A Dual-Readout F2 Assay That Combines Fluorescence Resonance Energy Transfer and Fluorescence Polarization for Monitoring Bimolecular Interactions

Forster (fluorescence) resonance energy transfer (FRET) and fluorescence polarization (FP) are widely used technologies for monitoring bimolecular interactions and have been extensively used in high-throughput screening (HTS) for probe and drug discovery. Despite their popularity in HTS, it has been recognized that different assay technologies may generate different hit lists for the same biochemical interaction. Due to the high cost of large-scale HTS campaigns, one has to make a critical choice to employee one assay platform for a particular HTS. Here we report the design and development of a dual-readout HTS assay that combines two assay technologies into one system using the Mcl-1 and Noxa BH3 peptide interaction as a model system. In this system, both FP and FRET signals were simultaneously monitored from one reaction, which is termed -Dual-Readout F2 assay- with F2 for FP and FRET. This dual-readout technology has been optimized in a 1,536-well ultra-HTS format for the discovery of Mcl-1 protein inhibitors and achieved a robust performance. This F2 assay was further validated by screening a library of 102,255 compounds. As two assay platforms are utilized for the same target simultaneously, hit information is enriched without increasing the screening cost. This strategy can be generally extended to other FP-based assays and is expected to enrich primary HTS information and enhance the hit quality of HTS campaigns.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/90469/1/adt-2E2010-2E0292.pd

Greenland tidewater glacier advanced rapidly during era of Norse Settlement

ACKNOWLEDGMENTS We thank the Greenland Institute of Natural Resources for providing logistical support in Nuuk. Martin Blicher, Thomas Juul-Pedersen, and Johanne Vad are thanked for their research and field assistance. We acknowledge the support of the National Museum of Greenland for permission to undertake excavations near Norse ruin sites (permit 2015/03). Project funding was provided by the Leverhulme Trust Research Project grant 2014-093, and J.M. Lea was supported by funding from the Quaternary Research Association, British Society for Geomorphology, and a UK Research and Innovation (UKRI) Future Leaders Fellowship (MR/S017232/1). We thank two anonymous reviewers and the editor for constructive comments, which helped to substantially improve this paper. D.M. Pearce would like to dedicate this paper to her father Richard M. Pearce.Peer reviewedPostprin

The design-by-adaptation approach to universal access: learning from videogame technology

This paper proposes an alternative approach to the design of universally accessible interfaces to that provided by formal design frameworks applied ab initio to the development of new software. This approach, design-byadaptation, involves the transfer of interface technology and/or design principles from one application domain to another, in situations where the recipient domain is similar to the host domain in terms of modelled systems, tasks and users. Using the example of interaction in 3D virtual environments, the paper explores how principles underlying the design of videogame interfaces may be applied to a broad family of visualization and analysis software which handles geographical data (virtual geographic environments, or VGEs). One of the motivations behind the current study is that VGE technology lags some way behind videogame technology in the modelling of 3D environments, and has a less-developed track record in providing the variety of interaction methods needed to undertake varied tasks in 3D virtual worlds by users with varied levels of experience. The current analysis extracted a set of interaction principles from videogames which were used to devise a set of 3D task interfaces that have been implemented in a prototype VGE for formal evaluation

Greenland tidewater glacier advanced rapidly during era of Norse settlement

Our ability to improve prognostic modeling of the Greenland Ice Sheet relies on understanding the long-term relationships between climate and mass flux (via iceberg calving) from marine-terminating tidewater glaciers (TWGs). Observations of recent TWG behavior are widely available, but long-term records of TWG advance are currently lacking. We present glacial geomorphological, sedimentological, archaeological, and modeling data to reconstruct the ~20 km advance of Kangiata Nunaata Sermia (KNS; the largest tidewater glacier in southwest Greenland) during the first half of the past millennium. The data show that KNS advanced ~15 km during the 12th and 13th centuries CE at a rate of ~115 m a–1, contemporaneous with regional climate cooling toward the Little Ice Age and comparable to rates of TWG retreat witnessed over the past ~200 years. Presence of Norse farmsteads proximal to KNS demonstrates their resilience to climate change, manifest as a rapidly advancing TWG in a cooling climate. The results place limits on the magnitude of ice-margin advance and demonstrate TWG sensitivity to climate cooling as well as warming. These data combined with our grounding-line stability analysis provide a long-term record that validates approaches to numerical modeling aiming to link calving to climate

