Effect of Ca2+ on the Steady-State and Time-Resolved Emission Properties of the Genetically Encoded Fluorescent Sensor CatchER

We previously designed a calcium sensor CatchER (a GFP-based Calcium sensor for detecting high concentrations in the high calcium concentration environment such as ER) with a capability for monitoring calcium ion responses in various types of cells. Calcium binding to CatchER induces the ratiometric changes in the absorption spectra, as well as an increase in fluorescence emission at 510 nm upon excitation at both 395 and 488 nm. Here, we have applied the combination of the steady-state and time-resolved optical methods and Hydrogen/Deuterium isotope exchange to understand the origin of such calcium-induced optical property changes of CatchER. We first demonstrated that calcium binding results in a 44% mean fluorescence lifetime increase of the indirectly excited anionic chromophore. Thus, CatchER is the first protein-based calcium indicator with the single fluorescent moiety to show the direct correlation between the lifetime and calcium binding. Calcium exhibits a strong inhibition on the excited-state proton transfer nonadiabatic geminate recombination in protic (vs deuteric) medium. Analysis of CatchER crystal structures and the MD simulations reveal the proton transfer mechanism in which the disrupted proton migration path in CatchER is rescued by calcium binding. Our finding provides important insights for a strategy to design calcium sensors and suggests that CatchER could be a useful probe for FLIM imaging of calcium in situ

Observation of interference between two distinct autoionizing states in dissociative photoionization of H 2

Dissociative photoionization (DPI) of randomly oriented H2 molecules has been studied using linearly polarized synchrotron radiation at selected photon energies of 31, 33, and 35 eV. Large amplitude oscillations in the photoelectron asymmetry parameter β, as a function of electron energy, have been observed. The phase of these β oscillations are in excellent agreement with the results of recent close coupling calculations [Fernández and Martín, New J. Phys. 11, 043020 (2009)]. We show that the oscillations are the signature of interferences between the 1Q1 Σu+1 and 1Q2 Πu1 doubly excited states decaying at different internuclear distances. The oscillations thus provide information about the classical paths followed by the nuclei. The presence of such oscillations is predicted to be a general phenomenon in DPIWe acknowledge NSERC, CLS, and UoW for their support. We thank Mare Nostrum BSC and CCC-UAM for allocation of computer time. Work partially supported by the MICINN Projects No. FIS2010-15127, No. ACI2008-0777, and No. CSD 2007-00010, the European MC-ITN CORINF, and the COST Action CM070

Optical photometric GTC/OSIRIS observations of the young massive association Cygnus OB2

In order to fully understand the gravitational collapse of molecular clouds, the star formation process and the evolution of circumstellar disks, these phenomena must be studied in different Galactic environments with a range of stellar contents and positions in the Galaxy. The young massive association Cygnus OB2, in the Cygnus-X region, is an unique target to study how star formation and the evolution of circumstellar disks proceed in the presence of a large number of massive stars. We present a catalog obtained with recent optical observations in r,i,z filters with OSIRIS, mounted on the $10.4\,m$ GTC telescope, which is the deepest optical catalog of Cyg OB2 to date. The catalog consist of 64157 sources down to M=0.15 solar masses at the adopted distance and age of Cyg OB2. A total of 38300 sources have good photometry in all three bands. We combined the optical catalog with existing X-ray data of this region, in order to define the cluster locus in the optical diagrams. The cluster locus in the r-i vs. i-z diagram is compatible with an extinction of the optically selected cluster members in the 2.64<AV<5.57 range. We derive an extinction map of the region, finding a median value of AV=4.33 in the center of the association, decreasing toward the north-west. In the color-magnitude diagrams, the shape of the distribution of main sequence stars is compatible with the presence of an obscuring cloud in the foreground at about 850+/-25 pc from the Sun.Comment: Accepted for publication ApJS 201

Sex‐specific alterations in whole body energetics and voluntary activity in heterozygous R163C malignant hyperthermia‐susceptible mice

