Research IT maturity models for academic health centers: Early development and initial evaluation

This paper proposes the creation and application of maturity models to guide institutional strategic investment in research informatics and information technology (research IT) and to provide the ability to measure readiness for clinical and research infrastructure as well as sustainability of expertise. Conducting effective and efficient research in health science increasingly relies upon robust research IT systems and capabilities. Academic health centers are increasing investments in health IT systems to address operational pressures, including rapidly growing data, technological advances, and increasing security and regulatory challenges associated with data access requirements. Current approaches for planning and investment in research IT infrastructure vary across institutions and lack comparable guidance for evaluating investments, resulting in inconsistent approaches to research IT implementation across peer academic health centers as well as uncertainty in linking research IT investments to institutional goals. Maturity models address these issues through coupling the assessment of current organizational state with readiness for deployment of potential research IT investment, which can inform leadership strategy. Pilot work in maturity model development has ranged from using them as a catalyst for engaging medical school IT leaders in planning at a single institution to developing initial maturity indices that have been applied and refined across peer medical schools

Transcription of the Salmonella Invasion Gene Activator, hilA, Requires HilD Activation in the Absence of Negative Regulators

Salmonella enterica serovar Typhimurium causes human gastroenteritis and a systemic typhoid-like infection in mice. Infection is initiated by entry of the bacteria into intestinal epithelial cells and is mediated by a type III secretion system that is encoded by genes in Salmonella pathogenicity island 1. The expression of invasion genes is tightly regulated by environmental conditions such as oxygen and osmolarity, as well as by many bacterial factors. The hilA gene encodes an OmpR/ToxR family transcriptional regulator that activates the expression of invasion genes in response to both environmental and genetic regulatory factors. HilD is an AraC/XylS regulator that has been postulated to act as a derepressor of hilA expression that promotes transcription by interfering with repressor binding at the hilA promoter. Our research group has identified four genes (hilE, hha, pag, and ams) that negatively affect hilA transcription. Since the postulated function of HilD at the hilA promoter is to counteract the effects of repressors, we examined this model by measuring hilA::Tn5lacZY expression in strains containing negative regulator mutations in the presence or absence of functional HilD. Single negative regulator mutations caused significant derepression of hilA expression, and two or more negative regulator mutations led to very high level expression of hilA. However, in all strains tested, the absence of hilD resulted in low-level expression of hilA, suggesting that HilD is required for activation of hilA expression, whether or not negative regulators are present. We also observed that deletion of the HilD binding sites in the chromosomal hilA promoter severely decreased hilA expression. In addition, we found that a single point mutation at leucine 289 in the C-terminal domain of the α subunit of RNA polymerase leads to very low levels of hilA::Tn5lacZY expression, suggesting that HilD activates transcription of hilA by contacting and recruiting RNA polymerase to the hilA promoter

Research IT maturity models for academic health centers: Early development and initial evaluation.

This paper proposes the creation and application of maturity models to guide institutional strategic investment in research informatics and information technology (research IT) and to provide the ability to measure readiness for clinical and research infrastructure as well as sustainability of expertise. Conducting effective and efficient research in health science increasingly relies upon robust research IT systems and capabilities. Academic health centers are increasing investments in health IT systems to address operational pressures, including rapidly growing data, technological advances, and increasing security and regulatory challenges associated with data access requirements. Current approaches for planning and investment in research IT infrastructure vary across institutions and lack comparable guidance for evaluating investments, resulting in inconsistent approaches to research IT implementation across peer academic health centers as well as uncertainty in linking research IT investments to institutional goals. Maturity models address these issues through coupling the assessment of current organizational state with readiness for deployment of potential research IT investment, which can inform leadership strategy. Pilot work in maturity model development has ranged from using them as a catalyst for engaging medical school IT leaders in planning at a single institution to developing initial maturity indices that have been applied and refined across peer medical schools

Opioids Bind to the Amino Acids 84 to 118 of UDP- Glucuronosyltransferase UGT2B7

ABSTRACT The UDP-glucuronosyltransferase UGT2B7 is an important human UGT isoform that catalyzes the conjugation of many endogenous and exogenous compounds, among them opioids, resulting in the formation of D-glucuronides. The binding site of the aglycone is located in the N-terminal half of the protein. Using NMR analysis, we demonstrate that the opioid binding site in UGT2B7 is within the 84 to 118 N-terminal amino acids. Three maltose binding protein-UGT2B7 fusion proteins, 2B7F3 and 2B7F4 incorporating the amino acids 24 to 118 and 24 to 96 of UGT2B7, respectively, and 2B7F5 incorporating amino acids 84 to 118 of UGT2B7 were expressed in Escherichia coli and purified by affinity chromatography. NMR analysis showed that morphine was bound to the fusion protein 2B7F3 with a K D value similar to the K D values obtained for the previously produced fusion proteins, which included amino acids 24 to 180. Morphine did not bind to 2B7F4, but it did bind to 2B7F5. Both NMR 1-D spectra and NOESY experiments indicated that the 2B7F5 protein was mediating magnetization transfer within the morphine. These results allowed us to predict and model a binding site within the amino acids 96 to 101 of UGT2B7. A mutant fusion protein 2B7F3 with the substitution D99A was produced, and the NMR spectroscopy analysis of the protein supported the model. A marked reduction of morphine binding was observed when the charged aspartate was substituted with alanine

Opioids Bind to the Amino Acids 84 to 118 of UDP-glucuronosyltransferase UGT2B7

The UDP-glucuronosyltransferase UGT2B7 is an important human UGT isoform that catalyzes the conjugation of many endogenous and exogenous compounds, among them opioids, resulting in the formation of D-glucuronides. The binding site of the aglycone is located in the N-terminal half of the protein. Using NMR analysis, we demonstrate that the opioid binding site in UGT2B7 is within the 84 to 118 N-terminal amino acids. Three maltose binding protein-UGT2B7 fusion proteins, 2B7F3 and 2B7F4 incorporating the amino acids 24 to 118 and 24 to 96 of UGT2B7, respectively, and 2B7F5 incorporating amino acids 84 to 118 of UGT2B7 were expressed in Escherichia coli and purified by affinity chromatography. NMR analysis showed that morphine was bound to the fusion protein 2B7F3 with a KD value similar to the KD values obtained for the previously produced fusion proteins, which included amino acids 24 to 180. Morphine did not bind to 2BTF4, but it did bind to 2B7F5. Both NMR 1-D spectra and NOESY experiments indicated that the 2B7F5 protein was mediating magnetization transfer within the morphine. These results allowed us to predict and model a binding site within the amino acids 96 to 101 of UGT2B7. A mutant fusion protein 2B7F3 with the substitution D99A was produced, and the NMR spectroscopy analysis of the protein supported the model. A marked reduction of morphine binding was observed when the charged aspartate was substituted with alanine