Atmospheric stilling leads to prolonged thermal stratification in a large shallow polymictic lake

To quantify the effects of recent and potential future decreases in surface wind speeds on lake thermal stratification, we apply the one-dimensional process-based model MyLake to a large, shallow, polymictic lake, Võrtsjärv. The model is validated for a 3-year period and run separately for 28 years using long-term daily atmospheric forcing data from a nearby meteorological station. Model simulations show exceptionally good agreement with observed surface and bottom water temperatures during the 3-year period. Similarly, simulated surface water temperatures for 28 years show remarkably good agreement with long-term in situ water temperatures. Sensitivity analysis demonstrates that decreasing wind speeds has resulted in substantial changes in stratification dynamics since 1982, while increasing air temperatures during the same period had a negligible effect. Atmospheric stilling is a phenomenon observed globally, and in addition to recent increases in surface air temperature, needs to be considered when evaluating the influence of climate change on lake ecosystems

Climate patterns during former periods of mountain glaciation in Britain and Ireland: Inferences from the cirque record

We map glacial cirques, and analyse spatial variability in their altitude and aspect to derive a long-term, time-integrated, perspective on climate patterns during former periods of mountain glaciation (likely spanning multiple Quaternary glaciations) in Britain and Ireland. The data reveal that, although air temperatures were important, exposure to moisture-bearing air masses was the key factor in regulating sites of former mountain glacier formation, and indicate that during such periods, moisture supply was largely controlled by North Atlantic westerlies, with notable inland precipitation gradients (precipitation decreasing inland), similar to present day. In places, trends in cirque altitude may also reflect regional differences in the extent of cirque deepening, controlled by the dimensions and dynamics of the glaciers that came to occupy them. Specifically, comparatively deep cirques in coastal locations may reflect the former presence of dynamic (fed by moisture from the North Atlantic), but comparatively small, glaciers (largely confined to their cirques). By contrast, decreasing cirque depth further inland, may reflect the former presence of larger and/or less dynamic ice masses, occupying comparatively continental climatic conditions

ATRX:put me on repeat

Mutations in the chromatin-remodeling protein ATRX cause alpha thalassaemia and mental retardation, but the severity of the disorder is independent of the specific mutation. In this issue of Cell, Law et al. (2010) demonstrate that ATRX alters gene expression by binding to G-rich tandem repeats, and the degree of transcriptional silencing caused by ATRX mutations correlates with the number of repeats

Neuroprotection by selective allosteric potentiators of the EP2 prostaglandin receptor

Activation of the Gαs-coupled EP2 receptor for prostaglandin E2 (PGE2) promotes cell survival in several models of tissue damage. To advance understanding of EP2 functions, we designed experiments to develop allosteric potentiators of this key prostaglandin receptor. Screens of 292,000 compounds identified 93 that at 20 μM (i) potentiated the cAMP response to a low concentration of PGE2 by > 50%; (ii) had no effect on EP4 or β2 adrenergic receptors, the cAMP assay itself, or the parent cell line; and (iii) increased the potency of PGE2 on EP2 receptors at least 3-fold. In aqueous solution, the active compounds are largely present as nanoparticles that appear to serve as active reservoirs for bioactive monomer. From 94 compounds synthesized or purchased, based on the modification of one hit compound, the most active increased the potency of PGE2 on EP2 receptors 4- to 5-fold at 10 to 20 μM and showed substantial neuroprotection in an excitotoxicity model. These small molecules represent previously undescribed allosteric modulators of a PGE2 receptor. Our results strongly reinforce the notion that activation of EP2 receptors by endogenous PGE2 released in a cell-injury setting is neuroprotective