sputum and bronchial lavage fluid from asthmatics: a new noninvasive tool in epidemiologic studies on air pollution and asthma

We investigated correlations between ozone-induced increases in inflammatory markers in induced sputum and in bronchial lavage fluid. Sixteen volunteers with intermittent asthma participated in a placebo-controlled parallel study with two exposures. Six days before and 16 h after the first exposure to ozone (0.4 ppm during 2 h) sputum was induced with hypertonic saline. This resulted in a significant increase in the sputum levels of eosinophil cationic protein (ECP; 1.8-fold; p = .03), neutrophil elastase (5.0-fold; p = .005) and the total cell number (1.6-fold; p = .02). After 4 weeks, a second exposure was randomized for air or ozone. Six days before and 16 h after the second exposure a bronchial lavage was performed. ECP values in sputum and in bronchial lavage fluid obtained after ozone correlated significantly (Rs = .79; p = .04), as did interleukin-8 (IL-8) values (Rs = .86; p = .01), and the percentage eosinophils (Rs = .89; p = .007). Moreover, the ozone-induced changes in percentage eosinophils observed in sputum and lavage fluid were highly correlated (Rs = .93; p = .003). In conclusion, changes in eosinophils, IL-8, and ECP markers induced by ozone and measured in sputum reflect the inflammatory responses in the lower airways of asthmatics, and may provide a noninvasive tool in epidemiologic studies on air pollution and asthma.     

Viscoelastic properties of bronchorrhoea sputum in bronchial asthmatics

Dynamic viscoelastic properties of 13 bronchorrhoea sputum samples from asthmatics with bronchorrhoea, defined as the production of watery sputum of 100 ml or more per day during asthmatic attacks, were examined and then compared with 7 saliva and 12 mucoid sputum samples obtained from patients during remission. Dynamic viscosity (eta') and elasticity (G') of bronchorrhoea sputum increased rapidly with time up to 2 hours after collection and slowly thereafter, whereas eta' and G' of saliva and mucoid sputum remained unchanged up to 6 hours after collection. Then, eta' and G' of saliva, bronchorrhoea and mucoid sputum samples were measured between 2 to 4 hours after expectoration. Bronchorrhoea sputum samples showed significantly larger values at frequencies of both 0.1 and 1.0 rad/sec than did saliva samples and also significantly smaller values than did mucoid sputum samples. Thus, bronchorrhoea sputum differed in dynamic viscoelastic properties from saliva, indicating that it does not result from hypersalivation. Based on data of viscoelastic changes with time, it can be assumed that the viscoelasticity of bronchorrhoea sputum in the airways is considerably less than the optimal range reported previously for mucociliary velocity, suggesting the possibility of impaired mucociliary transport.     

Airways responsiveness determined by consecutive histamine challenges in asymptomatic asthmatics

The effect of two consecutive histamine inhalation challenges on airways responsiveness was assessed in a group of eight nonsmoking nonmedicated asthmatics aged 19-27 yr. All subjects had a base-line forced expiratory volume in 1 s (FEV1) of greater than 80% of their predicted normal value before the initial challenge and were allowed to recover to greater than 95% of the initial base-line FEV1 value before the second challenge was initiated. The average airways recovery time after the first challenge was 44 min but ranged between 30 and 90 min. The mean +/- SD values of cumulative histamine dose units provoking a 20% decrease of the FEV1 from the buffer control value (PD20FEV1) were 10.79 +/- 5.95 determined with the first and 30.50 +/- 46.36 with the second challenge (P greater than 0.05). We conclude that sequential histamine challenges performed in mild asthmatics with closely controlled prechallenge airways function are well tolerated. Although some variance does exist in intersubject airways recovery time and in intra-subject histamine airways responsiveness determined by sequential challenges, our data do not support recent observations (J. Appl. Physiol. 63: 1572-1577, 1987) that histamine tolerance is a characteristic finding associated with bronchial asthma.     

