The ERAD Inhibitor Eeyarestatin I Is a Bifunctional Compound with a Membrane-Binding Domain and a p97/VCP Inhibitory Group

Protein homeostasis in the endoplasmic reticulum (ER) has recently emerged as a therapeutic target for cancer treatment. Disruption of ER homeostasis results in ER stress, which is a major cause of cell death in cells exposed to the proteasome inhibitor Bortezomib, an anti-cancer drug approved for treatment of multiple myeloma and Mantle cell lymphoma. We recently reported that the ERAD inhibitor Eeyarestatin I (EerI) also disturbs ER homeostasis and has anti-cancer activities resembling that of Bortezomib.Here we developed in vitro binding and cell-based functional assays to demonstrate that a nitrofuran-containing (NFC) group in EerI is the functional domain responsible for the cytotoxicity. Using both SPR and pull down assays, we show that EerI directly binds the p97 ATPase, an essential component of the ERAD machinery, via the NFC domain. An aromatic domain in EerI, although not required for p97 interaction, can localize EerI to the ER membrane, which improves its target specificity. Substitution of the aromatic module with another benzene-containing domain that maintains membrane localization generates a structurally distinct compound that nonetheless has similar biologic activities as EerI.Our findings reveal a class of bifunctional chemical agents that can preferentially inhibit membrane-bound p97 to disrupt ER homeostasis and to induce tumor cell death. These results also suggest that the AAA ATPase p97 may be a potential drug target for cancer therapeutics

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Reconnaissance Report of the May 28, 2009 Honduras Earthquake, M 7.3

A team sponsored by the Earthquake Engineering Research Institute (EERI) and the Geoengineering Extreme Event Reconnaissance (GEER) Association carried out a field investigation in conjunction with Honduran colleagues from June 18-23 to document effects of the May 28 earthquake. The EERI-GEER team was invited by Mr. Marco Sandoval, Executive Director of the Comisión Ejecutiva Valle de Sula (CEVS). Mr. Sandoval sent a team of engineers to accompany the EERI-GEER team. The team included experts in structural, geotechnical engineering, as well as in disaster response and recovery. The investigators were supported by EERI: Abdeldjelil Belarbi and GEER: Ronaldo Luna and Kermit Applegate, all from Missouri S&T, Rolla. The CEVS team consisted of Humberto Calderon, Osvaldo Rivera, and Luis Alonso Lopez. Observations of other individuals who visited the earthquake-affected region have also been incorporated in this report. This material is based upon work supported by the National Science Foundation through the GeoEnvironmental Engineering and GeoHazards Mitigation Program under Grant No. CMMI-0825734. Any opinions, findings, and conclusions or recommendations expressed in this material are those of the authors and do not necessarily reflect the views of the NSF. The GEER Association is made possible by the vision and support of the NSF Geoenvironmental Engineering and GeoHazards Mitigation Program Directors: Dr. Richard Fragaszy and the late Dr. Cliff Astill. GEER members also donate their time, talent, and resources to collect time-sensitive field observations of the effects of earthquakes. The EERI efforts were supported through the Learning From Earthquakes (LFE) program which is also funded by the National Science Foundation

Chemical Modulation of Endocytic Sorting Augments Adeno-associated Viral Transduction

Intracellular trafficking of viruses can be influenced by a variety of inter-connected cellular sorting and degradation pathways involving endo-lysosomal vesicles, the ubiquitin-proteasome system, and autophagy-based or endoplasmic reticulum-associated machinery. In the case of recombinant adeno-associated viruses (AAV), proteasome inhibitors are known to prevent degradation of ubiquitinated AAV capsids, thereby leading to increased nuclear accumulation and transduction. However, the impact of other cellular degradation pathways on AAV trafficking is not well understood. In the current study, we screened a panel of small molecules focused on modulating different cellular degradation pathways and identified eeyarestatin I (EerI) as a novel reagent that enhances AAV transduction. EerI improved AAV transduction by an order of magnitude regardless of vector dose, genome architecture, cell type, or serotype. This effect was preceded by sequestration of AAV within enlarged vesicles that were dispersed throughout the cytoplasm. Specifically, EerI treatment redirected AAV particles toward large vesicles positive for late endosomal (Rab7) and lysosomal (LAMP1) markers. Notably, MG132 and EerI (proteasomal and endoplasmic reticulum-associated degradation inhibitors, respectively) appear to enhance AAV transduction by increasing the intracellular accumulation of viral particles in a mutually exclusive fashion. Taken together, our results expand on potential strategies to redirect recombinant AAV vectors toward more productive trafficking pathways by deregulating cellular degradation mechanisms

