Using DSC (differential scanning calorimetry), we measure the kinetics of the cubic-HII phase transition of monoolein in bulk sucrose solutions. We find that the transition temperature is dramatically lowered, with each 1 mol/kg of sucrose concentration dropping the transition by 20 °C. The kinetics of this transition also slow greatly with increasing sucrose concentration. For low sucrose concentrations, the kinetics are asymmetric, with the cooling (HII-cubic) transition taking twice as long as the heating (cubic-HII) transition. This asymmetry in transition times is reduced for higher sucrose concentrations. The cooling transition (cubic-HII) exhibits Avrami exponents in the range of 2 to 2.5 and the heating transition shows Avrami exponents ranging from 1 to 3. A classical Avrami interpretation would be that these processes occur via a one or two dimensional pathway with variable nucleation rates. A non-classical perspective would suggest that these exponents reflect the time dependence of pore formation (cooling) and destruction (heating). New density measurements of monoolein show that the currently accepted value is about 5% too low; this has substantial implications for electron density modeling. Structural calculations indicate that the head group area and lipid length in the cubic-HII transition shrink by about 12 % and 4 % respectively; this reduction is practically the same as that seen in a lipid with a very different molecular structure (rac-di-12:0 β-GlcDAG) that makes the same transition. Thermodynamic considerations suggest there is a hydration shell about one water molecule thick in front of the lipid head groups in both the cubic and HII phases.
Accumulating evidence supports a positive correlation between the presence of antiphosphatidylethanolamine (aPE) autoantibodies and clinical symptoms of antiphospholipid syndroms (APS). However, there lacks a standardized ELISA-based method for detecting aPE. The current study was to investigate the dependence of aPE ELISA on lipid concentration and composition of PE antigens. Our data indicated that there are different optimal PE concentrations for conducting ELISA assays for cofactor dependent and independent aPE. In addition, using a two-component synthetic lipid system, we demonstrated that aPE ELISA readouts can be modulated to approach the performance level of egg PE, which is currently the most commonly used PE antigen. These data raised the possibility of ultimately replacing natural PE antigens with a blend of defined synthetic lipid species, thus overcoming a known variable factor in aPE detection. The outcome of this study will help pave the way to developing a standardized aPE test.
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