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Human lung organ-specific antigens on normal lung, lung tumors, and a lung tumor cell line.

An antiserum raised in rabbits against a lung tumor cell line (2563) was selected from a library of antisera against normal and malignant human lung, lung tumor cell lines, and fetal tissues and was found by complement fixation, immunofluorescence, and saturation binding assays to contain antibodies for antigens characteristic of those found in normal lung. Studies with the adsorbed antiserum (A49) revealed: 1) An antigen was shared by normal lung and normal kidney (NLK-1) 2) lung tissue-specific antigen(s) were present on normal lung tissue (NL-1); 3) NL-1 was found on both external and internal cell membranes; and 4) NL-1, in addition to being present on normal lung and the adenocarcinoma-derived cell line 2563, was present on 1 of 2 metastatic lung adenocarcinomas but on none of 4 metastatic lung tumors of other histologic types.

Adenocarcinoma

Transbronchial biopsy for the diagnosis of lung transplant rejection. Comparison with needle and open-lung biopsy techniques in canine lung allografts.

The accurate diagnosis of lung transplant rejection requires histologic examination of the grafted lung. Because transbronchial lung biopsy has been advocated as an effective diagnostic procedure for a variety of lung diseases, it was elected to assess this technique in rejecting transplanted lungs. Twenty-two dogs received allografts and underwent simultaneous lung biopsy by 3 techniques when signs of rejection occurred. Open lung biopsy was diagnostic of rejection in all instances. Transthoracic needle biopsy correlated with the open biopsy in 59 per cent of the cases. No specimen obtained by transbronchial lung biopsy provided sufficient material to permit fulfillment of the strict histologic criteria needed to diagnose allograft rejection. Although transbronchial lung biopsy is successful in many pulmonary infiltrative processes, it appears to be inadequate for the diagnosis of lung allograft rejection.

Animals

Regional lung expansion at total lung capacity in intact vs. excised canine lungs.

A computer-based biplane videoroentgenographic recording technique that determines the spatial coordinates of radiopaque lung parenchymal markers was used to compare regional lung expansion at total lung capacity (TLC) in the intact dog (prone and supine) and after removal from the chest. The reproducibility of the technique was examined by repeated determinations of intermarker distances at various static lung volumes during stepwise inflation and deflation of the lungs. Most of the variability in repeated determinations of intermarker distances at any lung volume was due to cardiogenic motion. When marker positions were determined repeatedly at the same phase of the cardiac cycle, the maximum coefficient of variation was less than 3% for a marker pair separated by 16.5 mm. At TLC, distances between all intralobar marker pairs in the intact thorax (prone and supine) and excised were highly linearly related (r = 0.96-0.99), whereas distances between interlobar marker pairs did not correlate as well (r = 0.77-0.86). We conclude that at TLC 1) the intact thorax does not distort the shape of the individual lobes from the state of isotropic expansion, and 2) in different body positions, overall lung shape may be different due to displacementof lobes relative to each other, but individual lobes remain uniformly expanded.

Animals

The weight of human lungs as a diagnostic criterium (distinction of normal lungs from shock lungs by histologic, morphometric and biochemical investigations).

Lung weight, extravascular water content, hemoglobin content as an index of blood congestion, and DNA content as an index of cellular content, were determined for 57 patients dying from multiple, extrathoracic trauma. This report, which confirms previous publications, concerns itself in particular with the problems of determination of the "normal" lung weight. Studies of the influence of survival time on the lung showed that lung weight may significantly increase to twice its value after very short times up to one hour after trauma as a result of blood congestion, and following longer survival may be elevated several times as a result of developing interstitial edema. Early appearing interstitial edema is morphometrically detectable already 10 minutes after the beginning of shock. Normal weights were obtained from 10 cases after immediate death following severe cranial trauma or decapitation by being run over. Our results show that normal lungs have an average weight of 234 g (s +/- 39). Literature values for "normal" lung weight are approximately twice as high.

DNA

The cat lung strip as an in vitro preparation of peripheral airways: a comparison of beta-adrenoceptor agonists, autacoids and anaphylactic challenge on the lung strip and trachea.

