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Biomedical subjects

W Mitzner

Publications and source records attributed to W Mitzner.

At least 19 recordsLinked to original sources

Effect of lung inflation and airway muscle tone on airway diameter in vivo.

How normal airway dimensions change with lung volume is of great importance in determining flow limitation during the normal forced vital capacity maneuver as well as in the manifestation of obstructive lung disease. The literature presents a confusing picture, with some results suggesting that airway diameter increases linearly with the cube root of lung volume and others showing a highly nonlinear relation. The effect of smooth muscle contraction on lung-airway interdependence is even less well understood. Recent morphological work explicitly assumes that airway basement membrane is nondistensible, although the lung volume at which this maximal airway size is reached is unknown. With smooth muscle contraction, folding of the epithelium and basement membrane accounts for the changes in luminal area. In this study, we measured the effect of lung inflation on relaxed and contracted airway areas by using high-resolution computed tomography at different transpulmonary pressures, each held for 2 min. We found that fully relaxed airways are quite distensible up to a pressure of 5-7 cmH2O (P < 0.001), where they reach a maximal size with no further distension up to an airway pressure of 30 cmH2O (P = 0.49). Thus relaxed airways clearly do not expand isotropically with the lung. With smooth muscle tone, the airways in different animals responded differently to lung inflation, with some animals showing minimal airway dilation up to an airway pressure of 20 cmH2O and others showing airways that were more easily dilated with lung expansion. However, maximal diameter of these moderately constricted airways was not usually achieved even up to an airway pressure of 30 cmH2O. Thus a transient deep inspiration in vivo would be expected to have only a small effect on contracted airways.

Animals

Effects of bronchial vascular engorgement on airway dimensions.

Airway vascular engorgement has been suggested to cause luminal narrowing and airflow obstruction. To determine the extent to which changes in bronchial vascular volume could influence airway dimensions, we studied the effects of left atrial pressure elevation on airway morphometry in sheep (n = 17). The bronchial branch of the bronchoesophageal artery was cannulated and perfused with autologous blood (0.6 ml.min-1.kg-1). A balloon-tipped catheter was inserted into the left atrial appendage to elevate left atrial pressure by 10 mmHg, and papaverine was infused into the bronchial artery to eliminate airway smooth muscle tone. Morphological measurements were made from rapidly frozen lungs excised in vivo. Left atrial pressure elevation caused a 79% increase in total vascular area (P = 0.0002). Average airway luminal area was significantly decreased from 86 to 71% of the airway maximal area (P < 0.0001). Noteworthy were the prominent bronchial vessels located within mucosal folds. However, when papaverine was infused during left atrial pressure elevation, despite a comparable total vascular area, luminal narrowing did not occur and remained at 87% of the maximal area (P = 0.6267). In conclusion, we found that engorgement of the bronchial vasculature leads to an increase in the vascular area in regions inside and outside the smooth muscle layer. The associated decrease in luminal area only occurs in the presence of airway smooth muscle tone. This suggests a reflex effect on the airway caused by the vascular engorgement. We conclude that vascular engorgement of the airway wall per se has a negligible effect on airway obstruction.

Airway Resistance

Airway hyperresponsiveness to acetylcholine: segregation analysis and evidence for linkage to murine chromosome 6.

