Determinants of alveolar nitrogen slope and closing volumes in healthy adolescents.

The alveolar nitrogen slope (PIII), closing volume (CV), and closing capacity (CC) were measured by the single-breath nitrogen washout method (SBN2) in a group of 187 healthy children and adolescents (92 boys, 95 girls), 10 to 16 yr old, from the general population of Lorraine, France. The test was performed using a computerized system, which also made the calculations. About one out of five healthy subjects in this population were unable to satisfactorily perform the test; the failure rate was the same for the two sexes (20% in boys, 21.5% in girls) and significantly higher in younger children (26.6 and 14.5% for children under and over the age of 13, respectively; p = 0.03). The distribution of results was skewed for PIII and practically normal for log PIII, CV, VC, and CV/VC or CC/TLC ratios. PIII was highly significantly, inversely related to anthropometric variables; the highest coefficient was that for the age-weight interaction term in boys (= r -0.57 for PIII, -0.62 for log PIII) and for weight in girls (r = -0.57 for both PIII and log PIII). Because the anthropometric variables were strongly interrelated (r between 0.45 and 0.79), multiple regressions did not materially improve the prediction of PIII. In simple regression, weight alone explained 36% of the variability of log PIII in boys and 32% in girls. The mean PIII was significantly higher in girls as compared to boys (1.14 +/- 0.38 versus 0.98 +/- 0.17% N2/L, p = 0.02); CV and CC in milliliters were related to body build as other lung volumes; the CV/VC in girls and CC/TLC ratio in both sexes were not related to anthropometric variables. In boys, CV/VC decreased significantly with height (p = 0.035 for CV/VC versus height3).

Halothane-sparing effect of benzodiazepines in ponies.

The halothane-sparing effect of 2 benzodiazepines, diazepam and temazepam, were investigated in ponies by measuring the minimum alveolar concentration (MAC) for halothane before and after drug administration. The MAC value for halothane decreased 29% and 16% when either 0.044 mg/kg of diazepam or 0.044 mg/kg of temazepam, respectively, was administered intravenously. Heart rate, respiratory rate, systolic and mean arterial blood pressure, and expired CO2 were also measured. No differences were present in these variables before and after drug administration nor were differences noted between the benzodiazepines.

[Antimetastatic effect of L-lysine-alpha oxidase]. / Antimetastaticheskii éffekt L-lizin-alfa-oksidazy.

The influence of a new antitumor enzyme L-lysine alpha-oxidase on Lewis lung carcinoma spreading was studied in mice in which primary tumor had been removed. The enzyme was found to significantly decrease the extent and number of lung metastases as compared to mice which hadn't received L-lysine alpha-oxidase. This was matched by recovery of alveolar macrophages functional activity, as assessed by adenosine deaminase and 5' nucleotidase levels in these cells. Moreover, antimetastatic and cytostatic effect was confirmed by the measuring of polyamine concentration in mice erythrocytes.

Surfactant-associated glycoproteins accumulate in alveolar cells and secretions during reparative stage of hyaline membrane disease.

Surfactant-associated (SA) glycoproteins are lung-specific proteins produced in the human lung by alveolar type II cells and Clara cells. The distribution of these proteins was studied immunohistochemically in lung tissue obtained postmortem from 12 stillborn fetuses and 49 infants with hyaline membrane disease (HMD). By 21 weeks of gestation, SA glycoproteins were detected in the fetal alveolar epithelium and within Clara cells. The staining increased in intensity and extent with advancing gestational age. Infants with HMD who survived less than 48 hours did not generally exhibit stainable material either within type II cells or secretions, but staining was often noted in Clara cells as well as focally beneath hyaline membranes. In infants surviving more than 48 hours, intense staining of hyaline membranes, alveolar secretions, proliferating alveolar type II cells, and Clara cells was evident. Immunoreactivity was intense in hypertrophic type II cells that formed a continuous alveolar epithelial lining in lungs with bronchopulmonary dysplasia. Included in the population of infants with HMD were 15 infants with pulmonary hypoplasia. The lungs of these infants showed minimal staining for SA glycoproteins regardless of postnatal survival time. The results provide an immunomorphologic basis for defining normal and abnormal lung maturation. They also indicate that enhanced SA glycoprotein production is a sustained response of regenerating and hypertrophic type II cells in premature infants.

Surfactant displaces particles toward the epithelium in airways and alveoli.

