Automated quantitative analysis of single and double label autoradiographs

A method for the analysis of silver grain content in both single and double label autoradiographs is presented. The total grain area is calculated by counting the number of pixels at which the recorded light intensity in transmission dark field illumination exceeds a selected threshold. The calibration tests included autoradiographs with low (3H- thymidin) and high (3H-desoxyuridin) silver grain density. The results are proportional to the customary visual grain count. For the range of visibly countable grain densities in single labeled specimens, the correlation coefficient between the computed values and the visual grain counts is better than 0.96. In the first emulsion of the two emulsion layer autoradiographs of double labeled specimens (3H-14C- thymidin) the correlation coefficient is 0.919 and 0.906. The method provides a statistical correction for the background grains not due to the isotope. The possibility to record 14C tracks by shifting the focus through the second emulsion of the double labeled specimens is also demonstrated. The reported technique is essentially independent of size, shape and density of the grains

Universal scaling of nonlocal and local resistance fluctuations in small wires

Resistance fluctuations in small metal samples result from coherent transport of the carriers. The wave functions of the carriers extend into regions which are not accessible classically. We have directly measured the length dependence of the nonlocal magnetoresistance fluctuations in Sb and Au wires by studying regions of our samples separated from the classical current path by a distance L which varied from 3Lcphi down to 0.2Lcphi (where Lcphi is the phase coherence length for the carriers in the metal). These fluctuations decay exponentially with L/Lcphi. Measurements along the classical current paths scale more slowly with L/Lcphi than predicted by the analytical theory but are in agreement with numerical simulations. We have also studied the length dependence of the magnetic field correlation scale BC, and we find that it is in qualitative agreement with a recent model calculation that accounts for the voltage probes

Probe-Configuration-Dependent Decoherence in an Aharonov-Bohm Ring

We have measured transport through mesoscopic Aharonov-Bohm (AB) rings with two different four-terminal configurations. While the amplitude and the phase of the AB oscillations are well explained within the framework of the Landaur-B\"uttiker formalism, it is found that the probe configuration strongly affects the coherence time of the electrons, i.e., the decoherence is much reduced in the configuration of so-called nonlocal resistance. This result should provide an important clue in clarifying the mechanism of quantum decoherence in solids.Comment: 4 pages, 4 figures, RevTe

The molecular organization of differentially curved caveolae indicates bendable structural units at the plasma membrane

Caveolae are small coated plasma membrane invaginations with diverse functions. Caveolae undergo curvature changes. Yet, it is unclear which proteins regulate this process. To address this gap, we develop a correlative stimulated emission depletion (STED) fluorescence and platinum replica electron microscopy imaging (CLEM) method to image proteins at single caveolae. Caveolins and cavins are found at all caveolae, independent of curvature. EHD2 is detected at both low and highly curved caveolae. Pacsin2 associates with low curved caveolae and EHBP1 with mostly highly curved caveolae. Dynamin is absent from caveolae. Cells lacking dynamin show no substantial changes to caveolae, suggesting that dynamin is not directly involved in caveolae curvature. We propose a model where caveolins, cavins, and EHD2 assemble as a cohesive structural unit regulated by intermittent associations with pacsin2 and EHBP1. These coats can flatten and curve to enable lipid traffic, signaling, and changes to the surface area of the cell

Interplay between pulsations and mass loss in the blue supergiant 55 Cygnus = HD 198478

