Mixture analysis of asymmetry: modelling directional asymmetry, antisymmetry and heterogeneity in fluctuating asymmetry

The occurrence of different forms of asymmetry complicates the analysis and interpretation of patterns in asymmetry. Furthermore, between-individual heterogeneity in developmental stability (DS) and thus fluctuating asymmetry (FA), is required to find relationships between DS and other factors. Separating directional asymmetry (DA) and antisymmetry (AS) from real FA and understanding between-individual heterogeneity in FA is therefore crucial in the analysis and interpretation of patterns in asymmetry. In this paper we introduce and explore mixture analysis to (i) identify FA, DA and AS from the distribution of the signed asymmetry, and (ii) to model and quantify between-individual heterogeneity in developmental stability and FA. In addition, we expand mixtures to the estimation of the proportion of variation in the unsigned FA that can be attributed to between-individual heterogeneity in the presumed underlying developmental stability (the so-called hypothetical repeatability). Finally, we construct weighted normal probability plots to investigate the assumption of underlying normality of the different components. We specifically show that (i) model selection based on the likelihood ratio test has the potential to yield models that incorporate nearly all heterogeneity in FA; (ii) mixtures appear to be a powerful and sensitive statistical technique to identify the different forms of asymmetry; (iii) restricted measurement accuracy and the occurrence of many zero observations results in an overestimation of the hypothetical repeatability on the basis of the model parameters; and (iv) as judged from the high correlation coefficients of the normal probability plots, the underlying normality assumption appears to hold for the empirical data we analysed. In conclusion, mixtures provide a useful statistical tool to study patterns in asymmetry.     

Low-volume ventilation causes peripheral airway injury and increased airway resistance in normal rabbits.

Lung mechanics and morphometry of 10 normal open-chest rabbits (group A), mechanically ventilated (MV) with physiological tidal volumes (8-12 ml/kg), at zero end-expiratory pressure (ZEEP), for 3-4 h, were compared with those of five rabbits (group B) after 3-4 h of MV with a positive end-expiratory pressure (PEEP) of 2.3 cmH(2)O. Relative to initial MV on PEEP, MV on ZEEP caused a progressive increase in quasi-static elastance (+36%) and airway (Rint; +71%) and viscoelastic resistance (+29%), with no change in the viscoelastic time constant. After restoration of PEEP, quasi-static elastance and viscoelastic resistance returned to control levels, whereas Rint remained elevated (+22%). On PEEP, MV had no effect on lung mechanics. Gas exchange on PEEP was equally preserved in groups A and B, and the lung wet-to-dry ratios were normal. Both groups had normal alveolar morphology, whereas only group A had injured respiratory and membranous bronchioles. In conclusion, prolonged MV on ZEEP induces histological evidence of peripheral airway injury with a concurrent increase in Rint, which persists after restoration of normal end-expiratory volumes. This is probably due to cyclic opening and closing of peripheral airways on ZEEP.     

Numerical modelling of nanoparticle deposition in the nasal cavity and the tracheobronchial airway

Recent advances in nanotechnology have seen the manufacture of engineered nanoparticles for many commercial and medical applications such as targeted drug delivery and gene therapy. Transport of nanoparticles is mainly attributed to the Brownian force which increases as the nanoparticle decreases to 1 nm. This paper first verifies a Lagrangian Brownian model found in the commercial computational fluid dynamics software Fluent before applying the model to the nasal cavity and the tracheobronchial (TB) airway tree with a focus on drug delivery. The average radial dispersion of the nanoparticles was 9x greater for the user-defined function model over the Fluent in-built model. Deposition in the nasal cavity was high for very small nanoparticles. The particle diameter range in which the deposition drops from 80 to 18% is between 1 and 10 nm. From 10 to 150 nm, however, there is only a small change in the deposition curve from 18 to 15%. A similar deposition curve profile was found for the TB airway.     

