==================================BSR41================================== 41. We need to quantitate surfactant in rat lung, (comparing Mg deficient and Mg fed). How can surfactant be measured: a) on slides of lung b) from lung tissue or secretion - histochemistry - immunochemistry - biochemistry. Surfactant lung, as in respiratory distress syndrome. 1 UI - 87091333 AU - Jackson JC ; Palmer S ; Truog WE ; Standaert TA ; Murphy JH ; Hodson WA TI - Surfactant quantity and composition during recovery from hyaline membrane disease. AB - The appearance of phosphatidylglycerol in the tracheal wash of infants with hyaline membrane disease (HMD) has been reported to be associated with clinical signs of recovery. We analyzed lung tissue and bronchoalveolar lavage surfactant in an animal model of HMD to determine whether phosphatidylglycerol or some other component is necessary for recovery. The amount and composition of phospholipid (PL) was determined in the premature Macaca nemestrina monkey (140 days' gestation) during an acute stage of HMD, and in two stages of recovery. These changes were compared to observations made in healthy premature controls (140 days), gestational age-matched fetuses (140 days), and fetuses of 150 days' gestation (term = 168 days). The amount of PL and its surfactant composition in lung homogenates of the right lower lobe and in lavage of the excised left lung was determined. Compared to 140-day fetuses, the healthy controls had a several-fold increase in lavage PL and disaturated phosphatidylcholine (DSPC) during the first few days of life (p less than 0.05). Prior to recovery, animals with HMD had no such increase in lavage PL or DSPC and demonstrated poor deflation stability. Recovery was associated with increased tissue and lavage PL (p less than 0.05) and increased fractions of phosphatidylinositol and DSPC (p less than 0.05), but not phosphatidylglycerol. The tissue compositional changes observed during recovery reflected maturational changes observed in the fetal animals studied at 10 days' greater gestational age.(ABSTRACT TRUNCATED AT 250 WORDS) MH - Animal ; Gestational Age ; Hyaline Membrane Disease/*METABOLISM/PATHOLOGY ; Infant, Newborn ; Lung/PATHOLOGY ; Macaca nemestrina ; Organ Weight ; Phospholipids/ANALYSIS ; Pulmonary Surfactants/*ANALYSIS ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. ; Time Factors SO - Pediatr Res 1986 Dec;20(12):1243-7 2 UI - 87059665 AU - Krieglsteiner HP ; Lohninger A ; Riedl R ; el Kalak H ; Kaiser E TI - The assessment of foetal lung maturity by chemical analysis of amniotic fluid. AB - Advances in the understanding of the neonatal respiratory distress syndrome have led to a proliferation of amniotic fluid tests. Measurement of pulmonary surfactant production is the most direct means of assessing pulmonary maturity. Assays of surfactant are subjected to certain pre-analysis sources of variation, such as variability in amniotic fluid volume, sample collection site, centrifugation speed and time, and contamination with blood and/or meconium. Amniotic fluid surfactant assays can be divided into biochemical and functional tests. When properly performed, both approaches yield results that correlate well with clinical findings. However, no single method has achieved the distinction of total reliability and universal applicability. In most tests the value for mature lungs is almost 99% accurate. On the other hand, immature values have very low accuracy. Therefore, it is advisable to perform an additional test or to repeat the determination. The determination of the lecithin/sphingomyelin ratio is characterized by sufficient accuracy for routine analyses. For scientific studies we recommend the use of a capillary gas-chromatographic method allowing an accurate assessment of dipalmitoylphosphatidylcholine, the most important surfactant constituent. RF - REVIEW ARTICLE: 206 REFS. MH - Amniotic Fluid/*ANALYSIS ; Chromatography ; Female ; Fetus/*PHYSIOLOGY ; Human ; Infant, Newborn ; Lung/*PHYSIOLOGY ; Phosphatidylcholines/ ANALYSIS ; Phosphatidylglycerols/ANALYSIS ; Phospholipids/ANALYSIS ; Pregnancy ; Pulmonary Surfactants/*ANALYSIS ; Respiratory Distress Syndrome/PREVENTION & CONTROL ; Review ; Support, Non-U.S. Gov't SO - J Clin Chem Clin Biochem 1986 Oct;24(10):705-17 3 UI - 87111961 AU - Forsman LM ; Hallman M ; Autero M ; Rapola J ; Andersson LC TI - Presence of apocrine epithelial antigen (AEA) in type II pneumocytes and in hyaline membranes of neonatal RDS. AB - We have investigated the distribution of an apocrine membrane antigen (AEA) in pulmonary tissue using a rabbit antiserum raised against fat globule glycoproteins isolated from human milk. In indirect immunostaining (PAP, IF) of sections from normal lung tissue, the membranes facing the alveolar lumen of cells corresponding to the type II pneumocytes in the alveolar walls were decorated. The selective distribution of AEA to the membranes of type II pneumocytes was confirmed in double immunostaining by identification of these cells with rat antibodies against surfactant apoprotein. In fetal lung tissue, the AEA antigen was detected by the 9th week of gestation. In lung samples from newborns which had died of respiratory distress syndrome (RDS) the intra-alveolar hyaline membranes stained for the AEA antigen. SDS-PAGE of the immunoprecipitate obtained with anti-AEA serum from radiolabelled glycoprotein fraction of normal lung tissue revealed a single band of 79,000 dalton apparent molecular weight. These findings indicate that the AEA constitutes a membrane marker of the type II pneumocytes and might be involved in the secretory process of surfactant. Immunohistological evidence for the presence of AEA in the hyaline membranes of neonatal RDS is also presented. MH - Antigens/*ANALYSIS ; Human ; Infant, Newborn ; Lung/*IMMUNOLOGY ; Membrane Proteins/*ANALYSIS ; Membranes/ANALYSIS ; Pulmonary Alveoli/ PATHOLOGY ; Pulmonary Surfactants/ANALYSIS ; Respiratory Distress Syndrome/*IMMUNOLOGY ; Support, Non-U.S. Gov't SO - J Pathol 1986 Dec;150(4):289-94 4 UI - 87101781 AU - Rice WR ; Singleton FM TI - P2-purinoceptors regulate surfactant secretion from rat isolated alveolar type II cells. AB - Rat isolated alveolar Type II cells were utilized to examine the effect of purine and pyrimidine analogues on secretion of pulmonary surfactant. ATP potently stimulated [3H]-phosphatidylcholine ([3H]-PC) secretion in a time- and dose-dependent manner. The effect of ATP was noted by one hour of exposure and persisted for three hours. The EC50 (concentration producing 1/2 the maximal response) for ATP-induced [3H]-PC secretion was 100 nM. ADP was also a potent secretagogue for surfactant secretion, but AMP and adenosine had no significant effect on surfactant secretion at concentrations less than or equal to 250 microM. The EC50 for ADP-induced [3H]-PC secretion was 250 nM. Other purine and pyrimidine nucleotides (ITP, GTP, CTP, TTP) were examined for their effect on [3H]-PC secretion. All purine and pyrimidine triphosphates examined significantly augmented [3H]-PC secretion, but were much less potent than ATP. The EC50s were ITP = 10 microM; GTP = 100 microM; CTP = 250 microM; TTP = 100 microM. Neither 8-phenyltheophylline (10 microM, a P1-purinoceptor antagonist), propranolol (100 microM, a beta-adrenoceptor antagonist), nor indomethacin (10 microM, a prostaglandin synthetase inhibitor) inhibited ATP-induced [3H]-PC secretion from isolated Type II cells. These data provide evidence for regulation of surfactant secretion from alveolar Type II cells by a P2-purinoceptor. MH - Animal ; In Vitro ; Male ; Phosphatidylcholines/SECRETION ; Pulmonary Alveoli/*SECRETION ; Pulmonary Surfactants/*SECRETION ; Purine Nucleotides/PHARMACOLOGY ; Pyrimidine Nucleotides/PHARMACOLOGY ; Rats ; Rats, Inbred Strains ; Receptors, Purinergic/DRUG EFFECTS/*PHYSIOLOGY ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. SO - Br J Pharmacol 1986 Nov;89(3):485-91 5 UI - 87091330 AU - Hallman M ; Merritt TA ; Pohjavuori M ; Gluck L TI - Effect of surfactant substitution on lung effluent phospholipids in respiratory distress syndrome: evaluation of surfactant phospholipid turnover, pool size, and the relationship to severity of respiratory failure. AB - The turnover and pool size of surfactant has been studied in animals, but there is little similar information in humans. In the present investigation lung effluent phospholipids were studied in 29 small preterm infants with severe RDS. Thirteen were treated with mechanical ventilation, and 16 additionally received natural human surfactant. The first dose (60 mg surfactant/kg body wt) was given between 2 and 10 h of age, and the surfactant was given again if there was an insufficient response. Together 260 aspirates, recovered during routine suctioning of the airways, were analyzed for phospholipids. Phosphatidylglycerol, present only in exogenous surfactant, was used as a specific marker to estimate the apparent pool size and the half-life of surfactant phospholipid. In addition, the saturated phosphatidylcholine/sphingomyelin ratios were correlated with the ventilatory index (mean airway pressure X fractional inspiratory oxygen/arterial oxygen tension). There was a linear correlation between the ventilatory index and the saturated phosphatidylcholine/sphingomyelin (r approximately -0.70) but no consistent correlation between the ventilatory index and the amount of phospholipids in the aspirate. The saturated phosphatidylcholine/sphingomyelin ratio increased during the surfactant-induced remission of respiratory failure, decreased during the recovery. The control infants tended to have lower saturated phosphatidylcholine/sphingomyelin ratios during the first week than the surfactant-treated infants.(ABSTRACT TRUNCATED AT 250 WORDS) MH - Half-Life ; Human ; Infant, Newborn ; Lung/*METABOLISM ; Phosphatidylcholines/METABOLISM ; Phosphatidylglycerols/METABOLISM ; Phospholipids/*METABOLISM ; Prospective Studies ; Pulmonary Surfactants/ METABOLISM/*THERAPEUTIC USE ; Random Allocation ; Respiratory Distress Syndrome/*DRUG THERAPY/METABOLISM ; Sphingomyelins/METABOLISM ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. SO - Pediatr Res 1986 Dec;20(12):1228-35 6 UI - 87053712 AU - McMahon JB ; Smith AC ; del Campo A ; Singh G ; Katyal SL ; Schuller HM TI - Characterization of rat alveolar type II cells in vitro by immunological, biochemical, and morphological criteria. AB - Using an anti-rat surfactant apoprotein antiserum which specifically reacts with cytoplasmic structures in alveolar type II cells on histopathology sections of rat lung, we have examined the immunoreactivity of pulmonary type II cells in vitro. Single cell suspensions of lung tissue were prepared from male Fischer 344 rats by intratracheal elastase digestion according to standard published methods. Cytocentrifuged preparations of the resulting cell suspensions revealed that approximately 40% of the cells stained positive for surfactant apoprotein using an immunoperoxidase staining technique. Without further cell fractionation steps, the cell suspensions were plated at colonial densities in growth medium. The cells that attached after 24 hours of incubation and at daily intervals were analyzed for surfactant apoprotein immunoreactivity as well as for proliferation, morphology, and phospholipid biosynthesis. The percentage of immunopositive cells increased with time from 75% at day 1 to 94% at 4 days after plating. This increase was paralleled by a linear increase in the number of immunopositive cells, which expanded into cell colonies. During the initial 5 days in vitro, the immunopositive cells retained their epithelial morphology and contained cytoplasmic osmiophilic bodies. Phospholipid biosynthesis by the isolated lung cells was analyzed and the data revealed that the rate of incorporation of 14C-choline into phosphatidylcholine increased with time in culture. These studies indicated that the anti-rat surfactant apoprotein antisera can be used to identify and quantitate functional alveolar type II cells in vitro. Thus the specific antisera may facilitate studies of type II cells undergoing various environmental alterations both in vivo and in vitro. MH - Animal ; Cell Division ; Cell Separation ; Immunoenzyme Technics ; In Vitro ; Male ; Phosphatidylcholines/BIOSYNTHESIS ; Pulmonary Alveoli/ *CYTOLOGY/IMMUNOLOGY/METABOLISM ; Pulmonary Surfactants/ANALYSIS/ IMMUNOLOGY ; Rats ; Rats, Inbred F344 ; Support, U.S. Gov't, Non-P.H.S. ; Support, U.S. Gov't, P.H.S. SO - Exp Lung Res 1986;11(4):263-75 7 UI - 87049776 AU - Rice WR ; Singleton FM TI - Regulation of surfactant secretion from isolated Type II pneumocytes by substance P. AB - Substance P, an eleven amino acid neuropeptide, significantly inhibited release of [3H]phosphatidylcholine from pulmonary Type II epithelial cells in vitro. Basal release and release in response to the beta-adrenergic agonist, terbutaline and 12-O-tetradecanoylphorbol 13-acetate (TPA) were significantly decreased in the presence of substance P. Inhibitory effects of substance P were noted following a 1 h exposure of primary cultures of Type II cells in vitro and persisted up to 3 h in the presence of the secretagogues, TPA and terbutaline. The IC50 values for substance P inhibition of [3H]PC release were 10 microM for basal release, 40 microM for TPA-induced release and 50 microM for terbutaline-induced release. The related neuropeptide, physalaemin and the stable active analog of substance P, [pGlu5, MePhe8, MeGly9]substance P [5-11], had no significant inhibitory effects on surfactant release whether in the presence or absence of TPA or terbutaline. These data support the hypothesis that NH2-terminal basic groups of substance P are necessary for inhibition of surfactant secretion from isolated Type II cells and support the concept that an inhibitory system contributes to mediation of surfactant secretion from Type II epithelial cells. MH - Animal ; Lung/DRUG EFFECTS/*SECRETION ; Physalaemin/PHARMACOLOGY ; Pulmonary Surfactants/*SECRETION ; Rats ; Rats, Inbred Strains ; Substance P/ANALOGS & DERIVATIVES/*PHARMACOLOGY ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. ; Terbutaline/PHARMACOLOGY ; Tetradecanoylphorbol Acetate/PHARMACOLOGY ; Time Factors SO - Biochim Biophys Acta 1986 Nov 28;889(2):123-7 