In: Septic Shock11 Nov 2014
The in vitro myocardial cell contractility assay employed in this study represented a modification of a previously described cell culture system.’ The addition of the latex microspheres improved the reliability of the system. Using this revised method, the amount of depression can be quantified reproducibly. Since the sensitivity of the cell culture system changes as the cells mature, it is imperative to choose cultures at the same level of maturation each time an assay is performed. With the initial system that did not use latex beads, small changes in the relationship of myocardial cell edge to the microscope optics caused technical difficulties.
These resulted in long delays while finding a cell in which the cell edge and the optical tracking system were very stable. However, the latex bead system is not altered by small changes in the relation of the myocardial cell to the microscope optics because the computer program tracking system continuously recenters the gate over the bead. Thus, this revised method decreases the technical difficulties and enhances the reproducibility and reliability of this in vitro bioassay of cardiac cell performance. there
Understanding the changes in the maturation of the heart cells grown in tissue culture, coupled with the improved reliability of the latex bead video tracking system, allowed us to systematically evaluate serial ultrafiltration experiments in sera from a variety of patient groups. The criteria for MDS activity in these experiments were identical to those of the initial experiments performed on the 50 patients enrolled into the septic shock protocol to determine the incidence of patients with circulating MDS. The MDS activity was consistently found in the supernatant fraction of the serum samples, suggesting that the molecular weight of the MDS activity is greater than 10,000 daltons and may be greater than 30,000 daltons.
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