Malignant hyperthermia (MH) is characterized by induction of skeletal muscle hyperthermia in response to a dysregulated increase in myoplasmic calcium. Although altered energetics play a central role in MH, MH‐susceptible humans and mouse models are often described as having no phenotype until exposure to a triggering agent. The purpose of this study was to determine the influence of the R163C ryanodine receptor 1 mutation, a common MH mutation in humans, on energy expenditure, and voluntary wheel running in mice. Energy expenditure was measured by indirect respiration calorimetry in wild‐type (WT) and heterozygous R163C (HET) mice over a range of ambient temperatures. Energy expenditure adjusted for body weight or lean mass was increased (P < .05) in male, but not female, HET mice housed at 22°C or when housed at 28°C with a running wheel. In female mice, voluntary wheel running was decreased (P < .05) in the HET vs WT animals when analyzed across ambient temperatures. The thermoneutral zone was also widened in both male and female HET mice. The results of the study show that the R163C mutations alters energetics even at temperatures that do not typically induce MH

The Distance and Size of the Red Hypergiant NML Cyg from VLBA and VLA Astrometry

We have measured the annual parallax and proper motion of NML Cyg from multiple epoch VLBA observations of the circulstellar H2O and SiO masers. The measured parallax of NML Cyg is 0.620+/-0.047 mas, corresponding to a distance of 1.61+/-0.12 kpc. The measured proper motion of NML Cyg is mu_x = -1.55+/-0.42 mas/yr eastward and mu_y= -4.59+/-0.41 mas/yr northward. Both Both the distance and proper motion are consistent with that of Cyg OB2, within their joint uncertainty, confirming their association. Taking into consideration molecular absorption signatures seen toward NML Cyg, we suggest that NML Cyg lies on the far side of the Cyg OB2 association. The stellar luminosity revised with our distance brings NML Cyg significantly below the empirical luminosity limit for a red supergiant. Using the VLA observation the radio photosphere and the SiO maser as a phase reference, we partially resolve the radio photosphere of NML Cyg at 43 GHz and find its diameter is about 44 mas, suggesting an optical stellar diameter of 22 mas, if the size of radio photosphere is 2 times the optical photosphere. Based on the position of circumstellar SiO masers relative to the radio photosphere, we estimate the absolute position of NML Cyg at epoch 2008.868 to be R.A. = 20h46m25.5382s +/- 0.0010s, Decl. = 40d06'59.379" +/- 0.015". The peculiar motions of NML Cyg, the average of stars in Cyg OB2, and four other star-forming regions rules out that an expanding "Stroemgren sphere" centered on Cyg OB2 is responsible for the kinematics of the Cygnus X region.Comment: 15 pages, 11 figures, accepted by A&

TeV gamma-rays and neutrinos from photo-disintegration of nuclei in Cygnus OB2

TeV gamma-rays may provide significant information about high energy astrophysical accelerators. Such gamma-rays can result from the photo-de-excitation of PeV nuclei after their parents have undergone photo-disintegration in an environment of ultraviolet photons. This process is proposed as a candidate explanation of the recently discovered HEGRA source at the edge of the Cygnus OB2 association. The Lyman-alpha background is provided by the rich O and B stellar environment. It is found that (1) the HEGRA flux can be obtained if there is efficient acceleration at the source of lower energy nuclei; (2) the requirement that the Lorentz-boosted ultraviolet photons can excite the Giant Dipole resonance implies a strong suppression of the gamma-ray spectrum compared to an E_\gamma^{-2} behavior at energies \alt 1 TeV (some of these energies will be probed by the upcoming GLAST mission); (3) a TeV neutrino counterpart from neutron decay following helium photo-disintegration will be observed at IceCube only if a major proportion of the kinetic energy budget of the Cygnus OB2 association is expended in accelerating nuclei.Comment: To be published in Phys. Rev.