8 UI - 87039069 AU - Infante JP TI - De novo sn-glycerol-3-phosphorylcholine synthetase activity in lung and muscle and its subcellular location. AB - The activity of glycerophosphorylcholine synthetase, a newly discovered enzyme involved in the synthesis of acyl-specific phosphatidylcholines, is reported in rat lung and muscle. Its subcellular location appears to be mitochondrial. The implication of these findings in the synthesis of lung surfactant and the pathology of muscular dystrophy are discussed. MH - Animal ; Lung/*ENZYMOLOGY/ULTRASTRUCTURE ; Male ; Mitochondria/ *ENZYMOLOGY ; Muscles/*ENZYMOLOGY/ULTRASTRUCTURE ; Muscular Dystrophy, Animal/PATHOLOGY ; Phosphatidylcholines/BIOSYNTHESIS ; Pulmonary Surfactants/BIOSYNTHESIS ; Rats ; Rats, Inbred Strains ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, Non-P.H.S. ; Transferases/ *METABOLISM SO - Mol Cell Biochem 1986 Aug;71(2):135-7 9 UI - 87026820 AU - Ikegami M ; Jobe A ; Berry D TI - A protein that inhibits surfactant in respiratory distress syndrome. AB - A protein that interferes with surfactant function is present in the airways and alveoli of infants with respiratory distress syndrome (RDS). This inhibitor is also found in serum and amniotic fluid and presumably appears in the alveoli because of the abnormal protein leak present in the lungs of infants with RDS. This protein has a molecular weight of about 110,000 and is resistant to boiling or lipid extraction. As measured by radioimmunoassay, the ratio of inhibitor to phosphatidylcholine decreased from 8.1 +/- 2.3 early in the course of RDS to 0.7 +/- 0.1 on the day of extubation. The value at extubation was the same as that measured for preterm infants without RDS. The inhibitor to phosphatidylcholine ratio in airway samples from infants with RDS correlated significantly with simultaneously recorded pO2/FiO2 ratios and the peak inspiratory pressures used to normalized pCO2 values. These results are consistent with the concept that the inhibitor contributes to surfactant dysfunction and thus the respiratory failure characteristic of RDS. MH - Amniotic Fluid/METABOLISM ; Blood Gas Analysis ; Human ; Infant, Newborn ; Molecular Weight ; Phosphatidylcholines/ANALYSIS ; Proteins/*ISOLATION & PURIFICATION ; Pulmonary Alveoli/METABOLISM ; Pulmonary Surfactants/ *ANTAGONISTS & INHIBITORS ; Respiratory Distress Syndrome/*METABOLISM ; Respiratory System/*METABOLISM ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. ; Surface Tension SO - Biol Neonate 1986;50(3):121-9 10 UI - 87000668 AU - Brown LA ; Longmore WJ TI - Altered phospholipid secretion in type II pneumocytes isolated from streptozotocin-diabetic rats. AB - To study the effect of diabetes on pulmonary surfactant secretion, type II pneumocytes from adult streptozotocin-induced diabetic rats were placed in short-term culture. As opposed to a linear secretory rate by control type II cells, the secretory rate of type II cells from diabetic animals was biphasic reaching a minimum at 1.5 h. When exogenous surfactant containing radioactive phosphatidylcholine was added to the incubation media for 1.5 h, the cells from diabetic animals incorporated more exogenous phosphatidylcholine into lamellar bodies than control cells. This suggests that in the type II cell from diabetic animals, the rate of reutilization is greater than the rate of secretion until 1.5 h, at which time the rate of secretion becomes greater. The altered secretory pattern was reversed by in vivo insulin treatment 30 min prior to killing but not by the addition of insulin to the incubation media. When challenged by isoproterenol, a beta-adrenergic agonist, the secretory pattern of cells from diabetic animals was biphasic as observed with basal secretion; however, secretion was stimulated 30% as opposed to 100% increase in control cells. These data suggest that basal and stimulated secretion are altered in the cultured type II cell from diabetic animals and restored by in vivo but not in vitro insulin treatment. MH - Animal ; Diabetes Mellitus, Experimental/*METABOLISM ; In Vitro ; Insulin/ PHARMACOLOGY ; Isoproterenol/PHARMACOLOGY ; Lung/*SECRETION/ ULTRASTRUCTURE ; Male ; Phosphatidylcholines/SECRETION ; Pulmonary Surfactants/*SECRETION ; Rats ; Rats, Inbred Strains ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. SO - Biochim Biophys Acta 1986 Sep 12;878(2):258-65 11 UI - 86321034 AU - Duncan AW ; Oh TE ; Hillman DR TI - PEEP and CPAP. AB - Positive end-expiratory pressure (PEEP) maintains airway pressure above atmospheric at the end of expiration, and may be used with mechanical ventilation or spontaneous breathing. CPAP, or continuous positive airway pressure, refers to spontaneous ventilation with a positive airway pressure being maintained throughout the whole respiratory cycle. PEEP/CPAP primarily improves oxygenation by increasing functional residual capacity, and may increase lung compliance and decrease the work of breathing. PEEP/CPAP may be applied using endotracheal tubes, nasal masks or prongs, or face masks or chambers to treat a wide range of adult and paediatric respiratory disorders. Complications associated with their use relate to the pressures applied and include pulmonary barotrauma, decreased cardiac output and raised intracranial pressure. RF - REVIEW ARTICLE: 120 REFS. MH - Airway Resistance ; Apnea/THERAPY ; Asthma/THERAPY ; Bronchiolitis, Viral/ THERAPY ; Burns, Inhalation/THERAPY ; Cardiovascular System/PHYSIOLOGY ; Heart Failure, Congestive/THERAPY ; Human ; Hyaline Membrane Disease/ THERAPY ; Infant, Newborn ; Intubation ; Kidney/PHYSIOLOGY ; Lung/ INJURIES ; Nomenclature ; Pneumonia/THERAPY ; Positive Pressure Respiration/ADVERSE EFFECTS/*METHODS ; Postoperative Period ; Pulmonary Edema/THERAPY ; Pulmonary Surfactants/METABOLISM ; Respiration ; Respirators ; Respiratory Distress Syndrome, Adult/PREVENTION & CONTROL/ THERAPY ; Review ; Sleep Apnea Syndromes/THERAPY ; Work of Breathing SO - Anaesth Intensive Care 1986 Aug;14(3):236-50 12 UI - 86315112 AU - Shimura S ; Aoki T ; Takishima T TI - Morphological aspects of the surfactant system in human lungs. A freeze-fracture study. AB - We examined alveolar type II cells and extracellular surfactant (tubular myelin) of human lungs and compared them with those of experimental animals (mouse, rat and dog) using a freeze-fracture technique for electron microscopy. Lamellar bodies (intracellular surfactant) in alveolar type II cells of human lung showed features different from those of the experimental animal lungs. Namely, multicentric foci of lamellae, an amorphous portion with some degree of lattice-like structure in the mature bodies, unpaired split membranes of lamellae and also our previously reported connection of the lamellar bodies to the endoplasmic reticulum (Am. Rev. resp. Dis. 127: 1983) were found in alveolar type II cells of human lungs but not in type II cells of experimental animal lungs. On the other hand, there were no differences noted in extracellular surfactant between human and experimental animal lungs. These findings indicate the possibility that a mode of surfactant production and/or storage in alveolar type II cells exists