Functional lung imaging with synchrotron radiation: Methods and preclinical applications.

Many lung disease processes are characterized by structural and functional heterogeneity that is not directly appreciable with traditional physiological measurements. Experimental methods and lung function modeling to study regional lung function are crucial for better understanding of disease mechanisms and for targeting treatment. Synchrotron radiation offers useful properties to this end: coherence, utilized in phase-contrast imaging, and high flux and a wide energy spectrum which allow the selection of very narrow energy bands of radiation, thus allowing imaging at very specific energies. K-edge subtraction imaging (KES) has thus been developed at synchrotrons for both human and small animal imaging. The unique properties of synchrotron radiation extend X-ray computed tomography (CT) capabilities to quantitatively assess lung morphology, and also to map regional lung ventilation, perfusion, inflammation and biomechanical properties, with microscopic spatial resolution. Four-dimensional imaging, allows the investigation of the dynamics of regional lung functional parameters simultaneously with structural deformation of the lung as a function of time. This review summarizes synchrotron radiation imaging methods and overviews examples of its application in the study of disease mechanisms in preclinical animal models, as well as the potential for clinical translation both through the knowledge gained using these techniques and transfer of imaging technology to laboratory X-ray sources.

High-power-load DCLM monochromator for a computed tomography program at BMIT at energies of 25-150 keV.

The research program at the biomedical imaging facility requires a high-flux hard-X-ray monochromator that can also provide a wide beam. A wide energy range is needed for standard radiography, phase-contrast imaging, K-edge subtraction imaging and monochromatic beam therapy modalities. The double-crystal Laue monochromator, developed for the BioMedical Imaging and Therapy facility, is optimized for the imaging of medium- and large-scale samples at high energies with the resolution reaching 4 µm. A pair of 2 mm-thick Si(111) bent Laue-type crystals were used in fixed-exit beam mode with a 16 mm vertical beam offset and the first crystal water-cooled. The monochromator operates at energies from 25 to 150 keV, and the measured size of the beam is 189 mm (H) × 8.6 mm (V) at 55 m from the source. This paper presents our approach in developing a complete focusing model of the monochromator. The model uses mechanical properties of crystals and benders to obtain a finite-element analysis of the complete assembly. The modeling results are compared and calibrated with experimental measurements. Using the developed analysis, a rough estimate of the bending radius and virtual focus (image) position of the first crystal can be made, which is also the real source for the second crystal. On the other hand, by measuring the beam height in several points in the SOE-1 hutch, the virtual focus of the second crystal can be estimated. The focusing model was then calibrated with measured mechanical properties, the values for the force and torque applied to the crystals were corrected, and the actual operating parameters of the monochromator for fine-tuning were provided.

Radiation therapy at compact Compton sources.

The principle of the compact Compton source is presented briefly. In collision with an ultrarelativistic electron bunch a laser pulse is back-scattered as hard X-rays. The radiation cone has an opening of a few mrad, and the energy bandwidth is a few percent. The electrons that have an energy of the order of a few tens of MeV either circulate in storage ring, or are injected to a linac at a frequency of 10-100 MHz. At the interaction point the electron bunch collides with the laser pulse that has been amplified in a Fabry-Perot resonator. There are several machines in design or construction phase, and projected fluxes are 10(12) to 10(14) photons/s. The flux available at 80 keV from the ThomX machine is compared with that used in the Stereotactic Synchrotron Radiation Therapy clinical trials. It is concluded that ThomX has the potential of serving as the radiation source in future radiation therapy programs, and that ThomX can be integrated in hospital environment.

Differences in the pattern of bronchoconstriction induced by intravenous and inhaled methacholine in rabbit.

We measured bronchoconstriction in central bronchi, and in small peripheral airways causing the emergence of ventilation defects (VD), through two delivery routes: intravenous (IV) and inhaled MCh, in 2 groups of rabbits (A: n=5; B: n=4), using synchrotron imaging of regional lung structure and ventilation. We assessed the effect an initial IV challenge on a subsequent inhaled challenge in group B. Inhaled MCh decreased central airway cross-sections (CA) by 13-22%, but increased VD area by 25-49%. IV MCh decreased CA by 44% but increased the area of ventilation defects (VD) by 13% only. An initial IV MCh challenge reduced regional ventilation heterogeneity following a subsequent inhaled MCh challenge, suggesting the role of agonist-receptor interaction in the response pattern. Heterogeneous agonist distribution due to uneven aerosol deposition could explain the different patterns of response between IV and inhaled routes. This mechanism could participate in the emergence of ventilation heterogeneities during bronchial challenge, or exposure to allergen in asthmatic patients.