1 A new in vitro preparation, the isolated lung strip of the cat, is described for investigating the direct effect of drugs on the smooth muscle of the peripheral airways of the lung. The preparation comprises a thin strip of lung parenchyma which can be mounted in a conventional organ bath for isometric tension recording. Its pharmacological responses have been characterized and compared with the isolated tracheal preparation of the cat. 2 The lung strip exhibited an intrinsic tone which was relaxed by catecholamines, aminophylline and flufenamate. It was contracted strongly by histamine, prostaglandin F2alpha, acetylcholine, compound 48/80, potassium depolarizing solution and alternating current field stimulation. In contrast, the cat trachea was unresponsive to histamine and prostaglandin F2alpha and did not exhibit an intrinsic tone. 3 (-)-Isoprenaline and (-)-adrenaline were much more potent in relaxing the lung strip than the trachea. The potency order of relaxation responses to isoprenaline, adrenaline and (+/-)-noradrenaline in the lung strip was isoprenaline greater than adrenaline greater than noradrenaline but in the trachea was isoprenaline greater than noradrenaline greater than or equal to adrenaline. 4 beta2-Adrenoceptor selective agonists salbutamol and terbutaline were more potent in the lung strip than the trachea, suggesting beta2-adrenoceptors predominated in the lung strip. Propranolol was equipotent in inhibiting isoprenaline relexations of the lung strip and trachea, whereas practolol was much less effective in inhibiting lung strip than trachea, further supporting a predominance of beta2-adrenoceptors in lung strip and beta1-adrenoceptors in trachea. 5 Strong Schultz-Dale type contractions were elicited in both lung strips and trachea by Ascaris lumbricoides antigen in actively sensitized cats. The initial phase of the contractile response of the lung strip following challenge was shown to be due to histamine release and was absent in the trachea. The delayed phase of the contraction which took several minutes to develop in both the mepyramine-treated lung strip and trachea was not due to prostaglandins E1, F2alpha or bradykinin, the probable mediator being slow reacting substance of anaphylaxis (SRS-A). 6 It is concluded that the isolated lung strip of the cat is useful as an in vitro model for investigating the effect of drugs on the smooth muscle of the peripheral airways of the lungs.

Adrenergic beta-Agonists

Quantitation of phosphatidylcholine secretion in lung slices and primary cultures of rat lung cells.

Rat lung slices and isolated rat lung cells were used to study the secretion of phosphatidylcholine by the lung in vitro. The rate of incorporation of [(3)H]choline by lung slices was 20-fold greater than by liver slices and 4-fold greater in lung cells compared to confluent skin fibroblasts. Labeling lung slices or cells with [(3)H]choline for up to 8 hr failed to reveal a significant amount of labeled phosphatidylcholine in the medium of either system compared to the medium from liver slice or fibroblast controls. Labeling of isolated lung cells for up to 24 hr, with or without 10% fetal calf serum, also showed no significant difference in the amount of labeled phosphatidylcholine in the medium compared to control fibroblast cultures. Washing labeled lung slices or cells with a nonlysing concentration of Triton X-100 (0.05%) did not selectively release labeled phosphatidylcholine, indicating that any secreted phosphatidylcholine did not adhere to the surface of the lung slices or cells. Experiments were performed to determine whether the small amount of phosphatidylcholine in the medium and detergent-released phosphatidylcholine was similar to the tissue and cell phosphatidylcholine. The saturated fatty acid composition of the phosphatidylcholine released by Triton X-100 and in the medium (from lung slices) was identical to that of the tissue phosphatidylcholine. In addition, the relative labeling rates of the phospholipids released by Triton X-100 and in the medium (labeled with [(14)C]glycerol) were identical to those of the tissue and cell phospholipids. Based on these results, we conclude that phosphatidylcholine is not secreted by lung slices and lung cells in large amounts compared to controls. The implication of these data is that pulmonary surfactant material may actually not be secreted by the lung in vitro, and perhaps in vivo, in the manner that is currently generally accepted.

Animals

Lung interdependence and lung-chest wall interaction of sublobar and lobar units in pigs.