Numerical modelling of nanoparticle deposition in the nasal cavity and the tracheobronchial airway

Recent advances in nanotechnology have seen the manufacture of engineered nanoparticles for many commercial and medical applications such as targeted drug delivery and gene therapy. Transport of nanoparticles is mainly attributed to the Brownian force which increases as the nanoparticle decreases to 1 nm. This paper first verifies a Lagrangian Brownian model found in the commercial computational fluid dynamics software Fluent before applying the model to the nasal cavity and the tracheobronchial (TB) airway tree with a focus on drug delivery. The average radial dispersion of the nanoparticles was 9x greater for the user-defined function model over the Fluent in-built model. Deposition in the nasal cavity was high for very small nanoparticles. The particle diameter range in which the deposition drops from 80 to 18% is between 1 and 10 nm. From 10 to 150 nm, however, there is only a small change in the deposition curve from 18 to 15%. A similar deposition curve profile was found for the TB airway.     

Effects of neutrophils on collateral ventilation and peripheral lung reactivity in dogs

We studied the effects of neutrophil activation on collateral ventilation and peripheral lung reactivity in anesthetized dogs. A fiberoptic bronchoscope was wedged into a segmental airway under direct vision. Ventilation beyond the obstruction thus occurred only through collateral channels. Through one lumen of a double-lumen catheter threaded through the suction port of a bronchoscope, 5% CO2 in air was infused at a known constant rate (V coll). Through the other lumen, pressure at the tip of the bronchoscope was monitored (Pb). For measurements of resistance to flow through the collateral system (Rcs), the ventilation was stopped at functional residual capacity (FRC). Histamine was delivered through the bronchoscope to the obstructed lung segment in the form of an aerosol mist generated by an ultrasonic nebulizer. Measurements of Rcs were used as a parameter of the peripheral lung reactivity to histamine challenge. Within one hour after intravenous infusion of phorbol myristate acetate (PMA), a neutrophil activator, the reactivity to histamine significantly increased. After this, Rcs increased even without histamine challenge. This increase may have been due to an edematous injury of lung caused by PMA. The nature of the injury was confirmed by wet to dry weight ratios. In the other group, the white cell count dropped below 1000 per cu. mm. after intravenous infusion of nitrogen mustard. The same experimental protocols were followed. The Rcs did not increase even with histamine challenge. Our results suggested that substances such as oxygen radicals and arachidonic acid metabolites, which can be released by activated neutrophils, may not not only increase peripheral lung reactivity, but may also induce pulmonary edema.     

Modeling the bifurcating flow in a human lung airway

The inspiratory flow characteristics in a three-generation lung airway have been numerically investigated using a control volume method to solve the fully three-dimensional laminar Navier-Stokes equations. The three-generation airway is extracted from the fifth to seventh branches of the model of Weibel (Morphometry of the Human Lung, Academic Press, New York, Springer, Berlin, 1963) with in-plane and 90 degrees off-plane configurations. Computations are carried out in the Reynolds number range of 200-1600, corresponding to mouth-air breathing rates ranging from 0.27 to 2.16l/s, or an averaged height of a man breathing from quiet to vigorous state. Particular attention is paid to establishing relations between the Reynolds number and the overall flow characteristics, including flow patterns and pressure drop. The ratio of airflow rate through the medial branch to that of the lateral branch for an in-plane airway increases as Re(0.227). However, the total pressure drop coefficient varies as Re(-0.497) for an in-plane airway and as Re(-0.464) for an off-plane airway. These pressure drop results are in good agreement with the experimentally measured behavior of Re(-0.5) and are more accurate than the numerically determined behavior of Re(-0.61) assuming the airways to be approximated by two-dimensional channels.     

Relationship of central and peripheral airway resistance to lung volume in dogs

We could not reconcile reported relationships between lung resistance measurements and lung volume with bronchographic and anatomic studies showing that airway diameters change monotonically with lung volume and that small airways change diameter proportionately at least as much as large ones. Accordingly we measured central and peripheral airways resistances with a new technique. The relevant pressures were measured with a tracheal cannula, a wedged retrograde catheter, and two parenchymal needles in seven open-chested dogs while pleural pressure was oscillated at 1 Hz. In contrast to previous studies, the volume dependency of peripheral resistance was at least as great as that of central resistance with vagi intact, the volume dependencies of central and peripheral resistances were not abolished by vagotomy, and neither resistance increased systematically at high volumes. Volume dependency of central resistance resembled predictions for isotropic expansion of airways with vagi cut but increased with bronchomotor tone. These results fit generally with bronchographic data. Previous studies may have been affected by volume dependency due to "tissue resistance" and catheter phase lags.     

