Project description:LPS (10 μg/50 μl PBS; E. coli Serotype 055:B5, Sigma-Aldrich) was administered onto the nares. Baseline levels of genes in mice treated with vehicle (Hanks Balanced Salt solution (HBSS) and PBS), Intranasal S100s (10 μg/50 μl HBSS) were given 2 h before LPS; control mice received equal volumes of PBS and HBSS. Mice were sacrificed 4 h post LPS inhalation. Expression of inflammatory genes was evaluated with the RT-qPCR array. Relative quantities of mRNA in duplicate samples were obtained using the LightCycler® 480 Software 1.5 and the Efficiency-Method.
Project description:Recombinant-murine S100A8 were used at 10 mg/50 ml in Hanks Balanced Salt solution (HBSS) or control HBSS administered onto the nares of BALB/C mice. To assess direct effects, mice were sacrificed 1, 4, 6, or 12 h post-inhalation of S100A8. Because S100A8 is reported to initiate proinflammatory responses by ligating TLR4 and/or RAGE, a quantitative PCR array was developed to analyze 49 genes, selected to reflect potential acute inflammatory changes induced by ligation of these receptors. Expression of inflammatory genes was evaluated with the RT-qPCR array. Relative quantities of mRNA in duplicate samples were obtained using the LightCycler® 480 Software 1.5 and the Efficiency-Method.
Project description:Recombinant-murine S100A9 were used at 10 mg/50 ml in Hanks Balanced Salt solution (HBSS) or control HBSS administered onto the nares of BALB/C mice. To assess direct effects, mice were sacrificed 1, 4, 6, or 12 h post-inhalation of S100A9. Because S100A9 is reported to initiate proinflammatory responses by ligating TLR4 and/or RAGE, a quantitative PCR array was developed to analyze 49 genes, selected to reflect potential acute inflammatory changes induced by ligation of these receptors. Expression of inflammatory genes was evaluated with the RT-qPCR array. Relative quantities of mRNA in duplicate samples were obtained using the LightCycler® 480 Software 1.5 and the Efficiency-Method.
Project description:Recombinant-murine S100A8 and the corresponding Cys42 to Ala42 mutant were used at 10 mg/50 ml HBSS administered onto the nares of BALB/C mice. To assess direct effects, mice were sacrificed 1, 4, 6, 12 or 20 h post-inhalation of S100A8 or 12 and 20 h post-inhalation of Ala42S100A8. For comparison with Dex inhalation (used at 10 mg/50 ml HBSS), lungs were harvested 6 and 12 h post administration. Because S100A8 is reported to initiate proinflammatory responses by ligating TLR4 and/or RAGE, a quantitative PCR array was developed to analyse 63 genes, selected to reflect potential acute inflammatory changes induced by ligation of these receptors. Expression of inflammatory genes was evaluated with the RT-qPCR array. Relative quantities of mRNA in duplicate samples were obtained using the LightCyclerM-BM-. 480 Software 1.5 and the Efficiency-Method.
Project description:Recombinant-murine S100A8 and the corresponding Cys42 to Ala42 mutant were used at 10 mg/50 ml HBSS administered onto the nares of BALB/C mice. To assess direct effects, mice were sacrificed 1, 4, 6, 12 or 20 h post-inhalation of S100A8 or 12 and 20 h post-inhalation of Ala42S100A8. For comparison with Dex inhalation (used at 10 mg/50 ml HBSS), lungs were harvested 6 and 12 h post administration. Because S100A8 is reported to initiate proinflammatory responses by ligating TLR4 and/or RAGE, a quantitative PCR array was developed to analyse 63 genes, selected to reflect potential acute inflammatory changes induced by ligation of these receptors. Expression of inflammatory genes was evaluated with the RT-qPCR array. Relative quantities of mRNA in duplicate samples were obtained using the LightCyclerM-BM-. 480 Software 1.5 and the Efficiency-Method.