The Website Accommodations Test: Applying the Americans with Disabilities Act to Websites

In 2017, 814 lawsuits were filed alleging discrimination under the Americans with Disabilities Act (ADA) due to website inaccessibility, up from 262 in the previous year. Beginning in July 2010, the federal Department of Justice (DOJ) considered issuing regulations under ADA Title III related to website accessibility. However, no changes have been made to date, leaving courts split over whether websites constitute places of public accommodation via the ADA. Dispositive to some jurisdictions’ holdings is whether a website has a nexus to a physical place, which may lend toward viewing the site as a public accommodation. Other jurisdictions provide that all websites are public accommodations under the ADA. The current approaches conflict with the Congressional intent of the ADA and provide little guidance to website owners. Thus, neither approach is sufficient for the uniform application of ADA protections. Absent Congressional action, courts should utilize a new “website accommodations test” (WAT) in applying the ADA to websites

THE ROLE OF DKSA IN THE STRINGENT RESPONSE IN THE LYME DISEASE SPIROCHETE BORRELIA BURGDORFERI

Borrelia burgdorferi, the bacterium that causes Lyme disease, is maintained in nature through an enzootic cycle that includes a tick vector and a vertebrate host. The bacterium is acquired by an Ixodes tick from an infected vertebrate. The bacterium alters its gene expression to adapt to different environments of the tick and vertebrate. Between tick feedings, B. burgdorferi must contend with nutrient stress. The stringent response is a physiological mechanism when bacteria switch from “thriving” to “surviving” mode in response to limited nutrient resources; it is mediated by an increase in the nucleotide alarmone guanosine penta- or tetraphosphate, abbreviated as (p)ppGpp. An important target of the stringent response is transcriptional regulation, which is mediated by the RNA polymerase-binding transcription factor DksA. We hypothesized that DksA potentiates the effects of the stringent response for B. burgdorferi during starvation, and regulates B. burgdorferi genes involved in tick persistence. We have taken a reverse genetic approach in which we have generated a dksA null mutant and the corresponding complemented strain. In vitro experiments indicate that DksA may play a role in growth, cell viability during starvation, and cell morphology. We also provide evidence that the presence of glycerol during starvation affects cell viability and morphology of B. burgdorferi

Targeting glutathione peroxidases : identifying environmental modulators, and screening for novel small molecule inhibitors