In anesthetized and paralyzed pigs in the left decubitus position we obstructed, at functional residual capacity (FRC), either the right middle and lower lobes, or a small posterior basal lung unit, and then passively inflated the unobstructed remaining lung. Measurements were made of alveolar pressure in the obstructed and unobstructed lung regions as well as of esophageal pressure. The tendency of the obstructed lung region to inflate as the remainder of the lung was inflated was assessed by an index (A), which was the end-inspiratory pressure difference between unobstructed and obstructed alveolar pressures, normalized by the change in elastic recoil of unobstructed lung. With the chest wall intact, inflation of unobstructed lung resulted in a tendency to inflate the obstructed regions. This tendency was abolished with the chest wall removed. In a second group of pigs with the basilar lung unit obstructed, the height of the unit was changed by turning a pig from right to left decubitus positions. In each position A was assessed with both spontaneous and positive pressure ventilation. The magnitude of A was found to vary directly with the magnitude of caudal diaphragmatic motion and was greatest with the lung unit dependent and with spontaneous ventilation. These results suggest that lung-chest wall interaction was a more important factor tending to preserve homogenous inflation than lung tissue interdependence.

Animals

Pulmonary insulin responsivitiy: in vivo effects of insulin on the diabetic rat lung and specific insulin binding to lung receptors in normal rats.

Adult rats were rendered diabetic by a single iv injection of streptozotocin (70 or 75 mg/kg). In these rats, serum insulin fell to minimal levels during the 48 h following drug treatment, and this was roughly paralleled by a progressive decrease in the ability of the lung to oxidize glucose. The addition of insulin to diabetic rat lung slices in vitro had no restorative effect on the depressed glucose oxidative rate during a 2 h incubation period; however, two daily treatments of the rats with 1 unit of protamine, zinc insulin completely restored lung glucose oxidation rate to normal, without significantly reducing the hyperglycemic state of the rats. An examination of the temporal changes in glucose utilization by the rat lung after acute insulin treatment revealed that the diabetic lung responded directly to serum levels of insulin, whereas the normal lung appeared to be unaffected by serum insulin levels as hihg as 87 ng/ml. The reduced rate of glucose oxidation in the diabetic lung was apparent after perfusion of the lung with glucose-free medium, and was characterized by a significant reduction in Vmax without an alteration in Km. This was attended by a depressed ability of the lung to incorporate [3H]leucine into protein and an increased ability to produce lactate, but hexose monophosphate shunt activity was normal. Specific receptors for insulin have been identified and partially characterized in crude membrane preparations of normal rat lung. The interaction of insulin with these receptors was rapid, reversible, saturable, and was dependent upon time and temperature. The binding of labeled insulin was inhibited by low concentrations of unlabeled insulin and by high concentrations of proinsulin, whereas it was unaffected by the presence of glucagon, gastrin, prolactin, ACTH, or growth hormone in microgram amounts. These observations suggest that insulin regulates the transport and utilization of glucose in the rat lung, and that this tissue contains specific receptors for insulin.

Animals

Acute changes of lung volumes and lung mechanics in asthma and in normal subjects.

Lung volumes, pulmonary mechanics, and specific airway conductance (sGaw) were studied before and 15 minutes after 200 microgram of aerosol salbutamol in nine asthmatics undergoing mild spontaneous exacerbations of their disease and in five normal subjects. In addition, three of the normal subjects were studied after voluntarily breathing at high lung volumes for one minute. The normal subjects and four of the asthmatics showed no overall changes in lung volumes or pressure-volume (PV) curves after salbutamol, even though airway dilatation was produced in the asthmatic subjects. In another five asthmatics, salbutamol induced a significant fall in the total lung capacity (TLC) in three, and in the residual volume and functional residual capacity in all five. There was a significant displacement of the PV curve downwards and to the right in all five, with increased lung compliance (Cl stat) in two. The fall in TLC could be accounted for by the increase in lung elasticity. There is reason to believe that this change in lung mechanics could be due to the reversal of asthma-induced stress relaxation. Sustained breathing at high lung volume is known to cause stress relaxation of the lung. However, this breathing pattern did not alter the PV curve or TLC in two of three normal subjects, which is consistent with stress relaxation being rapidly reversible. The other normal subject had an acute shift of the PV curve upwards and to the left after breathing at high lung volume. It has been concluded that stress relaxation is usually rapidly reversed but that in some individuals it may only be slowly reversed. This more persistent form of stress relaxation may contribute to the acute changes of TLC found in some asthmatics.

