Figure 4

(a, b) Leukocyte numbers/300 epithelial cells (a); TUNEL positive cells (b). No higher incidence of cell death or leukocyte influx in bronchial epithelial cells was observed after inhalation of sphingosine. Hemalaun staining was performed for paraffin sections from bronchial biopsies from EVLP minipig lungs that were inhaled with 125 µM sphingosine (SPH), 500 µM sphingosine (SPH) or with octylglucopyranoside (OGP) as control. Three sections with 5 visual fields were analyzed for each lung and leukocyte numbers per 300 epithelial cells were counted. TUNEL reaction was performed for paraffin sections from bronchial biopsies from EVLP minipig lungs that were inhaled with 125 µM sphingosine (SPH), 500 µM sphingosine (SPH) or with octylglucopyranoside (OGP) as control. Statistical analysis of leukocyte influx and TUNEL positive cells after inhalation of octylglucopyranoside (OGP), 125 µM (125 µM SPH) or 500 µM (500 µM SPH) sphingosine containing 5 ml suspension did not reveal a statistically significant difference. Given are numbers of leukocytes/300 epithelial cells (n) and percentage of TUNEL positive cells from 3 sections with 5 visual fields per animal (blinded tests), Given are mean ± SD, ANOVA.