Model simulations of gas mixing and ventilation distribution in the human lung

A new lung model that incorporates intra-acinar diffusion- and convection-dependent inhomogeneities (DCDI) and interregional and intraregional convection-dependent inhomogeneities (CDI) is described. The model is divided into two regions, each containing two subunits. Each of the four subunits in the model consists of a multi-branch-point structure, based on the anatomic data from Haefeli-Bleuer and Weibel (Anat. Record 220: 401-414, 1988). The subunit turnover (TO), i.e., the ratio of subunit tidal to resting volume, and the flow sequences (FS) between the subunits are used as model parameters. The model simulates the normalized alveolar slope (Sn), Fowler and Bohr dead space (VDF and VDB), and alveolar mixing efficiency (AME) as a function of breath number (n) during a multiple-breath N2 washout (MBNW). For the first breath of the MBNW, these indexes are poorly sensitive to the TO distribution or FS between the subunits. However, as the washout proceeds, the n dependence of both Sn and VDB becomes markedly distinct for simulations with different TO and FS. VDF increases only slightly with n during the MBNW for a large range of TO and FS combinations, and AME is independent of FS. Comparison of published experimental observations with model simulations gave a consistent picture of ventilation maldistribution in the human lung. MBNW simulations in conditions of weightlessness, which will be performed shortly in Spacelab, suggest that it will be possible to evaluate quantitatively the intraregional elastic inhomogeneities in the human lung.     

Modeling the bifurcating flow in a CT-scanned human lung airway

The inspiratory flow characteristics in a CT-scanned human lung model were numerically investigated using low Reynolds number (LRN) kappa-omega turbulent model. The five-generation airway is extracted from the trachea to segmental bronchi of a 60-year-old Chinese male patient. Computations were carried out in the Reynolds number range of 900-2100, corresponding to mouth-air breathing rates of 190-440ml/s. Flow patterns on the Re=2100 and flow rate distribution were presented. In this model, the flow pattern is very complex. To count the effect of laryngeal jet on trachea inlet, the trachea was extended and modified to simulate the larynx, consequently the inlet velocity profile is biased towards the rear wall. In the inferior lobar bronchi, there are two stems in which the axial velocity is stronger but secondary velocity is weaker. Secondary flow in the lateral bronchi is stronger than the medial ones. With increasing Re, the air flow increases in the middle, inferior lobes and left main bronchus, i.e., flow biases to left and downward.     

Modeling the bifurcating flow in an asymmetric human lung airway

In a former paper, the inspiratory flow characteristics in a three-generation symmetric bifurcation airway have been numerically investigated using a control volume method to solve the fully three-dimensional laminar Navier-Stokes equations. The present paper extends the work to deal with asymmetric airway extracted from the 5th-11th branches of the model of Weibel (Morphometry of the Human Lung. New York Academic Press, Verlag, 1963) in order to more appropriately model human air passage. Computations are carried out in the Reynolds number range 200-1600, corresponding to mouth-air breathing rates ranging from 0.27 to 2.16l/s, representative for an averaged height man breathing from quiet to vigorous state. Particular attentions are paid to establishing relations between the Reynolds number and the overall flow characteristics, including flow patterns and pressure drop. The study shows that the ratios of airflow rate through the medial branches to that of their mother branches are the same, and this is also true for the ratios of airflow rate through the lateral branches. This partially explains why regular human breathing is not affected by airways of different sizes.     