Ozone-induced ultrastructural changes in the plasma membrane of Chlorella sorokiniana

Abstract. Modifications in plasma membrane structure and permeability were observed in Chlorella sorokiniana following exposure to 0.2 gm⁻³(140 p.p.m.) O₃ for 30 min. Sixty-eight per cent of the cells were plasmolysed after 15 min O₃ exposure with disruption of organelles similar to that previously described in higher plants. Freeze-fracture exposed large areas of plasma membrane in 90% of the control cells and those exposed to O₃for short periods. After 20 min O₃ 90% of the cells cross-fracture, which indicates a change in molecular interactions in the membrane exposed to O₃ The earliest observed ultraslructural alteration is an aggregation of particles on the plasma membrane P face, statistically significant after 10 min O₃ Changes in ⁸⁶Rb influx occur during a similar time. After more extended exposure to O³ the plasma membrane P face shows regions of lipid phase transition to the crystalline state.     

Seasonal allergic asthma and non-specific bronchial responsiveness

Summary Bronchial responsiveness to methacoline (PD20 FEV1 mcg) was measured in 64 non smoker asthmatic patients with baseline FEV190% predicted. Patients underwent skin prick tests (SPT) and RAST.Allergic patients had: SPT3+ and RAST-score>II class to the same antigen and correlation with asthmatic symptoms; non allergic patients had negative SPT and RAST. We divided patients in four groups: 1st) allergic seasonal asthmatics before pollen season; 2nd) allergic seasonal asthmatics during pollen season; 3rd) allergic perennial asthmatics; 4th) non-allergic perennial asthmatics.A significant difference in log PD20 was observed between 1st and 2nd group (p<0.0005); between 1st and 3rd group (p<0.0005); between 1st and 4th group (p<0.0005). In allergic seasonal asthmatics before pollen season 10/20 subjects were non-responsive to methacholine (PD201600 mcg), while in 2nd, 3rd and 4th group no subjects were non-responsive.The authors conclude that non-specific bronchial hyperresponsiveness to methacoline is not constant in seasonal allergic asthmatics out of pollen season.     

Ozone-induced pulmonary inflammation and epithelial proliferation are partially mediated by PAF

Ozone(O₃) exposure stimulates airwayinflammation and epithelial sloughing in a number of species, includingmice. Platelet-activating factor (PAF) is a lipid mediator released byactivated mast cells, macrophages, and epithelial cells and causespulmonary inflammation and hyperpermeability. We hypothesized that theactivation of PAF receptors is central to the development ofinflammation and epithelial injury induced by acuteO₃ exposure in mice. To test thishypothesis, O₃-susceptibleC57BL/6J mice were treated with a PAF-receptor antagonist, UK-74505, orvehicle either before or immediately after 3-h exposure toO₃ (2 parts/million) or filtered air. Bronchoalveolar lavage (BAL) fluids were collected 6 and 24 hafter exposure. Differential cell counts and protein content of thelavage were used as indicators of inflammation in the airways. O₃-induced epithelial injury wasassessed by light microscopy, and DNA synthesis in epithelium ofterminal bronchioles was estimated by using abromodeoxyuridine-labeling index. Intercellular adhesion molecule 1 (ICAM-1) expression was also examined in the lung by immunohistochemical localization.O₃ caused significant increases inpolymorphonuclear leukocytes and protein in the BAL fluid, increasedpulmonary epithelial proliferation, and increased epithelial expressionof ICAM-1 compared with air-exposed, vehicle-treated control mice.Relative to O₃-exposed,vehicle-treated control mice, UK-74505 before exposure significantly(P < 0.05) decreased BAL protein,polymorphonuclear leukocytes, and epithelial cells. O₃-induced inflammation wassimilarly attenuated in mice treated with UK-74505 after exposure.These experiments thus support the hypothesis thatO₃-induced airways inflammationand epithelial damage in mice are partially mediated by activation ofPAF receptors, possibly through modulation of ICAM-1 expression.