Adult

Connective tissue proteins of the baboon lung: concentration, content and synthesis of collagen in the normal lung.

Lung connective tissue proteins have been assayed in 12 normal juvenile baboons (Papio cynocephalus). The collagen concentration (26.8 +/- 5.1 microgram hydroxyproline per mg dry weight) is comparable to values reported for adult human lung, while the rate of collagen synthesis (0.242 +/- 0.082 nmoles hydroxyproline per mg DNA per hour) is approximately half that of human lung. Proteins other than collagen are synthesized at approximately equal rates in human and baboon lung. Elastin concentration was estimated to be 33 microgram insoluble elastin protein per mg dry weight, within the range reported for normal human lung. Values have also been calculated for the lobar contents of protein, DNA, and collagen, as well as the rates of synthesis of collagen and other proteins in the entire upper lobe of the right lung of these animals. The collagen concentrations in the lung show wide differences among various animal species. However, the baboon lung is similar to human lung. This similarity may be important in the interpretation of alterations in lung connective tissue metabolism associated with experimental diseases.

Animals

Effect of lung surface tension on bronchial collapsibility in excised dog lungs.

Bronchial collapsibilities were studied in air- and saline-filled excised dog lungs. The intrapulmonary bronchi were isolated from the rest of the lung parenchyma with beads placed at their tributary bronchi as described previously by Takishima et al. (J. Appl. Physiol. 38: 875-881, 1975). Pressure-volume relations of the isolated bronchi were obtained while lung volume (VL) was kept constant. When lung recoil pressure (PL) was reduced by filling the lung with saline at a given VL, bronchial areas were smaller and bronchial collapsibilities were larger than in the air-filled lung. When bronchial areas and bronchial collapsibilities in air- and saline-filled lungs were compared at a given PL, they were approximately identical. We concluded that bronchial areas and collapsibilities were primarily determined by PL rather than VL, and lung surface tension itself made bronchial collapsibility equal to or even less than the degree of collapsibility due to forces applied from surrounding lung tissues that distended the bronchi.

Airway Resistance

Alveolar macrophage migration. Influence of lung lining material and acute lung insult.

Migration of alveolar macrophages collected by lavage from normal rhesus monkeys was tested in an under agarose migration system. Lung lining material and serum albumin obtained from normal rhesus monkeys enhanced the random migration of alveolar macrophages. A chemotactic effect for alveolar macrophages was demonstrated in response to lung lining material. Gel filtration of lung lining material using Sephadex G-200 indicated the presence of 4 distinct fractions. Fraction IV, which had a molecular weight of less than 5,000 daltons, had the greatest ability to enhance alveolar macrophage migration. Macrophages obtained from lungs of rhesus monkeys after they breathed an oxidant gas (ozone) for 7 days demonstrated decreases in both the number of cells randomly migrating and the distance they migrated. The addition of normal lung lining material to macrophages exposed to ozone enhanced their random mobility but did not restore migration values to control values. Ozonized lung lining material or rhesus monkey serum did not significantly alter alveolar macrophage migration from that of control lung lining material or serum. These data indicate that components of the acellular lining material of the lung can produce directional migration of alveolar macrophages and may serve to direct the central flow and clearance of macrophages from alveolar regions. Intraluminal alveolar macrophage accumulation during lung insult with ozone appeared to be related more to migration inhibition of resident cells than to recruitment of additional cells by chemotaxis.

Animals

Fetal lamb lung lavage and its effect on lung phosphatidylcholine.

