Background Sulfur mustard (SM) is a potent chemical vesicant warfare agent

Background Sulfur mustard (SM) is a potent chemical vesicant warfare agent that remains a significant military and civilian threat. the BenchMark XT IHC/ISH workstation of Ventana (Tucson, AZ) to react with the primary and secondary antibodies. The primary antibody used was mouse anti-iNOS MAb from BD Biosciences, and was diluted 1:1000 in TBS + 0.5% (w/v) Triton X-100 (TBST). The secondary antibody was pre-diluted biotinylated rabbit 83797-69-7 IC50 anti-mouse IgG from Ventana. iNOS signals were detected by quantum dot-streptavidin conjugates (streptavidin-Qdot655), which was diluted 1:200 with the Qdots incubation buffer from Invitrogen (Carlsbad, CA). The fluorescence signals were visualized, captured and processed as reported previously [25,26]. Data analysis For comparative studies, Student’s t-test (unpaired) or one-way ANOVA assessments (with Bonferroni post test if P < 0.05) were 83797-69-7 IC50 used for statistical analysis. Differences were considered statistically significant if a P value of < 0.05 was achieved. Abbreviations BTE, bronchial/tracheal epithelial calcein AM, calcein acetoxy methyl ester ELISA, enzyme-linked immunosorbent assay EthD-1, ethidium homodimer-1 IL, interleukin iNOS, inducible nitric oxide synthase MTS, 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt NO, nitric oxide RT-PCR, reverse transcriptase polymerase chain reaction NOS, nitric oxide synthase; RXM, roxithromycin SAE, small air passage epithelial; SE, standard error SM, sulfur mustard TNF, tumor necrosis factor TBS, tris-buffered saline TBST, tris-buffered saline with Triton. Authors' contributions PR originated the project, supervised the overall conduct of the research which was performed in her laboratory, and provided continuous evaluation of the experimental data. XG carried out all of the experimental work in this study, performed the statistical analyses, and drafted the manuscript. RR (along with PR) conceived of the study and carried out SM exposure experiment. YX and PEB carried out the immunocytochemical studies and analyzed the data. 83797-69-7 IC50 All authors read and approved the final manuscript. Acknowledgements We thank Drs. Hiroshi Ishida and Michael Zidanic of Walter Reed Army Institute STK3 of Research, Metallic Spring, MD for helpful discussions and technical guidance on fluorescence microscopy study, respectively. We thank Ms. Betty Benton of the US Army Medical Research Institute of Chemical Defense, Aberdeen Proving Ground, MD for technical assistance with sulfur mustard exposure. This research was supported by the Defense Threat Reduction Agency (Project # 3.F0003_05_WR_C). The opinions or assertions contained herein are the private views of the authors and are not to be construed as recognized or as reflecting true views of the US Department of the Army or the Department of Defense. Certain commercial gear or materials are identified in this paper in order to designate properly the experimental procedures. Such identification does not imply recommendation or endorsement by the National Institute of Standards and Technology, nor does it imply that the materials or gear identified are necessarily the best available for the purpose..

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