PMC3534390_01
Chest radiograph on admission.. Notes: Large consolidations and multiple lobular shadows were seen in the upper and middle fields of the right lung. The left lung was operated and collapsed due to old tuberculosis.
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A chest radiograph revealed right upper and lower lobe consolidation with collapse of the left chest wall caused by his previous thoracoplasty (Figure 1).
Formation of NETs in the sputum of this patient. (A) 4',6-diamidino-2-phenylindole staining at high magnification (1000x). (B) Immunohistochemistry against anti-human histone-H3 mouse monoclonal antibody. (C) Immunohistochemistry against anti-human neutrophil elastase rabbit polyclonal antibody. (D) Merged photograph of (A-C). These pictures indicate that the fibrous constituents are NETs. (E) Gram-staining of endotracheal-aspirated sputum at low magnification (1000x).. Note: Arrow indicates numerous fibrous constituents.. Abbreviation: NETs, neutrophil extracellular traps.
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Gram staining of endotracheal-aspirated sputum revealed a large number of neutrophils, and light microscopy at high magnification (1000x) showed numerous fibrous constituents (Figure 2A-E). The same specimens were analyzed using immunohistochemistry with DAPI (4',6-diamidino-2-phenylindole) staining (Invitrogen, Carlsbad, CA) for DNA detection (Figure 2A), anti-human histone H3 mouse monoclonal antibody (MABI0001, MAB Institute Inc, Sapporo, Japan) for histone (Figure 2B), and anti-human neutrophil elastase rabbit polyclonal antibody (GTX72042, GeneTex Inc, Irvine, CA) for neutrophil elastase (Figure 2C). Figure 2D is a merged photograph of Figure 2A-C.
Gram-staining of endotracheal-aspirated sputum.. Notes: A large number of Gram-negative coccobacilli (black arrowheads), including a proportion phagocytosed by neutrophils (white arrowheads) and numerous fibrous NETs (arrows) are observed at higher magnification (1000x).. Abbreviation: NETs, neutrophil extracellular traps.
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A number of Gram-negative coccobacilli, some of which had been phagocytosed by neutrophils, were observed at high magnification (1000x, Figure 3).