016). When ELF levels were lower than plasma levels there was a significant correlation (r = 0.47, P = 0.001) between plasma and except ELF endostatin at the onset of ALI (data not shown). However, in some patients, calculated ELF levels were actually higher than plasma suggesting intra-alveolar generation of endostatin. Interestingly, at day 4 ELF endostatin correlated with lung injury score (r = 0.499, P = 0.049) but not in ALI patients at day 0.OLV and LPS challenge induce a BAL neutrophilia and increase PPIOLV and LPS challenge induced a BAL neutrophilia and an increase in protein permeability index (PPI) compared with normal controls (PPI in OLV: median = 0.00636, IQR = 0.00389 to 0.01188; P = 0.013 vs normal; PPI in LPS: median = 0.0025, IQR = 0.0013 to 0.003, P = 0.011 versus normal; normal PPI = 0.
0014, IQR = 0.0008 to 0.0024). Percentage of neutrophils in BAL rose from 1.1% (IQR = 0.0 to 2.65) in normal BALF to median 11% (IQR = 2 to 60.5) in OLV (P = 0.011), and 41% (IQR = 24 to 57) in LPS challenge as described previously [22].OLV and LPS challenge increases BALF endostatinInhalation challenge with LPS increased BALF endostatin from a median of 0.08 ng/ml in normal individuals to 0.385 ng/ml (IQR = 0.285 to 0.448, P = 0.001) in LPS-challenged patients. OLV increased BAL endostatin (median compared with normal and LPs challenge; Figure Figure4).4). BALF endostatin correlated with PPI in the LPS challenge (r = 0.63, P = 0.006) but not in OLV patients. In contrast, plasma levels of endostatin were not altered by LPS challenge or OLV (data not shown).
Figure 4BALF endostatin in the different at-risk groups compared with normal individuals. BALF = bronchoalveolar lavage fluid; LPS = lipopolysaccharide; OLV = one lung ventilation.Type XVIII collagen precursors are elevated in both the plasma and BALF of patients with ALI compared with normal controlsIn exploratory experiments performed in five consecutive non-selected patients, plasma and BALF from patients with ALI demonstrated increased density and number of fragments of the circulating precursor forms of type XVIII collagen compared with normal controls. Whole lane densitometric analysis of the western blot by anti-LONG antibody confirmed increased plasma precursors in ALI (mean optical density (OD) = 198, SE 38) compared with normal plasma (mean OD 79.8, SE 7.9, 95% CI = 11.6 to 226, P = 0.
037) (representative gels are shown in Figure Figure5).5). Western blotting of the precipitated proteins with three different antibodies to human collagen XVIII (anti-ALL, anti-LONG and anti-endostatin) revealed a main 120 kDa band, which is presumably derived from the middle variant of collagen, containing the N-terminus and the collagenous Brefeldin_A central part of the molecule but lacking the endostatin domain (Figure (Figure55 and data not shown).
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