A genetic predisposition to nonspecific airway hyperresponsiveness (AHR) can be demonstrated in humans and in many animal models. The goal of the current study was to gain insight into the molecular mechanisms that determine AHR by mapping the genes that control this phenotype. We describe genetic studies in a mouse model of differential sensitivity to acetylcholine (ACh)-induced AHR. This model was used to ascertain the number, magnitude of effect, and chromosomal location of quantitative trait loci (QTL) providing susceptibility to ACh-induced AHR. Segregation analyses indicated that a major locus acting additively with a polygenic effect segregates with the airway pressure-time index (APTI) in the progeny of hyperresponsive A/J and hyporesponsive C3H/HeJ mice. Additionally, four loci segregate with respiratory system resistance (Rrs). Examination of the genome for markers linked to these phenotypes indicated that a QTL on chromosome 6 was common to both traits. QTL analysis in the [(C3H/HeJ x A/J)F1 x A/J] backcross generation revealed significant linkage for ACh-induced AHR within the interval spanning the chromosome 6 deoxyribonucleic acid (DNA) markers D6Mit16 and D6Mit13. A/J alleles in this interval were associated with significantly greater airway responsiveness than were C3H/HeJ alleles. Several important candidate genes map to this region, including the locus for the interleukin-5 (IL-5) receptor. This mapping information in the mouse may relate to human studies in which bronchial hyperresponsiveness links to the chromosomal region containing the gene for IL-5 (1).

Acetylcholine

Hyperpnea with dry air causes time-dependent alterations in mucosal morphology and bronchovascular permeability.

This study examines the morphological and physiological changes that occur in canine peripheral airways after hyperpnea with dry air. Peripheral airways were exposed to a 5-min 2,000 ml/min dry air challenge (DAC) at 24, 6, 2, or 1 h before or 60 s after (0 h) the injection of colloidal carbon. After recording the dry air-induced increase in peripheral airway resistance, the lungs were removed and prepared for morphometric analysis (n = 5). Light microscopy revealed that 50% of the airway perimeter appeared damaged at 0, 1, and 2 h after DAC, and repair was evident 6-24 h after the challenge. The average goblet-to-ciliated cell ratio decreased from 0.34 before DAC to 0.15 after DAC and recovered within 24 h. Dry air-induced bronchovascular leakage occurred immediately after DAC and persisted for > or = 24 h. DAC decreased mast cell number only in regions where the mucosa was damaged, and this decrease was inversely correlated with bronchovascular leakage. Finally, leukocyte infiltration was evident 1-2 h after DAC and continued throughout the 24-h period. We conclude that hyperpnea with dry air causes mucosal injury, inflammation, and microvascular leakage and that these dry air-induced effects persist for > or = 24 h after DAC.

Air

Visualization of airway obstruction in vivo during pulmonary vascular engorgement and edema.

Although pulmonary vascular engorgement has often been hypothesized to decrease airway caliber, leading to airway obstruction in asthma, direct evidence for this hypothesis is lacking. In the present study, we used high-resolution computed tomography to directly measure the changes in the caliber and wall thickness of conducting airways after volume loading with normal saline (NS) and homologous blood. Five anesthetized dogs received 0.2 mg/kg of atropine followed by either three sequential fluid challenges of 50 ml/kg of NS or two sequential challenges of 25 ml/kg of blood. Several weeks later, the same dogs received the other fluid challenge. Volume loading with 150 ml/kg of NS decreased the average airway luminal area to 68 +/- 3% (+/- SE) of baseline. Concomitantly, airway wall thickness increased to 150 +/- 6% of baseline. Volume loading with 50 ml/kg of blood decreased the average airway luminal area to 81 +/- 2% of baseline. Concomitantly, airway wall thickness increased to 108 +/- 2% of baseline. Therefore, for comparable changes in pulmonary vascular pressure, an infusion of NS caused a significantly greater decrease in airway luminal area and a larger increase in airway wall thickness than an infusion of blood. This suggests that the presence of edema fluid in or immediately surrounding the airway wall acts to decrease the airway lumen. However, since the degree of airway narrowing was only moderate, even with a most extreme fluid load, it seems unlikely that airway wall thickening or edema could be a primary cause of conducting airway obstruction in patients with asthma or impaired left ventricular function.(ABSTRACT TRUNCATED AT 250 WORDS)

Airway Obstruction

A beta 2-adrenergic agonist inhibits dry air-induced injury in canine peripheral airways.

