'Free' surfactant in gastric aspirates and bronchoalveolar lavage in children with and without reflux oesophagitis.

AIM: Dipalmitoylphosphatidycholine (DPPC) is the characteristic and main constituent of surfactant. Adsorption of surfactant to epithelial surfaces may be important in the masking of receptors. The aims of the study were to (i) compare the quantity of free DPPC in the airways and gastric aspirates of children with gastroesophageal reflux disease (GORD) to those without and (ii) describe the association between free DPPC levels with airway cellular profile and capsaicin cough sensitivity. METHODS: Children aged <14 years were defined as 'coughers' if a history of cough in association with their GORD symptoms was elicited before gastric aspirates and nonbronchoscopic bronchoalveolar lavage (BAL) were obtained during elective flexible upper gastrointestinal endoscopy. GORD was defined as histological presence of reflux oesophagitis. Spirometry and capsaicin cough-sensitivity test was carried out in children aged >6 years before the endoscopy. RESULTS: Median age of the 68 children was 9 years (interquartile range (IQR) 7.2). Median DPPC level in BAL of children with cough (72.7 microg/mL) was similar to noncoughers (88.5). There was also no significant difference in DPPC levels in both BAL and gastric aspirates of children classified according to presence of GORD. There was no correlation between DPPC levels and cellular counts or capsaicin cough-sensitivity outcome measures. CONCLUSION: We conclude that free DPPC levels in the airways and gastric aspirate is not influenced by presence of cough or GORD defined by histological presence of reflux oesophagitis. Whether quantification of adsorbed surfactant differs in these groups remain unknown. Free DPPC is unlikely to have a role in masking of airway receptors.

Comparison of surfactant lipids between pleural and pulmonary lining fluids.

Saturated phospholipids (PCs), particularly dipalmitoylphosphatidylcholine (DPPC), predominate in surfactant lining the alveoli, although little is known about the relationship between saturated and unsaturated PCs on the outer surface of the lung, the pleura. Seven healthy cats were anesthetized and a bronchoalveolar lavage (BAL) was performed, immediately followed by a pleural lavage (PL). Lipid was extracted from lavage fluid and then analyzed for saturated, primarily dipalmitoylphosphatidylcholine (DPPC), and unsaturated PC species using high-performance liquid chromatography (HPLC) with combined fluorescence and ultraviolet detection. Dilution of epithelial lining fluid (ELF) in lavage fluids was corrected for using the urea method. The concentration of DPPC in BAL fluid (85.3+/-15.7 microg/mL) was significantly higher (P=0.021) than unsaturated PCs ( approximately 40 microg/mL). However, unsaturated PCs ( approximately 34 microg/mL), particularly stearoyl-linoleoyl-phosphatidylcholine (SLPC; 17.4+/-6.8), were significantly higher (P=0.021) than DPPC (4.3+/-1.8 microg/mL) in PL fluid. These results show that unsaturated PCs appear functionally more important in the pleural cavity, which may have implications for surfactant replenishment following pleural disease or thoracic surgery.

Adsorption of surfactant to bronchial epithelium: possible role of receptor 'unmasking' in asthma.

