Exotoxin A is one of the virulence factors of Pseudomonas aeruginosa, a bacterium that can cause infections resulting in adverse health outcomes and increased burden to health care systems. Current methods of diagnosing P. aeruginosa infections are time consuming and can require significant preparation of patient samples. This study utilized a novel variation of the Systematic Evolution of Ligand by Exponential Enrichment, Decoy-SELEX, to identify an Exotoxin A specific single-stranded DNA (ssDNA) molecular recognition element (MRE). Its emphasis is on increasing stringency in directing binding toward free target of interest and at the same time decreasing binding toward negative targets. A ssDNA MRE with specificity and affinity was identified after fourteen rounds of Decoy-SELEX. Utilizing surface plasmon resonance measurements, the determined equilibrium dissociation constant (K d) of the MRE is between 4.2 µM and 4.5 µM, and is highly selective for Exotoxin A over negative targets. A ssDNA MRE modified sandwich enzyme-linked immunosorbent assay (ELISA) has been developed and achieved sensitive detection of Exotoxin A at nanomolar concentrations in human serum. This study has demonstrated the proof-of-principle of using a ssDNA MRE as a clinical diagnostic tool.

Achieving minimum borrowing cost for municipal bonds is an ongoing concern. Apostolou, Apostolou, and Dorminey (2014) demonstrated that a fund accounting measure, spending rate magnitude (i.e., the absolute value of general fund revenues minus general fund expenditures) proxied for management's ability to balance revenues and expenditures. Their findings indicated that achieving balance at spending rate equilibrium (i.e., where general fund revenues equal general fund expenditures) was associated with minimum borrowing cost, and that the magnitude of departure from spending rate equilibrium was associated with higher borrowing cost. Little is known regarding the information content of spending rate in the years during and after the financial crisis that started in 2007. Given the crash of the housing bubble and severe reductions to property tax revenues, municipalities experienced a prolonged recovery that was characterized by a change to borrowing patterns and deficit spending. In this study, I extended from Apostolou et al. (2014) and examined whether the information content of spending rate was conditional to levels of economic activity. I accomplished this by using state leading indexes compiled by the Federal Reserve Bank to proxy for local economic activity. Based on a sample of 1,970 general obligations bonds that were issued by U. S. counties between 1995 through 2014, I tested three conditional hypotheses for significant interaction between economic activity and spending rate magnitude. Results indicated that the association between spending rate magnitude and borrowing cost was positively moderated by economic activity. Positive moderation was also significant for spending rate surpluses (i.e., where general fund revenues exceed general fund expenditures). These results suggest that departures from spending rate equilibrium are more severely assessed on borrowing cost when economic activity is positive, and less severely assessed on borrowing cost when economic activity is negative. Overall, my findings provide preliminary evidence that managerial competence over the balance of revenues and expenditures is evaluated more severely when economic conditions are goodnot when they are bad.

Cytochrome P450 (P450) protein-protein interactions have been shown to alter their catalytic activity. Furthermore, these interactions are isoform specific and can elicit activation, inhibition, or no effect on enzymatic activity. Studies show that these effects are also dependent on the protein partner cytochrome P450 reductase (CPR) and the order of protein addition to purified reconstituted enzyme systems. In this study, we use controlled immobilization of P450s to a gold surface to gain a better understanding of P450-P450 interactions between three key drug-metabolizing isoforms (CYP2C9, CYP3A4, and CYP2D6). Molecular modeling was used to assess the favorability of homomeric/heteromeric P450 complex formation. P450 complex formation in vitro was analyzed in real time utilizing surface plasmon resonance. Finally, the effects of P450 complex formation were investigated utilizing our immobilized platform and reconstituted enzyme systems. Molecular modeling shows favorable binding of CYP2C9-CPR, CYP2C9-CYP2D6, CYP2C9-CYP2C9, and CYP2C9-CYP3A4, in rank order. K D values obtained via surface plasmon resonance show strong binding, in the nanomolar range, for the above pairs, with CYP2C9-CYP2D6 yielding the lowest K D , followed by CYP2C9-CYP2C9, CYP2C9-CPR, and CYP2C9-CYP3A4. Metabolic incubations show that immobilized CYP2C9 metabolism was activated by homomeric complex formation. CYP2C9 metabolism was not affected by the presence of CYP3A4 with saturating CPR concentrations. CYP2C9 metabolism was activated by CYP2D6 at saturating CPR concentrations in solution but was inhibited when CYP2C9 was immobilized. The order of addition of proteins (CYP2C9, CYP2D6, CYP3A4, and CPR) influenced the magnitude of inhibition for CYP3A4 and CYP2D6. These results indicate isoform-specific P450 interactions and effects on P450-mediated metabolism.