maintenance of the inflammatory response. Elevated MIF levels are linked to inflammatory diseases such as sepsis, arthritis and colitis, and also diseases with an inflammatory component such as atherosclerosis, diabetes and cancer. Efforts are being made to develop small molecule inhibitors for clinical use, but little is known about the endogenous regulation of MIF activity. the biological activity of MIF is mediated through interaction with the CD74 receptor, but interestingly, MIF also has a keto enol tautomerase activity that is catalyzed by its N-terminal proline. the importance of tautomerase activity is unclear, but inhibition is associated with impaired CD74 binding. We have previously shown that isothiocyanates found in cruciferous vegetables are effective MIF inhibitors through covalent modification of the N-terminal proline. We are currently investigating the ability of other dietary and endogenous electrophiles to modify the N-terminal proline and inhibit MIF activity. Green tea, cocoa and red wine are rich in catechins, and oxidation of the catechol moiety generated potent MIF inhibition. Furthermore, we are characterizing the modification introduced on MIF by LC/MS. This provides a putative mechanism for the anti-inflammatory activity of these compounds.

Mitochondria and Duox2 mediate ROS Generation as an Innate Immune Response against Influenza a virus infection in Human Nasal Epithelium Hyun Jik Kim1, Chang-Hoon Kim2, and Joo-Heon Yoon2 1Chung-Ang University College of Medicine, Republic of Korea, 2Yonsei University College of Medicine, Republic of Korea The regulated production of ROS has been considered a unique property of phagocytic cells which use this ROS system to induce innate defense system. Our goals of this study are to explore the role of ROS for the mechanism of antiviral innate immunity and to investigate the source of ROS after influenza a virus (IAV) infection in nasal epithelium. Our results demonstrated that intracellular ROS generation was increased after 1 hr and 2 day post of infection (PI) respectively. Interestingly, viral titer and mRNA level were significantly higher in case of scavenging ROS. Induced ROS generation at PI 1 hr was decreased after inhibition of mitochondria respiratory chain reaction and was also suppressed after knock-down of Duox2 gene expression in nasal epithelium. ROS generation at PI 2 day was totally decreased in case of suppression of mitochondria-generated ROS, not Duox2-generated ROS. Inhibition of mitochondrial ROS generation and knock-down of Duox2 gene expression highly induced IAV mRNA level and viral titer. Our findings suggest that production of ROS be primarily responsible for controlling IAV infection and both mitochondria and Duox2 might be important sources of ROS generation requiring for antiviral innate immune response in nasal epithelium

Oxidized Lipids and Eicosanoids Are Biomarkers of Inflammation in Sickle Cell Disease Sainath Kotha1, Leslie Witkoff1, Hemalatha Rao1, Travis Gurney1, Payal Desai1, Krishna Rao Maddipati2, Eric Kraut1, and Narasimham Parinandi1 1The Ohio State University College of Medicine, United States, 2Wayne State University School of Medicine, United States Acute chest syndrome has been identified as a severe inflammatory state in sickle cell disease (SCD) and elevated levels of the secretory phospholipase A2 (sPLA2) have been observed in circulation of SCD patients during acute inflammatory episodes. This suggests the role of sPLA2 in arachidonic acid release from the phospholipids and subsequent formation of the inflammatory eicosanoids catalyzed by the cyclooxygenase (COX) and lipoxygenase (LOX) pathways. However, a thorough analysis of the lipids and lipid-derived inflammatory biomarkers both before and during crisis is warranted. Hence, our initial study was conducted to determine the (a) fatty acid profiles, (b) triglyceride levels, (c) extent of lipid peroxidation, and (d) levels of eicosanoids in SCD patients without evidence of acute crisis. Outpatient SCD patients (n = 24; male) not in acute pain crisis and normal adult subjects (n = 5; male and female) were recruited for the study and blood samples were collected upon the institutional review board (IRB) approval. Esterified and non-esterified fatty acids in blood serum were determined by the gas chromatography-mass spectrometry (GS-MS) method. Eicosanoids in blood serum were determined by the liquid chromatography-mass spectrometry (LCMS) method. the results revealed that: (a) the expression and activity of secretory phospholipase A2 (sPLA2) were significantly higher in blood serum of SCD patients as compared to the same in normal subjects; (b) the levels of all the esterified fatty acids (saturated and unsaturated) and triglycerides in blood serum were significantly lower in SCD patients as compared to the same in the normal subjects; (c) the levels of non-esterified (free) fatty acid levels (Figure) were significantly higher including the unsaturated fatty acids in blood serum of SCD patients as compared to that in the normal subjects; (d) the levels of total lipid peroxides were significantly higher in the blood serum of SCD patients as compared to that in the normal subjects; and (e) the levels of eicosanoids (thromboxane A2, prostaglandin E3, and lipoxygenase-catalyzed hydroxyl arachidonic acid metabolites) were significantly elevated in the blood serum of SCD subjects as compared to the same in the normal subjects. This study demonstrates that the alterations in the sPLA2 expression and activity, fatty acid profiles, increased lipid peroxide levels, and elevated inflammatory eicosanoids levels in the blood serum of SCD patients could serve as the lipid biomarkers of inflammation. from the present study it is concluded that SCD patients have altered serum lipidome profile even when not in acute pain crisis. Whether these changes are magnified during acute pain crisis and could be targeted to alter development of crisis requires further evaluation.

doi: 10.1016/j.freeradbiomed.2013.10.470

doi: 10.1016/j.freeradbiomed.2013.10.471 doi: 10.1016/j.freeradbiomed.2013.10.472