This study was designed to investigate the early stages of particle deposition on airway and alveolar surfaces. To do this we used morphometric studies of aerosol deposition, in situ measurements of surface tension, and in vitro assays of particle displacement and mathematical modelling. We observed that latex particles, equal or less than 6 microns in diameter deposited in hamster lungs were submerged in the subphase of the alveolar lining layer and became completely coated with an osmiophilic film. Similar results were obtained for particles deposited in the conductive airways which were also covered with a surface active film, having a surface tension of 32 +/- 2 dyn.cm-1. In vitro experiments showed that pulmonary surfactant promotes the displacement of particles from air to the aqueous phase and that the extent of particle immersion depends on the surface tension of the surface active film. The lower the surface tension the greater is the immersion of the particles into the aqueous subphase. Mathematical analysis of the forces acting on a particle deposited on an air-fluid interface show that for small particles (less than 100 microns) the surface tension force is several orders of magnitude greater than forces related to gravity. Thus, even at the relatively high surface tension obtained in the airways (32 +/- 2 dyn.cm-1) particles will still be displaced into the aqueous subphase. Particles in peripheral airways and alveoli likely are below the surfactant film and submerged in the subphase. This may promote clearance by macrophages. In addition, particle displacement into the subphase is likely to increase the contact between the epithelial cell and particle. Toxic or allergenic particles would be available to interact with epithelial cells and this may be important in the pathophysiology of airway disease.

Quantitative X-ray microanalysis of alveolar macrophages after long-term treatment with amiodarone.

Treatment with the iodine-containing antiarrhythmic drug, amiodarone, can cause pulmonary toxicity. Alveolar macrophages are particularly susceptible to formation of lipidrich lamellar bodies in amiodarone-treated animals. Amiodarone and several of its metabolites accumulate in the cell. Previously, we have reported that the technique of X-ray microanalysis is useful in monitoring the distribution of iodine in freeze-dried cryosections of alveolar macrophages from Fischer 344 rats 24 hr after a single dose of amiodarone. In the present study, we examine the effects of longer term amiodarone treatment of 1 or 9 weeks. Substantial changes in iodine distribution occur in the cells with increasing length of drug treatment. High concentrations of iodine are found early in the lamellar bodies. The iodine levels in the nuclei slowly increase with the length of treatment, and after 9 weeks of treatment, approach those found in the lamellar bodies. It is possible that this accumulation of iodine in the nuclei is due to the presence of polar metabolites. In addition, the potassium concentration in the cell decreases and the sodium increases with treatment duration. These changes in cations are most likely due to altered ion transport in the macrophages by the inhibition of membrane Na-K-ATPase by the drug and its principal metabolite, desethylamiodarone.

The comparison of a fibrogenic and two nonfibrogenic dusts by bronchoalveolar lavage.

Analysis of bronchoalveolar lavage fluid (BALF) appears to be a sensitive approach to characterizing an acute inflammatory response within the lung. More work, however, is needed to determine if analyses of BALF endpoints can predict chronic responses (i.e., fibrosis). The objective of the present study was to compare the dose and temporal pulmonary response of a known fibrogenic agent, silica, and two known nonfibrogenic agents, aluminum oxide and titanium dioxide. Animals were instilled with silica (0, 0.2, 1.0, or 5.0 mg/100 g body wt), titanium dioxide (1.0 or 5 mg/100 g body wt), aluminium oxide (1.0 or 5.0 mg/100 g body wt) or saline. Animals (n = 5/group) were terminated 1, 7, 14, 28, and 63 days following instillation, and the BALF was characterized by biochemical and cellular assays. Histopathological changes were determined at 60 days after exposure. The biochemical results demonstrated BALF levels of lactate dehydrogenase (LDH), beta-glucuronidase (BG), N-acetylglucosaminidase (NAG), and total protein (TP) increased in a dose-related fashion at the earlier time points for all test materials, with the magnitude of change being greatest for silica. The temporal response for these parameters was significantly different for the two classes of materials. With time, the response for the fibrogenic dust steadily increased, while the levels for the nonfibrogenic dusts decreased toward normal values during the 2-month study period. Of the cellular changes, total cell numbers, neutrophils, and lymphocyte numbers were the most sensitive markers of the pulmonary response. As shown with the biochemical parameters, the cellular response to silica increased with time while that of the nuisance dusts did not. It was also found that, similar to inhalation studies, high doses of a nuisance dust may result in toxicity/inflammation. This toxicity at high dose levels emphasizes the importance of choosing relevant doses when comparing potentially fibrogenic and nonfibrogenic dusts. In conclusion, the persistent and progressive changes seen in the biochemical (LDH, TP, BG, NAG) and cellular parameters (total cells, neutrophils and lymphocytes) following silica administration correlated with the fibrotic response which occurred after exposure to this material. The less dramatic and transient changes seen with aluminum oxide and titanium dioxide correlated with the inert nature of these nuisance dusts. The results of this study indicate evaluation of BALF may provide a means to predict the chronic pulmonary response to a material.

