A National Dialogue on Health Information Technology and Privacy

Increasingly, government leaders recognize that solving the complex problems facing America today will require more than simply keeping citizens informed. Meeting challenges like rising health care costs, climate change and energy independence requires increased level of collaboration. Traditionally, government agencies have operated in silos -- separated not only from citizens, but from each other, as well. Nevertheless, some have begun to reach across and outside of government to access the collective brainpower of organizations, stakeholders and individuals.The National Dialogue on Health Information Technology and Privacy was one such initiative. It was conceived by leaders in government who sought to demonstrate that it is not only possible, but beneficial and economical, to engage openly and broadly on an issue that is both national in scope and deeply relevant to the everyday lives of citizens. The results of this first-of-its-kind online event are captured in this report, together with important lessons learned along the way.This report served as a call to action. On his first full day in office, President Obama put government on notice that this new, more collaborative model can no longer be confined to the efforts of early adopters. He called upon every executive department and agency to "harness new technology" and make government "transparent, participatory, and collaborative." Government is quickly transitioning to a new generation of managers and leaders, for whom online collaboration is not a new frontier but a fact of everyday life. We owe it to them -- and the citizens we serve -- to recognize and embrace the myriad tools available to fulfill the promise of good government in the 21st Century.Key FindingsThe Panel recommended that the Administration give stakeholders the opportunity to further participate in the discussion of heath IT and privacy through broader outreach and by helping the public to understand the value of a person-centered view of healthcare information technology

Spatio-temporal and neighborhood characteristics of two dengue outbreaks in two arid cities of Mexico.

Little is currently known about the spatial-temporal dynamics of dengue epidemics in arid areas. This study assesses dengue outbreaks that occurred in two arid cities of Mexico, Hermosillo and Navojoa, located in northern state of Sonora. Laboratory confirmed dengue cases from Hermosillo (N=2730) and Navojoa (N=493) were geocoded by residence and assigned neighborhood-level characteristics from the 2010 Mexican census. Kernel density and Space-time cluster analysis was performed to detect high density areas and space-time clusters of dengue. Ordinary Least Square regression was used to assess the changing socioeconomic characteristics of cases over the course of the outbreaks. Both cities exhibited contiguous patterns of space-time clustering. Initial areas of dissemination were characterized in both cities by high population density, high percentage of occupied houses, and lack of healthcare. Future research and control efforts in these regions should consider these space-time and socioeconomic patterns.National Institutes of Health - NIAID [R01-AI091843]12 month embargo; Available online 3 January 2017.This item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at [email protected]

Application of Multiplexed Kinase Inhibitor Beads to Study Kinome Adaptations in Drug-Resistant Leukemia

<div><p>Protein kinases play key roles in oncogenic signaling and are a major focus in the development of targeted cancer therapies. Imatinib, a BCR-Abl tyrosine kinase inhibitor, is a successful front-line treatment for chronic myelogenous leukemia (CML). However, resistance to imatinib may be acquired by BCR-Abl mutations or hyperactivation of Src family kinases such as Lyn. We have used <u>m</u>ultiplexed kinase <u>i</u>nhibitor <u>b</u>eads (MIBs) and quantitative mass spectrometry (MS) to compare kinase expression and activity in an imatinib-resistant (MYL-R) and -sensitive (MYL) cell model of CML. Using MIB/MS, expression and activity changes of over 150 kinases were quantitatively measured from various protein kinase families. Statistical analysis of experimental replicates assigned significance to 35 of these kinases, referred to as the MYL-R kinome profile. MIB/MS and immunoblotting confirmed the over-expression and activation of Lyn in MYL-R cells and identified additional kinases with increased (MEK, ERK, IKKα, PKCβ, NEK9) or decreased (Abl, Kit, JNK, ATM, Yes) abundance or activity. Inhibiting Lyn with dasatinib or by shRNA-mediated knockdown reduced the phosphorylation of MEK and IKKα. Because MYL-R cells showed elevated NF-κB signaling relative to MYL cells, as demonstrated by increased IκBα and IL-6 mRNA expression, we tested the effects of an IKK inhibitor (BAY 65-1942). MIB/MS and immunoblotting revealed that BAY 65-1942 increased MEK/ERK signaling and that this increase was prevented by co-treatment with a MEK inhibitor (AZD6244). Furthermore, the combined inhibition of MEK and IKKα resulted in reduced IL-6 mRNA expression, synergistic loss of cell viability and increased apoptosis. Thus, MIB/MS analysis identified MEK and IKKα as important downstream targets of Lyn, suggesting that co-targeting these kinases may provide a unique strategy to inhibit Lyn-dependent imatinib-resistant CML. These results demonstrate the utility of MIB/MS as a tool to identify dysregulated kinases and to interrogate kinome dynamics as cells respond to targeted kinase inhibition.</p></div

Targeted inhibition of kinases detected by MIB/MS leads to induction of apoptosis.

