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[12] Asthma was defined using British Thoracic Society criteria,[13] and exacerbation as a deterioration in symptoms of wheeze, dyspnoea or cough, an increased use of short-acting ��2-agonist medication or a deterioration in self-monitored peak expiratory flow.[14] Pneumonia was defined according to the British Thoracic Society.[15] Controls were studied either on presentation with pneumonia, or with signs and symptoms consistent with a respiratory tract infection. This was a prospective study examining subjects suffering from LY2109761 mouse an exacerbation of obstructive lung disease or, in the case of controls, undergoing symptoms of lower respiratory tract infection, and comparing them with the stable state. Patients were identified from presentations to the Emergency Department or clinics. Subsequently, they were invited to return when clinically stable to repeat investigations. Stability was defined as being at least 6 weeks after their most recent exacerbation and free from symptoms of infection or exacerbation. After history and examination, chest radiograph and venepuncture, blood was taken for full blood count, biochemistry, liver function tests, C-reactive protein and atypical serology. Subjects completed the St George's Respiratory Questionnaire.[16] Spirometry was performed using a MicroPlus spirometer (Micro Direct Inc, Lewiston, ME, USA). EBC and induced sputum were collected according to published recommendations.[17, EPZ5676 solubility dmso 18] A custom-designed curved glass tube was used for EBC. Viral polymerase chain reaction analysis was performed on nasopharyngeal and throat swabs. Sputum was cultured for respiratory pathogens and, if the patient was febrile, blood cultures were sent, as was atypical serology. EBC was immediately tested for pH and both EBC and sputum were processed and analysed for hydrogen peroxide (H2O2), protein, leukotriene B4 (LTB4), interleukin (IL)-6, IL-8, tumour necrosis factor (TNF)-��, IP-10 and neopterin. H2O2 was measured spectrophotometrically by horseradish peroxidase-catalysed oxidation of tetramethylbenzidine; lower limit of detection 0.2?��mol/L. Total protein was measured using a Quantipro BCA assay kit (Sigma Aldrich, Sydney, Australia); lower limit of detection 4?��g/mL. LTB4 was measured by a specific Oxacillin acetylcholinesterase competitive enzyme immunoassay (Cayman Chemical, Ann Arbor, MI, USA); lower limit of detection 4?��g/mL. IL-6 was measured using a specific sandwich enzyme immunoassay (Cayman Chemical); lower limit of detection 7.8?pg/mL. IL-8 was measured using a specific sandwich enzyme immunoassay (Amersham Biosciences, GE Healthcare, Piscataway, NJ, USA); lower limit of detection