in human lung which is different from that in experimental animal lungs. MH - Aged ; Animal ; Comparative Study ; Dogs ; Extracellular Space/METABOLISM ; Freeze Fracturing ; Human ; Lung/*METABOLISM ; Male ; Mice ; Microscopy, Electron ; Middle Age ; Myelin Proteins/METABOLISM ; Pulmonary Alveoli/METABOLISM/ULTRASTRUCTURE ; Pulmonary Surfactants/ *METABOLISM ; Rats ; Species Specificity SO - Respiration 1986;50(2):139-46 13 UI - 86292030 AU - Berggren P ; Lachmann B ; Curstedt T ; Grossmann G ; Robertson B TI - Gas exchange and lung morphology after surfactant replacement in experimental adult respiratory distress syndrome induced by repeated lung lavage. AB - Severe respiratory insufficiency was induced in adult guinea pigs by repeated lung lavage. The animals were then ventilated for 75 min with 100% O2, insufflation pressure 28/6-8 cmH2O (2.7/0.6-0.8 kPa), frequency 30/min, and 33% inspiration time. One group of animals (I) was treated with protein-depleted porcine surfactant, prepared by a combination of sucrose-gradient centrifugation, heating to 90 degrees C, and chloroform/methanol extraction. Another group (II) received the phospholipid fraction of porcine surfactant, isolated from minced lungs by chloroform/methanol extraction and liquid-gel chromatography. Surfactant was administered in two 1-ml doses (lipid concentration 90 mg/ml) instilled via the tracheal cannula about 15 and 45 min after the lavage procedure. Non-treated, lavaged animals served as controls. After 75 min of ventilation, control values for PaO2 and PaCO2 were 13.3 +/- 6.8 and 6.8 +/- 2.3 kPa (mean +/- s.d.), respectively. The corresponding values in Group I of surfactant-treated animals were 52.9 +/- 7.7 and 4.4 +/- 1.1 kPa, in Group II 53.5 +/- 7.3 and 4.8 +/- 1.3 kPa (P less than 0.02-0.002). The two groups of surfactant-treated animals also had significantly improved alveolar air expansion in histological sections, as reflected by increased alveolar volume density (0.67 +/- 0.05 and 0.62 +/- 0.11 vs 0.45 +/- 0.08 in controls; P less than 0.002). The benefits of surfactant replacement in this experimental model were thus similar to those previously observed in animal models of neonatal surfactant deficiency as well as in babies with respiratory distress syndrome (RDS). Our data suggest that surfactant replacement might have a therapeutic effect also in clinical adult RDS. MH - Animal ; Carbon Dioxide/BLOOD ; Guinea Pigs ; Irrigation ; Lung/ *PATHOLOGY ; Oxygen/BLOOD ; *Pulmonary Gas Exchange ; Pulmonary Surfactants/*THERAPEUTIC USE ; Respiratory Distress Syndrome, Adult/*DRUG THERAPY/ETIOLOGY/PATHOLOGY ; Support, Non-U.S. Gov't ; Tidal Volume SO - Acta Anaesthesiol Scand 1986 May;30(4):321-8 14 UI - 86286387 AU - Singh G ; Katyal SL ; Wong-Chong ML TI - A quantitative assay for a Clara cell-specific protein and its application in the study of development of pulmonary airways in the rat. AB - Rat lung lavage contains a 10 kDa protein that has been shown by immunocytochemistry to be specific for Clara cells. An inhibition enzyme-linked immunosorbent assay was established for this protein using rabbit antibody to the 10 kDa Clara cell protein. The assay has a sensitivity of about 3.0 ng/ml and a working range of about 5 to 50 ng/ml. Quantitation of the 10 kDa protein in amniotic fluid revealed an increase of about 4-fold at day 20 of gestation. The 10 kDa protein content of lung homogenate increased steadily from day 18 of gestation to 1 wk after birth, after which a decline was observed. Nearly 60-fold increase in the concentration of the 10 kDa Clara cell protein in lungs was noted from day 18 of gestation to birth and a further about 7-fold increase was noted from the day of birth to 1 wk of age. A progressive increase in the 10 kDa protein, with increasing age, was also noted on immunoblot analysis of lung homogenates. As judged from the immunoblots of lung homogenates, stained with rabbit antirat 10 kDa protein antiserum, the content of an antigenically similar 200 kDa Clara cell protein was negligible. The quantitative results for 10 kDa Clara cell protein parallel the results of immunocytochemistry and quantitation of the volume density of Clara cell granules indicating that quantitation for the 10 kDa protein could be used to monitor the development of Clara cells and that of the pulmonary airways. MH - Amniotic Fluid/ANALYSIS ; Animal ; Enzyme-Linked Immunosorbent Assay ; Gestational Age ; Immunosorbent Technics ; Lung/*CYTOLOGY/EMBRYOLOGY ; Molecular Weight ; Proteins/ANALYSIS/IMMUNOLOGY ; Pulmonary Surfactants/ ANALYSIS ; Rats ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, Non-P.H.S. ; Support, U.S. Gov't, P.H.S. SO - Pediatr Res 1986 Aug;20(8):802-5 15 UI - 86279552 AU - Walker SR ; Williams MC ; Benson B TI - Immunocytochemical localization of the major surfactant apoproteins in type II cells, Clara cells, and alveolar macrophages of rat lung. AB - The adsorptive properties of phospholipids of pulmonary surfactant are markedly influenced by the presence of three related proteins (26-38 KD, reduced) found in purified surfactant. Whether these proteins are pre-assembled with lipids before secretion is uncertain but would be expected for a lipoprotein secretion. We performed indirect immunocytochemistry on frozen thin sections of rat lung to identify cells and intracellular organelles that contain these proteins. The three proteins, purified from lavaged surfactant, were used to generate antisera in rabbits. Immunoblotting of rat surfactant showed that the IgG reacted with the three proteins and a 55-60 KD band which may be a polymer of the lower MW species. Specific gold labeling occurred over alveolar type II cells, bronchiolar Clara cells, alveolar macrophages, and tubular myelin. In type II cells labeling occurred in synthetic organelles and lamellar bodies, which contain surfactant lipids. Lamellar body labeling was increased fivefold by pre-treating tissue sections with a detergent. Multivesicular bodies and some small apical vesicles in type II cells were also labeled. Secondary lysosomes of alveolar macrophages were immunoreactive. Labeling in Clara cells exceeded that of type II cells, with prominent labeling in secretory granules, Golgi apparatus, and endoplasmic reticulum. These observations clarify the organelles and pathways utilized in the elaboration of surfactant. After synthesis, the proteins move, probably via multivesicular bodies, to lamellar bodies. Both lipids and proteins are present in tubular myelin. Immunologically identical or closely similar proteins are synthesized by Clara cells and secreted from granules which appear not to contain lipid. The role of these proteins in bronchiolar function is unknown. MH - Animal ; Antigen-Antibody Reactions ; Apoproteins/*ANALYSIS ; Bronchi/ ANALYSIS/ULTRASTRUCTURE ; Cytoplasmic Granules/ANALYSIS/ULTRASTRUCTURE ; Freezing ; Macrophages/*ANALYSIS/ULTRASTRUCTURE ; Male ; Organoids/ ANALYSIS/ULTRASTRUCTURE ; Proteolipids/*ANALYSIS/IMMUNOLOGY ; Pulmonary Alveoli/*ANALYSIS/ULTRASTRUCTURE ; Pulmonary Surfactants/*ANALYSIS/ IMMUNOLOGY ; Rabbits ; Rats ; Rats, Inbred Strains ; Stains and Staining ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. SO - J Histochem Cytochem 1986 Sep;34(9):1137-48 16 UI - 86265921 AU - Kuroki Y ; Dempo K ; Akino T TI - Immunohistochemical study of human pulmonary surfactant apoproteins with monoclonal antibodies. Pathologic application for hyaline membrane disease. AB - Three monoclonal antibodies, PC6, PE10, and PE12, were used for immunohistochemical studies of human lungs by immunoperoxidase staining. Monoclonal antibodies PC6 and PE10 against pulmonary surfactant apoproteins stained faint granules in the cytoplasm of some alveolar wall cells in adult lung. These stained cells appeared to be alveolar Type II cells. A fetal lung of 20 weeks' gestation had no any positive staining. However, a few scattered positive cells were observed in a newborn lung of 31 weeks' gestation, and the stained cells increased progressively with increasing gestational age. The positively stained cells were very few in the lungs of newborns who died of respiratory distress syndrome (RDS), but the lungs of newborns who died of other causes after recovery from RDS showed many positively stained cells. These results suggest that the immunohistochemical demonstration of the monoclonal antibodies PC6 and PE10 could be a good pathodiagnostic indicator reflecting the localization and development of pulmonary surfactant by alveolar Type II cells. On the other hand, monoclonal antibody PE12 was found to recognize the antigen that occurs on the surfaces of the alveoli of fetal, newborn, and adult lungs as one component of the alveolar lining layer, different from pulmonary surfactant. MH - Adult ; Antibodies, Monoclonal/*DIAGNOSTIC USE ; Apoproteins/ANALYSIS ; Fetus/PATHOLOGY ; Gestational Age ; Human ; Hyaline Membrane Disease/ DIAGNOSIS/*PATHOLOGY ; Immunoenzyme Technics ; Infant, Newborn ; Lung/ PATHOLOGY ; Pulmonary Surfactants/ANALYSIS/*BIOSYNTHESIS ; Respiratory Distress Syndrome/*DIAGNOSIS ; Support, Non-U.S. Gov't SO - Am J Pathol 1986 Jul;124(1):25-33 17 UI - 86258808 AU - Sosenko IR ; Lewis PL ; Frank L TI - Metyrapone delays surfactant and antioxidant enzyme maturation in developing rat lung. AB - The surfactant system and the antioxidant enzyme system of the fetal lung have chronologically similar developmental patterns and both can be accelerated by the administration of exogenous glucocorticoids. To test whether the antioxidant enzyme system, like the surfactant system, is regulated, at least in part, by endogenous glucocorticoids, we injected pregnant rats for 3 days prior to delivery with metyrapone, an adrenal 11-beta hydroxylase inhibitor which crosses the placenta and blocks endogenous glucocorticoid synthesis, or saline. Metyrapone offspring had significantly decreased lung tissue disaturated phosphatidylcholine/total phospholipids (p less than 0.05) compared to controls at days 21 and 22 of gestation. Activities of the antioxidant enzymes superoxide dismutase, catalase, and glutathione peroxidase were similarly significantly reduced (p less than 0.01) in the lungs of metyrapone offspring at both gestational days studied. One day premature metyrapone pups demonstrated poorer survival than control pups from 25 min after delivery (44% survival versus 83%, p less than 0.05) to 90 min (6% survival versus 78%, p less than 0.01). These findings of delayed maturation of the surfactant and antioxidant enzyme systems following adrenal glucocorticoid blockade suggest that both systems are regulated, at least in part, by an endogenous glucocorticoid mechanism. MH - Animal ; Antioxidants/ANALYSIS/METABOLISM ; Catalase/ANALYSIS/METABOLISM ; Glutathione Peroxidase/ANALYSIS/METABOLISM ; Lung/ANALYSIS/*DRUG EFFECTS ; Metyrapone/*PHARMACOLOGY ; Phosphatidylcholines/ANALYSIS/ METABOLISM ; Pulmonary Surfactants/*METABOLISM ; Rats ; Rats, Inbred Strains ; Superoxide Dismutase/ANALYSIS/METABOLISM ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. SO - Pediatr Res 1986 Jul;20(7):672-5 18 UI - 86258803 AU - Rieutort M ; Farrell PM ; Engle MJ ; Pignol B ; Bourbon JR TI - Changes in surfactant phospholipids in fetal rat lungs from normal and diabetic pregnancies. AB - The purposes of this study were to adapt and evaluate further a pulmonary surfactant isolation method applicable to unperfused fetal rat lung, to quantitate key phospholipids phosphatidylcholine (disaturated phosphatidylcholine, and phosphatidylglycerol) of the isolated material during the last 3 days of gestation, and to determine if abnormalities in surfactant phospholipids were present in fetuses of diabetic pregnancies. A simplified scheme of sucrose gradient centrifugation proved useful for small scale preparations of material enriched in the phospholipids most characteristic of pulmonary surfactant. It was shown that fetal blood phospholipids did not contaminate the surfactant fraction and therefore would not produce artifacts in assessment of lung maturational changes. Analyses of subcellular fractions isolated at 19.5, 20.5, and 21.5 days revealed that the percentages of disaturated phosphatidylcholine relative to total phospholipids were 23-44% in the surfactant preparations and 14-21% in the residual (nonsurfactant) fractions, while the disaturated phosphatidylcholine/phosphatidylcholine ratios were 0.62 +/- 0.06 and 0.41 +/- 0.03, respectively. Summation of the amounts of individual phospholipids in the two fractions yielded data that were nearly identical to the concentrations of these compounds in whole fetal lung samples analyzed independently, implying that losses during the surfactant isolation technique were negligible. The concentrations of phosphatidylcholine, disaturated phosphatidylcholine, phosphatidylglycerol, and total phospholipids increase markedly (more than 10-fold) and progressively in surfactant fractions prepared from normal fetal rat lung at 19.5, 20.5, and 21.5 days of gestation. In contrast, the residual fractions showed no changes from 19.5 to 20.5 days and then relatively modest increases from 20.5 to 21.5 days, except for phosphatidylglycerol, which increased markedly.(ABSTRACT TRUNCATED AT 250 WORDS) MH - Animal ; Diabetes Mellitus, Experimental/*METABOLISM ; Female ; Fetus/ *ANALYSIS ; Lung/*ANALYSIS ; Phospholipids/*ANALYSIS ; Pregnancy ; Pulmonary Surfactants/ANALYSIS ; Rats ; Rats, Inbred Strains ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. SO - Pediatr Res 1986 Jul;20(7):650-4 19 UI - 86246013 AU - Bhat R ; Zikos-Labropoulou E TI - Resuscitation and respiratory management of infants weighing less than 1000 grams. AB - Care of the tiny neonate with regard to assisted ventilation and possible surfactant therapy is discussed in this review. Management in the delivery room and after is also included. MH - Birth Weight ; Bronchopulmonary Dysplasia/THERAPY ; Ductus Arteriosus, Patent/THERAPY ; Female ; Fetal Organ Maturity/DRUG EFFECTS ; Human ; Hyaline Membrane Disease/THERAPY ; *Infant, Low Birth Weight ; Infant, Newborn ; Lung/EMBRYOLOGY ; Pneumothorax/THERAPY ; Pregnancy ; Prognosis ; Pulmonary Emphysema/THERAPY ; Pulmonary Surfactants/ADMINISTRATION & DOSAGE ; Respiration, Artificial/*METHODS ; Respiratory Distress Syndrome/ *THERAPY ; Resuscitation/*METHODS ; Support, Non-U.S. Gov't SO - Clin Perinatol 1986 Jun;13(2):285-97 20 UI - 86243483 AU - Dobbs LG ; Gonzalez RF ; Marinari LA ; Mescher EJ ; Hawgood S TI - The role of calcium in the secretion of surfactant by rat alveolar type II cells. AB - Beta adrenergic agonists, tetradecanoylphorbol acetate, and the ionophore A23187 all stimulate surfactant secretion in type II cells isolated from rats. We found that combinations of these agonists cause augmented secretion, suggesting that the agonists may effect different steps in the secretory process. Previous studies have shown that cAMP is likely to be an intracellular 'second messenger' in type II cells. A23187, which has been reported to increase cAMP in some cell systems, did not increase the cAMP content of type II cells. We investigated the possible role of Ca2+ as another 'second messenger' by studying cellular 45Ca fluxes and the effect of extracellular calcium depletion on secretion. Depletion of extracellular calcium for as long as 3 h did not alter stimulated secretion, although basal secretion was increased. Secretagogues did not stimulate 45Ca influx from extracellular sources. A23187 and, to a lesser extent, terbutaline caused an acceleration of 45Ca efflux from type II cells. The addition of terbutaline or tetradecanoylphorbol acetate to A23187 further accelerated 45Ca efflux, suggesting that these agonists may act on separate calcium pools or by different mechanisms on the same calcium pool. Although secretion from type II cells is not inhibited by extracellular calcium depletion, the studies on 45Ca efflux suggest that Ca2+ plays a role in the regulation of surfactant secretion from isolated type II cells. MH - A-23187/PHARMACOLOGY ; Adenosine Cyclic Monophosphate/METABOLISM ; Animal ; Calcium/METABOLISM/*PHYSIOLOGY ; In Vitro ; Pulmonary Alveoli/ *SECRETION ; Pulmonary Surfactants/*SECRETION ; Rats ; Support, U.S. Gov't, P.H.S. ; Tetradecanoylphorbol Acetate/PHARMACOLOGY SO - Biochim Biophys Acta 1986 Jun 27;877(2):305-13 21 UI - 86225859 AU - Webb DS ; Jeska EL TI - Enhanced luminol-dependent chemiluminescence of stimulated rat alveolar macrophages by pretreatment with alveolar lining material. AB - Previous reports indicate that the in vitro bactericidal activity of rat alveolar macrophages (AM) is dependent on the lipid fraction (ALM-L) of the alveolar lining material (ALM). The present study demonstrates that luminol-dependent chemiluminescence of stimulated rat AM is increased when rat AM are preincubated in the ALM or in the ALM-L. Evidence is presented that oxidation of the unsaturated lipids is responsible for the increase. In addition to pretreatment with the ALM, cells were also pretreated in commercial preparations of several lipids found to be present in the ALM. Preincubation in these lipids produced a significant increase in the luminol-dependent chemiluminescence response. However, when a saturated lipid, dipalmitoyl phosphatidylcholine, was used no increase was found. Pretreatment in ALM did not increase the nitro blue tetrazolium dye reduction by the AM, nor was the phagocytosis of latex beads by the AM altered by the addition of the ALM. MH - Animal ; Luminescence ; Luminol ; *Macrophage Activation ; Macrophages/ *PHYSIOLOGY ; Nitroblue Tetrazolium/METABOLISM ; Oxidation-Reduction ; Phagocytosis ; Phospholipids/PHYSIOLOGY ; Pulmonary Alveoli/CYTOLOGY/ *PHYSIOLOGY ; Pulmonary Surfactants/PHYSIOLOGY ; Rats ; Support, U.S. Gov't, Non-P.H.S. SO - J Leukocyte Biol 1986 Jul;40(1):55-64 22 UI - 86223680 AU - Young SL ; Silbajoris R TI - Dexamethasone increases adult rat lung surfactant lipids. AB - Prenatal administration of glucocorticoids stimulates epithelial cell maturation and induces a precocious development of pulmonary surfactant. The response of the adult lung to steroid administration is less well understood. We administered dexamethasone (2 mg X kg-1 X day-1) to adult male rats for 1 wk by daily subcutaneous injection. After pentobarbital anesthesia we lavaged the lungs and also isolated lamellar bodies from the tissue. Lipid analyses of the extracellular and intracellular surfactant compartments showed two- to fourfold greater amounts of total phospholipids and disaturated phosphatidylcholine compared with control. These changes were not found in kidney nor liver and were not present in plasma membrane, mitochondrial, or microsomal fractions from lungs. Morphometric analyses of the type II cells showed that anatomic measures of the lamellar body pool did not increase. We conclude that glucocorticoids have a significant effect to increase lung surfactant lipid pools of adult rat lungs by changing the phospholipid content of lamellar bodies, without changing lamellar body volume. MH - Animal ; Dexamethasone/*PHARMACOLOGY ; Lung/*DRUG EFFECTS/METABOLISM/ ULTRASTRUCTURE ; Male ; Phospholipids/METABOLISM ; Pulmonary Surfactants/ *METABOLISM ; Rats ; Rats, Inbred Strains ; Subcellular Fractions/ METABOLISM ; Support, U.S. Gov't, Non-P.H.S. SO - J Appl Physiol 1986 May;60(5):1665-72 23 UI - 86176498 AU - Taeusch HW ; Keough KM ; Williams M ; Slavin R ; Steele E ; Lee AS ; Phelps D ; Kariel N ; Floros J ; Avery ME TI - Characterization of bovine surfactant for infants with respiratory distress syndrome. AB - Exogenous surfactant treatment of surfactant-deficient disease states is now under study in a number of centers, using a variety of surfactant preparations. We have chosen one preparation because of its current and potential clinical usefulness, and we have characterized it using selected tests and assays that we thought would be necessary (although not necessarily sufficient) to justify extended clinical use. We found its lipid composition to resemble that of other surfactants derived from lung mince. There is little variation among several batches with regard to lipid composition or surface tension-lowering capability. Morphologic heterogeneity occurs in individual samples of pelleted material studied by electron microscopy. Arterial oxygenation is improved when the material is administered to animals depleted of surfactant. A low molecular weight protein was identified that reacted with antibody that specifically binds nonserum surfactant proteins in a number of animal species (including human and cow). The characteristics of this surfactant preparation should be useful for comparison as newer and simpler products become available. MH - Animal ; Cattle ; Human ; Infant, Newborn ; Lipids/*ANALYSIS ; Lung/ PHYSIOLOGY ; Male ; Methods ; Proteins/*ANALYSIS ; Pulmonary Surfactants/ *ANALYSIS/THERAPEUTIC USE ; Rats ; Respiratory Distress Syndrome/*THERAPY ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. ; Surface Tension SO - Pediatrics 1986 Apr;77(4):572-81 24 UI - 86232254 AU - Stark AR ; Frantz ID 3d TI - Respiratory distress syndrome. AB - Increasing knowledge of the pathophysiology of respiratory distress syndrome has led to improvements in clinical management. Future advances in prevention and therapy, including administration of agents to prevent prematurity or to accelerate lung maturation, provision of surfactant replacement, and new techniques of mechanical