A hypothetical index of hypothalamic-pituitary-adrenal (HPA) axis function.

The short ACTH stimulation test (SST) is commonly used by clinicians to determine the integrity of the hypothalamic-pituitary-adrenal (HPA) axis in cases of suspected adrenal insufficiency. Concern has arisen for under-diagnosis of central hypocortisolism using the standard dose (250 mcg) SST with the recent introduction of low-dose (1 mcg) SST as a potentially more sensitive test for central hypocortisolism. Amidst this uncertainty, I will present a novel index hypothesized to extend the sensitivity of the standard dose SST as an adjunctive diagnostic tool for any level of hypocortisolism.     

Effect of Redox Balance Alterations on Cellular Localization of LAT and Downstream T-Cell Receptor Signaling Pathways

The integral membrane protein linker for activation of T cells (LAT) is a central adapter protein in the T-cell receptor (TCR)-mediated signaling pathways. The cellular localization of LAT is extremely sensitive to intracellular redox balance alterations. Reduced intracellular levels of the antioxidant glutathione (GSH), a hallmark of chronic oxidative stress, resulted in the membrane displacement of LAT, abrogated TCR-mediated signaling and consequently hyporesponsiveness of T lymphocytes. The membrane displacement of LAT is accompanied by a considerable difference in the mobility of LAT upon native and nonreducing denaturing polyacrylamide gel electrophoresis analysis, a finding indicative of a conformational change. Targeted mutation of redox-sensitive cysteine residues within LAT created LAT mutants which remain membrane anchored under conditions of chronic oxidative stress. The expression of redox-insensitive LAT mutants allows for restoration of TCR-mediated signal transduction, whereas CD28-mediated signaling pathways remained impaired. These results are indicative that the membrane displacement of LAT as a result of redox balance alterations is a consequence of a conformational change interfering with the insertion of LAT into the plasma membrane. Conclusively, the data suggest a role for LAT as a crucial intermediate in the sensitivity of TCR signaling and hence T lymphocytes toward chronic oxidative stress.