Calcium Transient Assays for Compound Screening with Human iPSC-derived Cardiomyocytes: Evaluating New Tools

Calcium (Ca2+) plays a central role in regulating many biological processes in the cell from muscle contraction to neurotransmitter release. The need for reliable fluorescent calcium indicator dyes is of vast importance for studying many aspects of cell biology as well as screening compounds using phenotypic high throughput assays. We have assessed two of the latest generation of calcium indicator dyes, FLIPR Calcium 6 and Cal-520 AM for studying calcium transients (CaTs) in induced pluripotent stem cell (iPSC) -derived human cardiomyocytes. FLIPR Calcium 6 and Cal-520 dyes both displayed robust CaTs with a high signal-to-noise ratio (SNR) and were non-toxic to the cells. The analysis showed that CaT amplitudes were stable between measurements, but CaT duration was more variable and tended to increase between reads. Two methods were compared for drug-screening hit-selection; difference in average (unstandardized) and standardized difference. The unstandardized difference was better for assessing CaT amplitude, whereas standardized difference was equal to or better for assessing CaT duration. In summary, FLIPR Calcium 6 and Cal-520 are suitable dyes for drug-screening using iPSC-derived human cardiomyocytes

Global Projections of Intense Tropical Cyclone Activity for the Late Twenty-First Century from Dynamical Downscaling of CMIP5/RCP4.5 Scenarios

Global projections of intense tropical cyclone activity are derived from the Geophysical Fluid Dynamics Laboratory (GFDL) High Resolution Atmospheric Model (HiRAM; 50-km grid) and the GFDL hurricane model using a two-stage downscaling procedure. First, tropical cyclone genesis is simulated globally using HiRAM. Each storm is then downscaled into the GFDL hurricane model, with horizontal grid spacing near the storm of 6 km, including ocean coupling (e.g., cold wake generation). Simulations are performed using observed sea surface temperatures (SSTs) (1980-2008) for a control run with 20 repeating seasonal cycles and for a late-twenty-first-century projection using an altered SST seasonal cycle obtained from a phase 5 of CMIP (CMIP5)/representative concentration pathway 4.5 (RCP4.5) multimodel ensemble. In general agreement with most previous studies, projections with this framework indicate fewer tropical cyclones globally in a warmer late-twenty-first-century climate, but also an increase in average cyclone intensity, precipitation rates, and the number and occurrence days of very intense category 4 and 5 storms. While these changes are apparent in the globally averaged tropical cyclone statistics, they are not necessarily present in each individual basin. The interbasin variation of changes in most of the tropical cyclone metrics examined is directly correlated to the variation in magnitude of SST increases between the basins. Finally, the framework is shown to be capable of reproducing both the observed global distribution of outer storm size-albeit with a slight high bias-and its interbasin variability. Projected median size is found to remain nearly constant globally, with increases in most basins offset by decreases in the northwest Pacific

Dynamical Downscaling Projections of Twenty-First-Century Atlantic Hurricane Activity: CMIP3 and CMIP5 Model-Based Scenarios

Twenty-first-century projections of Atlantic climate change are downscaled to explore the robustness of potential changes in hurricane activity. Multimodel ensembles using the phase 3 of the Coupled Model Intercomparison Project (CMIP3)/Special Report on Emissions Scenarios A1B (SRES A1B; late-twenty-first century) and phase 5 of the Coupled Model Intercomparison Project (CMIP5)/representative concentration pathway 4.5 (RCP4.5; early- and late-twenty-first century) scenarios are examined. Ten individual CMIP3 models are downscaled to assess the spread of results among the CMIP3 (but not the CMIP5) models. Downscaling simulations are compared for 18-km grid regional and 50-km grid global models. Storm cases from the regional model are further downscaled into the Geophysical Fluid Dynamics Laboratory (GFDL) hurricane model (9-km inner grid spacing, with ocean coupling) to simulate intense hurricanes at a finer resolution.A significant reduction in tropical storm frequency is projected for the CMIP3 (-27%), CMIP5-early (-20%) and CMIP5-late (-23%) ensembles and for 5 of the 10 individual CMIP3 models. Lifetime maximum hurricane intensity increases significantly in the high-resolution experimentsby 4%-6% for CMIP3 and CMIP5 ensembles. A significant increase (+87%) in the frequency of very intense (categories 4 and 5) hurricanes (winds 59 m s(-1)) is projected using CMIP3, but smaller, only marginally significant increases are projected (+45% and +39%) for the CMIP5-early and CMIP5-late scenarios. Hurricane rainfall rates increase robustly for the CMIP3 and CMIP5 scenarios. For the late-twenty-first century, this increase amounts to +20% to +30% in the model hurricane\u27s inner core, with a smaller increase (similar to 10%) for averaging radii of 200 km or larger. The fractional increase in precipitation at large radii (200-400 km) approximates that expected from environmental water vapor content scaling, while increases for the inner core exceed this level

