Lung function tests, methacholine challenge tests and analysis
of eicosanoids
The lung function tests were performed with the spirometer-Lungest 1000
(MES, Krakow, Poland) according to ERS/ATS14standards. These children had not received any anti-asthmatic
medication, including oral steroids, for at least two days.
Utilizing established solid phase extraction approach for the collection
and purification of eicosanoids in EBCs, a panel of 6 eicosanoid species
derived from arachidonic acids, representing products from major
enzymatic pathways, was selected for initial discovery phase of the
study population consisting of 60 asthmatics and 20 healthy controls,
who were randomly selected from among the study populations. Eicosanoids
were extracted from EBCs15 and measured with the
respective enzyme immunoassay kit (Cayman Chemical, Ann Arbor, Michigan
and Neogen, Lexington, Ky) as described previously11.