ventilation, will further decrease mortality and morbidity. RF - REVIEW ARTICLE: 51 REFS. MH - Animal ; Critical Care ; Diuresis ; Diuretics/THERAPEUTIC USE ; Ductus Arteriosus, Patent/PHYSIOPATHOLOGY ; Female ; Fibroblast Growth Factor/ PHYSIOLOGY ; Human ; Infant, Newborn ; Lung/*PHYSIOPATHOLOGY ; Male ; Neuromuscular Blocking Agents/THERAPEUTIC USE ; Oxygen Inhalation Therapy ; Phosphatidylcholines/ANALYSIS ; Prenatal Diagnosis ; Pulmonary Edema/ PHYSIOPATHOLOGY ; Pulmonary Surfactants/PHYSIOLOGY/THERAPEUTIC USE ; Rats ; Respiration, Artificial/METHODS ; Respiratory Distress Syndrome/ DIAGNOSIS/*PHYSIOPATHOLOGY/THERAPY ; Review ; Risk ; Sphingomyelins/ ANALYSIS SO - Pediatr Clin North Am 1986 Jun;33(3):533-44 25 UI - 86175916 AU - Garite TJ ; Freeman RK ; Nageotte MP TI - Fetal maturity cascade: a rapid and cost-effective method for fetal lung maturity testing. AB - One hundred ninety-three amniotic fluid samples were tested for fetal lung maturity using a maturity cascade scheme involving the sequential use of, in order, the shake test, fluorescence polarimetry, and lecithin: sphingomyelin (L:S) ratio. If any of these tests indicated maturity, the sequence was terminated and no further test was performed, and the fetus was considered mature. Seventy percent of the tests yielded mature values and of these, 85 (63%) required a shake test only, 37 (27%) had a shake test and a fluorescence polarimetry, and only 14 (10%) required all three tests. From these 193 amniocenteses, 111 patients delivered within 72 hours of the procedure. One of 94 infants had respiratory distress syndrome after a mature test (1% false maturity) and ten of 17 had respiratory distress syndrome after an immature cascade (41% falsely immature). This approach saves time and cost and by confirming immaturity with multiple tests only when necessary and may improve predictability of neonatal respiratory distress syndrome. MH - Amniocentesis/*ECONOMICS ; Amniotic Fluid/*ANALYSIS ; Cesarean Section ; False Positive Reactions ; Female ; *Fetal Organ Maturity ; Fluorescence ; Gestational Age ; Human ; Infant, Newborn ; Labor ; Lung/EMBRYOLOGY ; Phosphatidylcholines/ANALYSIS ; Pregnancy ; Pulmonary Surfactants/ ANALYSIS ; Respiratory Distress Syndrome/*DIAGNOSIS ; Sphingomyelins/ ANALYSIS SO - Obstet Gynecol 1986 May;67(5):619-22 26 UI - 86131700 AU - Weaver TE ; Hull WM ; Ross G ; Whitsett JA TI - In vitro acetylation of rat pulmonary surfactant-associated glycoprotein(s) A primary translation products. AB - The primary translation products of pulmonary surfactant-associated glycoprotein(s) A, the major apolipoprotein in mammalian surfactants, exhibit extensive charge heterogeneity. After in vitro translation of poly(A)+ mRNA from rat lung, the primary translation products of glycoprotein(s) A were identified as a charge train of five proteins of 26 kDa (pI 4.6-5.0), the predominant forms being the more acidic members (pI less than 4.8). Inhibition of acetylation during in vitro translation of rat lung poly(A)+ mRNA resulted in a predominance of the more basic isoforms (pI greater than or equal to 4.8). Intracellular forms of glycoprotein(s) A were immunoprecipitated from rat Type II epithelial cells after treatment with tunicamycin or after deglycosylation with endoglycosidase H. Five intracellular precursors consisting primarily of acidic members of the charge train were identified, this being consistent with the intracellular acetylation of the protein. In contrast, canine glycoprotein(s) A translation products consisted of only three proteins of 26 kDa (pI 4.8-5.0), in which most of the radiolabel was concentrated in the more basic components. Acetylation may account for some, but not all, of the charge heterogeneity in the primary translation products and processed forms of surfactant-associated glycoprotein(s) A in the rat. MH - Acetylation ; Animal ; Cells, Cultured ; Dogs ; Electrophoresis, Polyacrylamide Gel ; Epithelium/METABOLISM ; Immunosorbent Technics ; Isoelectric Focusing ; Lung/ANALYSIS/METABOLISM ; Male ; Molecular Weight ; Poly A/METABOLISM ; *Protein Processing, Post-Translational ; Pulmonary Surfactants/*METABOLISM ; Rats ; Rats, Inbred Strains ; RNA, Messenger/ METABOLISM ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. ; Translation, Genetic SO - Biochim Biophys Acta 1986 Feb 14;869(3):330-6 27 UI - 86128537 AU - Coalson JJ ; Winter VT ; Martin HM ; King RJ TI - Colloidal gold immunoultrastructural localization of rat surfactant. AB - Using a polyclonal antiserum against the nonserum proteins in purified rat surfactant, we have localized protein antigen within the lamellar bodies of rat alveolar Type II cells perfusion-fixed with 2% cacodylate-buffered paraformaldehyde and postfixed with 0.5% osmium. A postembedment indirect immunogold ultrastructural localization was used and 20 nm gold particles were localized over the lamellae in Type II cell lamellar bodies, in tubular myelin, and in some of the secondary lysosomes of alveolar macrophages. Occasional labeling was seen in the rough endoplasmic reticulum and multivesicular bodies in some Type II cells, but the amount of this staining was not different from nonspecific background. There was, however, an invariant lack of labeling over all other lung cell types. These results demonstrate the presence of surfactant proteins within the lamellar body secretory product and support the idea that the surfactant lipoprotein complex is formed within intracellular sites prior to its secretion into the alveolar space. MH - Acinetobacter ; Animal ; Cellular Inclusions/METABOLISM ; Cytoplasm/ METABOLISM ; Gold/*DIAGNOSTIC USE ; Immunologic Technics ; Lysosomes/ METABOLISM ; Macrophages/METABOLISM ; Microscopy, Electron ; Pulmonary Alveoli/CYTOLOGY/METABOLISM/ULTRASTRUCTURE ; Pulmonary Surfactants/ *METABOLISM ; Rats ; Rats, Inbred Strains ; Support, U.S. Gov't, P.H.S. ; Tissue Distribution SO - Am Rev Respir Dis 1986 Feb;133(2):230-7 28 UI - 86127629 AU - Strayer DS ; Merritt TA ; Lwebuga-Mukasa J ; Hallman M TI - Surfactant-anti-surfactant immune complexes in infants with respiratory distress syndrome. AB - The authors sought to determine whether treatment of respiratory distress syndrome (RDS) with human surfactant resulted in the formation of detectable circulating immune complexes. Preterm infants with severe RDS were divided into two groups: one group received human surfactant by intratracheal instillation and the other group did not. Both groups received ventilatory management involving intermittent mandatory ventilation. Plasma samples were drawn from these babies prior to treatment and at intervals thereafter. The authors developed an ELISA assay specific for surfactant-anti-surfactant immune complexes and analyzed the plasma samples for such immune complexes. Complement levels were also measured. They found that with time plasma from RDS infants in both groups showed evidence of surfactant-anti-surfactant immune complex formation. The concentrations of immune complexes generally peaked within the first week of life and then appeared to diminish over 1-4 weeks after birth in RDS infants. There was no evidence at any time in either