Glutathione peroxidases (GPXs) are a family of selenoproteins that are critical regulators of reactive oxygen species (ROS) in the cell, specifically hydroperoxides like hydrogen peroxide (H2O2). ROS are important for normal cell signaling and are tightly controlled to promote cell growth, proliferation, and survival. Without the antioxidant activity of enzymes like GPXs, however, the oxidative burden in cells can reach a point that leads to DNA damage, carcinogenesis, and eventually cell death. Although this can be catastrophic in early development, as evidenced by knock-out studies, many cancer therapeutics function through manipulating redox balance, suggesting that targeting these enzymes could have therapeutic potential. The modified metabolism of cancer cells can result in increased hydroperoxide production, and GPXs are often overexpressed to compensate for the increased oxidative stress, as well as in cell lines resistant to chemotherapeutics. The studies comprising this thesis examine several aspects of GPX inhibition to better understand how to utilize GPX-targeting agents as potential anti-cancer therapeutics and to identify novel inhibitors for future development. Paper I addresses the effects of environmental heavy metal exposures on the erythrocyte GPX activity in 9-year-old children from the Matlab region of Bangladesh. Samples from 100 children were initially analyzed for concentrations of selenium, arsenic, mercury, and cadmium, as well as C-reactive protein (CRP) using inductively coupled plasma mass spectrometry (ICPMS). GPX1 expression levels in lysed samples were measured using high-throughput immunoblotting, and total GPX activity was measured using a GPX activity assay in lysates. After finding only a slight positive correlation between GPX1 expression and GPX activity, trace elements and CRP levels were considered, and a multivariable-adjusted linear regression analyses was used to assess predictors of GPX activity. Arsenic and CRP levels were significantly negatively associated with active GPX, while not correlated with each other. These results suggest that independently both arsenic exposure and increased CRP levels due to inflammation can suppress GPX activity in erythrocytes. Paper II characterized the off-target inhibition of selenoprotein thioredoxin reductases (TXNRDs) by the ferroptosis inducers, (1S, 3R)RSL3 and ML162. Identified originally in a synthetic lethal screen for compounds specifically cytotoxic to oncogenic RAS, these compounds were then found to induce an iron-dependent cell death via increased lipid peroxidation associated with GPX4-specific inhibition. However, neither compound showed inhibitory activity in biochemical assays using recombinant GPX4, but both show potent inhibition of TXNRD1 in both biochemical and cellular assays. In three cell lines with varying susceptibility to ferroptosis, the cell death induced by RSL3 differed from the cell death caused by more specific TXNRD1 inhibitors, TRi-1 and TRi-2. Specifically, while RSL3 cytotoxicity could be rescued by co-treatment with the ferroptosis suppressor, Fer-1, the cytotoxicity of the Tri compounds was not rescued. Additionally, selenium supplementation diminished the efficacy of RSL3 while the TRi compounds remained unchanged, or slightly more cytotoxic. In all, these studies indicate that the interconnectedness of TXNRD1 and ferroptosis, and furthermore TXNRD1 and GPX4, is complex, but that this important off-target effect needs to be understood to fully characterize the use of these ferroptosis inducers. Paper III established a discovery pipeline for the identification of novel specific inhibitors of GPX1 and GPX4. GR-coupled activity assays using recombinant GPX1 and GPX4 were optimized and miniaturized to 1536-well formats and screened against 12,000 small molecules with annotated mechanisms of action. A suite of confirmational assays were used to ensure specificity: a GR counter-assay was used to identify false-positives in the primary screens; orthogonal endpoint GPX assays were used to confirm inhibitory activity; GPX2 assay was used to further probe specificity of the confirmed active compounds between isoforms; a TXNRD1 assay was used to differentiate small molecules with broad Sec-targeting activity from GPXspecific inhibition; and nano Differential Scanning Fluorimetry (DSF) was used to confirm direct binding. Interestingly, all GPX1 inhibitors identified showed crossinhibition of GPX2. Ultimately, five novel GPX1/GPX2 inhibitors, 13 GPX4 inhibitors, and 2 novel pan-GPX inhibitors. This series of assays and the resulting compounds identified provide a basis for future development of GPX-specific inhibitors. Paper IV profiled the cytotoxicity of a library of >10,000 small molecules with annotated mechanisms of action (MOA) and >100,000 small molecule scaffolds in a diversity library in both normal and cancer cell lines. These screens revealed a low overall cytotoxicity rate of the diversity library. Importantly, cytotoxicity was assessed in four normal cell lines (HEK293, immortalized human embryonic kidney cells; NIH3T3, an embryonic mouse fibroblast cells; HaCat, immortalized human keratinocytes; and CRL-7250, a primary human foreskin fibroblast cells). The top enriched MOA categories showing broad cell killing in normal cells were proteosome inhibitors, heat shock protein 90 (HSP90) inhibitors, anaplastic lymphoma kinase (ALK) inhibitors, mammalian target of rapamycin (mTOR) inhibitors, and cyclindependent kinase (CDK) inhibitors. Cytotoxic compounds with specific activity against the human adenosarcoma cancer line, KB 3-1, that showed no activity in the normal cell lines were also highlighted. This work will be used as an additional triage step in lead-selection for chemotherapeutic development, removing compounds that show significant cytotoxicity in normal cell lines

Analisis dan Perancangan Sistem Akuntansi dengan Model Driven Development (MDD) Pada Konveksi Anemarie

Informasi Akuntansi merupakan salah satu alat untuk mengambil keputusan dalam perusahaan.Sistem Informasi Akuntansi dapat memberikan kemudahan dalam menghasilkan informasi yang akurat, tepat waktu dapat dipahami dan teruji.Selain itu, sistem informasi akuntansi dapat memudahkan siklus operasional perusahaan baik dari pembelian, produksi, sampai penjualan.Maka dari uraian itu, peneliti ingin menganalisis dan merancang sistem informasi akuntansi pada usaha yang tidak memiliki sistem informasi akuntansi dan masih melakukan pencatatan secara manual. Peneliti menggunakan data sistem akuntansi Konveksi Anemarie periode April 2017 dan Model Driven Development untuk merancang Sistem Informasi Akuntansi konveksi Anemarie. Hasil pengembangan Sistem Informasi Akuntansi dengan Model Driven Development ini ialah Desain Model Data dengan Entity Relathionship diagram baik secara keseluruhan maupun per subsistem, Desain model Proses dengan Data Flow Diagram, dan desain model output dengan pembuatan interface dari form yang dibutuhka

Canvass: a crowd-sourced, natural-product screening library for exploring biological space