Particle deposition in a CT-scanned human lung airway

The particle deposition in a computerized tomography (CT)-scanned human lung was numerically investigated. The five-generation airway is extracted from the trachea to segmental bronchi of a 60-year-old Chinese male patient. Computations were carried out in the flow rate range of 210–630 ml/s (Reynolds number range of 1000–3000) and particle size of 2–10 μm (Stokes number range of 0.0007–0.049). To count the effect of laryngeal jet on trachea inlet, the trachea was extended and modified to simulate the larynx, consequently the inlet velocity profile is biased towards the rear wall. The laryngeal jet-induced turbulence was simulated using low Reynolds number (LRN) κ–ω turbulent model. Particle deposition patterns, deposition efficiency and deposition factor were studied in detail. The turbulent flow has significant effect on the particle deposition, and the present deposition factor is compared well with the available data.     

Effects of emboli, positive-pressure ventilation, and airway water on lung water measurements

We tested the effects of microemboli, continuous positive-pressure ventilation (CPPV), and aspirated airway water on measurements of extravascular lung water by use of the technique of thermal indicator dilution (ETVL). A control group of dogs and a group of dogs in which dextran was infused created all levels of pulmonary edema. In an emboli group 0.125 g/kg of starch microemboli (63-74 micron diam) were infused. In groups with emboli and CPPV, starch emboli were infused and CPPV was then applied at 15 cmH2O. In an airway saline group measured amounts of saline were poured into the airway. In all groups postmortem pulmonary extravascular tissue weight (PETW) was determined and compared with the last ETVL. Emboli created an increased scatter when the last ETVL is compared with PETW because 1) blood trapped distal to emboli was included in the ETVL measurement, and/or 2) diffusion limitations for the thermal indicator were exceeded. Emboli and CPPV decreased ETVL/PETW. Airway saline (80 +/- 5%) was measured by ETVL. In conclusion, the ETVL technique is reliable in well-perfused lungs but loses accuracy in measuring lung water after emboli of any size or with large amounts of airway fluid.     

Leukotriene generation and small airway smooth muscle responsiveness in human lung

The hypothesis was tested that endogenous leukotriene (LT) production in the lung causes desensitisation of airway smooth muscle to LT. The synthesis of LTB4, C4, D4 and E4 by human lung tissue, obtained at thoracotomies, after stimulation with Ca-ionophore was assessed by HPLC. Functional studies of small airway smooth muscle from the same tissue specimens were carried out using LTC4 and methacholine as the contracting agents. Generation of LTB4, C4, D4 and E4 was 453 +/- 82, 84 +/- 15, 71 +/- 27 and 40 +/- 16 pmol/g fresh tissue respectively (mean +/- S.E.M., n = 10). All airway smooth muscle preparations responded to LTC4 in a concentration dependent way with a -log EC20 of 8.56 +/- 0.13, a -log EC50 of 7.95 +/- 0.08 and a Tmax of 82 +/- 11 mg force/mg tissue weight, corresponding to 79 +/- 4% of the maximal response to methacholine (mean +/- S.E.M.; 27 preparations from 10 patients). No correlations were found between any of the functional parameters (-logEC20, -logEC50, Tmax to LTC4 and methacholine) and the amounts of LT's generated by the lung tissue. Furthermore airway smooth muscle contractility was not significantly reduced after repeated exposure of bronchiolar strips to LTC4 in vitro. These findings suggest that the responsiveness of human peripheral airway smooth muscle to LT is not related to the capacity of the lung tissue to synthetize LT.     

Characterisation of urokinase plasminogen activator receptor variants in human airway and peripheral cells

Background  

Expression of the urokinase plasminogen activator receptor (UPAR) has been shown to have clinical relevance in various cancers. We have recently identified UPAR as an asthma susceptibility gene and there is evidence to suggest that uPAR may be upregulated in lung diseases such as COPD and asthma. uPAR is a key receptor involved in the formation of the serine protease plasmin by interacting with uPA and has been implicated in many physiological processes including proliferation and migration. The current aim was to determine key regulatory regions and splice variants of UPAR and quantify its expression in primary human tissues and cells (including lung, bronchial epithelium (HBEC), airway smooth muscle (HASM) and peripheral cells).     

