Expression and reconstitution of the bioluminescent Ca2+ reporter aequorin in human embryonic stem cells, and exploration of the presence of functional IP3 and ryanodine receptors during the early stages of their differentiation into cardiomyocytes

© The Author(s), 2016. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Science China Life Sciences 59 (2016): 811-824, doi:10.1007/s11427-016-5094-6.In order to develop a novel method of visualizing possible Ca2+ signaling during the early differentiation of hESCs into cardiomyocytes and avoid some of the inherent problems associated with using fluorescent reporters, we expressed the bioluminescent Ca2+ reporter, apo-aequorin, in HES2 cells and then reconstituted active holo-aequorin by incubation with f-coelenterazine. The temporal nature of the Ca2+ signals generated by the holo-f-aequorin-expressing HES2 cells during the earliest stages of differentiation into cardiomyocytes was then investigated. Our data show that no endogenous Ca2+ transients (generated by release from intracellular stores) were detected in 1–12-day-old cardiospheres but transients were generated in cardiospheres following stimulation with KCl or CaCl2, indicating that holo-f-aequorin was functional in these cells. Furthermore, following the addition of exogenous ATP, an inositol trisphosphate receptor (IP3R) agonist, small Ca2+ transients were generated from day 1 onward. That ATP was inducing Ca2+ release from functional IP3Rs was demonstrated by treatment with 2-APB, a known IP3R antagonist. In contrast, following treatment with caffeine, a ryanodine receptor (RyR) agonist, a minimal Ca2+ response was observed at day 8 of differentiation only. Thus, our data indicate that unlike RyRs, IP3Rs are present and continually functional at these early stages of cardiomyocyte differentiation.This work was supported by the Hong Kong Theme-based Research Scheme award (T13-706/11-1), the Hong Kong Research Grants Council (RGC) General Research Fund awards (662113, 16101714, 16100115), the ANR/RGC joint research scheme award (A-HKUST601/13), and the Innovation and Technology Commission (ITCPD/17-9). HYSC was supported by a Hong Kong University Grants Council post-graduate studentship (T13-706/11- 11PG)

Computing in String Field Theory Using the Moyal Star Product

Using the Moyal star product, we define open bosonic string field theory carefully, with a cutoff, for any number of string oscillators and any oscillator frequencies. Through detailed computations, such as Neumann coefficients for all string vertices, we show that the Moyal star product is all that is needed to give a precise definition of string field theory. The formulation of the theory as well as the computation techniques are considerably simpler in the Moyal formulation. After identifying a monoid algebra as a fundamental mathematical structure in string field theory, we use it as a tool to compute with ease the field configurations for wedge, sliver, and generalized projectors, as well as all the string interaction vertices for perturbative as well as monoid-type nonperturbative states. Finally, in the context of VSFT we analyze the small fluctuations around any D-brane vacuum. We show quite generally that to obtain nontrivial mass and coupling, as well as a closed strings, there must be an associativity anomaly. We identify the detailed source of the anomaly, but leave its study for future work.Comment: 77 pages, LaTeX. v3: corrections of signs or factors (for a list of corrections see beginning of source file

WW domain-mediated interaction with Wbp2 is important for the oncogenic property of TAZ

The transcriptional co-activators YAP and TAZ are downstream targets inhibited by the Hippo tumor suppressor pathway. YAP and TAZ both possess WW domains, which are important protein–protein interaction modules that mediate interaction with proline-rich motifs, most commonly PPXY. The WW domains of YAP have complex regulatory roles as exemplified by recent reports showing that they can positively or negatively influence YAP activity in a cell and context-specific manner. In this study, we show that the WW domain of TAZ is important for it to transform both MCF10A and NIH3T3 cells and to activate transcription of ITGB2 but not CTGF, as introducing point mutations into the WW domain of TAZ (WWm) abolished its transforming and transcription-promoting ability. Using a proteomic approach, we discovered potential regulatory proteins that interact with TAZ WW domain and identified Wbp2. The interaction of Wbp2 with TAZ is dependent on the WW domain of TAZ and the PPXY-containing C-terminal region of Wbp2. Knockdown of endogenous Wbp2 suppresses, whereas overexpression of Wbp2 enhances, TAZ-driven transformation. Forced interaction of WWm with Wbp2 by direct C-terminal fusion of full-length Wbp2 or its TAZ-interacting C-terminal domain restored the transforming and transcription-promoting ability of TAZ. These results suggest that the WW domain-mediated interaction with Wbp2 promotes the transforming ability of TAZ