Role of cellular effectors in the emergence of ventilation defects during allergic bronchoconstriction.

It is not known whether local factors within the airway wall or parenchyma may influence the emergence and spatial distribution of ventilation defects (VDs), thereby modulating the dynamic system behavior of the lung during bronchoconstriction. We assessed the relationship between the distribution of cellular effectors and the emergence of defects in regional ventilation distribution following allergen challenge. We performed high-resolution K-edge subtraction (KES) synchrotron imaging during xenon inhalation and measured the forced oscillatory input impedance in ovalbumin (OVA)-sensitized Brown-Norway rats (n = 12) at baseline and repeatedly following OVA challenge. Histological slices with best anatomic matching to the computed tomographic images were stained with a modified May-Grunwald Giemsa and immunohistochemical staining with monoclonal anti-rat CD68, in six rats. Slides were digitized and total cells and eosinophils were counted in the walls of bronchi and vessels randomly selected within and outside of VDs on the basis of xenon-KES images. Ventilated alveolar area decreased and ventilation heterogeneity, Newtonian resistance, tissue damping, and elastance increased following OVA challenge. Eosinophil, total cell, and CD68+ counts were significantly higher in the bronchial and vascular walls within vs. outside of the VDs. The minimal central airway diameters during OVA-induced bronchoconstriction were correlated with eosinophil (R = -0.85; P = 0.031) and total cell densities (R = -0.82; P = 0.046) in the airway walls within the poorly ventilated zones. Our findings suggest that allergic airway inflammation is locally heterogeneous and is topographically associated with the local emergence of VDs following allergen challenge.

Effect of positive end-expiratory pressure on regional ventilation distribution during mechanical ventilation after surfactant depletion.

BACKGROUND: Ventilator-induced lung injury occurs due to exaggerated local stresses, repeated collapse, and opening of terminal air spaces in poorly aerated dependent lung, and increased stretch in nondependent lung. The aim of this study was to quantify the functional behavior of peripheral lung units in whole-lung lavage-induced surfactant depletion, and to assess the effect of positive end-expiratory pressure. METHODS: The authors used synchrotron imaging to measure lung aeration and regional specific ventilation at positive end-expiratory pressure of 3 and 9 cm H2O, before and after whole-lung lavage in rabbits. Respiratory mechanical parameters were measured, and helium-washout was used to assess end-expiratory lung volume. RESULTS: Atelectatic, poorly, normally aerated, hyperinflated, and trapped regions could be identified using the imaging technique used in this study. Surfactant depletion significantly increased atelectasis (6.3±3.3 [mean±SEM]% total lung area; P=0.04 vs. control) and poor aeration in dependent lung. Regional ventilation was distributed to poorly aerated regions with high (16.4±4.4%; P<0.001), normal (20.7±5.9%; P<0.001 vs. control), and low (5.7±1.2%; P<0.05 vs. control) specific ventilation. Significant redistribution of ventilation to normally aerated nondependent lung regions occurred (41.0±9.6%; P=0.03 vs. control). Increasing positive end-expiratory pressure level to 9 cm H2O significantly reduced poor aeration and recruited atelectasis, but ventilation redistribution persisted (39.2±9.5%; P<0.001 vs. control). CONCLUSIONS: Ventilation of poorly aerated dependent lung regions, which can promote the local concentration of mechanical stresses, was the predominant functional behavior in surfactant-depleted lung. Potential tidal recruitment of atelectatic lung regions involved a smaller fraction of the imaged lung. Significant ventilation redistribution to aerated lung regions places these at risk of increased stretch injury.

X-ray phase-contrast imaging: from pre-clinical applications towards clinics.

Phase-contrast x-ray imaging (PCI) is an innovative method that is sensitive to the refraction of the x-rays in matter. PCI is particularly adapted to visualize weakly absorbing details like those often encountered in biology and medicine. In past years, PCI has become one of the most used imaging methods in laboratory and preclinical studies: its unique characteristics allow high contrast 3D visualization of thick and complex samples even at high spatial resolution. Applications have covered a wide range of pathologies and organs, and are more and more often performed in vivo. Several techniques are now available to exploit and visualize the phase-contrast: propagation- and analyzer-based, crystal and grating interferometry and non-interferometric methods like the coded aperture. In this review, covering the last five years, we will give an overview of the main theoretical and experimental developments and of the important steps performed towards the clinical implementation of PCI.