BACKGROUND: In a recent study in animals it has been shown how surface-active phospholipid (SAPL) in the form of a commercially available micronized (5 micromphi) dry powder (ALECT/PumactantT) was able to reduce afferent neural feedback to the brainstem in response to a methacholine challenge by the same order of magnitude as drugs commonly prescribed for asthma. The underlying theory assumed that adsorption of SAPL to bronchial epithelium masked irritant receptors eliciting the bronchoconstrictor reflex, thus providing a barrier to noxious stimuli entering the lungs. OBJECTIVE: To test the underlying assumption that SAPL was actually adsorbed (i.e., bound to bronchial epithelium), especially the major and most surface-active component of lung surfactant, namely dipalmitoyl phosphatidylcholine (DPPC). A secondary objective was to investigate any role of phosphatidylglycerol (PG) in promoting the adsorption of DPPC. METHODS: Radiolabeled DPPC dispersed ultrasonically in saline was used to incubate excised sections of porcine bronchial epithelium. The adsorbed DPPC was then quantified by rigorously rinsing the tissue of adhering fluid and then digesting it for beta-scintillation counting. Each test (n=8 runs) was repeated for ratios of DPPC:PG of 9:1, 7:3 (as per ALECT/PumactantT) and 1:1 for both dipalmitoyl PG (DPPG) and EggPG (as incorporated in ALECT/PumactantT). RESULTS: Despite rigorous rinsing postincubation, bronchial epithelium was found to adsorb DPPC at a level roughly equivalent to one close-packed monolayer; whereas both DPPG and EggPG promoted the adsorption of DPPC in a dose-dependent manner, reaching an approximate threefold increase for 7:3 DPPC:PG. CONCLUSION: DPPC adsorbs to bronchial epithelium in amounts necessary for the masking of receptors, and this adsorption (probably chemisorption) is quite strongly promoted by PG either in its indigenous state (DPPG) or in the form (EggPG) used in ALECT to suppress the sensitivity of bronchial irritant receptors in our previous study and in clinical trials just completed.

Raised bile acid concentrations in SIDS lungs at necropsy.

Previous studies of the abnormal physical properties of lung surfactant derived from infants experiencing prolonged expiratory apnoea, or who have died of sudden infant death syndrome (SIDS), have led to a search for the agent responsible. Bronchoalveolar lavage (BAL) has been performed upon 12 infants under 12 months at necropsy and the rinsings analysed for up to 26 bile acids using high performance liquid chromatography, which requires nanomolar quantities. They were also analysed for simultaneously retrieved phospholipid and proteolipid-a minor component of lung surfactant-as markers of lavage efficiency. Total bile acid (TBA) was found to be higher in six SIDS cases, reaching a mean (SE) 8.54 (2.24) mumol/l in the BAL fluid compared with 4.66 (1.47) mumol/l in the six controls of similar age. When related to the concomitant surfactant yield, the TBA/ proteolipid and TBA/phospholipid ratios both showed highly significant differences between index lungs and controls, providing another postmortem marker of SIDS with potential for development as a test of risk. Since the bile:phospholipid ratio determines whether phospholipase A2 synthesises or hydrolyses phospholipid, the raised TBA/phospholipid ratio could be highly significant, causing this enzyme to function more like its role in the gut than in the lung.

Abnormalities in surfactant in sudden infant death syndrome as a postmortem marker and possible test of risk.

OBJECTIVE: To determine whether physical abnormalities in lung surfactant, particularly inversion of the hysteresis between surface tension and surface area, are an effective postmortem marker for sudden infant death syndrome (SIDS). METHODOLOGY: Bronchoalveolar lavage (BAL) was employed to obtain lung rinsings from 55 infants under 24 months of age at autopsy, comprising 34 index cases and 21 controls. Folch reagent was used to extract the lipoid content which was then applied at two surface concentrations, fixed and BAL-determined (BALD), to the pool of a Langmuir trough in which surface tension (gamma) was measured continuously by the Wilhelmy method as surface area (A) was cycled. RESULTS: The gamma:A loops from SIDS samples were inverted relative to controls (i.e. they were hysteresis reversed, this inversion being quantified by an empirical surface tension: area reversal (STAR) score). There was a wide scatter of STAR scores, but a critical value was found which offered a significant (P = 0.017) separation of SIDS cases from controls for a fixed surface concentration and a highly significant separation (P = 1.0 x 10(-4)) for BALD surface concentrations. Differences in the yields of phospholipid and proteolipid, or their correlation to STAR scores, did not reach statistical significance. CONCLUSIONS: Inversion of the gamma :A loops (i.e. hysteresis inversion) would appear to offer a better postmortem marker of SIDS than any reported previously, the procedure having potential for development as a prospective test indicating the risk of this disease.