To facilitate computational toxicology, we developed an approach for generating high-resolution lung-anatomy and particle-deposition mouse models. Major processing steps of our method include mouse preparation, serial block-face cryomicrotome imaging, and highly automated image analysis for generating three-dimensional (3D) mesh-based models and volume-based models of lung anatomy (airways, lobes, sublobes, and near-acini structures) that are linked to local particle-deposition measurements. Analysis resulted in 34 mouse models covering 4 different mouse strains (B6C3F1: 8, BALB/C: 11, C57Bl/6: 8, and CD-1: 7) as well as both sexes (16 male and 18 female) and different particle sizes [2 μm ( n = 15), 1 μm ( n = 16), and 0.5 μm ( n = 3)]. On average, resulting mouse airway models had 1,616.9 ± 298.1 segments, a centerline length of 597.6 ± 59.8 mm, and 1,968.9 ± 296.3 outlet regions. In addition to 3D geometric lung models, matching detailed relative particle-deposition measurements are provided. All data sets are available online in the lapdMouse archive for download. The presented approach enables linking relative particle deposition to anatomical structures like airways. This will in turn improve the understanding of site-specific airflows and how they affect drug, environmental, or biological aerosol deposition. NEW & NOTEWORTHY Computer simulations of particle deposition in mouse lungs play an important role in computational toxicology. Until now, a limiting factor was the lack of high-resolution mouse lung models and measured local particle-deposition information, which are required for developing accurate modeling approaches (e.g., computational fluid dynamics). With the developed imaging and analysis approach, we address this issue and provide all of the raw and processed data in a publicly accessible repository.

Allograft survival is dependent on allorecognition due to peptide display differences by a major histocompatibility complex (MHC) distinct from self (major), or presentation of a novel peptide by identical MHC (minor). In the dog leukocyte antigen (DLA)-identical setting without immune suppression, robust lung allograft rejection based on minor antigen differences occurs within 2 weeks in normal recipients. We previously observed an increase in peripheral regulation in DLA-identical mixed hematopoietic chimeras (MC) as compared with normal dogs and saw a further regulation increase in MC with functioning solid organ transplants from their hematopoietic donors.1 Because tolerance induced thru regulation of minor antigens via MC could replace immune suppression, but would require a toxic conditioning regimen, we investigated the administration of ex vivo generated regulatory T-cell preparations (Treg preps) as a safer route to tolerance. In vitro generated, MHC-mismatched alloantigen-primed peripheral blood mononuclear cells contain highly suppressive CD25+ cells expressing 10-fold more FOXP3 than CD25+ cells in blood.2 We hypothesized that "off-the-shelf"3 recipient-derived major alloantigen-primed CD25-selected cells could regulate incoming minor antigen presentation differences in a DLA-identical lung transplantation model and would extend the duration of allografts compared with that of a historic control group.1 Dogs in both groups received orthotopic lung allografts4 without any immune suppression and underwent pulmonary function tests (PFT) to determine allograft duration. Tested controls had early function (median, 5 days; range, 2-7; n = 4) and were electively, serially sacrificed (median, 19 days; range, 6-33 days; n = 7) with allografts from day 8 onward found engorged at necropsy.1 The experimental group (n = 7) underwent the same surgery with only the addition of an intraoperative, intravenous Treg prep infusion immediately after allograft reperfusion. PFT were performed a week posttransplant and monthly thereafter; all evaluable recipients had early allograft function (median, 8 days; range, 5-8; n = 6), whereas 3 had extended duration with functional allografts on days 29, 48, and 183. Treg preps were variable in composition with a higher percentage of CD8+CD25+ cells found in preps with allograft rejection times as observed in the control group, whereas the highest CD4+CD25+ percentages were seen in the 2 preps with the longest allograft duration. There was no correlation between allograft duration and cell number infused; the longest duration was with the least number of cells, total, and per kg (Table 1). The in vitro suppressive capacity of a Treg prep correlated with the in vivo lung allograft duration (R2 = 0.81; P = 0.01; n = 6). Taken together, this suggests that quality of a Treg preparation is paramount to quantity.5 Improvements could employ CD8-depletion before infusion.TABLE 1: Lung Transplantation with Treg infusionBecause alloantigen-primed Treg preps can suppress a DLA-mismatched MLR, a DLA-mismatched primed Treg prep of recipient origin might prolong DLA-mismatched allograft function if educated against the incoming organ donor's antigens. This technique would require donor access for over a week and would be clinically difficult. However, if the few cells presenting minor antigen differences in an MHC-matched allograft can be regulated in an organ that contains many passenger leukocytes (lung), then perhaps all cells bearing major antigen differences in organs with few passenger leukocytes (kidney, heart) can be regulated using "off-the-shelf" Treg preps.