Comparison of in vitro breast cancer visibility in analyser-based computed tomography with histopathology, mammography, computed tomography and magnetic resonance imaging.

High-resolution analyser-based X-ray imaging computed tomography (HR ABI-CT) findings on in vitro human breast cancer are compared with histopathology, mammography, computed tomography (CT) and magnetic resonance imaging. The HR ABI-CT images provided significantly better low-contrast visibility compared with the standard radiological images. Fine cancer structures indistinguishable and superimposed in mammograms were seen, and could be matched with the histopathological results. The mean glandular dose was less than 1 mGy in mammography and 12-13 mGy in CT and ABI-CT. The excellent visibility of in vitro breast cancer suggests that HR ABI-CT may have a valuable role in the future as an adjunct or even alternative to current breast diagnostics, when radiation dose is further decreased, and compact synchrotron radiation sources become available.

Effect of positive end-expiratory pressure on regional ventilation distribution during bronchoconstriction in rabbit studied by synchrotron radiation imaging.

OBJECTIVE: To assess the effects of positive end-expiratory pressure on regional ventilation distribution in normal lung and after histamine-induced bronchoconstriction. DESIGN: Experimental study. SETTING: International research laboratory. SUBJECTS: Six healthy New Zealand rabbits weighing 2.5 ± 0.1 kg. INTERVENTIONS: Rabbits were anesthetized, tracheostomized, paralyzed, and mechanically ventilated. Synchrotron radiation computed tomography images of tissue density and specific ventilation were acquired using K-edge subtraction imaging with inhaled stable xenon gas in middle and caudal thoracic levels on 0 and 5 cm H(2)O positive end-expiratory pressure at baseline and twice after histamine inhalation. MEASUREMENTS AND MAIN RESULTS: At baseline, a positive end-expiratory pressure of 5 cm H(2)O significantly increased lung volume. Histamine inhalation caused patchy areas of decreased specific ventilation, including some areas with no ventilation. After histamine, positive end-expiratory pressure significantly increased the area of well-ventilated lung regions and decreased the heterogeneity of specific ventilation. This improvement went together with a significant but limited increase in the area of hyperinflated lung zones. CONCLUSIONS: The findings of this study suggest that in mechanically ventilated rabbit with severely heterogeneous bronchoconstriction, a positive end-expiratory pressure of 5 cm H(2)O significantly improves regional ventilation homogeneity through dilation of flow-limited airways and recruitment of closed airways.

Phase-contrast X-ray imaging of breast.

When an X-ray wave traverses an object, its amplitude and phase change, resulting in attenuation, interference, and refraction, and in phase-contrast X-ray imaging (PCI) these are converted to intensity changes. The relative change of the X-ray phase per unit path length is even orders of magnitude larger than that of the X-ray amplitude, so that the image contrast based on variation of the X-ray phase is potentially much stronger than the contrast based on X-ray amplitude (absorption contrast). An important medical application of PCI methods is soft-tissue imaging, where the absorption contrast is inherently weak. It is shown by in vitro examples that signs of malignant human breast tumor are enhanced in PCI images. Owing to the strong contrast, the radiation dose can be greatly reduced, so that a high-resolution phase-contrast X-ray tomography of the breast is possible with about 1 mGy mean glandular dose. Scattered radiation carries essential information on the atomic and molecular structure of the object, and particularly small-angle X-ray scattering can be used to trace cancer. The imaging methods developed at the synchrotron radiation facilities will become available in the clinical environment with the ongoing development of compact radiation sources, which produce intense X-ray beams of sufficient coherence. Several developments that are under way are described here.

Acute cigarette smoke inhalation blunts lung responsiveness to methacholine and allergen in rabbit: differentiation of central and peripheral effects.