We examined the effects of a beta 2-agonist on dry air-induced injury in canine peripheral airways. Dry air-induced bronchoconstriction (AIB) was assessed by measuring peripheral airway resistance in anesthetized dogs. Salbutamol reduced AIB by approximately 75% compared with control values. Colloidal carbon was used to detect bronchovascular leakage in contralateral sublobar segments that were pretreated with saline or salbutamol. About 87% of the perimeter of bronchi was damaged after dry air challenge in saline-treated segments. Salbutamol reduced mucosal damage by approximately 30% (P < 0.05). The mucosa of bronchioles was not injured. The average goblet-to-ciliated cell ratio (which reflects mucosal perturbation) in bronchi decreased from 0.38 in control bronchi to 0.15 in challenged bronchi, and this effect was also evident in bronchioles. Salbutamol did not affect this decrement. Dry air challenge also caused degranulation of mast cells located below damaged mucosa, dilation of bronchial vessels, and leakage from capillaries and venules located below normal ciliated and damaged mucosa of bronchi. Thus, we conclude that salbutamol attenuates epithelial damage and AIB but fails to inhibit mast cell degranulation and vascular hyperpermeability.

Air

Respiratory system mechanics in mice measured by end-inflation occlusion.

Characterization of pulmonary function parameters in mice will facilitate the dissection of genetic mechanisms underlying airway hyperresponsiveness. We evaluated acetylcholine (ACh)-induced respiratory system resistance (Rrs) and elastance (Ers) in A/J and C3H/HeJ mice and compared these results with the previously used airway pressure-time index (APTI). A low-dead-space ventilatory system was designed to ventilate anesthetized mice with constant inspiratory flow. The end-inflation occlusion method was used to measure Rrs and Ers at baseline and after intravenous ACh (12.5-75.0 micrograms/kg) challenge. ACh induced a dose-dependent rise in Rrs and Ers in A/J mice, whereas minimal changes were observed in C3H/HeJ mice. A/J mice had a higher baseline Rrs, yet the response to ACh was independent of baseline Rrs. Additionally, sequential ACh challenges led to augmented responses. Rrs, Ers, and APTI were strongly correlated, and each was useful to detect differences in interstrain cholinergic-induced airway responsiveness. The Rrs detected the smallest differences between the strains of mice studied.

Acetylcholine

Airway edema potentiates airway reactivity.

Thickening of the airway wall has been hypothesized to be one of the mechanisms contributing to airway hyperresponsiveness in asthma. If such thickening of the wall is internal to the airway smooth muscle or otherwise causes a decrease in baseline airway caliber, it should also cause exaggerated airway responsiveness. In the present study, we used high-resolution computed tomography to directly measure the changes in the caliber and wall thickness of conducting airways after aerosol histamine challenge before and after normal saline volume loading. On separate days, five anesthetized dogs received either a baseline aerosol challenge of 3 mg/ml of histamine for five breaths or the same aerosol challenge immediately after a 100 ml/kg bolus of normal saline infused over a 10-min period. Baseline aerosol histamine challenge decreased airway area to 71 +/- 2% (SE) of the control value (P < 0.05). Intravenous administration of 100 ml/kg of normal saline increased wall area by decreasing airway luminal area to 78 +/- 3% of the control value (P < 0.01), with no change in outer airway area. Aerosol histamine challenge superimposed on this engorgement with normal saline challenge further decreased airway luminal area to 54 +/- 3% of the control value (P < 0.01). Quantitative modeling indicated that the edema in the airway wall was mostly outside the smooth muscle and that the smooth muscle shortening with histamine was similar with and without edema. We conclude that a moderate degree of acute airway wall thickening can lead to a potentiated constrictor response to histamine.

Aerosols

Measurement of three-dimensional lung tree structures by using computed tomography.

