Medicine of the Future in America

Oxidative Changes of Bronchoalveolar Proteins in Cystic Fibrosis: Methods

A more relevant measure of oxidative stress in the lungs may be derived from assessments of samples containing epithelial lining fluid of the distal airways including the alveoli as obtained by BAL. Such measurements have been reported only in young CF patients.
The goal of this study was to investigate protein oxidation in CF patients with a wide range of endobronchial inflammation in relation to their lung function and degree of pulmonary inflammation. We hypothesized that the level of protein oxidation may be increased even in the presence of normal pulmonary function tests. Our findings demonstrated a close relation between the degree of protein oxidation, neutrophil granulocyte (PMN) count in BAL fluid (BALF), and lung function of the subjects, suggesting a causal relationship among these variables, BALF, which was available as a cell-free supernatant, was analyzed from 55 patients with CF. These patients had participated in the Bronchoalveolar Lavage for the Evaluation of Antiinflammatory Treatment study and the Glutathione study in the Munich, Cologne, and Berlin study centers.1112 The clinical characteristics of the patients are given in Table 1. canadian health care mall

Protease inhibitors were added, and aliquots of the BALF were stored at — 80°C. Preliminary work demonstrated that one or two thawing and refreezing cycles had no significant impact on protein carbonylation in the samples (data not shown). BALF from 11 healthy subjects (Table 1) was obtained in a study13 on vaccine development prior to the intervention and from healthy children who participated in a study on assessing bronchial inflammation by tracheostomy. BAL was performed with similar methodologies in the right middle lobe with a flexible bronchoscope as described previously. The study protocols were approved by the institutional review boards, and written informed consent was obtained from adult subjects and parents of the children before the study.

Table 1—Clinical Characteristics of the Patients

Characteristics CF, Median (Range) Control, Median (Range) p Value
Subjects, No. 55 11
Age, yr 15.3 (5.2-36.7) 24.0 (9-27) NS
Body mass index, kg/m2 18.8 (13.9-33.6)
FEV1, % of predicted 87.4 (36.0-125.3)
FVC, % of predicted 89.1 (59.7-127.5)
MEF25-75, % of predicted 64.3 (9.0-142.1)
Pseudomonas aeruginosa 19/53
Staphylococcus aureus 21/53
BALF recovery, % 50 (83.0-7.5) 61.3(75.0-42.0) NS
Total cells in BAL, X 104/mL 30 (0.02-974) 1.67 (0.59-3.25) 0.002
% PMNs 51.0 (0.2-96) 0.95 (0.5-2.7) 0.002
% Macrophages 45.0 (1.6-97) 93.9 (88.2-97.5) 0.0007
% Lymphocytes 4 (0-39.1) 4.1 (0.7-10.5) NS
% Eosinophils 0.4 (0-15) 0.65 (0.0-1.0) NS
This entry was posted in Cystic Fibrosis and tagged cystic fibrosis, lung disease, protein carbonyls.
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