Subject(s)
Airway Obstruction/metabolism , Antioxidants/analysis , Bronchoalveolar Lavage Fluid/chemistry , Oxidative Stress , Sputum/cytology , Airway Obstruction/pathology , Ascorbic Acid/analysis , Glutathione/analysis , Humans , Lung Neoplasms/metabolism , Lung Neoplasms/pathology , Nitrites/analysis , Proteins/analysis , Regression Analysis , Uric Acid/analysisABSTRACT
Proteomics is the large-scale analysis of protein profiles. This approach has not yet been reported in the study of acute lung injury (ALI). This study details protein profiles in plasma and pulmonary edema fluid (EF) from 16 ALI patients and plasma and bronchoalveolar lavage fluid (BALF) from 12 normal subjects. More than 300 distinct protein spots were evident in the EF and BALF of both normal subjects and ALI patients. Of these, 158 were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. In the plasma and EF protein profile of ALI patients, there were multiple qualitative changes. For instance, in all normal subjects, but in only one of the ALI patients, seven distinct surfactant protein A isoforms were evident. Nearly all ALI patients also had protein spots that indicate truncation or other posttranslational modifications. Several of these novel changes could serve as new biomarkers of lung injury.
Subject(s)
Proteomics , Pulmonary Edema/metabolism , Respiratory Distress Syndrome/metabolism , Adult , Blood Proteins/analysis , Blood Proteins/metabolism , Bronchoalveolar Lavage Fluid , Electrophoresis, Gel, Two-Dimensional , Extravascular Lung Water/metabolism , Female , Humans , Male , Middle Aged , ProteomeABSTRACT
The usual methods for preparing lungs for morphologic study involve the instillation of fixatives that modifyproteins and RNA such that the tissue is unsuitable for molecular studies. To develop a technique suitable for molecular studies, pieces of adult rat lungs were infiltrated with agarose, glutaraldehyde, or paraformaldehyde and the consistency of alveolar inflation was compared to lungs inflated with 10% formalin. Only direct injection with 1% agarose resulted in comparable inflation of lung tissue and preserved RNA and protein. Thus, this technique enables simultaneous molecular and morphometric analysis of the lung on small pieces of lung tissue in heterogeneous lung diseases.
Subject(s)
Lung , Organ Preservation Solutions , Organ Preservation/methods , Sepharose , Tissue Fixation/methods , Animals , Formaldehyde , Glutaral , Image Processing, Computer-Assisted , Immunohistochemistry , Injections , Male , Polymers , Proteins/analysis , Pulmonary Alveoli/anatomy & histology , Pulmonary Alveoli/chemistry , RNA/analysis , Rats , Rats, Sprague-DawleyABSTRACT
OBJECTIVE: To test the hypothesis that low concentrations of distal airspace water-soluble antioxidants are associated with acute lung injury. DESIGN: Prospective, cohort study. SETTING: Medical intensive care unit of two tertiary care hospitals. SUBJECTS: Subjects were 29 patients with acute lung injury and 23 normal, healthy, volunteers. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Pulmonary edema fluid from subjects with acute lung injury was aspirated immediately after intubation. Compared with the bronchoalveolar lavage from normal subjects (corrected for dilution using urea concentrations), undiluted edema fluid from acute lung injury subjects had significantly lower concentrations of the antioxidants urate (757 +/- 232 microM vs. 328 +/- 75 microM), glutathione (138 +/- 25 microM vs. 7 +/- 4 microM), and ascorbate (85 +/- 21 microM vs. 27 +/- 10 microM). CONCLUSIONS: Acute lung injury is associated with decreased concentrations of water-soluble antioxidants in the distal airspaces. In acute lung injury, the distal airspace antioxidants ascorbate, urate, and glutathione may play a role in attenuating lung injury.