Synchrony and Physiological Arousal Increase Cohesion and Cooperation in Large Naturalistic Groups

Separate research streams have identified synchrony and arousal as two factors that might contribute to the effects of human rituals on social cohesion and cooperation. But no research has manipulated these variables in the field to investigate their causal – and potentially interactive – effects on prosocial behaviour. Across four experimental sessions involving large samples of strangers, we manipulated the synchronous and physiologically arousing affordances of a group marching task within a sports stadium. We observed participants’ subsequent movement, grouping, and cooperation via a camera hidden in the stadium’s roof. Synchrony and arousal both showed main effects, predicting larger groups, tighter clustering, and more cooperative behaviour in a free-rider dilemma. However, synchrony and arousal interacted on measures of clustering and cooperation: such that synchrony only encouraged closer clustering — and encouraged greater cooperation—when paired with physiological arousal. The research has implications for understanding the nature and co-occurrence of synchrony and physiological arousal in rituals around the world. It also represents the first use of real-time spatial tracking as a precise and naturalistic method of simulating collective rituals

Fifty Years of IMF Variation: The Intermediate-Mass Stars

I track the history of star count estimates of the Milky Way field star and open cluster IMFs, concentrating on the neglected mass range from 1 to 15 M${_\odot}$. The prevalent belief in a universal IMF appears to be without basis for this mass range. Two recent estimates of the field star IMF using different methods and samples give values of the average logarithmic slope $\Gamma$ between -1.7 and -2.1 in the mass range 1.1 to 4 M${_\odot}$. Two older estimates between 2 and 15 M${_\odot}$ disagree severely; the field IMF in this range is essentially unknown from star counts. Variations in $\Gamma$ among open cluster IMFs in this mass range have not decreased despite numerous detailed studies, even for studies using homogeneous data and reduction procedures and including only clusters with a significant mass range. These cluster variations \textit{might} be due to the combined effects of sampling, systematic errors, stellar evolution uncertainties, dynamical evolution, and unresolved binaries. If so, then the cluster data are consistent with a universal IMF, but are also consistent with sizeable variations. The cluster data do not allow an estimate of an average IMF or $\Gamma$ because the average depends on the choice of weighting procedure and other effects. If the spread in cluster IMFs is in excess of the effects listed above, real IMF variations must occur that do not depend much on physical conditions explored so far. The complexity of the star formation process seen in observations and simulations suggests that large realization-to-realization differences might be expected, in which case an individual cluster IMF would be in part the product of evolutionary contingency in star formation, and the function of interest is the probability distribution of IMF parameters.Comment: 18 pages, including 4 figures: invited talk presented at the conference on "IMF@50: The Stellar Initial Mass Function Fifty Years Later" held at Abbazia di Spineto, Siena, Italy, May 2004; to be published by Kluwer Academic Publishers, edited by E. Corbelli, F. Palla, and H. Zinnecke

CHANDRA/VLA Follow-up of TeV J2032+4131, the Only Unidentified TeV Gamma-ray Source

The HEGRA Cherenkov telescope array group recently reported a steady and extended unidentified TeV gamma-ray source lying at the outskirts of Cygnus OB2. This is the most massive stellar association known in the Galaxy, estimated to contain ~2600 OB type members alone. It has been previously argued that the large scale shocks and turbulence induced by the multiple interacting supersonic winds from the many young stars in such associations may play a role in accelerating Galactic cosmic rays. Indeed, Cyg OB2 also coincides with the non-variable MeV-GeV range unidentified EGRET source, 3EG 2033+4118. We report on the near-simultaneous follow-up observations of the extended TeV source region with the CHANDRA X-ray Observatory and the Very Large Array (VLA) radio telescope obtained in order to explore this possibility. Analysis of the CO, HI, and IRAS 100 micron emissions shows that the TeV source region coincides with an outlying sub-group of powerful OB stars which have evacuated or destroyed much of the ambient atomic, molecular and dust material, and which may be related to the very high-energy emissions. An interesting SNR-like structure is also revealed near the TeV source region in the CO, HI and radio emission maps. Applying a numerical simulation which accurately tracks the radio to gamma-ray emission from primary hadrons as well as primary and secondary e+/-, we find that the broadband spectrum of the TeV source region favors a predominantly nucleonic - rather than electronic - origin of the high-energy flux, though deeper X-ray and radio observations are needed to confirm this. A very reasonable, ~0.1%, conversion efficiency of Cyg OB2's extreme stellar wind mechanical luminosity to nucleonic acceleration to ~PeV (10^15 eV) energies is sufficient to explain the multifrequency emissions.Comment: ApJ accepte