Quantitative Cell-based Protein Degradation Assays to Identify and Classify Drugs That Target the Ubiquitin-Proteasome System

We have generated a set of dual-reporter human cell lines and devised a chase protocol to quantify proteasomal degradation of a ubiquitin fusion degradation (UFD) substrate, a ubiquitin ligase CRL2^(VHL) substrate, and a ubiquitin-independent substrate. Well characterized inhibitors that target different aspects of the ubiquitin-proteasome system can be distinguished by their distinctive patterns of substrate stabilization, enabling assignment of test compounds as inhibitors of the proteasome, ubiquitin chain formation or perception, CRL activity, or the UFD-p97 pathway. We confirmed that degradation of the UFD but not the CRL2^(VHL) or ubiquitin-independent substrates depends on p97 activity. We optimized our suite of assays to establish conditions suitable for high-throughput screening and then validated their performance by screening against 160 cell-permeable protein kinase inhibitors. This screen identified Syk inhibitor III as an irreversible p97/vasolin containing protein inhibitor (IC_(50) = 1.7 μm) that acts through Cys-522 within the D2 ATPase domain. Our work establishes a high-throughput screening-compatible pipeline for identification and classification of small molecules, cDNAs, or siRNAs that target components of the ubiquitin-proteasome system

Select liquefaction case histories from the 2010-2011 Canterbury earthquake sequence

The 2010–2011 Canterbury earthquake sequence began with the 4 September 2010, Mw7.1 Darfield earthquake and includes up to ten events that induced liquefaction. Most notably, widespread liquefaction was induced by the Darfield and Mw6.2 Christchurch earthquakes. The combination of well-documented liquefaction response during multiple events, densely recorded ground motions for the events, and detailed subsurface characterization provides an unprecedented opportunity to add well-documented case histories to the liquefaction database. This paper presents and applies 50 high-quality cone penetration test (CPT) liquefaction case histories to evaluate three commonly used, deterministic, CPT-based simplified liquefaction evaluation procedures. While all the procedures predicted the majority of the cases correctly, the procedure proposed by Idriss and Boulanger (2008) results in the lowest error index for the case histories analyzed, thus indicating better predictions of the observed liquefaction response

Web-Based Benchmark for Keystroke Dynamics Biometric Systems: A Statistical Analysis

Most keystroke dynamics studies have been evaluated using a specific kind of dataset in which users type an imposed login and password. Moreover, these studies are optimistics since most of them use different acquisition protocols, private datasets, controlled environment, etc. In order to enhance the accuracy of keystroke dynamics' performance, the main contribution of this paper is twofold. First, we provide a new kind of dataset in which users have typed both an imposed and a chosen pairs of logins and passwords. In addition, the keystroke dynamics samples are collected in a web-based uncontrolled environment (OS, keyboards, browser, etc.). Such kind of dataset is important since it provides us more realistic results of keystroke dynamics' performance in comparison to the literature (controlled environment, etc.). Second, we present a statistical analysis of well known assertions such as the relationship between performance and password size, impact of fusion schemes on system overall performance, and others such as the relationship between performance and entropy. We put into obviousness in this paper some new results on keystroke dynamics in realistic conditions.Comment: The Eighth International Conference on Intelligent Information Hiding and Multimedia Signal Processing (IIHMSP 2012), Piraeus : Greece (2012

The macroseismic survey of the 27 February 2008 Market Rasen earthquake

Immediately following the occurrence of the Market Rasen earthquake on 27 February 2008 (5.2 ML, 4.5 Mw), an online questionnaire was opened on the BGS web site to collect felt reports. In addition, questionnaire data were collected automatically by USGS as part of the “Did You Feel It?” (DYFI) programme (Wald et al. 1999), and also by EMSC as part of its European monitoring. Some additional data were also gathered by agencies on the fringe of the felt area, notably ROB in Brussels, and DIAS in Dublin. This report summarises the findings

Haiti Earthquake January 2010: What Actions and Policies Can the Government of Haiti Implement to Improve Emergency Management Response

In 2010, Haiti experienced a devastating earthquake that destroyed much of its capital city and the governmental offices that should have guided the response to the disaster. This research focuses on how Haiti can benefit from the Caribbean Disaster Management Agency’s standards for disaster resilience as it works to recover from the earthquake. Unfortunately, Haiti has long been dependent on assistance from non-governmental organizations due to its extreme poverty; its recovery is complicated by the need to integrate disaster assistance and on-going economic and social assistance into its development of a more resilient society