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Biomedical subjects

M A Matthay

Publications and source records attributed to M A Matthay.

At least 145 records · Page 8Linked to original sources
PubMed1995

Resistance of the alveolar epithelium to injury from septic shock in sheep.

Experimentally, the intravenous administration of a bolus dose of Escherichia coli endotoxin in sheep or a bolus dose of live Pseudomonas aeruginosa in rats is insufficient to cause injury to the alveolar epithelial barrier. Therefore, the first objective of these studies was to maximize the injury caused by live bacteria to the lung by administering a large dose of live P. aeruginosa into the lung perfusate of goat lungs in situ. P. aeruginosa (2.4 x 10(10) colony-forming units [cfu]) and [131I]albumin (vascular protein tracer) were added to the lung perfusate. Even though the bacterial inoculum remained very high in this isolated perfused lung system, there was no change in the permeability to protein or clearance of fluid across the alveolar epithelium, although there was an increase in lung endothelial protein permeability. Therefore, since systemic factors have been implicated in the severity and pathogenesis of septic lung injury, the second objective was to administer a continuous intravenous infusion of live P. aeruginosa over 8 h in intact anesthetized sheep. The eight sheep so treated exhibited an intact, functional alveolar barrier, even though there was an increase in lung endothelial permeability to protein and an increase in extravascular lung water. In fact, in these eight sheep, alveolar epithelial fluid transport was significantly greater than in control sheep. In the three other septic sheep there was injury to the alveolar epithelial barrier with an increase in permeability of the barrier to protein, an inability to transport fluid out of the airspaces, and an even greater increase in extravascular lung water.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1995

Identification of patients with acute lung injury. Predictors of mortality.

A recent North-American-European Consensus Conference proposed new, uniform criteria for the definition of acute lung injury, in part to facilitate earlier identification of patients for clinical trials. However, these criteria have not been evaluated prospectively. We designed a prospective cohort study of 123 consecutive patients with acute lung injury prospectively identified on admission to the adult intensive care units of a tertiary care university hospital. The objectives were to determine if selection of patients using the new criteria for acute lung injury results in a significant change in the clinical characteristics, risk factors, or predictors of mortality when compared with prior studies of patients with adult respiratory distress syndrome (ARDS); and to determine if a quantitative index of the severity of acute lung injury has prognostic value in identifying nonsurvivors of acute lung injury. We used three methods: (1) prospective identification of patients with acute lung injury using a PaO2/FIO2 ratio < 300 and bilateral infiltrates on chest radiograph in the absence of left heart failure; (2) evaluation of the severity of lung injury using a four-point scoring system; and (3) stepwise logistic regression analysis to identify variables significantly associated with hospital mortality. Overall hospital mortality was 58%. Sepsis was the most common clinical disorder (50/123 or 41%) associated with the development of acute lung injury. Using the new definition for acute lung injury, 66 of the 123 patients were enrolled with a PaO2/FIO2 ratio between 150 and 299; 57 of the 123 patients had a PaO2/FIO2 < 150 at the time of entry into the study.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1995

Acid aspiration-induced lung injury in rabbits is mediated by interleukin-8-dependent mechanisms.

Acid aspiration lung injury may be mediated primarily by neutrophils recruited to the lung by acid-induced cytokines. We hypothesized that a major acid-induced cytokine was IL-8 and that a neutralizing anti-rabbit-IL-8 monoclonal antibody (ARIL8.2) would attenuate acid-induced lung injury in rabbits. Hydrochloric acid (pH = 1.5 in 1/3 normal saline) or 1/3 normal saline (4 ml/kg) was instilled into the lungs of ventilated, anesthetized rabbits. The rabbits were studied for 6 or 24 h. In acid-instilled rabbits without the anti-IL-8 monoclonal antibody, severe lung injury developed in the first 6 h; in the long-term experiments, all rabbits died with lung injury between 12 and 14 h. In acid-instilled rabbits given the anti-IL-8 monoclonal antibody (2 mg/kg, intravenously) either as pretreatment (5 min before the acid) or as treatment (1 h after the acid), acid-induced abnormalities in oxygenation and extravascular lung water were prevented and extravascular protein accumulation was reduced by 70%; in the long-term experiments, anti-IL-8 treatment similarly protected lung function throughout the 24-h period. The anti-IL-8 monoclonal antibody also significantly reduced air space neutrophil counts and IL-8 concentrations. This study establishes IL-8 as a critical cytokine for the development of acid-induced lung injury. Neutralization of IL-8 may provide the first useful therapy for this clinically important form of acute lung injury.