Blue supergiant stars are known to display photometric and spectroscopic variability that is suggested to be linked to stellar pulsations. Pulsational activity in massive stars strongly depends on the star's evolutionary stage and is assumed to be connected with mass-loss episodes, the appearance of macroturbulent line broadening, and the formation of clumps in the wind. To investigate a possible interplay between pulsations and mass-loss, we carried out an observational campaign of the supergiant 55 Cyg over a period of five years to search for photospheric activity and cyclic mass-loss variability in the stellar wind. We modeled the H, He I, Si II and Si III lines using the nonlocal thermal equilibrium atmosphere code FASTWIND and derived the photospheric and wind parameters. In addition, we searched for variability in the intensity and radial velocity of photospheric lines and performed a moment analysis of the line profiles to derive frequencies and amplitudes of the variations. The Halpha line varies with time in both intensity and shape, displaying various types of profiles: P Cygni, pure emission, almost complete absence, and double or multiple peaked. The star undergoes episodes of variable mass-loss rates that change by a factor of 1.7-2 on different timescales. We also observe changes in the ionization rate of Si II and determine a multiperiodic oscillation in the He I absorption lines, with periods ranging from a few hours to 22.5 days. We interpret the photospheric line variations in terms of oscillations in p-, g-, and strange modes. We suggest that these pulsations can lead to phases of enhanced mass loss. Furthermore, they can mislead the determination of the stellar rotation. We classify the star as a post-red supergiant, belonging to the group of alpha Cyg variables.Comment: 20 pages, 18 figures, 3 tables, accepted to Astronomy & Astrophysic

Unveiling the evolutionary state of three B supergiant stars: PU Gem, ϵ\epsilon CMa and η\eta CMa

We aim to combine asteroseismology, spectroscopy, and evolutionary models to establish a comprehensive picture of the evolution of Galactic blue supergiant stars (BSG). To start such an investigation, we selected three BSG candidates for our analysis: HD 42087 (PU Gem), HD 52089 ($\epsilon$ CMa) and HD 58350 ($\eta$ CMa). These stars show pulsations and were suspected to be in an evolutionary stage either preceding or succeding the red supergiant (RSG) stage. For our analysis, we utilized the 2-min cadence TESS data to study the photometric variability and obtained new spectroscopic observations at the CASLEO observatory. We calculated CMFGEN non-LTE radiative transfer models and derived stellar and wind parameters using the iterative spectral analysis pipeline XTGRID. The spectral modeling was limited to changing only the effective temperature, surface gravity, CNO abundances, and mass-loss rates. Finally, we compared the derived metal abundances with predictions from Geneva stellar evolution models. The frequency spectra of all three stars show either stochastic oscillations, nonradial strange modes, or a rotational splitting. We conclude that the rather short sectoral observing windows of TESS prevent establishing a reliable mode identification of low frequencies connected to mass-loss variabilities. The spectral analysis confirmed gradual changes in the mass-loss rates and the derived CNO abundances comply with the values reported in the literature. We were able to achieve a quantitative match with stellar evolution models for the stellar masses and luminosities. However, the spectroscopic surface abundances turned out to be inconsistent with theoretical predictions. The stars show N enrichment, typical for CNO cycle processed material, but the abundance ratios do not reflect the associated levels of C and O depletion.Comment: 29 pages, 18 figures, Accepted for publication in Galaxie

Phosphatidylinositol-(4,5)-bisphosphate regulates sorting signal recognition by the clathrin-associated adaptor complex AP2

The alpha,beta2,mu2,sigma2 heterotetrameric AP2 complex is recruited exclusively to the phosphatidylinositol-4,5-bisphosphate (PtdIns4,5P(2))-rich plasma membrane where, amongst other roles, it selects motif-containing cargo proteins for incorporation into clathrin-coated vesicles. Unphosphorylated and mu2Thr156-monophosphorylated AP2 mutated in their alphaPtdIns4,5P(2), mu2PtdIns4,5P(2), and mu2Yxxvarphi binding sites were produced, and their interactions with membranes of different phospholipid and cargo composition were measured by surface plasmon resonance. We demonstrate that recognition of Yxxvarphi and acidic dileucine motifs is dependent on corecognition with PtdIns4,5P(2), explaining the selective recruitment of AP2 to the plasma membrane. The interaction of AP2 with PtdIns4,5P(2)/Yxxvarphi-containing membranes is two step: initial recruitment via the alphaPtdIns4,5P(2) site and then stabilization through the binding of mu2Yxxvarphi and mu2PtdIns4,5P(2) sites to their ligands. The second step is facilitated by a conformational change favored by mu2Thr156 phosphorylation. The binding of AP2 to acidic-dileucine motifs occurs at a different site from Yxxvarphi binding and is not enhanced by mu2Thr156 phosphorylation