UNLABELLED: The lungs of intrauterine 135--136-day-old lambs were lavaged with amnionic fluid, with or without meconium, to determine the effect on lung phosphatidylcholine (PC) concentration, synthesis, and function. No differences were apparent between animals lavaged with amnionic fluid or amnionic fluid with meconium. When lavaged lungs were compared to nonlavaged controls, no detectable differences were observed in histology or the quantity of saturated (SPC) and unsaturated phosphatidylcholine (UPC). However, the lavaged lungs retained a larger fraction of maximal lung volume at 5 cm H2O distending pressure and the incorporation of (32P) orthophosphate into lung PC was significantly reduced. In addition, two lavaged animals who became acidotic (pH less than 7.20) exhibited decreased incorporation of 14C) palmitate into whole lung unsaturated, and saturated phosphatidylcholine. SPECULATION: These data indicate that amnionic fluid can reduce lamb lung de novo synthesis of PC and may contribute to the alterations in lung PC found in neonatal syndromes of respiratory distress. Acidosis may accentuate this effect.

Amniotic Fluid

Methods to evaluate the effects of toxic materials deposited in the lung on immunity in lung-associated lymph nodes.

Intratracheal instillation of sheep red blood cells (SRBC) induced a primary immune response in lung-associated lymph nodes of Chinese hamsters and Fischer-344 rats. Few or no anti-SRBC antibody forming cells (AFC) were found in the spleen or cervical lymph nodes after intratracheal immunization. The number of AFC was significantly suppressed in the lung-associated lymph nodes from hamsters exposed by inhalation of 239PuO2 and in rats exposed by intratracheal instillation of benzo(a)pyrene (BaP). Although the immunologic function of the lung-associated lymph nodes was suppressed by 239PuO2 and BaP exposure, there was no observable increase in the number of AFC in distant lymphoid tissues. These data indicate that damage to lung cells and/or cells in lung-associated lymph nodes can suppress immunity induced by deposition of antigen in the lung. Even though lung-associated lymph nodes in exposed animals contained lower numbers of AFC, antigen instilled into the lungs of exposed animals did not significatnly translocate to distant lymphoid tissues. This indicates that the antigen filtering capacity of the lung-associated lymph nodes was not altered.

Aerosols

[Method and technique of experimental autotransplantation of a lung and a lung lobe].

The authors have accumulated an experience of 350 canine autotransplantations of a lung and its lobe. The peculiarities of the operative technique are described, and special emphasis is laid on its technical details that largely determine the success of a lung autotransplantation. The described method and technique permit the performance of orthotopic autotransplantation of a whole lung and of its lobe, and of heterotopic autotransplantation of the lower lobe of the right lung into the left pleural cavity in place of a removed lower lobe or whole lung. The first stage of autotransplantation of a lung or its lobe consists in applying an atrio-venous or venous anastomosis and that of the pulmonary artery with continuous U-shaped eversion sutures, with a re-establishment of the circulation, thus reducing the period of lung ischaemia. Upon restoring the blood flow in the lung, a bronchial anastomosis is applied with interrupted or continuous sutures, and the ventilation is restored. The vascular and bronchial anastomoses are applied with synthetic suture material (Orsylon) on atraumatic needles. The extensive experience of the authors in long-term follow-up of the animals after transplantation (up to 5 years) demonstrates that the described technique and method of lung autotransplantation ensures good competence and patency of the vascular and bronchial anastomoses, without any stenoses observed in the late postoperative period. As shown by histological examinations, the inflammatory reaction around the suture material is minimal.

Animals

Static lung mechanics of intact and excised rhesus monkey lungs and lobes.

Subdivisions of lung volume and pressure-volume (PV) curves of the lung and chest wall (CW) were measured in 12 rhesus monkeys (Macacca mulatta) under pentobarbital anesthesia. In addition, volumes and PV curves were obtained on the excised lungs and lobes of 12 cynomolgus monkeys (M. fasicularis). Boyle's law was used to determine functional residual capacity (FRC) in the intact animals and water displacement to determine minimal volume (MV) in the excised lungs. Total lung capacity (TLC = lung volume at a transpulmonary pressure of 30 cmH2O) was similar in vivo and in vitro (90 + 83 ml/kg) but residual volume (RV = volume at airway pressure of -50 cmH2O) and MV differed markedly (16.5 + 5.9 ml/kg). In the intact animals a very stiff CW appeared to determine RV, whereas airway closure determined MV in excised lungs. PV curves of upper and lower lobes were not different when expressed as %TLC but when expressed as milliliters of gas per gram of lung, the upper lobes contained significantly more gas per unit weight.

Animals