A method was devised to computationally segment and measure three-dimensional pulmonary trees in situ. Bronchi and pulmonary vessels were computationally extracted from volumetric computed tomography data based on radiopacity differences between airway wall and airway lumen and between blood and parenchyma, respectively. The tree was reduced to a central axis to facilitate measurement of branch segment length and angle. Cross-sectional area was measured on a reconstructed computed tomography slice perpendicular to this central axis. The method was validated by scanning two Plexiglas phantoms and an intact lung. Reconstructed diameters in the phantoms were accurate for branches > 2 mm. In the lung airway branches between 1 and 2 mm in diameter were often unresolved when their angle of orientation with respect to the axis of the scanner was > 45 degrees. However, if a branch was resolved, its reconstructed diameter was little affected by orientation. This method represents a significant improvement in the analysis of complex pulmonary structures in three dimensions.

Algorithms

Variability in the size of individual airways over the course of one year.

Several studies have examined the reproducibility over time of standard clinical measurements of pulmonary airways parameters, such as FEV1 and FVC. These studies indicate fairly consistent reproducibility of such measurements over periods of months and years. Although such traditional pulmonary function tests can provide quantitative measures of lung function, they do not provide any insight into the local or regional variation of individual airways. Therefore, in a longitudinal study in dogs, we directly measured the baseline size (cross-sectional area) of the same individual canine airways multiple times over a 1-yr period, using high-resolution computed tomography (HRCT). We compared the variability of individual airway size both within and between subjects to the various sizes of airways and to seasonal differences in size. Five anesthetized, intubated, and ventilated dogs were studied between five and eight times over a 15-mo period. HRCT scans were acquired at end expiration with the dogs at FRC on all occasions. All individual airways showed considerable variability in size over time as measured by the coefficient of variation (CV) (range: 9.2 to 69.3). The sizes of individual airways within a dog were seen to change by as much as twofold over time. Some dogs also showed significantly greater overall variability in airway size than did other dogs (p < 0.01), but this variability was related neither to the size of the airway (p = 0.34) nor to the season. These findings indicate a substantial local and regional variability in airway caliber that would not be detectable with conventional pulmonary function tests.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

The effects of alpha-adrenergic agonists on hyperpnea-induced airway obstruction in dogs.

Two alpha-adrenergic agonists that inhibit hyperpnea-induced airway obstruction (HIAO) in asthmatic subjects were used to examine the role of bronchial blood flow in the development of HIAO in canine periphery airways. A bronchoscope was used to record peripheral airway resistance (Rp) in anesthetized dogs before and after hyperpnea with dry air. Hyperpnea increased Rp 64 +/- 8% (mean +/- SE) above baseline. Treatment with norepinephrine (NOR) either before or at various times after hyperpnea inhibited HIAO (p < 0.01). We also found that NOR inhibited acetylcholine-induced bronchoconstriction. However, beta-adrenergic blockade with propranolol completely eliminated these effects. Thus, NOR inhibited HIAO in canine peripheral airways via the stimulation of beta-adrenergic receptors and the attenuation of airway smooth muscle contractility. In contrast, pretreatment with methoxamine (MX) decreased HIAO by approximately 25% when compared with the vehicle control, and this effect was completely eliminated by alpha-adrenergic blockade with phentolamine. Relative to NOR, MX provides weak protection against HIAO via the direct stimulation of alpha-adrenergic receptors and their subsequent effect on either mucus secretion or bronchovascular tone. We conclude that bronchial blood flow plays at best a minor role in the development of HIAO.

Acetylcholine

Effects of increased bronchial blood flow on airway morphometry, resistance, and reactivity.

It has been suggested that airway obstruction may be mediated in part by airway vascular engorgement or airway wall edema. However, there are few data that support this conjecture. In this study we examined the effects of increased bronchial blood flow (Qba) on airway wall dimensions, conducting airway resistance, peripheral airway resistance, and airway reactivity assessed by methacholine aerosol challenge. The bronchial artery was perfused with autologous blood (control Qba = 0.6 ml.min-1.kg-1) in anesthetized ventilated sheep. The artery was perfused at either control (C) Qba or at high (H) Qba (300% of C Qba) for 3 h. Morphometry showed a doubling of the vascular area in airways exposed to H Qba (n = 4) compared with C Qba (n = 4). However, the significant increase in wall area could be accounted for only partially by the vascular changes, with edema fluid accumulation accounting for the major increase. Despite these changes, baseline airway resistance (n = 16) and peripheral airway resistance were both unaltered. Airway reactivity to methacholine before and after H Qba was also examined (n = 12). The 3 h of H Qba had no effect on airway reactivity regardless of whether challenge occurred with C or H Qba. The lack of effect of vascular engorgement on airway resistance or reactivity does not support a primary role for these factors in mediating airway obstruction.

