IL-5 was measured using a specific enzyme linked immunoassay (BioSource, Camarillo, CA).
8-isoprostane concentrations were measured in duplicate using a specific enzyme immunoassay kit (Cayman Chemicals, Ann Arbor, MI, USA).
Second strand cDNA was synthesized by DNA pol I and RNase H (Fermentas), and the poly(A) tail was removed using a specific enzyme (Gsul).
The assay measured only intact human osteocalcin and was performed on human osteoblast-conditioned media using a specific enzyme immunoassay (EIA) kit with a sensitivity of 0.5 ng/ml (Biomedical Technologies Inc., Stoughton, MA, USA).
Fasting serum Pepsinogen I was measured in all patients by radioimmunoassay (Pepsik; Sorin Biomedica, Saluggia, Italy) according to the manufacturer's instructions, while basal Pepsinogen II levels were determined using a specific enzyme immunoassay (BIOHIT Plc, Helsinki, Finland).
Tail vein blood was collected from the nu/nu mice in each group at the different time points and the PAF concentrations were measured using a specific enzyme immunoassay kit (Groundwork Biotechnology Diagnosticate, San Diego, CA, USA).
Soluble TNFα levels were determined using a specific enzyme-linked immunosorbent assay (ELISA) kit according to the manufacturer's instructions (BD Biosciences).
At the end of the incubation period, culture medium was collected and MMP-13 levels were determined using a specific enzyme-linked immunosorbent assay (ELISA).
By using a specific enzyme-linked immunosorbent assay (ELISA) test (R&D Systems, Abingdon, United Kingdom), we evaluated SPARC levels in the serum of patients both at diagnosis and at different time points during treatment with IM.
PGE2 release was determined using a specific PGE2 enzyme immunoassay (Cayman Chemical, Ann Arbor, Michigan, USA) according to the manufacturer's instructions.