     

Relationship between ozone exposure and pulmonary function changes

A detailed comparison of literature-reported averaged decrements in pulmonary function of normal subjects exposed to O3 has been undertaken. The data base was formed by including data published during the past 20 yr from studies that reported at least one of the pulmonary function variables (forced vital capacity, forced expiratory volume at 1 s, mean forced expiratory flow between 25 and 75% of forced vital capacity, and airway resistance) acquired at 2 h of exposures utilizing either original or modified Bates-Hazucha (intermittent exercise) protocol and that satisfied selection criteria. The final set of data (24 studies involving 299 subjects) was divided by ventilation rate (exercise loads) into four categories: light, moderate, high, and very high ventilation level. For each pulmonary function variable and ventilation level a quadratic function has been fitted to the data using regression procedures. The curve parameter estimates have been computed, tabulated, and statistically evaluated. The slope (quadratic coefficient) for each variable within a group and almost all variables between groups were significantly different from zero and from each other at P less than or equal to 0.0001.     

Emphysema distribution and annual changes in pulmonary function in male patients with chronic obstructive pulmonary disease

Background

The progression of chronic obstructive pulmonary disease (COPD) considerably varies among patients. Those with emphysema identified by quantitative computed tomography (CT) are associated with the rapid progression assessed by forced expiratory volume in one second (FEV₁). However, whether the rate of the decline in lung function is independently affected by the regional distribution or the severity of emphysema in the whole lung is unclear.

Methods

We followed up 131 male patients with COPD for a median of 3.7 years. We measured wall area percent (WA%) in right apical segmental bronchus, total lung volume, percent low attenuation volume (LAV%), and the standard deviation (SD) of LAV% values from CT images of 10 isovolumetric partitions (SD-LAV) as an index of cranial-caudal emphysema heterogeneity. Annual changes in FEV₁ were then determined using a random coefficient model and relative contribution of baseline clinical parameters, pulmonary function, and CT indexes including LAV%, SD-LAV, and WA% to annual changes in FEV₁ were examined.

Results

The mean (SD) annual change in FEV₁ was −44.4 (10.8) mL. Multivariate random coefficient model showed that higher baseline FEV₁, higher LAV%, current smoking, and lower SD-LAV independently contributed to an excessive decline in FEV₁, whereas ratio of residual volume to total lung capacity, ratio of diffusing capacity to alveolar ventilation, and WA% did not, after adjusting for age, height, weight, and ratio of CT-measured total lung volume to physiologically-measured total lung capacity.

Conclusions

A more homogeneous distribution of emphysema contributed to an accelerated decline in FEV₁ independently of baseline pulmonary function, whole-lung emphysema severity, and smoking status. In addition to whole-lung analysis of emphysema, CT assessment of the cranial-caudal distribution of emphysema might be useful for predicting rapid, progressive disease and for developing a targeted strategy with which to prevent disease progression.     

In vitro bronchial responsiveness in two highly inbred rat strains

Wang, C. G., J. J. Almirall, C. S. Dolman, R. J. Dandurand,and D. H. Eidelman. In vitro bronchial responsiveness in twohighly inbred rat strains. J. Appl.Physiol. 82(5): 1445-1452, 1997.We investigatedmethacholine (MCh)-induced bronchoconstriction in explanted airwaysfrom Fischer and Lewis rats. Lung explants, 0.5- to 1.0-mm thick, wereprepared from agarose-inflated lungs of anesthetized 8- to 12-wk-oldmale rats. After overnight culture, videomicroscopy was used to recordbaseline images of the individual airways. Dose-response curves to MChwere then constructed by repeated administration of MCh; airways werereimaged 10 min after each MCh administration. Airway internal luminalarea(A_i)was measured at successive MCh concentrations from10⁹ to10¹ M. Inaddition to the effective concentration leading to 50% of the achievedmaximal response, we also determined the effective concentrationleading to a 40% reduction inA_i.Both the effective concentration leading to 50% of the achievedmaximal response and the concentration leading to a 40% reduction inA_iwere significantly lower among Fischer rat airways(P < 0.05). Airway closure was morecommon among Fischer rat airways (17%) than among those of Lewis rats(7.5%). Responsiveness of Fischer rat airways was more heterogeneousthan among Lewis airways; a larger number of Fischer rat airwaysexhibited high sensitivity to MCh. There was no relationship betweenresponsiveness and baselineA_iin either strain. In a second experiment, we measured the rate ofcontraction of explanted airways from lungs inflated to 50, 75, and100% of total lung capacity. The average rate of contraction in thefirst 15 s was higher in Fischer rat airways at each inflation volume.These data indicate that the hyperresponsiveness of the Fischer rat reflects the responsiveness of individual airways throughout the airwaytree and are consistent with the notion that in this model hyperresponsiveness is an intrinsic property of airway smooth muscle.