NCATS thanks Dingyin Tao for assistance with compound characterization. This research was supported by the Intramural Research Program of the National Center for Advancing Translational Sciences, National Institutes of Health (NIH). R.B.A. acknowledges support from NSF (CHE-1665145) and NIH (GM126221). M.K.B. acknowledges support from NIH (5R01GM110131). N.Z.B. thanks support from NIGMS, NIH (R01GM114061). J.K.C. acknowledges support from NSF (CHE-1665331). J.C. acknowledges support from the Fogarty International Center, NIH (TW009872). P.A.C. acknowledges support from the National Cancer Institute (NCI), NIH (R01 CA158275), and the NIH/National Institute of Aging (P01 AG012411). N.K.G. acknowledges support from NSF (CHE-1464898). B.C.G. thanks the support of NSF (RUI: 213569), the Camille and Henry Dreyfus Foundation, and the Arnold and Mabel Beckman Foundation. C.C.H. thanks the start-up funds from the Scripps Institution of Oceanography for support. J.N.J. acknowledges support from NIH (GM 063557, GM 084333). A.D.K. thanks the support from NCI, NIH (P01CA125066). D.G.I.K. acknowledges support from the National Center for Complementary and Integrative Health (1 R01 AT008088) and the Fogarty International Center, NIH (U01 TW00313), and gratefully acknowledges courtesies extended by the Government of Madagascar (Ministere des Eaux et Forets). O.K. thanks NIH (R01GM071779) for financial support. T.J.M. acknowledges support from NIH (GM116952). S.M. acknowledges support from NIH (DA045884-01, DA046487-01, AA026949-01), the Office of the Assistant Secretary of Defense for Health Affairs through the Peer Reviewed Medical Research Program (W81XWH-17-1-0256), and NCI, NIH, through a Cancer Center Support Grant (P30 CA008748). K.N.M. thanks the California Department of Food and Agriculture Pierce's Disease and Glassy Winged Sharpshooter Board for support. B.T.M. thanks Michael Mullowney for his contribution in the isolation, elucidation, and submission of the compounds in this work. P.N. acknowledges support from NIH (R01 GM111476). L.E.O. acknowledges support from NIH (R01-HL25854, R01-GM30859, R0-1-NS-12389). L.E.B., J.K.S., and J.A.P. thank the NIH (R35 GM-118173, R24 GM-111625) for research support. F.R. thanks the American Lebanese Syrian Associated Charities (ALSAC) for financial support. I.S. thanks the University of Oklahoma Startup funds for support. J.T.S. acknowledges support from ACS PRF (53767-ND1) and NSF (CHE-1414298), and thanks Drs. Kellan N. Lamb and Michael J. Di Maso for their synthetic contribution. B.S. acknowledges support from NIH (CA78747, CA106150, GM114353, GM115575). W.S. acknowledges support from NIGMS, NIH (R15GM116032, P30 GM103450), and thanks the University of Arkansas for startup funds and the Arkansas Biosciences Institute (ABI) for seed money. C.R.J.S. acknowledges support from NIH (R01GM121656). D.S.T. thanks the support of NIH (T32 CA062948-Gudas) and PhRMA Foundation to A.L.V., NIH (P41 GM076267) to D.S.T., and CCSG NIH (P30 CA008748) to C.B. Thompson. R.E.T. acknowledges support from NIGMS, NIH (GM129465). R.J.T. thanks the American Cancer Society (RSG-12-253-01-CDD) and NSF (CHE1361173) for support. D.A.V. thanks the Camille and Henry Dreyfus Foundation, the National Science Foundation (CHE-0353662, CHE-1005253, and CHE-1725142), the Beckman Foundation, the Sherman Fairchild Foundation, the John Stauffer Charitable Trust, and the Christian Scholars Foundation for support. J.W. acknowledges support from the American Cancer Society through the Research Scholar Grant (RSG-13-011-01-CDD). W.M.W.acknowledges support from NIGMS, NIH (GM119426), and NSF (CHE1755698). A.Z. acknowledges support from NSF (CHE-1463819). (Intramural Research Program of the National Center for Advancing Translational Sciences, National Institutes of Health (NIH); CHE-1665145 - NSF; CHE-1665331 - NSF; CHE-1464898 - NSF; RUI: 213569 - NSF; CHE-1414298 - NSF; CHE1361173 - NSF; CHE1755698 - NSF; CHE-1463819 - NSF; GM126221 - NIH; 5R01GM110131 - NIH; GM 063557 - NIH; GM 084333 - NIH; R01GM071779 - NIH; GM116952 - NIH; DA045884-01 - NIH; DA046487-01 - NIH; AA026949-01 - NIH; R01 GM111476 - NIH; R01-HL25854 - NIH; R01-GM30859 - NIH; R0-1-NS-12389 - NIH; R35 GM-118173 - NIH; R24 GM-111625 - NIH; CA78747 - NIH; CA106150 - NIH; GM114353 - NIH; GM115575 - NIH; R01GM121656 - NIH; T32 CA062948-Gudas - NIH; P41 GM076267 - NIH; R01GM114061 - NIGMS, NIH; R15GM116032 - NIGMS, NIH; P30 GM103450 - NIGMS, NIH; GM129465 - NIGMS, NIH; GM119426 - NIGMS, NIH; TW009872 - Fogarty International Center, NIH; U01 TW00313 - Fogarty International Center, NIH; R01 CA158275 - National Cancer Institute (NCI), NIH; P01 AG012411 - NIH/National Institute of Aging; Camille and Henry Dreyfus Foundation; Arnold and Mabel Beckman Foundation; Scripps Institution of Oceanography; P01CA125066 - NCI, NIH; 1 R01 AT008088 - National Center for Complementary and Integrative Health; W81XWH-17-1-0256 - Office of the Assistant Secretary of Defense for Health Affairs through the Peer Reviewed Medical Research Program; P30 CA008748 - NCI, NIH, through a Cancer Center Support Grant; California Department of Food and Agriculture Pierce's Disease and Glassy Winged Sharpshooter Board; American Lebanese Syrian Associated Charities (ALSAC); University of Oklahoma Startup funds; 53767-ND1 - ACS PRF; PhRMA Foundation; P30 CA008748 - CCSG NIH; RSG-12-253-01-CDD - American Cancer Society; RSG-13-011-01-CDD - American Cancer Society; CHE-0353662 - National Science Foundation; CHE-1005253 - National Science Foundation; CHE-1725142 - National Science Foundation; Beckman Foundation; Sherman Fairchild Foundation; John Stauffer Charitable Trust; Christian Scholars Foundation)Published versionSupporting documentatio