Aerosols

Role of cationic proteins in the airway. Hyperresponsiveness due to airway inflammation.

Major basic protein (MBP) is a highly cationic protein found in the granules of eosinophils. It has been postulated that MBP may participate in the pathogenesis of airway hyperresponsiveness exhibited by asthmatic patients. Accordingly, we have employed a rat system to investigate the effect of human MBP instillation on airway responsiveness and the possible role of cationic charge in the determination of this effect. Major basic protein caused a significant increase in airway responsiveness to inhaled methacholine. Two polycations, poly-L-arginine and poly-L-lysine, also increased airway responsiveness to inhaled methacholine. Moreover, two other very different cationic proteins, platelet factor 4 (PF4) and cathepsin G were also capable of inducing airway hyperresponsiveness. These effects were dependent on their positive charge, since the charge--and, hence the effect--of these proteins was neutralized with low molecular weight heparin. In addition, other polyanions, such as low molecular weight heparin, albumin, or dextran sulfate, were also effective. We investigated whether two synthetic cationic proteins, poly-L-arginine and poly-L-lysine, could modify epithelial-dependent responses using a perfused guinea pig tracheal tube preparation. With an intact epithelium, methacholine was some 150 times less potent when applied intraluminally than when applied extraluminally. Perfusion of the luminal surface with cationic proteins increased the potency of intraluminally applied methacholine without modifying the responses to extraluminally applied methacholine. Cationic proteins also attenuated the relaxant effects of intraluminally applied KCl. These effects occurred in the absence of any overt epithelial cell damage. Our data demonstrates that cationic proteins can modify epithelial-dependent responses in the airways.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Dry air-induced mucosal cell injury and bronchovascular leakage in canine peripheral airways.

The purpose of this study was to examine the relationship between hyperpnea-induced mucosal injury, bronchovascular hyperpermeability, and airway reactivity. Hyperpnea-induced bronchoconstriction was assessed by measuring peripheral airway resistance (Rp) in anesthetized mechanically ventilated male mongrel dogs. Either colloidal carbon or monastral blue was used to localize bronchovascular leakage after a 5-min exposure to either a 1000 ml/min dry, 2000 ml/min wet, or 2000 ml/min dry air challenge. Morphometric analyses of cross-sectioned bronchi revealed that hyperpnea with dry air stimulated goblet cell degranulation, damaged the bronchial mucosa, and increased bronchovascular permeability. Exposure to only a 2000 ml/min dry air challenge produced marked mucosal injury when compared with control. Regardless of treatment, bronchial vessels lying below normal mucosa characterized by goblet/ciliated cell (G/C) ratios > or = 0.3 did not leak. A G/C transition zone between 0 and 0.3 separated normal from damaged mucosa. Within this zone, vascular permeability was inversely correlated with G/C ratio. In addition, airflow-induced changes in Rp were inversely related to G/C ratio and positively correlated with bronchovascular leakage. Although these correlations are consistent with the speculation that bronchovascular leakage and edema formation are responsible for the dry air-induced changes in Rp, it is equally plausible that bronchovascular leakage is not the cause of but occurs in concert with airway narrowing to protect cells in the bronchial mucosa from excessive losses of heat and water.

Airway Resistance

Spontaneous airways constrict during breath holding studied by high-resolution computed tomography.

