Our purpose was to evaluate the effect of the mechanical force of a sneeze on sinonasal cilia function and determine the molecular mechanism responsible for eliciting the ciliary response to a sneeze. A novel model was developed to deliver a stimulation simulating a sneeze (55 mmHg for 50 ms) at 26°C to the apical surface of mouse and human nasal epithelial cells. Ciliary beating was visualized, and changes in ciliary beat frequency (CBF) were determined. To interrogate the molecular cascades driving sneeze-induced changes of CBF, pharmacologic manipulation of intra- and extracellular calcium, purinergic, PKA, and nitric oxide (NO) signaling were performed. CBF rapidly increases by ≥150% in response to a sneeze, which is dependent on the release of adenosine triphosphate (ATP), calcium influx, and PKA activation. Furthermore, apical release of ATP is independent of calcium influx, but calcium influx and subsequent increase in CBF are dependent on the ATP release. Lastly, we observed a blunted ciliary response in surgical specimens derived from patients with chronic rhinosinusitis compared to control patients. Apical ATP release with subsequent calcium mobilization and PKA activation are involved in sinonasal ciliary response to sneezing, which is blunted in patients with upper-airway disease.

Using primary murine nasal and tracheal airway cultures we show inherent differences in cilia function and increased susceptibility of the upper airway to attachment by Pseudomonas. Understanding the differences between upper and subglottic airway mucociliary clearance should lead to novel approaches in the management of upper airway infection.

Objective: Tobacco smoke exposure has been correlated to recalcitrant chronic rhinosinustis. Prior work has demonstrated tobacco smoke mediated sinonasal biofilm formation. Thus our objectives were: 1) Construct a series of reporter constructs for pseudomonas biofilm forming genes. 2) Determine which biofilm forming genes are regulated by tobacco smoke. Method: Promoter regions of 6 genes involved in pseudomonas biofilm formation (LasI, LasR, RhlA, PilF, FlgK, and AlgC) were subcloned into the luciferase reporter plasmid pMini-CTX-Lux. Subcloning was confirmed with PCR with subsequent conjugation into Pseudomonas (PAO1). Each subclone was then challenged with serial tobacco smoke exposure and luciferase activity quantified. Results: Elevated levels of luminescence, indicating specific promoter activation was identified after daily tobacco-smoke exposure in the LasI, PilF, FlgK, and AlgC genes. The promoter activation of these genes was significantly elevated ( P < .05) after 2 consecutive exposures with no additional activation following a third exposure (Kruskall-Wallis non-parametric test). The level of promoter activation of the LasB gene was not affected by tobacco smoke exposure, while the RhlA gene decreased progressively by daily exposure. Conclusion: Utilizing the luciferase reporter construct mLuxI we demonstrate activation of several key biofilm forming genes by exposure to tobacco smoke. This confirms our prior finding that tobacco smoke induces biofilms in sinonasal microbes. Interestingly, not all biofilm pathways interrogated demonstrated activation, suggesting unique tobacco mediated biofilm forming pathways in pseudomonas.