Despite substantial development of sophisticated subject-specific computational models of aerosol transport and deposition in human lungs, experimental validation of predictions from these new models is sparse. We collected aerosol retention and exhalation profiles in seven healthy volunteers and six subjects with mild-to-moderate COPD (FEV1=50-80%predicted) in the supine posture. Total deposition was measured during continuous breathing of 1 and 2.9 mu m-diameter particles (tidal volume of I L, flow rate of 0.3 L/s and 0.75 L/s). Bolus inhalations of 1 mu m particles were performed to penetration volumes of 200, 500 and 800 mL (flow rate of 0.5 L/s). Aerosol bolus dispersion (H), deposition, and mode shift (MS) were calculated from these data. There was no significant difference in total deposition between healthy subjects and those with COPD. Total deposition increased with increasing particle size and also with increasing flow rate. Similarly, there was no significant difference in aerosol bolus deposition between subject groups. Yet, the rate of increase in dispersion and of decrease in MS with increasing penetration volume was higher in subjects with COPD than in healthy volunteers (H: 0.798 +/- 0.205 vs. 0.527 +/- 0.122 mL/mL, p=0.01; MS: -0.271 +/- 0.129 vs. -0.145 +/- 0.076 mL/mL, p=0.05) indicating larger ventilation inhomogeneities (based on H) and increased flow sequencing (based on MS) in the COPD than in the healthy group. In conclusion, in the supine posture, deposition appears to lack sensitivity for assessing the effect of lung morphology and/or ventilation distribution alteration induced by mild-to-moderate lung disease on the fate of inhaled aerosols. However, other parameters such as aerosol bolus dispersion and mode shift may be more sensitive parameters for evaluating models of lungs with moderate disease. (C) 2016 Elsevier Ltd. All rights reserved.

PURPOSE:Computer models for inhalation toxicology and drug-aerosol delivery studies rely on ventilation pattern inputs for predictions of particle deposition and vapor uptake. However, changes in lung mechanics due to disease can impact airflow dynamics and model results. It has been demonstrated that non-invasive, in vivo, 4DCT imaging (3D imaging at multiple time points in the breathing cycle) can be used to map heterogeneities in ventilation patterns under healthy and disease conditions. The purpose of this study was to validate ventilation patterns measured from CT imaging by exposing the same rats to an aerosol of fluorescent microspheres (FMS) and examining particle deposition patterns using cryomicrotome imaging.MATERIALS AND METHODS:Six male Sprague-Dawley rats were intratracheally instilled with elastase to a single lobe to induce a heterogeneous disease. After four weeks, rats were imaged over the breathing cycle by CT then immediately exposed to an aerosol of ∼ 1 μm FMS for ∼ 5 minutes. After the exposure, the lungs were excised and prepared for cryomicrotome imaging, where a 3D image of FMS deposition was acquired using serial sectioning. Cryomicrotome images were spatially registered to match the live CT images to facilitate direct quantitative comparisons of FMS signal intensity with the CT-based ventilation maps.RESULTS:Comparisons of fractional ventilation in contiguous, non-overlapping, 3D regions between CT-based ventilation maps and FMS images showed strong correlations in fractional ventilation (r = 0.888, p < 0.0001).CONCLUSION:We conclude that ventilation maps derived from CT imaging are predictive of the 1 μm aerosol deposition used in ventilation-perfusion heterogeneity inhalation studies.