Airway constriction during a breath hold could not be examined previously using standard methods. We used high-resolution computed tomography (HRCT) in vivo to assess the temporal changes in airway area and the effects of a deep inspiration with and without vagal suppression. Five dogs were anesthetized, intubated, and their lungs ventilated with 100 percent oxygen. Fifteen HRCT slices were obtained at functional residual capacity (FRC) either immediately after stopping ventilation at end expiration after either a tidal volume breath or three deep inspirations. Subsequently the dogs were given atropine, 0.2 mg/kg, and the scans were repeated. The cross-sectional areas of 33 airways ranging in size from 1.6 to 9.7 mm in diameter were measured. Airways were separated in three groups based on size: small (< 3 mm in diameter); medium (3 to 6-mm in diameter); and large (> 6 mm in diameter). The small, medium, and large airways showed a spontaneous constriction over time to 49 +/- 8 percent, 83 +/- 4 percent, and 82 +/- 4 percent of initial airway size, respectively (p < 0.01), (p < 0.0001). The deep inspiration caused an initial dilation only in the smallest airways to 133.3 +/- 4 percent. The subsequent constrictions were even greater than after the tidal volume breath averaging 67 +/- 15 percent, 61 +/- 6 percent, and 60 +/- 9 percent of initial airway area in the small, medium, and large airways, respectively (p = 0.001). Atropine caused an average increase in baseline airway area of 115 +/- 5 percent and 121 +/- 6 percent after a tidal volume breath and deep inspiration, respectively, compared with the preatropine controls, with no difference between the three groups. Atropine also completely abolished the spontaneous airway constriction observed after either a tidal volume breath or a deep inspiration in all three groups equally. In conclusion, using direct airway imaging in vivo, we found that airways spontaneously constrict during a prolonged expiratory pause, and a deep inspiration significantly augments this airway constriction. These responses are mediated via vagal afferent pathways, likely arising from progressively decreasing slow-adapting receptor activity.

Analysis of Variance

Direct in vivo visualization of bronchodilation induced by inhalational anesthesia using high-resolution computed tomography.

BACKGROUND: Volatile anesthetics are effective at preventing and reversing bronchospasm, but their effects on baseline airway tone are controversial. While tantalum bronchography has been used in the past to measure one-dimensional airway diameter changes, this method has inherent problems associated with the irritant effects of tantalum. Until recently, no other direct noninvasive in vivo method to assess airway caliber was available. The present investigation assesses the effects of the inhalation anesthetic halothane on individual unstimulated airways in vivo. METHODS: Ten studies were performed in seven dogs. All dogs were initially anesthetized with 15 mg/kg thiopental followed by a 10-mg.kg-1 x h-1 maintenance dose. Following tracheal intubation the lungs were mechanically ventilated (15 ml/kg, 15 beats/min). The dogs subsequently received increasing doses of halothane (range 0.5-1.5%). On a separate day, the dogs were pretreated with atropine (0.2 mg/kg) and the study was repeated. Fifty sequential high-resolution computed tomography scans were obtained using a 1-s scan time, 137 kVp, 220 mA, 2-mm slice thickness, and 1-mm table feed. Airway areas ranging in size from 3 to 22 mm in diameter were measured and analyzed by one way analysis of variance and Bonferroni pair-wise comparisons of means. RESULTS: Halothane in concentrations of 0.5%, 1.0%, and 1.5% showed significant dose-dependent dilation of the airways (percent increase from control) that averaged 90 +/- 19% (mean +/- SEM), 128 +/- 20%, and 182 +/- 27%, respectively (P = .017). Atropine pretreatment alone significantly dilated the airways to 151 +/- 25% (P = .002) of their baseline value. Halothane caused no further airway dilation in atropine pretreated dogs. CONCLUSIONS: Halothane dilates baseline airways by blocking baseline vagal tone. Since baseline airway tone, airway wall thickness, and initial airway diameter are major determinants of airway reactivity, the observed dilation by halothane may be one of the mechanisms by which inhalational anesthetics decrease airway reactivity.

Anesthesia, Inhalation