Primary human airway epithelial cell-dependent inhibition of human lung mast cell degranulation

Introduction

Chronic mast cell activation is a characteristic feature of asthma. BEAS-2B human airway epithelial cells (AEC) profoundly inhibit both constitutive and IgE-dependent human lung mast cell (HLMC) histamine release. The aim of this study was to examine the regulation of HLMC degranulation by primary AEC from healthy and asthmatic subjects, and investigate further the inhibitory mechanism.

Methods

HLMC were co-cultured with both BEAS-2B and primary AEC grown as monolayers or air-liquid interface (ALI) cultures.

Results

Both constitutive and IgE-dependent HLMC histamine release were attenuated by BEAS-2B, primary AEC monolayers and ALI cultures. This occurred in the absence of HLMC-AEC contact indicating the presence of a soluble factor. Unlike healthy ALI AEC, asthmatic ALI-AEC did not significantly reduce constitutive histamine release. AEC inhibitory activity was transferable in primary AEC monolayer supernatant, but less active than with Transwell co-culture, suggesting that the inhibitory factor was labile. The AEC inhibitory effects were attenuated by both AEC wounding and pertussis toxin, indicating the involvement of a G₀/G_i receptor coupled mechanism. Solid phase extraction of lipids (<10 kDa) removed the AEC inhibitory activity. The lipid derivatives resolvin D1 and D2 and lipoxin A₄ attenuated HLMC histamine release in a dose-dependent fashion but were not detectable in co-culture supernatants.

Conclusions

Primary AEC suppress HLMC constitutive and IgE-dependent histamine secretion through the release of a soluble, labile lipid mediator(s) that signals through the G₀/G_i receptor coupled mechanism. Manipulation of this interaction may have a significant therapeutic role in asthma.     

Numerical investigation of the three-dimensional flow in a human lung model

The flow field at inspiration and expiration in the upper human airways consisting of the trachea down to the sixth generation of the bronchial tree is numerically simulated. The three-dimensional steady flow at a hydraulic diameter-based Reynolds number Re(D)=1250 is computed via a lattice-Boltzmann method (LBM). The simulation is validated by the experimental data based on particle-image velocimetry (PIV) measurements. The good agreement between numerical and experimental results is evidenced by comparing velocity contours and distributions in a defined reference plane. The results show the LBM to be an accurate tool to numerically predict flow structures in the human lung. Using an automatic Cartesian grid generator, the overall process time from meshing to a steady-state solution is <12h. Moreover, the numerical simulation allows a closer analysis of the secondary flow structures than in the experimental investigation. The three-dimensional streamline patterns reveal some insight on the air exchange mechanism at inspiration and expiration. At inspiration, the slower near-wall tracheal flow enters through the right principal bronchus into the right upper lobar bronchus. The bulk mass flux in the trachea is nearly evenly distributed over the left upper, center and lower lobar bronchi and the right center and lower bronchi. At expiration, the air from the right upper lobar bronchus enters the right center of the trachea and displaces the airflow from the lower and center right bronchi such that the tracheal positions of the streamlines at inspiration and expiration are switched. The flow in the left bronchi does not show this kind of switching. The findings emphasize the impact of the asymmetry of the lung geometry on the respiratory air exchange mechanism.     

Endocrine-like cells of the pulmonary epithelium of the human adult lung

Summary The morphological and histochemical characteristics of endocrinelike cells of the pulmonary epithelium of the right lower lobe of 12 human adult lungs were studied.Few cells were reactive to the argyrophil silver method of Grimelius and of Sevier and Munger and cells with a similar morphology and distribution emitted a green or yellow fluorescence after treatment of the lung epithelium with the amine precursors L-DOPA or L-HTP, respectively. A greater number of cells seems to be demonstrated by electron microscopy. The cells were characterized by small, round secretory granules showing a central dense core and a very thin clear halo between the core and the surrounding membrane.The cells are thought to be related to the endocrine-like cells of the pulmonary epithelium of the human foetal lung and to cells of carcinoids of larger bronchi.