Studies of h/e Aharonov-Bohm Photovoltaic Oscillations in Mesoscopic Au Rings

We have investigated a mesoscopic photovoltaic (PV) effect in micron-size Au rings in which a dc voltage Vdc is generated in response to microwave radiation. The effect is due to the lack of inversion symmetry in a disordered system. Aharonov-Bohm PV oscillations with flux period h/e have been observed at low microwave intensities for temperatures ranging from 1.4 to 13 K. For moderate microwave intensities the h/e PV oscillations are completely quenched providing evidence that the microwaves act to randomize the phase of the electrons. Studies of the temperature dependence of Vdc also provide evidence of the dephasing nature of the microwave field. A complete theoretical explanation of the observed behavior seems to require a theory for the PV effect in a ring geometry.Comment: 10 pages (RevTex twocolumn style), 8 figures-2 pages (one postscript file) To be published in Phys. Rev.

Analytical solutions for radiation-driven winds in massive stars - II: The δ-slow regime

Accurate mass-loss rates and terminal velocities from massive stars winds are essential to obtain synthetic spectra from radiative transfer calculations and to determine the evolutionary path of massive stars. From a theoretical point of view, analytical expressions for the wind parameters and velocity profile would have many advantages over numerical calculations that solve the complex non-linear set of hydrodynamic equations. In a previous work, we obtained an analytical description for the fast wind regime. Now, we propose an approximate expression for the line-force in terms of new parameters and obtain a velocity profile closed-form solution (in terms of the Lambert W function) for the δ-slow regime. Using this analytical velocity profile, we were able to obtain the mass-loss rates based on the m-CAK theory. Moreover, we established a relation between this new set of line-force parameters with the known stellar and m-CAK line-force parameters. To this purpose, we calculated a grid of numerical hydrodynamical models and performed a multivariate multiple regression. The numerical and our descriptions lead to good agreement between their values.Fil: Araya, I.. Universidad Mayor; ChileFil: Christen, A.. Universidad de Valparaíso; ChileFil: Cure, M.. Universidad de Valparaíso; ChileFil: Cidale, Lydia Sonia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Astrofísica La Plata. Universidad Nacional de La Plata. Facultad de Ciencias Astronómicas y Geofísicas. Instituto de Astrofísica La Plata; ArgentinaFil: Venero, Roberto Oscar José. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Astrofísica La Plata. Universidad Nacional de La Plata. Facultad de Ciencias Astronómicas y Geofísicas. Instituto de Astrofísica La Plata; ArgentinaFil: Arcos, C.. Universidad de Valparaíso; ChileFil: Gormaz Matamala, A.. Universidad de Valparaíso; Chile. Universidad Adolfo Ibañez; ChileFil: Haucke, Maximiliano. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Astrofísica La Plata. Universidad Nacional de La Plata. Facultad de Ciencias Astronómicas y Geofísicas. Instituto de Astrofísica La Plata; ArgentinaFil: Escarate, P.. Universidad Austral de Chile; ChileFil: Clavería, H.. Pontificia Universidad Católica de Valparaíso; Chil

Fast neurotransmitter release regulated by the endocytic scaffold intersectin.

Sustained fast neurotransmission requires the rapid replenishment of release-ready synaptic vesicles (SVs) at presynaptic active zones. Although the machineries for exocytic fusion and for subsequent endocytic membrane retrieval have been well characterized, little is known about the mechanisms underlying the rapid recruitment of SVs to release sites. Here we show that the Down syndrome-associated endocytic scaffold protein intersectin 1 is a crucial factor for the recruitment of release-ready SVs. Genetic deletion of intersectin 1 expression or acute interference with intersectin function inhibited the replenishment of release-ready vesicles, resulting in short-term depression, without significantly affecting the rate of endocytic membrane retrieval. Acute perturbation experiments suggest that intersectin-mediated vesicle replenishment involves the association of intersectin with the fissioning enzyme dynamin and with the actin regulatory GTPase CDC42. Our data indicate a role for the endocytic scaffold intersectin in fast neurotransmitter release, which may be of prime importance for information processing in the brain