Despite the prevalence of active smoking in asthmatics, data on the short-term effect of acute mainstream tobacco smoke exposure on airway responsiveness are very scarce. The aim of this study was to assess the immediate effect of acute exposure to mainstream cigarette smoke on airway reactivity to subsequent nonspecific and allergenic challenges in healthy control (n = 5) and ovalbumin-sensitized rabbits (n = 6). We combined low-frequency forced oscillations and synchrotron radiation CT imaging to differentiate central airway and peripheral airway and lung parenchymal components of the response to airway provocation. Acute exposure to smoke generated by four successive cigarettes (CS) strongly inhibited the central airway response to subsequent IV methacholine (MCh) challenge. In the sensitized animals, although the response to ovalbumin was also inhibited in the central airways, mainstream CS did not blunt the peripheral airway response in this group. In additional groups of experiments, exposure to HEPA-filtered CS (n = 6) similarly inhibited the MCh response, whereas CO (10,000 ppm for 4 min, n = 6) or nitric oxide inhalation instead of CS (240 ppm, 4 x 7 min, n = 5) failed to blunt nonspecific airway responsiveness. Pretreatment with alpha-chymotrypsin to inhibit endogenous VIP before CS exposure had no effect (n = 4). Based on these observations, the gas phase of mainstream cigarette smoke may contain one or more short-term inhibitory components acting primarily on central airways and inhibiting the response to both specific and nonspecific airway provocation, but not on the lung periphery where both lung mechanical parameters, and synchrotron-imaging derived parameters, showed large changes in response to allergen challenge in sensitized animals.

Directional lithium hydride Compton profiles of 0.1% statistical accuracy.

The present status and accuracy of determining the electron momentum density from experimental Compton profiles is reviewed. The new spectrometers operating at third-generation synchrotron radiation sources have made possible measurements with 0.1% statistical accuracy at the Compton peak. A comparable accuracy of the Compton profiles is achieved only after careful corrections for departures from the impulse approximation, effects of multiple scattering, and variations in the analyser response function. Detailed descriptions are given of the correction procedures applied to the data collected by the Johann-type scanning spectrometer that is one of the Compton spectrometers in use at the ESRF. Special attention is paid to the calculation and correction of the glitches that are caused by extra reflections of the analyser crystal. The Fourier transform of the Compton profile, the reciprocal form factor, is calculated, and its use in data treatment and presentation is discussed.

Methacholine and ovalbumin challenges assessed by forced oscillations and synchrotron lung imaging.

RATIONALE: Methacholine (Mch) is routinely used to assess bronchial hyperreactivity; however, little is known about the differences in the lung response pattern between this provocation and that observed with ovalbumin (Ova) after allergic sensitization. OBJECTIVES: To compare (1) the central versus peripheral effects of Mch and Ova within the lung by combining measurements of airway and tissue mechanics with synchrotron radiation (SR) imaging, and (2) to assess the extent to which mechanical and imaging parameters are correlated. METHODS: We used the low-frequency forced oscillation technique and SR imaging in control (n = 12) and ovalbumin-sensitized (n = 13) rabbits, at baseline, during intravenous Mch infusion (2.5 microg/kg/min, 5.0 microg/kg/min, or 10.0 microg/kg/min), after recovery from Mch, and after intravenous Ova injection (2.0 mg). We compared intravenous Mch challenge with inhaled Mch (125 mg/ml, 90 s) in a separate group of control animals (n = 5). MEASUREMENTS AND MAIN RESULTS: Airway conductance and tissue elastance were measured by low-frequency forced oscillation technique. The central airway cross-sectional area, the ventilated alveolar area, and the heterogeneity of specific ventilation were quantified by SR imaging. Mch infusion induced constriction predominantly in the central airways, whereas Ova provocation affected mainly the peripheral airways, leading to severe ventilation heterogeneities in sensitized animals. Mch inhalation affected both conducting and peripheral airways. The correlations between airway conductance and central airway cross-sectional area (R = 0.71) and between tissue elastance and ventilated alveolar area (R = -0.72) were strong. CONCLUSIONS: The pattern of lung response caused by intravenous Mch and Ova are fundamentally different. Although inhaled Mch induces a heterogeneous lung response similar to that observed with intravenous allergen, these similar patterns are due to different mechanisms.

Paradoxical conducting airway responses and heterogeneous regional ventilation after histamine inhalation in rabbit studied by synchrotron radiation CT.