A highly automated method for the segmentation of airways in the serial block-face cryomicrotome images of rat lungs is presented. First, a point inside of the trachea is manually specified. Then, a set of candidate airway centerline points is automatically identified. By utilizing a novel path extraction method, a centerline path between the root of the airway tree and each point in the set of candidate centerline points is obtained. Local disturbances are robustly handled by a novel path extraction approach, which avoids the shortcut problem of standard minimum cost path algorithms. The union of all centerline paths is utilized to generate an initial airway tree structure, and a pruning algorithm is applied to automatically remove erroneous subtrees or branches. Finally, a surface segmentation method is used to obtain the airway lumen. The method was validated on five image volumes of Sprague-Dawley rats. Based on an expert-generated independent standard, an assessment of airway identification and lumen segmentation performance was conducted. The average of airway detection sensitivity was 87.4% with a 95% confidence interval (CI) of (84.9, 88.6)%. A plot of sensitivity as a function of airway radius is provided. The combined estimate of airway detection specificity was 100% with a 95% CI of (99.4, 100)%. The average number and diameter of terminal airway branches was 1179 and 159 μm, respectively. Segmentation results include airways up to 31 generations. The regression intercept and slope of airway radius measurements derived from final segmentations were estimated to be 7.22 μm and 1.005, respectively. The developed approach enables the quantitative studies of physiology and lung diseases in rats, requiring detailed geometric airway models.

Prior studies exploring the spatial distributions of ventilation and perfusion have partitioned the lung into discrete regions not constrained by anatomical boundaries and may blur regional differences in perfusion and ventilation. To characterize the anatomical heterogeneity of regional ventilation and perfusion, we administered fluorescent microspheres to mark regional ventilation and perfusion in five Sprague-Dawley rats and then using highly automated computer algorithms, partitioned the lungs into regions defined by anatomical structures identified in the images. The anatomical regions ranged in size from the near-acinar to the lobar level. Ventilation and perfusion were well correlated at the smallest anatomical level. Perfusion and ventilation heterogeneity were relatively less in rats compared to data previously published in larger animals. The more uniform distributions may be due to a smaller gravitational gradient and/or the fewer number of generations in the distribution trees before reaching the level of gas exchange, making regional matching of ventilation and perfusion less extensive in small animals.

In the current study, we used a canine model of radiation-induced lung injury to test the effect of a single i.v. infusion of 10×10(6)/kg of marrow fibroblasts on the progression of damage following 15 Gy exposure to the right lung. The fibroblasts, designated DS1 cells, are a cloned population of immortalized cells isolated from a primary culture of marrow stromal cells. DS1 cells were infused at week 5 post-irradiation when lung damage was evident by imaging with high-resolution computed tomography (CT). At 13 weeks post-irradiation we found that 4 out of 5 dogs receiving DS1 cells had significantly improved pulmonary function compared to 0 out of 5 control dogs (p = 0.047, Fisher's Exact). Pulmonary function was measured as the single breath diffusion capacity-hematocrit (DLCO-Hct), the total inspiratory capacity (IC), and the total lung capacity (TLC), which differed significantly between control and DS1-treated dogs; p = 0.002, p = 0.005, and p = 0.004, respectively. The DS1-treated dogs also had less pneumonitis detected by CT imaging and an increased number of TTF-1 (thyroid transcription factor 1, NKX2-1) positive cells in the bronchioli and alveoli compared to control dogs. Endothelial-like progenitor cells (ELC) of host origin, detected by colony assays, were found in peripheral blood after DS1 cell infusion. ELC numbers peaked one day after infusion, and were not detectable by 7 days. These data suggest that infusion of marrow fibroblasts stimulates mobilization of ELC, which is associated with a reduction in otherwise progressive radiation-induced lung injury. We hypothesize that these two observations are related, specifically that circulating ELC contribute to increased angiogenesis, which facilitates endogenous lung repair.

We aimed to investigate the role of endothelin‐mediated vasoconstriction following acute pulmonary thromboembolism (APTE). Thirteen anesthetized piglets (~25 kg) were ventilated with 0 PEEP. Cardiac output (Qt) and wedge pressure (Pw) were measured by a Swan Ganz catheter, along with arterial and venous blood gases. APTE was induced by autologous blood clots (~0.8 g/kg, 12–16 pieces) via a jugular venous catheter at time = 0 minutes until the mean pulmonary arterial pressure (Ppa) was about 2.5 times the baseline at 30 minutes. Eight control animals (Group 1) received only normal saline afterward, while the remaining five (Group 2) received at time = 40‐minute saline plus Tezosentan, a nonspecific endothelin antagonist. The drug was initially given as an intravenous bolus (10 mg/kg), followed by an infusion (2 mg/min) until the end of the experiment at 2 hours. Hemodynamic data were measured before APTE and then at 30‐minute intervals. Pulmonary vascular resistance index (PVRI) was calculated as (Ppa‐Pw)/CI, where CI was cardiac index or Qt/W (body weight). Fluorescent microspheres (FMS) were used to mark regional blood flows and ventilation for cluster analysis. PVRI acutely increased within minutes and remained high despite some recovery over time. With Tezosentan treatment, the results showed that endothelin‐mediated vasoconstriction persisted significantly up to 2 hours and accounted for about 25% of the increase in PVRI while clot obstruction accounted for the remaining 75%. CI remained relatively constant throughout. Tezosentan also affected PVRI indirectly by mitigating the shift of regional blood flow back to the embolized areas over time, possibly by attenuating vasoconstriction in the nonembolized areas. We conclude that following APTE, although the increased PVRI is mostly due to mechanical embolic obstruction, secondary factors such as vasoconstriction and pattern of regional blood flow over time also play important roles.