Holographic and Wilsonian Renormalization Groups

We develop parallels between the holographic renormalization group in the bulk and the Wilsonian renormalization group in the dual field theory. Our philosophy differs from most previous work on the holographic RG; the most notable feature is the key role of multi-trace operators. We work out the forms of various single- and double-trace flows. The key question, `what cutoff on the field theory corresponds to a radial cutoff in the bulk?' is left unanswered, but by sharpening the analogy between the two sides we identify possible directions.Comment: 31 pages, 3 figures. v2: Minor clarifications. Added reference

The role of potential vorticity anomalies in the Somali Jet on Indian summer monsoon intraseasonal variability

The climate of the Indian subcontinent is dominated by rainfall arising from the Indian summer monsoon (ISM) during the summer season (June to September). Intraseasonal variability during the monsoon is characterized by periods of heavy rainfall interspersed by drier periods, known as active and break events respectively. Understanding and predicting such events is of vital importance for forecasting human impacts such as water resources. The Somali Jet is a key regional feature of this circulation. In the present study, we find that the spatial structure of Somali Jet potential vorticity (PV) anomalies varies considerably during active and break periods. Analysis of these anomalies shows a mechanism whereby sea surface temperature (SST) anomalies propagate north/northwestwards through the Arabian Sea, caused by a positive feedback loop joining anomalies in SST, convection, modification of PV by diabatic heating and mixing in the atmospheric boundary layer, wind stress curl, and upwelling processes. The feedback mechanism is consistent with observed coupled ocean-atmosphere system variability timescales of approximately 20 days. This research suggests that better understanding and prediction of monsoon subseasonal variability in the South Asian monsoon may be gained by analysis of the day-to-day dynamical evolution of PV in the Somali Jet

Dynamical downscaling of tropical cyclones from CCSM4 simulations of the Last Glacial Maximum

Dynamical downscaling of simulations of the Last Glacial Maximum (LGM) and late twentieth century (20C) were conducted using the Weather Research and Forecasting (WRF) model with the aim of (1) understanding how the downscaled kinematic and thermodynamic variables influence simulated tropical cyclone (TC) activity over the western North Pacific during the LGM and the 20C periods and (2) to test the relevance of TC genesis factors for the colder LGM climate. The results show that, despite the lower temperatures during the LGM, the downscaled TC climatology over the western North Pacific in the LGM simulation does not differ significantly from that in the 20C simulation. Among the TC environmental factors, the TC potential intensity, mid‐tropospheric entropy deficit, and vertical wind shear during the LGM were consistent with previous analyses of TC genesis factors in LGM global climate model simulations. Changes in TC genesis density between the LGM and the 20C simulations seem to be well represented by the ventilation index, a nondimensional measure of the combined effects of vertical wind shear, and thermodynamic properties, suggesting the potential applicability of those factors for TC activity evaluation during the LGM and possibly other climates

Induction of endothelial cell apoptosis by the antivascular agent 5,6-dimethylxanthenone-4-acetic acid

5,6-Dimethylxanthenone-4-acetic acid, synthesised in this laboratory, reduces tumour blood flow, both in mice and in patients on Phase I trial. We used TUNEL (TdT-mediated dUTP nick end labelling) assays to investigate whether apoptosis induction was involved in its antivascular effect. 5,6-Dimethylxanthenone-4-acetic acid induced dose-dependent apoptosis in vitro in HECPP murine endothelial cells in the absence of up-regulation of mRNA for tumour necrosis factor. Selective apoptosis of endothelial cells was detected in vivo in sections of Colon 38 tumours in mice within 30 min of administration of 5,6-Dimethylxanthenone-4-acetic acid (25 mg kg−1). TUNEL staining intensified with time and after 3 h, necrosis of adjacent tumour tissue was observed. Apoptosis of central vessels in splenic white pulp was also detected in tumour-bearing mice but not in mice without tumours. Apoptosis was not observed in liver tissue. No apoptosis was observed with the inactive analogue 8-methylxanthenone-4-acetic acid. Positive TUNEL staining of tumour vascular endothelium was evident in one patient in a Phase I clinical trial, from a breast tumour biopsy taken 3 and 24 h after infusion of 5,6-Dimethylxanthenone-4-acetic acid (3.1 mg m−2). Tumour necrosis and the production of tumour tumour necrosis factor were not observed. No apoptotic staining was seen in tumour biopsies taken from two other patients (doses of 3.7 and 4.9 mg m−2). We conclude that 5,6-Dimethylxanthenone-4-acetic acid can induce vascular endothelial cell apoptosis in some murine and human tumours. The action is rapid and appears to be independent of tumour necrosis factor induction