Comparison of non-collagenous type IV collagen subunits in human glomerular basement membrane, alveolar basement membrane, and placenta.

This study examines the similarities and differences in the noncollagenous domain (NC1) of type IV collagen from human glomerular basement membrane (hGBM), alveolar basement membrane (hABM), and placenta (hPBM). Following collagenase digestion, NC1 domain was isolated on Bio-Gel A-0.5m or by cation exchange chromatography on S-Sepharose. NC1 from each source was characterized by SDS PAGE, and two dimension NEPHGE/SDS PAGE. Immunoblotting and ELISA inhibition was performed using antibody probes specific for M28 , M28+, M26 and M24 monomer subunits of human NC1. It was observed that all NC1 subunits were present in hGBM and hABM derived material, however M28 and M28+ monomers were absent in hPBM NC1. These findings indicate that while alpha 1(IV) and alpha 2(IV) collagen chains are present in hGBM, hABM and hPBM, alpha 3(IV) and alpha 4(IV) collagen chains are only found in hGBM and hABM but are absent in hPBM. It can now be appreciated that heterogeneity of alpha (IV) chain composition exists in basement membranes from various organs.

Procoagulant (thromboplastin) activity in human bronchoalveolar lavage fluids is derived from alveolar macrophages.

Fibrin deposition in the alveolar space and the lung interstitium is a prominent feature of many types of inflammatory pulmonary diseases. Cells of the monocyte/macrophage line are the primary cells supplying procoagulant activity in inflammatory lesions. In the present study we found that both lung alveolar macrophages (LAM) and bronchoalveolar lavage fluids (BALF) from humans contained procoagulant activities. The procoagulant in BALF was associated with membrane vesicles which sedimented at 100,000 g for 1 h. By electron microscopy the BALF ultrasediment was seen to consist almost exclusively of membrane material and this was confirmed by monitoring the content of different marker enzymes for specific subcellular structures. Using macrophage membrane markers, at least part of the BALF-ultrasediment was shown to be derived from LAM. On the basis of phospholipase C sensitivity, antibody neutralization and the site of action of the procoagulant in the sequential activation of coagulation factors, both the LAM-associated and the BALF-associated procoagulant activity was identified as thromboplastin (tissue factor) or thromboplastin-factor VII complexes. This suggests that alveolar macrophages and the LAM-derived thromboplastin-containing microvesicles may contribute to intraalveolar and interstitial fibrin deposition in vivo and probably also have consequences for the development of pulmonary fibrosis.

Changes in alveolar-arterial oxygen partial pressure difference during orthotopic liver transplantation.

Changes in the alveolar-arterial oxygen partial pressure difference (PAO2-PaO2) were measured in 39 patients undergoing orthotopic liver transplantation without veno-arterial or veno-venous bypass. The operation can be divided into an initial dissection phase, an anhepatic phase when the hepatic artery, portal vein and vena cava are clamped, and a post-anhepatic phase after the vascular clamps are released. There was an initial increase in (PAO2-PaO2) during the dissection phase, followed by an immediate decrease when the liver was removed. This decrease continued throughout the anhepatic period, but a further increase in (PAO2-PaO2) occurred after release of all the vascular clamps and during abdominal closure.

Pulmonary bombesin and calcitonin in hamsters during exposure to hyperoxia and diethylnitrosamine.

Combined exposure of hamsters to 60% hyperoxia and the carcinogen diethylnitrosamine for 6 wk resulted in the development of lung tumors. This was associated with progressive loss of body weight as well as increases in the pulmonary-associated peptides, mammalian bombesin (MB) and immunoreactive calcitonin (iCT). After 3 wk of exposure, multiple bronchial epithelial hyperplastic foci were noted, along with increased lung levels of MB and iCT as well as increased serum levels of MB. At this time, immunocytochemistry revealed the presence of MB and iCT within hyperplastic pulmonary neuroendocrine (PNE) cells. In addition, the localization of MB to alveolar type II cells was noted, along with the presence of lamellar bodies and secretion granules in these cells on electron microscopy. After 6 wk of exposure, distinctive microscopic pulmonary tumorlets were seen. These tumorlets were associated with a marked increase in lung and serum MB, and to a lesser extent lung and serum iCT. At this time, MB and iCT were localized exclusively to these abnormal PNE cell sites. These results, which may have relevance in humans, suggest that endogenous peptides may be important components in the process of development of neuroendocrine cancer.