Pulmonary computational fluid dynamics models require that three-dimensional images be acquired over multiple points in the dynamic breathing cycle without breath holds or changes in ventilatory mechanics. With small animals, these requirements can result in long imaging times (∼90 minutes), over which lung mechanics, such as compliance, may gradually change if not carefully monitored and controlled. These changes, caused by derecruitment of parenchymal tissue, are manifested as an upward drift in peak inspiratory pressure (PIP) or by changes in the pressure waveform and/or lung volume over the course of the experiment. We demonstrate highly repeatable mechanical ventilation in anesthetized rats over a long duration for dynamic lung x-ray computed tomography (CT) imaging. We describe significant updates to a basic commercial ventilator that was acquired for these experiments. Key to achieving consistent results was the implementation of periodic deep breaths, or sighs, of extended duration to maintain lung recruitment. In addition, continuous monitoring of breath-to-breath pressure and volume waveforms and long-term trends in PIP and flow provide diagnostics of changes in breathing mechanics.

OBJECTIVES:To adapt an animal model of acute lung injury for use as a standard protocol for a screening initial evaluation of limited function, or "surge," ventilators for use in mass casualty scenarios.DESIGN:Prospective, experimental animal study.SETTING:University research laboratory.SUBJECTS:Twelve adult pigs.INTERVENTIONS:Twelve spontaneously breathing pigs (six in each group) were subjected to acute lung injury/acute respiratory distress syndrome via pulmonary artery infusion of oleic acid. After development of respiratory failure, animals were mechanically ventilated with a limited-function ventilator (simplified automatic ventilator [SAVe] I or II; Automedx, Germantown, MD) for 1 hr or until the ventilator could not support the animal. The limited-function ventilator was then exchanged for a full-function ventilator (Servo 900C; Siemens-Elema, Solna, Sweden).MEASUREMENTS AND MAIN RESULTS:Reliable and reproducible levels of acute lung injury/acute respiratory distress syndrome were induced. The SAVe I was unable to adequately oxygenate five animals with Pao2 (52.0±11.1 torr) compared to the Servo (106.0±25.6 torr; p=.002). The SAVe II was able to oxygenate and ventilate all six animals for 1 hr with no difference in Pao2 (141.8±169.3 torr) compared to the Servo (158.3±167.7 torr).CONCLUSIONS:We describe a novel in vivo model of acute lung injury/acute respiratory distress syndrome that can be used to initially screen limited-function ventilators considered for mass respiratory failure stockpiles and that is intended to be combined with additional studies to definitively assess appropriateness for mass respiratory failure. Specifically, during this study we demonstrate that the SAVe I ventilator is unable to provide sufficient gas exchange, whereas the SAVe II, with several more functions, was able to support the same level of hypoxemic respiratory failure secondary to acute lung injury/acute respiratory distress syndrome for 1 hr.

Real-time imaging of cellular and subcellular dynamics in vascularized organs requires image resolution and image registration to be simultaneously optimized without perturbing normal physiology. This problem is particularly pronounced in the lung, in which cells may transit at speeds >1 mm s(-1) and in which normal respiration results in large-scale tissue movements that prevent image registration. Here we report video-rate, two-photon imaging of a physiologically intact preparation of the mouse lung that is stabilizing and nondisruptive. Using our method, we obtained evidence for differential trapping of T cells and neutrophils in mouse pulmonary capillaries, and observed neutrophil mobilization and dynamic vascular leak in response to stretch and inflammatory models of lung injury in mice. The system permits physiological measurement of motility rates of >1 mm s(-1), observation of detailed cellular morphology and could be applied in the future to other organs and tissues while maintaining intact physiology.

