Exhaled and non-exhaled non-invasive markers for assessment of respiratory inflammation in patients with stable COPD and healthy smokers.

We aimed at comparing exhaled and non-exhaled non-invasive markers of respiratory inflammation in patients with chronic obstructive pulmonary disease (COPD) and healthy subjects and define their relationships with smoking habit. Forty-eight patients with stable COPD who were ex-smokers, 17 patients with stable COPD who were current smokers, 12 healthy current smokers and 12 healthy ex-smokers were included in a cross-sectional, observational study. Inflammatory outcomes, including prostaglandin (PG) E2 and 15-F2t-isoprostane (15-F2t-IsoP) concentrations in exhaled breath condensate (EBC) and sputum supernatants, fraction of exhaled nitric oxide (FENO) and sputum cell counts, and functional (spirometry) outcomes were measured. Sputum PGE2 was elevated in both groups of smokers compared with ex-smoker counterpart (COPD: P < 0.02; healthy subjects: P < 0.03), whereas EBC PGE2 was elevated in current (P = 0.0065) and ex-smokers with COPD (P = 0.0029) versus healthy ex-smokers. EBC 15-F2t-IsoP, a marker of oxidative stress, was increased in current and ex-smokers with COPD (P < 0.0001 for both) compared with healthy ex-smokers, whereas urinary 15-F2t-IsoP was elevated in both smoker groups (COPD: P < 0.01; healthy subjects: P < 0.02) versus healthy ex-smokers. FENO was elevated in ex-smokers with COPD versus smoker groups (P = 0.0001 for both). These data suggest that the biological meaning of these inflammatory markers depends on type of marker and biological matrix in which is measured. An approach combining different types of outcomes can be used for assessing respiratory inflammation in patients with COPD. Large studies are required to establish the clinical utility of this strategy.

A fast one-step extraction and UPLC-MS/MS analysis for E2/D 2 series prostaglandins and isoprostanes.

Prostaglandins (PG) and isoprostanes (iso-PG) may be derived through cyclooxygenase or free radical pathways and are important signaling molecules that are also robust biomarkers of oxidative stress. Their quantification is important for understanding many biological processes where PG, iso-PG, or oxidative stress are involved. One of the common methods for PG and iso-PG quantifications is LC-MS/MS that allows a highly selective, sensitive, simultaneous analysis for prostanoids without derivatization. However, the currently used LC-MS/MS methods require a multi-step extraction and a long (within an hour) LC separation to achieve simultaneous separation and analysis of the major iso-PG. The developed and validated for brain tissue analysis one-step extraction protocol and UPLC-MS/MS method significantly increases the recovery of the PG extraction up to 95 %, and allows for a much faster (within 4 min) major iso-PGE2 and -PGD2 separation with 5 times narrower chromatographic peaks as compared to previously used methods. In addition, it decreases the time and cost of analysis due to the one-step extraction approach performed in disposable centrifuge tubes. All together, this significantly increases the sensitivity, and the time and cost efficiency of the PG and iso-PG analysis.

A sensitive direct ELISA for detection of prostaglandin E2.

In order to improve the indirect ELISA for detection of PGE(2), a modified direct ELISA technique was developed to measure PGE(2) in cell culture supernatants. An evaluation of three types of coating buffer showed that PGE(2) was adsorbed efficiently to the solid phase using the gelatin phosphate buffer. The sensitivity of the assay was increased by employing polyclonal rabbit anti-PGE(2) antibody dilution of 1/100 and 1% skimmed milk as a blocking solution, with the detection limit of 7.8-500 ng/well. The within-run and between-run coefficients of variation (CV) ranges were 3.2-3.7% and 3.4-3.8%, respectively. A linear standard curve was observed over the range of 0.078-5 microg/mL with a coefficient of determination (r(2)) of 0.99. Our results indicated that the developed direct ELISA was sensitive and suitable for a quick determination of PGE(2) levels from cell culture supernatants.

Detection of implant crevicular fluid prostaglandin E2 levels for the assessment of peri-implant health: a pilot study.

Inflammatory changes in the peri-implant tissues may lead to peri-implantitis and bone loss. Prostaglandin E2 has been shown to have proinflammatory effects on peri-implant tissues, including mediation of bone resorption. The aim of this study was to assess prostaglandin E2 levels in implant crevicular fluid and the possibility of using this method in diagnosing peri-implant mucositis. Twenty-four dental implants with 3 mm or greater probing depths comprised the test group and another 24 implants with probing depths less than 3 mm served as the control group. Plaque index (PI), gingival index (GI), and probing pocket depths (PPD) were recorded. Implant crevicular fluid was obtained by collection onto periopapers. Then, prostaglandin E2 levels were evaluated using a commercially available enzyme immuno-assay kit. PI, GI, PPD, and implant crevicular fluid (ICF) levels of prostaglandin E2 were found to be statistically significantly higher in the test group (P < 0.05). In the test group, gingival index and probing depths were found to be statistically significantly related with ICF prostaglandin E2 levels (P < 0.05). In the control group, there was no statistically significant positive correlation between clinical parameters and ICF prostaglandin E2 levels (P > 0.05). It may be speculated that biochemical tests, such as the detection of prostaglandin E2 levels in the crevicular fluid are useful diagnostic methods for the maintenance of functional dental implants.

Effects of soluble interleukin-1 type II receptor on rabbit antigen-induced arthritis: clinical, biochemical and histological assessment.

OBJECTIVES: To investigate the effects of soluble interleukin-1 (IL-1) type II receptor (sIL-1RII) on a number of clinical, biochemical and histological parameters in rabbit antigen-induced arthritis. METHODS: Arthritis was induced by intra-articular injection of methylated bovine serum albumin (mBSA) into rabbits pre-sensitized to the same antigen. An initial i.v. bolus of sIL-1RII was administered, followed by s.c. mini-pump dosing for 14 days, starting at the time of the arthritis induction. Animals received vehicle (saline 500 microl + 5 microl/h), low-dose sIL-1RII (13.4 microg + 1.34 microg/h) or high-dose sIL-1RII (40.2 microg + 4.02 microg/h). RESULTS: Marked, dose-related inhibition of joint diameter, plasma prostaglandin E2 (PGE2), and synovial fluid IL-1alpha and IL-1beta concentrations were seen after administration of sIL-1RII. However, synovial fluid PGE2 concentrations and synovial fluid cell counts were not affected. A significant inhibitory effect was also seen histologically on soft-tissue swelling and joint damage with high-dose sIL-1RII. CONCLUSIONS: These results demonstrate that IL-1 plays an important role in the pathogenesis of rabbit antigen-induced arthritis, thus confirming it as an excellent animal model with respect to evaluating anti-cytokine therapies for rheumatoid arthritis.