[Alveolocellular carcinoma diagnosed by bronchoalveolar lavage]. / Alveolecellekarcinom diagnosticeret ved bronkoalveolaer lavage.

Alveolocellular carcinoma is localized to the bronchioles and alveoli and the diagnostic procedures usually employed will only rarely contribute to the diagnosis. A case is presented where performance of fiberbronchoscopy and bronchoalveolar lavage with cytological examination of the cyto-centrifuged preparation led to the diagnosis of alveolocellular carcinoma in a patient with diffuse infiltrates.

Estimation of minimum alveolar concentration (MAC) for halothane, enflurane and isoflurane in spontaneously breathing guinea pigs.

MAC for halothane, enflurane and isoflurane was determined in guinea pigs (Cavia porcellus) exposed to constant anesthetic concentrations (2.5 hours each) in a flow-through glass chamber. The following values were obtained (N = 8 for each anesthetic): 1.01 +/- 0.03 vol% for halothane, 2.17 +/- 0.04 vol% for enflurane, and 1.15 +/- 0.05 vol% for isoflurane. In guinea pigs, MAC for halothane and enflurane are similar to those reported for other rodents, while MAC for isoflurane is lower. The data indicate that guinea pigs possibly are more susceptible to isoflurane's anesthetic actions than other rodents.

Distribution of elastin and collagen in canine lung alveolar parenchyma.

We have quantified the fibrous collagen (predominantly type I) and elastin in four locations of perceived mechanical importance: one quasi-planar feature, the alveolar septum or wall (W), and three linear features, the junction (J) of three septa, the free edges (E) of septa, and the line along which two septa join at a distinct angle or bend (B). The frequencies of these four features on light micrographs and the areas of transections through collagen and elastin seen on electron micrographs were combined to give the volumes of collagen and elastin within each feature. We find that E and B have similar compositions and contain most (4/5) of the parenchymal elastin in their relatively heavy cables. The E and B are interconnected and similar in location and composition, and they may constitute a functional entity in which elastin provides tension over a range of lung volumes, opposing septal tensions. In J and W, elastin is typically sparse and fine. Calculations, however, suggest it contributes the dominant portion of septal tension at lower lung volumes. Elastin may be essential to stabilizing septal configuration. Collagen, on the other hand, is distributed relatively evenly throughout E, B, J, and W, consistent with the role of protecting all components against rupture.

Changes in lung elastic fiber structure and concentration associated with hyperoxic exposure in the developing rat lung.

Prolonged hyperoxic exposure is associated with impaired alveolarization of the lung in both the rat and the human neonate. Elastin is currently thought to play a pivotal role in the alveolarization of the lung by providing the structural framework around which new alveoli will develop. Previous studies in both the rat and the human neonate have demonstrated a risk for proteolytic destruction of lung elastin associated with prolonged hyperoxic exposure. The present study was undertaken to determine whether continuous exposure to 100% oxygen during the period of alveolar development in the rat (Days 4 to 13) would alter lung elastin. Parenchymal lung elastic fiber length, volume density of parenchyma, mean linear intercept, and internal surface area were quantitated using morphometric techniques, and the values were compared in control, oxygen-exposed, and malnourished rat pups. Stereologic measurements indicated that total elastic fiber length was significantly greater in lungs of control pups than in lungs of either the oxygen-exposed or the malnourished pups. Examination of sections of lung tissue 20 to 30 microns thick indicated altered elastic fiber structure and numerous alveolar fenestrae only in the hyperoxic pups. The results of these studies demonstrated that hyperoxic exposure during alveolarization alters both total length and structure of lung elastic fibers and suggest that impaired lung development might be due in part to these observed changes.

The topography of particle deposition in the human lung.

The effect of varying particle size on the site of deposition of inhaled particles in the human lung was measured in 11 young healthy male subjects. The simultaneous inhalation of two chemically inert, radiolabelled particles, differing in size but in no other respect, controlled for all other variables including airways geometry, breathing pattern and posture. Under conditions of quiet respiration the larger particles (3.5 microns) were preferentially deposited in the upper rather than the lower zones of the lungs as compared with the smaller particles (1.1 microns). Furthermore, the penetrance of the larger particles beyond the mucociliary escalator was greater for 3.5 microns particles in all lung zones and particularly at the apex. These findings may be of significance in the pathogenesis of those diseases induced by the inhalation of particles, vapours or fumes.

Ozone-induced alterations of lamellar body lipid and protein during alveolar injury and repair.