Previous studies reported that regional CO(2) tension might affect regional ventilation (V) following acute pulmonary thromboembolism (APTE). We investigated the pathophysiology and magnitude of these changes. Eight anesthetized and ventilated piglets received autologous clots at time = 0 min until mean pulmonary artery pressure was 2.5 times baseline. The distribution of V and perfusion (Q) at four different times (-5, 30, 60, 120 min) was mapped by fluorescent microspheres. Regional V and Q were examined postmortem by sectioning the air-dried lung into 900-1,000 samples of approximately 2 cm(3) each. After the redistribution of regional Q by APTE, but in the scenario assuming that no V shift had yet occurred, CO(2) tension in different lung regions at 30 min post-APTE (P(X)CO(2)) was estimated from the V/Q data and divided into four distinct clusters: i.e., P(X)CO(2) < 10 Torr; 10 < P(X)CO(2) < 25 Torr; 25 < P(X)CO(2) < 50 Torr; P(X)CO(2) > 50 Torr. Our data showed that the clusters in higher V/Q regions (with a P(X)CO(2) < 25 Torr) received approximately 35% less V when measured within 30 min of APTE, whereas, in contrast, the lower V/Q regions showed no statistically significant increases in their V. However, after 30 min, there was minimal further redistribution of V. We conclude that there are significant compensatory V shifts out of regions of low CO(2) tension soon following APTE, and that these variations in regional CO(2) tension, which initiate CO(2)-dependent changes in airway resistance and lung parenchymal compliance, can lead to improved gas exchange.

We performed a multicenter study to test the hypothesis that tidal liquid ventilation (TLV) would improve cardiopulmonary, lung histomorphological, and inflammatory profiles compared with conventional mechanical gas ventilation (CMV). Sheep were studied using the same volume-controlled, pressure-limited ventilator systems, protocols, and treatment strategies in three independent laboratories. Following baseline measurements, oleic acid lung injury was induced and animals were randomized to 4 hours of CMV or TLV targeted to "best PaO2" and PaCO2 35 to 60 mm Hg. The following were significantly higher (p < 0.01) during TLV than CMV: PaO2, venous oxygen saturation, respiratory compliance, cardiac output, stroke volume, oxygen delivery, ventilatory efficiency index; alveolar area, lung % gas exchange space, and expansion index. The following were lower (p < 0.01) during TLV compared with CMV: inspiratory and expiratory pause pressures, mean airway pressure, minute ventilation, physiologic shunt, plasma lactate, lung interleukin-6, interleukin-8, myeloperoxidase, and composite total injury score. No significant laboratories by treatment group interactions were found. In summary, TLV resulted in improved cardiopulmonary physiology at lower ventilatory requirements with more favorable histological and inflammatory profiles than CMV. As such, TLV offers a feasible ventilatory alternative as a lung protective strategy in this model of acute lung injury.

Previous studies reported that the degree of hypoxemia following acute pulmonary thromboembolism (APTE) was highly variable and that its mechanism was mainly due to the creation of many high and low ventilation/perfusion (V/Q) units, as a result of the heterogeneous regional blood flow (Q) caused by embolic obstruction. We studied the effect of changing cardiac output (Q t ) on gas exchange after APTE in 5 embolized piglets (23 ± 3 Kg), using Dobutamine intermittently at approximately 20 μg/kg/min for 120 minutes. The distribution of ventilation (V) and perfusion (Q) at various times was mapped using fluorescent microspheres in 941 ± 60 lung regions. After APTE, increase in Q t by Dobutamine improved venous oxygen tension (PvO 2 ) but arterial PaO 2 did not change consistently. On the other hand, cluster analysis showed that the V/Q ratio of most lung regions was lowered due to increases in Q at the same time. We concluded that the effect of changing cardiac output on gas exchange following APTE was affected by the simultaneous and varying balance between the changing V/Q mismatch and the concomitantly changing PvO 2 , which might explain the unpredictability of PaO 2 in the clinical setting.