Dynamic modeling of mean-reverting spreads for statistical arbitrage

Statistical arbitrage strategies, such as pairs trading and its generalizations, rely on the construction of mean-reverting spreads enjoying a certain degree of predictability. Gaussian linear state-space processes have recently been proposed as a model for such spreads under the assumption that the observed process is a noisy realization of some hidden states. Real-time estimation of the unobserved spread process can reveal temporary market inefficiencies which can then be exploited to generate excess returns. Building on previous work, we embrace the state-space framework for modeling spread processes and extend this methodology along three different directions. First, we introduce time-dependency in the model parameters, which allows for quick adaptation to changes in the data generating process. Second, we provide an on-line estimation algorithm that can be constantly run in real-time. Being computationally fast, the algorithm is particularly suitable for building aggressive trading strategies based on high-frequency data and may be used as a monitoring device for mean-reversion. Finally, our framework naturally provides informative uncertainty measures of all the estimated parameters. Experimental results based on Monte Carlo simulations and historical equity data are discussed, including a co-integration relationship involving two exchange-traded funds.Comment: 34 pages, 6 figures. Submitte

The role of self-gentrification in sustainable tourism: Indigenous entrepreneurship at Honghe Hani Rice Terraces World Heritage Site, China

This article examines three forms of tourism gentrification occurring within the newly inscribed (2013) Honghe Hani Rice Terraces UNESCO World Heritage Site in Yunnan, China. The indigenous Hani and Yi communities who populate this remote mountainous area, possess distinct cultural practices that have supported the rice terrace ecosystem for centuries. This article draws on interviews and non-participant observation conducted with inhabitants and newcomers to analyse the types of gentrification occurring within the site. We argue that indigenous cultural practices, and consequently rice cultivation in the area, are threatened by gentrifier-led and state-led gentrification combined with high levels of outward migration of indigenous persons. This could pose a significant threat to the sustainability of tourism at this site and may ultimately compromise the site’s World Heritage Status. In the midst of these dangers, some indigenous people are shown to be improving their socioeconomic standing – and becoming “middle class” or “gentry” – particularly through adopting entrepreneurial strategies gleaned from their encounters with outside-gentrifiers and tourists. This article proposes the concept of “self-gentrification” as a way to describe individuals who seek to improve themselves and their own community, while under threat of gentrification

Measurement of the Total Active 8B Solar Neutrino Flux at the Sudbury Neutrino Observatory with Enhanced Neutral Current Sensitivity

The Sudbury Neutrino Observatory (SNO) has precisely determined the total active (nu_x) 8B solar neutrino flux without assumptions about the energy dependence of the nu_e survival probability. The measurements were made with dissolved NaCl in the heavy water to enhance the sensitivity and signature for neutral-current interactions. The flux is found to be 5.21 +/- 0.27 (stat) +/- 0.38 (syst) x10^6 cm^{-2}s^{-1}, in agreement with previous measurements and standard solar models. A global analysis of these and other solar and reactor neutrino results yields Delta m^{2} = 7.1^{+1.2}_{-0.6}x10^{-5} ev^2 and theta = 32.5^{+2.4}_{-2.3} degrees. Maximal mixing is rejected at the equivalent of 5.4 standard deviations.Comment: Submitted to Phys. Rev. Let

In-situ characterization of the Hamamatsu R5912-HQE photomultiplier tubes used in the DEAP-3600 experiment

The Hamamatsu R5912-HQE photomultiplier-tube (PMT) is a novel high-quantum efficiency PMT. It is currently used in the DEAP-3600 dark matter detector and is of significant interest for future dark matter and neutrino experiments where high signal yields are needed. We report on the methods developed for in-situ characterization and monitoring of DEAP's 255 R5912-HQE PMTs. This includes a detailed discussion of typical measured single-photoelectron charge distributions, correlated noise (afterpulsing), dark noise, double, and late pulsing characteristics. The characterization is performed during the detector commissioning phase using laser light injected through a light diffusing sphere and during normal detector operation using LED light injected through optical fibres

Low Multiplicity Burst Search at the Sudbury Neutrino Observatory

Results are reported from a search for low-multiplicity neutrino bursts in the Sudbury Neutrino Observatory (SNO). Such bursts could indicate detection of a nearby core-collapse supernova explosion. The data were taken from Phase I (November 1999 - May 2001), when the detector was filled with heavy water, and Phase II (July 2001 - August 2003), when NaCl was added to the target. The search was a blind analysis in which the potential backgrounds were estimated and analysis cuts were developed to eliminate such backgrounds with 90% confidence before the data were examined. The search maintained a greater than 50% detection probability for standard supernovae occurring at a distance of up to 60 kpc for Phase I and up to 70 kpc for Phase II. No low-multiplicity bursts were observed during the data-taking period.Comment: 11 pages, 4 figures, submitted to Ap