group of immune-complex-mediated injury or of decreased serum complement levels. It is concluded that circulating immune complexes between surfactant and antibodies to surfactant are probably found in most neonates with respiratory distress syndrome, that they do not produce pulmonary damage detectable by clinical and serologic means, and that treatment of neonatal RDS with human surfactant similarly does not produce lung injury as determined with these techniques. MH - Antibody Specificity ; Antigen-Antibody Complex/*ANALYSIS ; Complement/ IMMUNOLOGY ; Enzyme-Linked Immunosorbent Assay ; Fluorescent Antibody Technic ; Human ; Immune Sera/IMMUNOLOGY ; Infant, Newborn ; *Infant, Premature ; Lung/IMMUNOLOGY ; Pulmonary Surfactants/*IMMUNOLOGY/ THERAPEUTIC USE ; Respiratory Distress Syndrome/DRUG THERAPY/*IMMUNOLOGY ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. SO - Am J Pathol 1986 Feb;122(2):353-62 29 UI - 86127506 AU - Aberg A ; Gislen L TI - Use of the drop volume of amniotic fluid in estimating the risk for respiratory distress syndrome in the newborn infant. AB - The present study describes the testing and function of the drop-volume method in the analysis of fetal lung maturity with use of amniotic fluid. Elevated surface tension resulting from a lack of surface-active phospholipids (surfactant) is the primary etiologic defect in the development of respiratory distress syndrome. The drop-volume method quantifies the surface tension of amniotic fluid with use of the fact that the volume of a falling drop of liquid is proportional to the quantity of surfactant in the solution. The drop-volume method requires only 2 minutes and 2 ml of amniotic fluid and predicts fetal lung maturity with an accuracy equal to or greater than that of other tests currently in use. MH - Adult ; Amniotic Fluid/*ANALYSIS ; Equipment and Supplies ; False Negative Reactions ; Female ; Fetal Organ Maturity ; Human ; Infant, Newborn ; Lung/EMBRYOLOGY ; Mathematics ; Phosphatidylcholines/ANALYSIS ; Pregnancy ; Pulmonary Surfactants/*ANALYSIS ; Respiratory Distress Syndrome/*DIAGNOSIS ; Risk ; Sphingomyelins/ANALYSIS ; Surface Tension SO - Am J Obstet Gynecol 1986 Jan;154(1):68-74 30 UI - 86111911 AU - Post M ; Barsoumian A ; Smith BT TI - The cellular mechanism of glucocorticoid acceleration of fetal lung maturation. Fibroblast-pneumonocyte factor stimulates choline-phosphate cytidylyltransferase activity. AB - The cellular mechanism by which glucocorticoids stimulate phosphatidylcholine biosynthesis has been studied in the fetal rat lung in vivo and in cultured fetal rat lung cells of varying levels of complexity. Administration of dexamethasone to pregnant rats at 18 days gestation resulted in a significant increase in saturated phosphatidylcholine content in fetal lung 24 h after injection. Dexamethasone administration increased the activity of fetal lung choline-phosphate cytidylyltransferase by 34%. It had no effect on the activities of fetal lung choline kinase and choline phosphotransferase. Exposure of fetal lung type II cells in organotypic cultures (which contain both type II cells and fibroblasts) to cortisol resulted in a 1.6-fold increase in the incorporation of [Me-3H]choline into saturated phosphatidylcholine. The activities of the enzymes in the choline pathway for the de novo biosynthesis of phosphatidylcholine were not significantly altered except for a 105% increase in choline-phosphate cytidylyltransferase activity. Treatment of monolayer cultures of fetal type II cells with cortisol-conditioned medium from fetal lung fibroblasts resulted in a 1.5-fold increase in saturated phosphatidylcholine production. This effect correlated with a doubling of choline-phosphate cytidylyltransferase activity. Additional evidence that this stimulatory action is mediated by fibroblast-pneumonocyte factor, produced by fetal lung fibroblasts in response to cortisol, was obtained. The factor was partially purified from cortisol-conditioned medium of fetal lung fibroblasts by gel filtration and affinity chromatography. Based on biological activity, a 3000-fold purification was obtained. Stimulation of saturated phosphatidylcholine synthesis in type II cells by fibroblast-pneumonocyte factor was maximal within 60 min of incubation. Pulse-chase experiments indicated that the stimulatory effect was correlated with an increased conversion of choline phosphate into CDP choline. Moreover, the enhanced phosphatidylcholine formation by fetal type II cells in response to fibroblast-pneumonocyte factor was accompanied by decreased levels of cellular choline phosphate. These findings further support the concept that glucocorticoid action on surfactant-associated phosphatidylcholine synthesis occurs ultimately at the level of the alveolar type II cell and involves fibroblast-pneumonocyte factor which stimulates the activity of choline-phosphate cytidylyltransferase. MH - Animal ; Cells, Cultured ; Choline Kinase/ANALYSIS ; Cholinephosphotransferase/ANALYSIS ; Dexamethasone/*PHARMACOLOGY ; Female ; Fetal Organ Maturity/DRUG EFFECTS ; Fibroblast Growth Factor/ISOLATION & PURIFICATION/PHARMACOLOGY ; Fibroblasts/DRUG EFFECTS ; Hydrocortisone/ *PHARMACOLOGY ; Lung/ANALYSIS/DRUG EFFECTS/*EMBRYOLOGY ; Nucleotidyltransferases/ANALYSIS ; Phosphatidylcholines/ANALYSIS ; Pregnancy ; Pulmonary Surfactants/BIOSYNTHESIS ; Rats ; Rats, Inbred Strains ; Stimulation, Chemical ; Support, U.S. Gov't, P.H.S. SO - J Biol Chem 1986 Feb 15;261(5):2179-84 31 UI - 86085921 AU - Floros J ; Phelps DS ; Kourembanas S ; Taeusch HW TI - Primary translation products, biosynthesis, and tissue specificity of the major surfactant protein in rat. AB - Rat lung tissue was labeled with [35S]methionine and the major surfactant-associated proteins immunoprecipitated using a specific antiserum. The protein pattern obtained was very similar to that seen in rat bronchoalveolar lavage. Rat lung mRNA was subsequently translated in an in vitro rabbit reticulocyte system, and surfactant-associated protein-related polypeptides were immunoprecipitated. A 26-kDa polypeptide was identified and characterized as follows. (a) Unlabeled surfactant proteins added to the immunoprecipitation mixture completely inhibited its immunoprecipitation. (b) Two-dimensional gel electrophoresis of the 26-kDa protein resolved it into 3 isoforms. (c) Inclusion of dog pancreatic microsomes in the translation mixture resulted in the formation of two distinct higher molecular weight groups of isoforms, suggesting that the 26-kDa protein is destined to become a glycoprotein. Immunoprecipitation of [35S]methionine-labeled rat lung tissue proteins after tunicamycin treatment resulted in 3 isoforms, identical to the ones seen in the primary translation products. In addition, expression of the surfactant proteins appears specific to the lung. MH - Animal ; Electrophoresis, Polyacrylamide Gel ; Lung/ANALYSIS ; Methionine/ METABOLISM ; Proteolipids/*BIOSYNTHESIS/GENETICS ; Pulmonary Surfactants/ *BIOSYNTHESIS/GENETICS ; Rats ; RNA, Messenger/METABOLISM ; Support, Non-U.S. Gov't ; Support, U.S. Gov't, P.H.S. ; *Translation, Genetic SO - J Biol Chem 1986 Jan 15;261(2):828-31