Alveolar Type II cells in the rat respond to severe, acute ozone injury (3 ppm ozone for eight hours) by increasing their intracellular pool of surfactant; however, the newly stored surfactant is abnormal in composition. Lamellar bodies isolated between 24 and 96 hours after ozone exposure contained significantly more cholesterol in relation to phosphatidylcholine than did controls. By contrast, the cholesterol content of surfactant isolated from alveolar lavage remained unchanged throughout an 8-day post-ozone period. The total protein content of lamellar bodies in relation to phosphatidylcholine was significantly decreased at 24 and 48 hours post-ozone. Analysis of lamellar body proteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that the amount of a 14 kDa proteolipid was greatly reduced at the end of the eight-hour ozone exposure and remained low for at least 48 hours. This proteolipid appeared to be a specific lamellar body component since it was not detected in extracellular surfactant. The findings indicate that oxidative alveolar stress initiates characteristic alterations in both lipid and protein constituents of stored surfactant, without perturbation in the composition of extracellular surfactant.

Respiratory adaptation in the highest inhabitants and highest Sherpa mountaineers.

Arterial blood gases, acid-base and hematocrit of six highest inhabitants on Aucanquilcha (5950 m) in Chile were studied. These blood gases were compared with the alveolar gases of highest mountain climbers in Nepal, Sherpas and acclimatized lowlanders, and on average high altitude natives in the Chilean and Peruvian Andes and in the Nepal Himalayas. The mean arterial PCO2 (27.5 Torr) was lower than the standard sea level normal values, indicating a modest hypoxic hyperventilation. The mean arterial pH was 7.400, showing a complete renal compensation of respiratory alkalosis. The mean hematocrit (62%) and hemoglobin (20.7 g/dl) values were greater than the standard sea level values. These blood data showed that the highest inhabitants were acclimatized to hypoxia of their residential altitude. The respiratory gases showed less hyperventilation in the highest inhabitants and Sherpa mountaineers of high altitudes relative to the acclimatized lowlanders. Also, the average high altitude natives in the Andes and Himalayas showed less hyperventilation compared to the acclimatized lowlanders. We conclude that the attenuated hyperventilation is an appropriate respiratory adaptation to high altitude hypoxia in the native high altitude residents, allowing them to conserve metabolic energy expended for hyperventilation and to use the ventilatory reserve for a better performance at greater altitudes.

Immunohistochemical study of lung adenocarcinoma using monoclonal antibody for 60-kilodalton antigen in type II pneumocytes and nonciliated bronchiolar epithelial cells. Comparison with two antisurfactant apoprotein antibodies.

Monoclonal antibody KP16D3 was produced by immunizing mice with monkey bronchoalveolar lavage. KP16D3 revealed the immunohistochemical reactivity in the cytoplasm of some nonciliated bronchiolar epithelial cells and type II pneumocytes and thereby recognized specifically a protein with an apparent molecular weight of 60 kD with the use of Western blotting and immunoaffinity column chromatography followed by SDS-PAGE. Examination of 76 primary and 4 metastatic lung carcinomas in primary lung carcinoma KP16D3 showed immunohistochemical positivity only to mucin-nonproducing papillary adenocarcinoma (27/28) and bronchioloalveolar carcinoma (2/2), except for one case of large cell carcinoma. All other primary lung carcinomas such as squamous cell carcinoma, acinar adenocarcinoma, and small cell carcinoma had negative results. From these findings, KP16D3 seems to be an effective immunohistochemical marker of mucin-nonproducing papillary adenocarcinoma and bronchioloalveolar carcinoma of the lung and it appears to be useful to investigate both the histogenesis and functional expression of primary lung adenocarcinoma.

Estimation of pH in individual alveolar macrophage phagolysosomes.

A method for measurement of phagolysosomal pH in individual alveolar macrophages based on a cytofluorometric technique with fluorescein-labeled silica particles (FSP) as a probe was developed. The size of the FSP, 3.0 or 5.0 microns, did not affect the result of the pH measurements. The average pH values, range 5.1-5.5, of individual particles in macrophages from three rabbits agreed well with the pH values obtained in a macrophage population using a fluorescence spectrometer and the FSP. The variation of pH in phagolysosomes in alveolar macrophages from five rabbits was investigated. Measurements were performed 3, 6, and 24 h after addition of the FSP in cells containing one particle as well as in cells containing two particles. The variation in pH was small, with a coefficient of variation less than 10% in all rabbits at all times. There was a significant correlation between values obtained from two phagolysosomes in the same cell, indicating a cell factor responsible for 10-30% of the total variance. The fact that the variation of pH is small in normal, untreated rabbit alveolar macrophages should be of importance when estimating alveolar clearance of inorganic particles due to dissolution in the acid phagolysosomal milieu.