Is keiretsu really a source of competitive advantage for Japanese automotive suppliers?

Purpose - The purpose of this paper is to examine how firms create and sustain competitive advantage in the inter-firm business relationships from a supplier's perspective. It also investigates what factors affect their competitiveness and relationship between buyers and suppliers. Design/methodology/approach - This is an exploratory study on keiretsu partnerships composed of four main phases: analysis of theoretical perspectives, construction of a conceptual framework, interview of a CEO, and finally, a survey questionnaire with Japanese automotive suppliers. Findings - As a result, this paper classified these 11 companies into four supplier groups (affiliated or independent Tier 1 suppliers; affiliated or independent Tier 2 suppliers) and analysed their competitiveness developing the research propositions further. The benefits of affiliation under a keiretsu partnership are discussed, showing that there may be little benefit in being an affiliated Tier 1 supplier. Even more critical, the results show that independent Tier 2 supplier may be more competitive than affiliated tier ones. Originality/value - These intriguing results reveal an urgent need of investigating Japanese automotive supply chains from the suppliers' perspectives in the future research. This paper extended the literatures on competitive advantage and business relationships at both theory and managerial practice

Peripheral versus central cannulation of VA-ECMO for primary graft dysfunction after heart transplantation: A systematic review and meta-analysis

Heart transplantation; Mechanical circulatory support; Primary graft dysfunctionTrasplante cardíaco; Soporte circulatorio mecánico; Disfunción primaria del injertoTrasplantament cardíac; Suport circulatori mecànic; Disfunció primària de l'empeltBackground Severe primary graft dysfunction (PGD) after heart transplantation (HT) is a major cause of death and requires veno-arterial extracorporeal membrane oxygenation (VA-ECMO). Methods We conducted a systematic review and meta-analysis including studies of adult HT recipients who required VA-ECMO for PGD to determine whether a peripheral or central configuration was associated with higher mortality. The primary endpoints were short-term and one-year mortality. Secondary endpoints were VA-ECMO-related complications. Results Overall, we included 16 studies comprising 874 patients from 33 centers. Using a random-effects model, peripheral cannulation was associated with a nonsignificant reduction in short-term mortality (odds ratios [OR] = 0.73, 95% confidence interval [CI] = 0.41-1.28, I2 = 55.8%) and a significant reduction in 1-year mortality (OR = 0.60, 95%CI = 0.37-0.97, I2 = 35.9%). Peripheral cannulation decreased the risk of bleeding but increased the risk of limb ischemia and infection, with similar rates of stroke and need for renal replacement therapy. Overall, certainty of evidence was low. Conclusions With low certainty evidence, peripheral VA-ECMO cannulation may reduce short-term and 1-year mortality with lower bleeding rates but higher limb-related complications, supporting peripheral configuration in HT recipients with severe PGD