Animals↗
PubMed1994

Transcellular water transport in lung alveolar epithelium through mercury-sensitive water channels.

The movement of water between the air space and capillary compartments is important for the maintenance of air space hydration during respiration and for reabsorption of excess alveolar fluid. We have obtained immunocytochemical and functional evidence that plasma-membrane water channels are responsible for water transport in the intact lung. Northern and quantitative immunoblot analysis showed high expression of CHIP28 (channel-forming integral membrane protein of 28 kDa) water channels in rat lung; immunocytochemistry showed CHIP28 localization to epithelial cell plasma membranes. Stopped-flow light scattering measurements of osmotic water permeability (Pf) in freshly isolated rat alveolar type II epithelial cells indicated a high Pf of 0.015 +/- 0.002 cm/s (10 degrees C) that was weakly temperature-dependent (activation energy, 4 kcal/mol) and reversibly inhibited by 78 +/- 4% by 0.5 mM HgCl2. An in situ-perfused sheep lung model was used to determine the route for water movement in intact lung. Blood-to-air-space water transport was measured by sampling air space fluid after instillation into distal air spaces of hyperosmolar saline (900 mOsm) containing radioiodinated albumin and [14C]mannitol. In seven sets of experiments, air space osmolality and radioiodinated albumin equilibrated with a t1/2 of 0.85 +/- 0.1 min. In the contralateral lung perfused with 0.5 mM HgCl2, t1/2 increased to 2.7 +/- 0.4 min; the inhibitory effect of HgCl2 was fully reversed by 5 mM 2-mercaptoethanol. These results provide direct evidence for transcellular movement of water across the alveolar epithelium in intact lung through mercury-sensitive water channels.

Animals↗
PubMed1994

Molecular cloning of a mercurial-insensitive water channel expressed in selected water-transporting tissues.

Two mercurial-inhibitable water-transporting proteins have been identified: CHIP28, an erythrocyte water channel also expressed in kidney tubules and selected extrarenal epithelia, and WCH-CD, a kidney collecting duct water channel. In searching for a protein responsible for the high transcellular water permeability in lung alveolus, we cloned a 32-kDa water channel (mercurial-insensitive water channel (MIWC)) from a rat lung cDNA library with several novel features. Water permeability was strongly increased in Xenopus oocytes expressing MIWC in a mercurial-insensitive manner, in contrast to known water channels. By in situ hybridization, MIWC showed an unique distribution in cells that do not express CHIP28, including kidney papillary vasa recta, cells lining the subarachnoid space and ventricles in brain, the inner nuclear layer in retina, and the conjunctival epithelium. An alternatively spliced form of MIWC with a 165-base pair deletion in the coding sequence was also identified; relative expression of the spliced mRNA was tissue-specific. The MIWC water channel may participate in the urinary concentrating mechanism, the absorption of cerebrospinal fluid, and other physiological processes.

Amino Acid Sequence↗
PubMed1994

Effect of endocytosis inhibitors on alveolar clearance of albumin, immunoglobulin G, and SP-A in rabbits.

