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Assays were performed to evaluate DNA methylation patterns on SAECs exposed to PEPs (administered doses of 0.5 and 30 [micro]g/mL) for 24 hr.
RNA and DNA were extracted simultaneously from SAECs using an AllPrep Mini Kit (Qiagen) according to the manufacturer's protocol.
Supernatants from treated SAECs were assayed by Eve Technologies Corporation, which used a Human Primary Cytokine Array/Chemokine Array 41-Plex Panel (Millipore) according to the manufacturer's protocol.
In particular, SAECs experienced > 40% cell death after exposure to PEPs ([PM.sub.0.1], 100 [micro]g/mL administered dose) when compared with untreated cells.
To evaluate the potential of PEPs to induce ROS production in epithelial cells (SAECs) and macrophages (THP-1 cells), two types of cells that are in direct contact with inhaled foreign material, the levels of superoxide ions were measured.
In order to evaluate the effects of PEPs on such biological reactions, levels of a wide variety of these mediators were measured in SAECs following a 24-hr exposure to PEPs (5 and 40 [micro]g/mL administered doses).
The objective of this study was to evaluate the potential toxicity of various doses of PEPs in human small airway epithelial cells (SAECs), macrophages (THP-1 cells) and lymphoblasts (TK6 cells).
The results showed that both the epithelial cells (SAECs, at a 100-[micro]g/mL delivered dose) and the macrophages (THP-1 cells, at a 2.59-[micro]g/mL delivered dose) were negatively affected by treatment with PEPs and experienced > 40% cell death.
As outlined in select experiments, R3/1 or SAECs were transfected with tat or human siRNA for RAGE (siRAGE) or a scrambled control siRNA sequence (siControl) generated by Santa Cruz 24 hr before DPM exposure.
DPM induces RAGE mRNA and protein in R3/1 cells and SAECs. To determine whether RAGE is up-regulated by exposure to DPM, we assessed RAGE mRNA levels in R3/1 cells and SAECs by quantitative real-time RT-PCR and compared cells exposed to DPM or fresh media.
However, similar observations in human SAECs suggest that RAGE up-regulation is likely associated with cellular responses to DPM exposure.
Safmarine's agency functions are performed in the UK by ASECO (UK) Ltd and in Continental Europe by its SAECS consortium partners as follows: DAL in Germany, Nedlloyd in the Netherlands and France, CMBT in Belgium, Lloyd Triestino in Italy, Witty in Spain and ASECO in Portugal.
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