We studied both central conducting airway response and changes in the distribution of regional ventilation induced by inhaled histamine in healthy anesthetized and mechanically ventilated rabbit using a novel xenon-enhanced synchrotron radiation computed tomography (CT) imaging technique, K-edge subtraction imaging (KES). Images of specific ventilation were obtained using serial KES during xenon washin, in three axial lung slices, at baseline and twice after inhalation of histamine aerosol (50 or 125 mg/ml) in two groups of animals (n = 6 each). Histamine inhalation caused large clustered areas of poor ventilation, characterized by a drop in average specific ventilation (sV(m)), but an increase in sV(m) in the remaining lung zones indicating ventilation redistribution. Ventilation heterogeneity, estimated as coefficient of variation (CV) of sV(m) significantly increased following histamine inhalation. The area of ventilation defects and CV were significantly larger with the higher histamine dose. In conducting airways, histamine inhalation caused a heterogeneous airway response combining narrowing and dilatation in individual airways of different generations, with the probability for constriction increasing peripherally. This finding provides further in vivo evidence that airway reactivity in response to inhaled histamine is complex and that airway response may vary substantially with location within the bronchial tree.

Toward high-contrast breast CT at low radiation dose.

This study was approved by the local research ethics committee, and patient informed consent was obtained. The purpose of this study was to demonstrate that high-spatial-resolution low-dose analyzer-based x-ray computed tomography (CT) can substantially improve the radiographic contrast of breast tissue in vitro when compared with that attained by using diagnostic mammography and CT. An excised human breast tumor was examined by using analyzer-based x-ray imaging with synchrotron radiation. The correspondence between analyzer-based x-ray images and diagnostic mammograms, CT images, and histopathologic findings was determined. Calcifications and fine details of soft tissue, which are at the contrast detection limit on diagnostic mammograms, are clearly visible on planar analyzer-based x-ray images. Analyzer-based x-ray CT yields high contrast from smoothly varying internal structures, such as tumorous mass lesions, corresponding to information on actual structures seen at histopathologic analysis. The mean glandular dose of 1.9 mGy in analyzer-based x-ray CT is approximately equivalent to the dose administered during single-view screening mammography. The improved visibility of mammographically indistinguishable lesions in vitro suggests that analyzer-based x-ray CT may be a valuable method in radiographic evaluation of the breast, thereby justifying further investigations.

Imaging of lung function using synchrotron radiation computed tomography: what's new?

There is a growing interest in imaging techniques as non-invasive means of quantitatively measuring regional lung structure and function. Abnormalities in lung ventilation due to alterations in airway function such as those observed in asthma and COPD are highly heterogeneous, and experimental methods to study this heterogeneity are crucial for better understanding of disease mechanisms and drug targeting strategies. In severe obstructive diseases requiring mechanical ventilation, the optimal ventilatory strategy to achieve recruitment of poorly ventilated lung zones remains a matter of considerable debate. We have used synchrotron radiation computed tomography (SRCT) for the in vivo study of regional lung ventilation and airway function. This imaging technique allows direct quantification of stable Xenon (Xe) gas used as an inhaled contrast agent using K-edge subtraction imaging. Dynamics of Xe wash-in can be used to calculate quantitative maps of regional specific lung ventilation. More recently, the development of Spiral-CT has allowed the acquisition of 3D images of the pulmonary bronchial tree and airspaces. This technique gives access to quantitative measurements of regional lung volume, ventilation, and mechanical properties. Examples of application in an experimental model of allergic asthma and in imaging lung recruitment as a function of mechanical ventilation parameters will be presented. The future orientations of this technique will be discussed.

Refraction and scattering of X-rays in analyzer-based imaging.

A new algorithm is introduced for separation of the scattered and non-scattered parts of a monochromatic and well collimated synchrotron radiation beam transmitted through a sample and analyzed by reflection from a perfect crystal in the non-dispersive setting. The observed rocking curve is described by the Voigt function, which is a convolution of Lorentzian and Gaussian functions. For the actual fitting, pseudo-Voigtians are used. The fit yields the scaled integrated intensity (the effect of absorption), the center of the rocking curve (the effect of refraction), and the intensity of the transmitted beam is divided into the scattered and non-scattered parts. The algorithm is tested using samples that exhibit various degrees of refraction and scattering. Very close fits are achieved in an angular range that is 15 times the full width at half-maximum of the intrinsic rocking curve of the analyzer. The scattering part has long tails of Lorentzian shape owing to the ;long-slit geometry' of the set-up. Quantitative images of absorption, refraction and scattering are constructed and compared with results of earlier treatments. The portion of scattering and the second moment of the observed rocking curve both increase linearly with the sample thickness and yield identical maps of the effects of scattering. The effects of refraction are calculated using the geometrical optics approximation, and a good agreement with experiment is found. The fits with reduced number of data points (minimum number is five) yield closely the same results as fits to the full data set.