Protein in the alveolar space may be cleared by endocytosis and degradation inside alveolar epithelial cells, by transcytosis across the alveolar epithelium, or by restricted diffusion through the epithelium. The relative contributions of these three pathways to clearance of large quantities of protein from the air spaces is not known. This study investigated the effects of monensin and nocodazole, agents which inhibit endocytosis in cell culture, on alveolar epithelial protein transport in anesthetized rabbits. There was evidence that monensin and nocodazole inhibited endocytosis by the alveolar epithelium in vivo. Nocodazole increased the number of vesicles in the alveolar epithelium and capillary endothelium. Monensin increased vesicle density in the endothelium. These results suggested that the inhibitors disrupted microtubules or interrupted cellular membrane traffic in the lung. Both inhibitors decreased lung parenchymal uptake of immunoreactive human albumin from the air spaces. Monensin and nocodazole inhibited albumin uptake in cultured alveolar type II cells. Monensin increased the amount of 125I-labeled surfactant protein A associated with the lungs, compared with the quantity remaining in the air space 2 h after instillation. Although the drugs decreased alveolar epithelial protein uptake, they did not decrease alveolar clearance of 125I-labeled immunoglobulin G or 131I-labeled albumin in anesthetized rabbits. Thus monensin- and nocodazole-sensitive protein-uptake pathways do not account for most alveolar protein clearance when the distal air spaces are filled with a protein solution.

Animals↗
PubMed1994

Transforming growth factor-alpha enhances alveolar epithelial cell repair in a new in vitro model.

Alveolar epithelial type II cells are essential for regenerating an intact alveolar barrier after destruction of type I cells in vivo. The first objective of these experimental studies was to develop an in vitro model to quantify alveolar epithelial cell wound repair. The second objective was to investigate mechanisms of alveolar epithelial cell wound healing by studying the effects of serum and transforming growth factor-alpha (TGF-alpha) on wound closure. Primary cultures of rat alveolar type II cells were prepared by standard methods and grown to form confluent monolayers in 48 h. Then a wound was made by denuding an area (mean initial area of 2.1 +/- 0.6 mm2) of the monolayer. Re-epithelialization of the denuded area over time in the presence or absence of serum was measured using quantitative measurements from time-lapse video microscopy. The half time of wound healing was significantly enhanced in the presence of serum compared with serum-free conditions (2.4 +/- 0.2 vs. 17.4 +/- 0.8 h, P < 0.001). We then tested the hypothesis that TGF-alpha is an important growth factor for stimulating wound repair of alveolar epithelial cells. Exogenous addition of TGF-alpha in serum-free medium resulted in a significantly more rapid wound closure, and, furthermore, the addition of a monoclonal antibody to TGF-alpha in the presence of serum significantly decreased fourfold the rate of wound closure. Measurement of internuclear cell distance confirmed that both cell motility and cell spreading were responsible for closure of the wound. These data demonstrate that 1) the mechanisms of alveolar cell repair can be studied in vitro and that 2) TGF-alpha is a potent growth factor that enhances in vitro alveolar epithelial cell wound closure.

Animals↗
PubMed1994

Alveolar liquid and protein clearance in anesthetized ventilated rats.

Alveolar and lung liquid clearances were studied over 1, 4, and 6 h in intact anesthetized ventilated rats by instillation of 5% albumin solution with 1.5 microCi of 125I-labeled albumin (3 ml/kg into 1 lung or 6 ml/kg into both lungs). Alveolar protein clearance as measured by residual 125I-albumin in the lung over 6 h was similar to the slow rates measured in other species. Alveolar liquid clearance was estimated by the concentration of albumin in the air spaces. After 1 h, this concentration was 7.8 +/- 0.7 g/dl, which was significantly greater than the initial protein concentration of 5.3 +/- 0.2 g/dl (P < 0.05). Amiloride (10(-3) M) inhibited 45% of the basal alveolar liquid clearance, and ouabain (10(-3) M), instilled and intravenously infused (0.004 mg), inhibited 30% of the clearance. beta-Adrenergic agonist instillation increased alveolar liquid clearance to the fastest 1-h rate (48 +/- 3% of instilled volume) that we observed in any intact species. The removal of the instilled fluid from the lung (expressed as lung liquid clearance; 0.96 +/- 0.3 ml/h) was twice as fast as the rate of alveolar and lung liquid clearance reported in the isolated or in situ rat lung models. The rate of alveolar and lung liquid clearance in these intact rats was significantly faster than those in prior studies in dogs and sheep and was similar to the rates in rabbits.

