Withaferin A Alters Intermediate Filament Organization, Cell Shape and Behavior

Withaferin A (WFA) is a steroidal lactone present in Withania somnifera which has been shown in vitro to bind to the intermediate filament protein, vimentin. Based upon its affinity for vimentin, it has been proposed that WFA can be used as an anti-tumor agent to target metastatic cells which up-regulate vimentin expression. We show that WFA treatment of human fibroblasts rapidly reorganizes vimentin intermediate filaments (VIF) into a perinuclear aggregate. This reorganization is dose dependent and is accompanied by a change in cell shape, decreased motility and an increase in vimentin phosphorylation at serine-38. Furthermore, vimentin lacking cysteine-328, the proposed WFA binding site, remains sensitive to WFA demonstrating that this site is not required for its cellular effects. Using analytical ultracentrifugation, viscometry, electron microscopy and sedimentation assays we show that WFA has no effect on VIF assembly in vitro. Furthermore, WFA is not specific for vimentin as it disrupts the cellular organization and induces perinuclear aggregates of several other IF networks comprised of peripherin, neurofilament-triplet protein, and keratin. In cells co-expressing keratin IF and VIF, the former are significantly less sensitive to WFA with respect to inducing perinuclear aggregates. The organization of microtubules and actin/microfilaments is also affected by WFA. Microtubules become wavier and sparser and the number of stress fibers appears to increase. Following 24 hrs of exposure to doses of WFA that alter VIF organization and motility, cells undergo apoptosis. Lower doses of the drug do not kill cells but cause them to senesce. In light of our findings that WFA affects multiple IF systems, which are expressed in many tissues of the body, caution is warranted in its use as an anti-cancer agent, since it may have debilitating organism-wide effects

A modern guide to quantitative spectroscopy of massive OB stars

Quantitative spectroscopy is a powerful technique from which we can extract information about the physical properties and surface chemical composition of stars. In this chapter, I guide the reader through the main ideas required to get initiated in the learning process to become an expert in the application of state-of-the-art quantitative spectroscopic techniques to the study of massive OB stars. NB: This chapter is intended to serve to young students as a first approach to a field which has attracted my attention during the last 20 years. I should note that, despite its importance, at present, the number of real experts in the field around the world is limited to less than 50 people, and about one third of them are close to retirement. Hence, I consider that this is a good moment to write a summary text on the subject to serve as guideline for the next generations of students interested in joining the massive star crew. If you are one of them, please, use this chapter as a first working notebook. Do not stop here. Dig also, for further details, into the literature I quote along the text. And, once there, dig even deeper to find all the original sources explaining in more detail the physical and technical concepts that are presently incorporated into our modern (almost) automatized tools.Comment: Accepted for publication in the book "Reviews in Frontiers of Modern Astrophysics: From Space Debris to Cosmology" (eds Kabath, Jones and Skarka; publisher Springer Nature) funded by the European Union Erasmus+ Strategic Partnership grant "Per Aspera Ad Astra Simul" 2017-1-CZ01-KA203-03556

The Tarantula Massive Binary Monitoring IV. Double-lined photometric binaries

A high fraction of massive stars are found to be binaries but only a few of them are reported as photometrically variable. By studying the populations of SB2 in the 30 Doradus region, we found a subset of them that have photometry from the OGLE project and that display variations in their light curves related to orbital motions. The goal of this study is to determine the dynamical masses and radii of the 26 binary components to investigate the mass-discrepancy problem and to provide an empirical mass-luminosity relation for the LMC. We use the PHOEBE programme to perform a systematic analysis of the OGLE V and I light curves obtained for 13 binary systems in 30 Dor. We adopt Teff, and orbital parameters derived previously to obtain the inclinations of the systems and the parameters of the individual components. Three systems display eclipses in their light curves, while the others only display ellipsoidal variations. We classify two systems as over-contact, five as semi-detached, and four as detached. The two remaining systems have uncertain configurations due to large uncertainties on their inclinations. The fact that systems display ellipsoidal variations has a significant impact on the inclination errors. From the dynamical masses, luminosities, and radii, we provide LMC-based empirical mass-luminosity and mass-radius relations, and we compare them to other relations given for the Galaxy, the LMC, and the SMC. These relations differ for different mass ranges, but do not seem to depend on the metallicity regimes. We also compare the dynamical, spectroscopic, and evolutionary masses of the stars in our sample. While the dynamical and spectroscopic masses agree with each other, the evolutionary masses are systematically higher, at least for stars in semi-detached systems. This suggests that the mass discrepancy can be partly explained by past or ongoing interactions between the stars.status: publishe

The VLT-FLAMES Tarantula Survey: XXIV. Stellar properties of the O-type giants and supergiants in 30 Doradus