<p>(<b>A</b>) MYL-R cells were treated for 48 hours with AZD6244 (AZD, 5 µM), BAY 65-1942 (BAY, 10 µM), AZD (5 µM) plus BAY (10 µM), or dasatinib (1 nM) and cell viability was assessed by MTS assay. <i>Error bars</i>, SE (N = 3). (<b>B</b>) MYL-R cells were treated for 24 hours with AZD (5 µM), BAY (10 µM), AZD (5 µM) plus BAY (10 µM), or dasatinib (1 nM) and caspase 3/7 activity was assessed by fluorometric assay. <i>Error bars</i>, SE (N = 2). (<b>C</b>) MYL-R cells were treated for 48 hours with AZD (5 µM), BAY (10 µM), AZD (5 µM) plus BAY (10 µM), or dasatinib (1 nM) and cell lysates were analyzed by immunoblot using the antibodies indicated. Data are representative of two separate experiments.</p

Kinome response to targeted MEK and IKK inhibitor treatment.

<p>(<b>A</b>) MYL-R cells were treated for 24 hours with AZD6244 (AZD, 5 µM), BAY 65-1942 (BAY, 10 µM) or AZD (5 µM) plus BAY (10 µM), and kinases were isolated and quantified by MIB/MS in two independent experiments. The relative abundances (Drug/DMSO) of kinases in the MEK/ERK pathway are shown. <i>Error bars</i>, SE (N = 2). (<b>B</b>) MYL-R cells were treated for 24 hours with AZD (5 µM), BAY(10 µM) or AZD (5 µM) plus BAY (10 µM), and were analyzed by immunoblot using the antibodies indicated. Data are representative of three separate experiments. (<b>C</b>) MYL-R cells were treated for 12 hours with AZD (5 µM), BAY (10 µM), AZD (5 µM) plus BAY (10 µM), or dasatinib (1 nM). Total RNA was isolated and the expression of IκBα and IL-6 were evaluated by qRT-PCR. <i>Error bars</i>, SE (N = 2).</p

Validation of the MYL-R kinome profiles by immunoblotting.

<p>(<b>A and B</b>) MYL and MYL-R cell lysates were analyzed by immunoblot with the antibodies indicated. Representative data are shown. (<b>A</b>) ATM, BCR-Abl and c-Kit were decreased in MYL-R relative to MYL cells as predicted from the kinome profile above. (<b>B, left panel</b>) Both total and activated Lyn and PKCβ were increased in MYL-R cells as shown using antibodies to detect phosphorylation of the activation loop of Lyn (p-Y416) or the autophosphorylation site of PKCβ. (<b>B, right panel</b>) Total amounts of IKKα, MEK2 and ERK1/2 were similar in MYL and MYL-R cells, however, phospho-specific antibodies demonstrated these kinases were more active in MYL-R cells. (<b>C</b>) Kinases from MYL and MYL-R cell lysates were captured by MIBs pull-down and analyzed by immunoblot using antibodies directed against total protein. The relative amounts of kinases captured from each cell lysate correlated with the abundance ratios predicted by the MYL-R kinome profile. MIBs exposed to MYL-R cell lysate (<b>C, left panel</b>) captured less ASK1, Jak1, MLTK, Yes, GSK3α, CDK2 and dCK; and (<b>C, right panel</b>) captured more NEK9, IKKα, RIPK2, Lyn and ERK. See also, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0066755#pone.0066755.s002" target="_blank">Figures S2</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0066755#pone.0066755.s003" target="_blank">S3</a>.</p

Lyn drives activation of MEK and increases activation of IKKα in MYL-R cells.

<p>(<b>A</b>) MYL-R cells were treated for 1 hour with dasatinib, as indicated, and lysates were analyzed by immunoblot with the antibodies indicated. Lyn phosphorylation was detected by p-SFK (Y416). For densitometry, the band densities of p-SFK, p-MEK and p-IKKα were normalized against the β-actin loading control and were plotted relative to the DMSO treatment. <i>Error bars</i>, SE (N = 3). (<b>B</b>) MYL-R cells were transduced with non-targeting shRNA (shCtrl) or shRNA directed against Lyn (shLyn) and lysates were analyzed by immunoblot with the antibodies indicated. Lyn phosphorylation was detected by p-SFK (Y416) antibody. For densitometry, the band densities of p-IKKα and p-MEK were plotted relative to total IKKα and MEK protein expression. <i>Error bars</i>, SE (N = 2).</p