Adrenergic beta-Agonists↗
PubMed1994

Alveolar fluid clearance in the resected human lung.

Although the mechanisms responsible for alveolar liquid clearance have been studied in several species, there has not been any information regarding the effect of ion transport agonists or antagonists on alveolar liquid clearance in the human lung. Therefore, we studied alveolar liquid clearance in the recently resected human lung from patients who underwent surgery for lung cancer. A test solution of 40 ml of isosmolar albumin solution was instilled into one segment of a resected lobe within 10 min of resection. Because protein leaves the air spaces very slowly, the concentration of alveolar protein over 4 h was used to quantify alveolar liquid clearance. Basal alveolar liquid clearance was 12 +/- 2% over 4 h. Amiloride (10(-5) M), an inhibitor of apical Na+ uptake, and ouabain (10(-3) M), an inhibitor of Na,K-ATPase activity, reduced alveolar liquid clearance by 40 and 49%, respectively (p < 0.005). Terbutaline (10(-3) or 10(-4) M) doubled alveolar liquid clearance to 28 +/- 9% over 4 h (p < 0.05). Propranolol (10(-4) M) and amiloride (10(-5) M) inhibited the terbutaline-induced increase in alveolar liquid clearance. In conclusion, (1) alveolar liquid clearance in the human lung can be markedly reduced by inhibition of apical sodium channel uptake or Na,K-ATPase activity, and (2) beta-adrenergic stimulation markedly increases the rate of alveolar liquid clearance in the resected human lung without pulmonary perfusion.

Aged↗
PubMed1994

The effect of salt water on alveolar epithelial barrier function.

The effect of hyperosmolar fluid aspiration (seawater) on lung fluid balance has not been well studied. Therefore, the effect of this clinically relevant form of acute lung injury on the alveolar epithelial and lung endothelial barriers was examined in ventilated, anesthetized rabbits. Seawater (4 ml/kg body weight, 881 +/- 29 mOsm/kg) with 3 microCi of 125I-albumin was instilled into the lower trachea of ventilated, anesthetized rabbits. Osmotic equilibration with plasma was completed within the first 5 min after seawater instillation. In parallel with the osmotic equilibration of the seawater in the air spaces, there was a 3-fold dilution of the alveolar protein tracer 125I-albumin, indicating an initial large (300%) increase in alveolar fluid volume. There was a marked decline in arterial oxygenation at the same time that the alveolar fluid volume markedly increased. The initial dilution of the alveolar protein tracer was followed by a progressive increase in the alveolar protein tracer concentration that continued until 6 h after seawater instillation. As the alveolar protein tracer concentrated, arterial oxygenation improved, indicting net alveolar liquid clearance. There was only a mild increase in the epithelial and endothelial permeability to protein within the first 2 h after seawater instillation. Thus, a large osmotically induced increase in alveolar fluid volume with severe pulmonary edema did not cause sustained injury to the endothelial or epithelial barriers of the lung. In fact, normal alveolar liquid clearance occurred, indicating the resistance of the epithelial barrier to hyperosmolar injury as well as its capacity to rapidly reabsorb excess alveolar fluid.

Analysis of Variance↗
PubMed1994

Stimulation of lung epithelial liquid clearance by endogenous release of catecholamines in septic shock in anesthetized rats.

Exogenous administration of beta-adrenergic agonists has previously been reported to increase lung liquid clearance by stimulation of active sodium transport across the alveolar epithelium. We hypothesized for this study that endogenous release of epinephrine in septic shock would stimulate liquid clearance from the airspaces in rats. Liquid clearance from the air spaces was measured by the concentration of protein over 4 h in a test solution of 5% albumin instilled into one lung. Bacteremic rats developed severe systemic hypotension and metabolic acidosis that was associated with a 100-fold rise in plasma epinephrine levels. There was a 100% increase in liquid clearance from the airspaces of the lung in the bacteremic compared with control rats. To determine the mechanisms responsible for this accelerated lung liquid clearance, amiloride (10(-3) M), a sodium transport inhibitor, was added to the air spaces. Amiloride prevented the increase in liquid clearance from the airspaces, indicating that this effect depended on increased uptake of sodium across the lung epithelium. The addition of propranolol (10(-4) or 10(-5) M) to the instillate also prevented the acceleration in alveolar liquid clearance in the bacteremic rats. We conclude that the release of endogenous catecholamines associated with septic shock markedly stimulates fluid clearance from the distal airspaces of the lung by a beta-adrenergic mediated stimulation of active sodium transport across the epithelial barrier. This data provides evidence for a previously unrecognized mechanism that can protect against or hasten the resolution of alveolar edema in pathological conditions, such as septic shock, that are associated with the endogenous release of catecholamines.