Context. The Tarantula region in the Large Magellanic Cloud (LMC) contains the richest population of spatially resolved massive O-Type stars known so far. This unmatched sample offers an opportunity to test models describing their main-sequence evolution and mass-loss properties. Aims. Using ground-based optical spectroscopy obtained in the framework of the VLT-FLAMES Tarantula Survey (VFTS), we aim to determine stellar, photospheric and wind properties of 72 presumably single O-Type giants, bright giants and supergiants and to confront them with predictions of stellar evolution and of line-driven mass-loss theories. Methods. We apply an automated method for quantitative spectroscopic analysis of O stars combining the non-LTE stellar atmosphere model fastwind with the genetic fitting algorithm pikaia to determine the following stellar properties: effective temperature, surface gravity, mass-loss rate, helium abundance, and projected rotational velocity. The latter has been constrained without taking into account the contribution from macro-Turbulent motions to the line broadening. Results. We present empirical effective temperature versus spectral subtype calibrations at LMC-metallicity for giants and supergiants. The calibration for giants shows a +1kK offset compared to similar Galactic calibrations; a shift of the same magnitude has been reported for dwarfs. The supergiant calibrations, though only based on a handful of stars, do not seem to indicate such an offset. The presence of a strong upturn at spectral type O3 and earlier can also not be confirmed by our data. In the spectroscopic and classical Hertzsprung-Russell diagrams, our sample O stars are found to occupy the region predicted to be the core hydrogen-burning phase by state-of-The-Art models. For stars initially more massive than approximately 60 M, the giant phase already appears relatively early on in the evolution; the supergiant phase develops later. Bright giants, however, are not systematically positioned between giants and supergiants at M 25 M. At masses below 60 M, the dwarf phase clearly precedes the giant and supergiant phases; however this behavior seems to break down at M 18 M. Here, stars classified as late O III and II stars occupy the region where O9.5-9.7 V stars are expected, but where few such late O V stars are actually seen. Though we can not exclude that these stars represent a physically distinct group, this behavior may reflect an intricacy in the luminosity classification at late O spectral subtype. Indeed, on the basis of a secondary classification criterion, the relative strength of Si iv to He i absorption lines, these stars would have been assigned a luminosity class IV or V. Except for five stars, the helium abundance of our sample stars is in agreement with the initial LMC composition. This outcome is independent of their projected spin rates. The aforementioned five stars present moderate projected rotational velocities (i.e., ν e sini < 200kms -1 ) and hence do not agree with current predictions of rotational mixing in main-sequence stars. They may potentially reveal other physics not included in the models such as binary-interaction effects. Adopting theoretical results for the wind velocity law, we find modified wind momenta for LMC stars that are ~0.3 dex higher than earlier results. For stars brighter than 10 5 L, that is, in the regime of strong stellar winds, the measured (unclumped) mass-loss rates could be considered to be in agreement with line-driven wind predictions if the clump volume filling factors were fV 1/8 to 1/6

The Tarantula Massive Binary Monitoring. I. Observational campaign and OB-type spectroscopic binaries

ontext. Massive binaries play a crucial role in the Universe. Knowing the distributions of their orbital parameters is important for a wide range of topics from stellar feedback to binary evolution channels and from the distribution of supernova types to gravitational wave progenitors, yet no direct measurements exist outside the Milky Way. Aims. The Tarantula Massive Binary Monitoring project was designed to help fill this gap by obtaining multi-epoch radial velocity (RV) monitoring of 102 massive binaries in the 30 Doradus region. Methods. In this paper we analyze 32 FLAMES/GIRAFFE observations of 93 O- and 7 B-type binaries. We performed a Fourier analysis and obtained orbital solutions for 82 systems: 51 single-lined (SB1) and 31 double-lined (SB2) spectroscopic binaries. Results. Overall, the binary fraction and orbital properties across the 30 Doradus region are found to be similar to existing Galactic samples. This indicates that within these domains environmental effects are of second order in shaping the properties of massive binary systems. A small difference is found in the distribution of orbital periods, which is slightly flatter (in log space) in 30 Doradus than in the Galaxy, although this may be compatible within error estimates and differences in the fitting methodology. Also, orbital periods in 30 Doradus can be as short as 1.1 d, somewhat shorter than seen in Galactic samples. Equal mass binaries (q> 0.95) in 30 Doradus are all found outside NGC 2070, the central association that surrounds R136a, the very young and massive cluster at 30 Doradus’s core. Most of the differences, albeit small, are compatible with expectations from binary evolution. One outstanding exception, however, is the fact that earlier spectral types (O2–O7) tend to have shorter orbital periods than later spectral types (O9.2–O9.7). Conclusions. Our results point to a relative universality of the incidence rate of massive binaries and their orbital properties in the metallicity range from solar (Z⊙) to about half solar. This provides the first direct constraints on massive binary properties in massive star-forming galaxies at the Universe’s peak of star formation at redshifts z ~ 1 to 2 which are estimated to have Z ~ 0.5 Z⊙

Vimentin organization modulates the formation of lamellipodia

The disassembly and withdrawal of vimentin intermediate filaments (VIF) from the plasma membrane induces membrane ruffling and the formation of a lamellipodium. Conversely, lamellipodium formation is inhibited when VIF are present