     

T cells of atopic asthmatics preferentially infiltrate into human bronchial xenografts in SCID mice

T cells play an important role in the pathogenesis of bronchial asthma. However, it is not completely known how circulating lymphocytes infiltrate into the airways of asthmatic patients. Because SCID mice are unable to reject xenogenic transplants, many xenotransplant models using various human tissues have been developed. Therefore, to examine the interaction between bronchi and T lymphocytes of asthma, it may be possible to use the human bronchial xenograft and PBMC xenograft in SCID mice. We transplanted human bronchi into the subcutaneum of SCID mice and i.p. injected PBMCs that were obtained from patients with atopic asthma, atopic dermatitis and rheumatoid arthritis, and normal subjects (asthmatic, dermatitis, rheumatic, and normal huPBMC-SCID mice). There was no difference in the percentage of CD3-, CD4-, CD8-, CD25-, CD45RO-, CD103-, and cutaneous lymphocyte Ag-positive cells in PBMCs among the patients with asthma, dermatitis, rheumatoid arthritis, and normal subjects, and CD3-positive cells in peripheral blood of asthmatic, dermatitis, rheumatic, and normal huPBMC-SCID mice. The number of CD3-, CD4-, and CD8-positive cells in the xenografts of asthmatic huPBMC-SCID mice was higher than those of dermatitis, rheumatic, and normal huPBMC-SCID mice. IL-4 mRNA and IL-5 mRNA were significantly higher in the xenografts of asthmatic huPBMC-SCID mice than those in the xenografts of normal huPBMC-SCID mice, but there were no significant differences in the expressions of IL-2 mRNA or IFN-gamma mRNA between them. These findings suggest that T cells, especially Th2-type T cells, of asthmatics preferentially infiltrate into the human bronchi.     

Ligation of the bronchial artery in sheep attenuates early pulmonary changes following exposure to smoke.

Smoke inhalation can produce acute pulmonary edema. Previous studies have shown that the bronchial arteries are important in acute pulmonary edema occurring after inhalation of a synthetic smoke containing acrolein, a common smoke toxin. We hypothesized that inhalation of smoke from burning cotton, known to contain acrolein, would produce in sheep acute pulmonary edema that was mediated by the bronchial circulation. We reasoned that occluding the bronchial arteries would eliminate smoke-induced pulmonary edema, whereas occlusion of the pulmonary artery would not. Smoke inhalation increased lung lymph flow from baseline from 2.4 +/- 0.7 to 5.6 +/- 1.2 ml/0.5 h at 30 min (P < 0.05) to 9.1 +/- 1 ml/0.5 h at 4 h (P < 0.05). Bronchial artery ligation diminished and delayed the rise in lymph flow with baseline at 2.8 +/- 0.7 ml/0.5 h rising to 3.1 +/- 0. 8 ml/0.5 h at 30 min to 6.5 +/- 1.5 ml/0.5 h at 240 min (P < 0.05). Wet-to-dry ratio was 4.1 +/- 0.2 in control, 5.1 +/- 0.3 in smoke inhalation (P < 0.05), and 4.4 +/- 0.4 in bronchial artery ligation plus smoke-inhalation group. Smoke inhalation after occlusion of the right pulmonary artery resulted in a wet-to-dry ratio after 4 h in the right lung of 5.5 +/- 0.8 (P < 0.05 vs. control) and in the left nonoccluded lung of 5.01 +/- 0.7 (P < 0.05). Thus the bronchial arteries may be major contributors to acute pulmonary and airway edema following smoke inhalation because the edema occurs in the lung with the pulmonary artery occluded but not in the lungs with bronchial arteries ligated.