Amiloride↗
PubMed1993

Soluble E-selectin is found in supernatants of activated endothelial cells and is elevated in the serum of patients with septic shock.

A quantitative sandwich ELISA for E-selectin in the fluid phase (soluble E-selectin, sEs) has been developed that is sensitive to 100 pg/ml. The assay shows no reactivity with either L- or P-selectins. We have used this to determine the fate of E-selectin after cell-surface expression and to test whether levels measured in vivo may represent the state of endothelial activation. E-selectin was first detectable in supernatants of IL-1-stimulated endothelial cells at 24 h, and increased slowly up until 72 h. However, over this time period the total E-selectin detectable in the system (cells plus supernatants) declined dramatically. 125I-surface-labeled endothelial cells cultured for 24 h show an E-selectin of reduced m.w. in the supernatant, indicating that the molecule is shed from the surface. The shed form also appears to be slightly smaller than the intact membrane form as determined from immunoprecipitation and molecular sieving studies. In addition, the cytoplasmic domain of the molecule found in supernatants of activated endothelial cells and in serum is not intact as determined by loss of reactivity with an antipeptide antibody specific for the cytoplasmic domain. We have examined the sera of 71 normal individuals. Without exception, sEs was found in serum in the range of 0.13 to 2.8 ng/ml, suggesting that even in the absence of overt inflammatory processes E-selectin is being synthesized and released into the bloodstream. In addition, bacteremic patients with hypotension, but not those without, showed markedly elevated sEs values. As determined by cell-binding studies, the blood-derived form of E-selectin is biologically active.

Adult↗
PubMed1993

Neutrophil-activating peptide-2 in patients with pulmonary edema from congestive heart failure or ARDS.

We carried out studies to determine whether the neutrophil-activation peptide-2 (NAP-2) plays a role in the recruitment and/or degranulation of neutrophils into the lungs of patients with the adult respiratory distress syndrome (ARDS) or congestive heart failure (CHF). NAP-2 precursors plus NAP-2 (beta-thromboglobulin-like antigen) were measured in lung fluids and plasmas with a radioimmunoassay, and NAP-2 was separated from its precursors by high-performance liquid chromatography. Pulmonary edema fluids (PEFs) from patients with CHF contained higher concentrations of the beta-thromboglobulin-like antigen than PEFs from patients with ARDS, and bronchoalveolar lavage fluids (BALs) from patients with ARDS contained higher concentrations of beta-thromboglobulin-like antigen than BALs from normal subjects. beta-Thromboglobulin-like antigen concentration was 4.1-fold greater in PEFs from patients with CHF than in their plasmas. Chemotactically active NAP-2 was also demonstrated in PEFs but not in plasmas from patients with CHF and ARDS. These data suggest that significant platelet degranulation occurred into the lungs of the patients with CHF and that NAP-2 and other platelet constituents may contribute to fluid formation in patients with CHF.

Bronchoalveolar Lavage Fluid↗
PubMed1993

Alveolar liquid and protein clearance in the absence of blood flow or ventilation in sheep.