Studies in whole animals, isolated lungs and pulmonary tissue strips have shown that the pulmonary vascular resistance (PVR) to hypoxia is temporally biphasic in nature. We studied the regional temporal response to hypoxia in prone pigs. The animals were ventilated with an FIO2 of 0.21 (control), followed by an FIO2 of 0.12 for 180min. A biphasic response in Ppa to hypoxia was seen with the first peak between 10 and 20min and a second rise in Ppa starting after 30min, which was due to an increase in cardiac output. Regional blood flow (Q˙) and ventilation (V˙A) were measured using i.v. infusion of 15μm and inhalation of 1μm fluorescent microspheres, respectively. We grouped the lung pieces according to their temporal relative flow response to hypoxia. The five groups were each spatially distributed similarly, but not identically, among the animals. The corresponding relative ventilation to each group did not vary much. We conclude that in the prone pig, the PVR response to sustained hypoxia varies among regions of the lungs. Following an initial rise in PVR in most lung pieces, we found unexpectedly that some regions continue to increase PVR progressively and while other regions decrease PVR after the initial increase. The net effect is little change of overall PVR to hypoxia with time. Normoxic control animals had little change in their hemodynamics and the large majority of the lung pieces did not change their resistance over 3h. We speculate that the differential response of regions may be due to a differential role of nitric oxide, endothelin-1 release or K+ channels.

STUDY OBJECTIVES:Hypercapnic acidosis has antiinflammatory effects in animal models of acute lung injury (ALI) and improves ventilation-perfusion (V/Q) matching in normal lungs. The effect of hypercapnia on V/Q matching in ALI is conflicting. Hypercapnic acidosis produced by reduced tidal volumes (Vts) was associated with an increased shunt fraction (QS/QT) in patients with ALI compared with control subjects. Vt differences between groups make the assessment of hypercapnic acidosis on V/Q matching difficult. Adding CO2 to the inhaled gas allows the comparison of gas exchange under identical Vt conditions. We hypothesized the presence of hypercapnic acidosis from inspired carbon dioxide (ICD) would improve gas exchange in ALI and would be superior to that of low minute ventilation (LVe) produced by reduced respiratory rate, rather than Vt.DESIGN:University laboratory study of anesthetized New Zealand White rabbits.INTERVENTIONS:Assessment of V/Q relationships using the multiple inert gas elimination technique was performed in 10 saline solution-lavaged animals, which were ventilated with 6 mL/kg Vts and a positive end-expiratory pressure of 8 cm H2O. Each rabbit was studied while it was in eucapnia, followed by hypercapnia (Pa(CO2), 95 to 100 mm Hg) induced by LVe from decreased respiratory rate and by 10% ICD, in random order.MEASUREMENTS AND RESULTS:The Pa(O2) was greater in ICD and LVe compared to eucapnia, but no significant differences in alveolar-arterial oxygen pressure difference or Pa(O2)/fraction of inspired oxygen ratio occurred. LVe statistically reduced the mean V/Q distributions compared with ICD and eucapnia. Log SDs of ventilation and combined retention and excretion curves of the dispersion index were both increased during LVe, indicating the presence of unfavorable changes in ventilation distribution. Neither LVe nor ICD altered the QS/QT.CONCLUSIONS:LVe slightly impairs overall gas exchange and ventilation distribution, but does not increase QS/QT compared with eucapnia and ICD. While ICD does not significantly improve gas exchange, it may be superior to LVe in achieving the antiinflammatory effects of "therapeutic" hypercapnia, since it does not adversely alter gas exchange and has the potential to make the lung more uniformly acidotic.

We studied the roles of endothelins in determining ventilation (Va) and perfusion (Q) mismatch in a porcine model of acute pulmonary thromboembolism (APTE), using a nonspecific endothelin antagonist, tezosentan. Nine anesthetized piglets (approximately 23 kg) received autologous clots (approximately 20 g) via a central venous catheter at time = 0 min. The distribution of Va and Q at five different time points (-30, -5, 30, 60, 120 min) was mapped by fluorescent microspheres of 10 different colors. Five piglets (group 1) received tezosentan (courtesy of Actelion) starting at time = 40 min for 2 h, and four piglets (group 2) received only saline and served as control. Our results showed that, in all of the animals at 30 min following APTE but before tezosentan, the mean Va/Q was increased, as was Va/Q heterogeneity (log SD Va/Q), which represented a widening of its main peak. Afterwards, tezosentan attenuated the pulmonary hypertension in group 1 but also produced moderate systemic hypotension. However, it did not improve arterial PO2 or Va/Q mismatch. We concluded that endothelin antagonism had minimal impact on gas exchange following APTE and confirmed our earlier observation that the main mechanism for hypoxemia in APTE was due to the mechanical redistribution of pulmonary regional blood flow away from the embolized vessels, resulting in the creation of many divergent low and high Va/Q regions.