Intercellular fluids, analysis

Figure 9.10 shows an example of a metabolomics sample separated by ambient pressure IMS coupled with TOF-MS. In this 2D spectrum, mass is plotted along the x-axis, and mobility data are plotted along the y-axis. A hot methanol extract of E. coli cells that had been separated from the extracellular fluid was used for this analysis. The hot methanol lysed the cells and produced an extract of the intercellular metabolome. This methanol extract was then diluted with a water solution of acetic acid to produce the final electrospray solution that was sprayed into the IMS for separation and detection of the metabolites by IMS-TOF-MS. 

Silica seems to be a potent activator of endothelial cells in vitro, too. Incubation of human dermal microvascular endothelial cells (HDMEC) with silica at non-toxic concentrations increased the steady-state levels of the messenger RNA (mRNA) for intercellular adhesion molecule 1 (ICAM-i), and also increased the corresponding levels of this cell-surface protein as shown by fluorescence-activated cell-sorter (FACS) analysis the incubation also increased the level of soluble protein in the culture fluid, as shown by means of ELISA, in a dose- and time-dependent manner. Additionally, increased levels of IL-6 in the culture supernatants have been found. In addition, a significant increase in coUagenase I mRNA in HDMEC has been demonstrated (Anderegg et al. 1997). 

---