The primary objective of these studies was to test the contribution of ventilation and blood flow to the removal of excess liquid from the air spaces and interstitium of the lung. First, after eliminating ventilation by clamping the left main bronchus in anesthetized sheep, alveolar and lung liquid clearance was not altered over 4 h compared with control sheep that were ventilated normally. Thus, removal of excess liquid across the alveolar epithelium was independent of the change in the transalveolar hydrostatic pressure gradient produced by ventilation. Second, to determine the effect of removing all blood flow to the lung, we developed a new in situ sheep lung model in which lung lymph flow was measured over 4 h with or without ventilation after the sheep had been exsanguinated. Alveolar liquid clearance, as measured by the percent increase in alveolar protein concentration over 4 h, was similar between sheep without blood flow (31 +/- 18%) compared with sheep with normal blood flow to the lungs (31 +/- 17%). Lung lymph flow contributed to only 10-15% of the clearance of the excess alveolar liquid that was transported to the interstitium, indicating that nonlymphatic pathways accounted for most of the excess lung liquid clearance in the absence of microvascular filtration. Third, because ouabain completely inhibited alveolar liquid clearance in this in situ sheep lung model, these data provide evidence that alveolar liquid clearance depends on an intact Na(+)-K(+)-ATPase-dependent pump mechanisms. Finally, this in situ model represents a unique experimental preparation that can be used to study the alveolar epithelial barrier without blood flow or ventilation for a short time (4 h) interval.

Anesthesia↗
PubMed1993

Effect of temperature on alveolar liquid and protein clearance in an in situ perfused goat lung.

To study the mechanisms of alveolar liquid and protein clearance, we investigated the effect of four different temperatures (7, 18, 30, and 38 degrees C) on the rate of liquid, [14C]mannitol, and 125I-labeled albumin (125I-albumin) transport from the air spaces of isolated perfused in situ goat lungs over 4 h under isogravimetric conditions. The concentrations of both native proteins and 125I-albumin in the alveolar fluid after 4 h were used to estimate alveolar liquid clearance. We instilled autologous serum (2 ml/kg) with 125I-albumin and [14C]mannitol into the distal air spaces of one lung and 131I-labeled albumin into the perfusate to measure the capillary-to-alveolar permeability to protein. Alveolar liquid clearance progressively declined from a control value of 24 +/- 6% at 38 degrees C to 12 +/- 7% at 30 degrees C, to 3 +/- 3% at 18 degrees C, and to -15 +/- 11% at 7 degrees C. 125I-albumin clearance into the perfusate was 0.3%/h of the instilled 125I-albumin at 38 degrees C. The clearance decreased abruptly to 0.06%/h at 30 degrees C and then slightly increased to 0.08%/h at 18 degrees C and to 0.15%/h at 7 degrees C. There were no changes in transport except for a twofold increase in mannitol at 7 degrees C. Increased permeability of the alveolar barrier to protein and small-molecular-weight tracers at 7 degrees C indicates that the barrier function of the alveolar epithelium is disrupted at this low temperature.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

Alveolar epithelial injury and pleural empyema in acute P. aeruginosa pneumonia in anesthetized rabbits.

We developed an experimental model of acute Pseudomonas aeruginosa pneumonia in anesthetized ventilated rabbits to determine whether bacterial-induced injury to the alveolar epithelium would occur and the effect of the injury on the pleural space. Dose-response studies established that 10(9) colony-forming units of P. aeruginosa (wild-type strain, PAO-1) were required to injure the epithelial barrier and to cause pleural empyema with exudative pleural effusions that contained both the instilled alveolar protein tracer and P. aeruginosa. We explored the mechanisms of P. aeruginosa-induced lung and pleural injury by using three isogenic bacterial strains to compare several extracellular virulence products. PAO-S21, which carries an insertion mutation in a regulatory gene that prevents the production of exoenzyme S, resulted in no lung or pleural injury. PAO-R1, which carries a deletion in a regulatory gene that controls the production of elastase and alkaline protease, caused the same degree of lung and pleural injury as PAO-1 did. Instillation of PLC-SRN, which has both structural genes encoding phospholipase C activity deleted, resulted in a moderate reduction in alveolar epithelial injury. Although other products may be involved, exoenzyme S and phospholipase C are important in mediating injury to the alveolar epithelial barrier in acute P. aeruginosa pneumonia in rabbits.

Anesthesia↗