Figures and data
Lung exudate extraction procedure and visualization.
A) Schematics of organic extraction to obtain the lipidic phase in the form of a dry film after solvent evaporation and subsequent lipid self-assembly upon rehydration of the dry film in a capillary tube. B) Laser confocal fluorescence microscopy images of fluorescently labeled pulmonary lipid extracts in water. C) Transmission electron microscopy images of BAL lung extracts from a dolphin with clinically healthy lungs exhibiting multivesicular structures (MVS) and tubular structures.
Lipidomic analysis.
A) Principal linear discriminant analysis of lipids from lung exudates. Replicates are represented by similar symbols. The values for components 1, 2, and 3 are shown. B) Top 30 species that discriminate between groups and account for variation within component 1. Cardiolipin species are indicated by red triangles. C) Cardiolipin abundances depicted as a Z-score. D) Cardiolipin species analysis in samples from acute/chronic pulmonary infection. Cardiolipin fatty acyl-chain length is shown on the x-axis, and cardiolipin fatty acyl unsaturation level (double bonds) is shown on the y-axis.
Categories assigned to Tursiops truncatus lung exudates with corresponding number of animal samples (n)
Microscopy analysis of lung extracts and lung tissue.
A) AFM topography maps of multilayered lung fluid extracts from dolphins with no disease and with lung disease. B) One-way ANOVA analysis of surface coverage with a significant p-value. C) Oneway ANOVA analysis of domain circularity by lung disease condition, resulting from topography map analysis with a significant p-value. The number of domains analyzed is shown.
Microscopy analysis of lung extracts and lung tissue.
A) AFM adhesion maps of multilayered lung fluid extracts from dolphins with no disease and with lung disease. B) One-way ANOVA analysis of cardiolipin abundance by lung disease condition. Cardiolipins’ abundance values correspond to the normalized signal counts to internal standards and sample weight. C) Bivariate fit of Young modulus by cardiolipin abundance. Linear fit and corresponding equation are shown. Elasticity means of lung extracts were calculated from fast force mapping.
Structural characterization of pulmonary membrane complexes.
SAXS (A) and WAXS (B) Intensity vs q data. C) One-way ANOVA analysis of lattice parameter d response to lung disease condition. d-spacing was calculated from SAXS peak positions in A. D) Cryo-EM data where scale bars correspond to 100 nm. The Cryo-EM images depict lung extracts from dolphins with chronic or acute lung disease (left) and dolphins with no lung disease (right). Nanodomain size is highlighted in blue (> 5 nm) and in orange (< 5 nm), and was estimated with rim analysis.