Energy-dispersive diffraction with synchrotron radiation and a germanium detector.

The response of an intrinsic Ge detector in energy-dispersive diffraction measurements with synchrotron radiation is studied with model calculations and diffraction from perfect Si single-crystal samples. The high intensity and time-structure of the synchrotron radiation beam leads to pile-up of the output pulses, and the energy distribution of the pile-up pulses is characteristic of the fill pattern of the storage ring. The pile-up distribution has a single peak and long tail when the interval of the radiation bunches is small, as in the uniform fill pattern, but there are many pile-up peaks when the bunch distance is a sizable fraction of the length of the shaping amplifier output pulse. A model for the detecting chain response is used to resolve the diffraction spectrum from a perfect Si crystal wafer in the symmetrical Laue case. In the 16-bunch fill pattern of the ESRF storage ring the spectrum includes a large number of ;extra reflections' owing to pile-up, and the model parameters are refined by a fit to the observed energy spectrum. The model is used to correct for the effects of pile-up in a measurement with the 1/3 fill pattern of the storage ring. Si reflections (2h,2h,0) are resolved up to h = 7. The pile-up corrections are very large, but a perfect agreement with the integrated intensities calculated from dynamical diffraction theory is achieved after the corrections. The result also demonstrates the convergence of kinematical and dynamical theories at the limit where the extinction length is much larger than the effective thickness of the perfect crystal. The model is applied to powder diffraction using different fill patterns in simulations of the diffraction pattern, and it is demonstrated that the regularly spaced pile-up peaks might be misinterpreted to arise from superlattices or phase transitions. The use of energy-dispersive diffraction in strain mapping in polycrystalline materials is discussed, and it is shown that low count rates but still good statistical accuracy are needed for reliable results.

High-resolution CT by diffraction-enhanced x-ray imaging: mapping of breast tissue samples and comparison with their histo-pathology.

The aim of this study was to introduce high-resolution computed tomography (CT) of breast tumours using the diffraction-enhanced x-ray imaging (DEI) technique and to compare results with radiological and histo-pathological examinations. X-ray CT images of tumour-bearing breast tissue samples were acquired by monochromatic synchrotron radiation (SR). Due to the narrow beam and a large sample-to-detector distance scattering is rejected in the absorption contrast images (SR-CT). Large contrast enhancement is achieved by the use of the DEI-CT method, where the effects of refraction and scatter rejection are analysed by crystal optics. Clinical mammograms and CT images were recorded as reference material for a radiological examination. Three malignant and benign samples were studied in detail. Their radiographs were compared with optical images of stained histological sections. The DEI-CT images map accurately the morphology of the samples, including collagen strands and micro-calcifications of dimensions less than 0.1 mm. Histo-pathological examination and reading of the radiographs were done independently, and the conclusions were in general agreement. High-resolution DEI-CT images show strong contrast and permit visualization of details invisible in clinical radiographs. The radiation dose may be reduced by an order of magnitude without compromising image quality, which would make possible clinical in vivo DEI-CT with future compact SR sources.

Differences in the time course of proximal and distal airway response to inhaled histamine studied by synchrotron radiation CT.

We studied the kinetics of proximal and distal bronchial response to histamine aerosol in healthy anesthetized and mechanically ventilated rabbits up to 60 min after histamine administration using a novel xenon-enhanced synchrotron radiation computed tomography imaging technique. Individual proximal airway constriction was assessed by measuring the luminal cross-sectional area. Distal airway obstruction was estimated by measuring the ventilated alveolar area after inhaled xenon administration. Respiratory system conductance was assessed continuously. Proximal airway cross-sectional area decreased by 57% of the baseline value by 20 min and recovered gradually but incompletely within 60 min. The ventilated alveolar area decreased immediately after histamine inhalation by 55% of baseline value and recovered rapidly thereafter. The results indicate that the airway reaction to inhaled histamine and the subsequent recovery are significantly slower in proximal than in distal bronchi in healthy rabbit. The findings suggest that physiological reaction mechanisms to inhaled histamine in the airway walls of large and small bronchi are not similar.