Endogenous VEGF isoform expression regulates tumour cell motility

Background: Vascular endothelial growth factor-A (VEGF) is produced by most cancer cells as multiple alternatively spliced isoforms, which display distinct receptor and matrix binding characteristics. In addition to being a major inducer of tumour angiogenesis, VEGF also has complex functions in angiogenesis-independent aspects of tumour growth,but the role of individual VEGF isoforms remains poorly understood. Here we investigated the effects of endogenous VEGF isoform expression on tumour cell migration and invasion. Method: We used a panel of mouse fibrosarcoma cells we developed (fs188, fs164 and fs120) that express single VEGF isoforms (188, 164 or 120 respectively), under endogenous promoter control. We investigated adhesion to different matrices, 2D migration and invasion through 3D collagen. Results: Fs188 cells, are typically mesenchymal, form ruffles, display strong integrin-dependent adhesion and express high levels of pERK1/2 and pAKT. In contrast, fs164 and fs120 cells are not typically mesenchymal in morphology; they display weak binding to collagen, lack ruffles and align longitudinally forming long multicellular chains and abundant cell-cell contacts. On 3D collagen, fs188 cells remain mesenchymal while fs164 and fs120 cells adopt a rounded/amoeboid and a mix of rounded/mesenchymal morphologies respectively. Cell morphology and migration are dependent on the cytoskeleton and actinomyosin contractility, to provide traction force in mesenchymal movement, and cortical contraction for rounded amoeboid motility. Consistent with their mesenchymal characteristics, fs188 cells migrated faster in 2D and invaded 3D collagen more efficiently than fs164 or fs120 cells. Contractility inhibitors caused fs164/fs120 cells to switch to a mesenchymal morphology and accelerated their migration but not that of fs188 cells. Conclusion: VEGF isoforms are emerging as potential biomarkers for anti-VEGF therapies. Our results suggest that individual VEGF isoforms influence the migration and invasion strategies of tumour cells thus adding to the complexity of VEGF signaling within the tumor microenvironment. Acknowledgements This work was funded by Cancer Research UK

Stellar Rotation in Young Clusters. II. Evolution of Stellar Rotation and Surface Helium Abundance

We derive the effective temperatures and gravities of 461 OB stars in 19 young clusters by fitting the H-gamma profile in their spectra. We use synthetic model profiles for rotating stars to develop a method to estimate the polar gravity for these stars, which we argue is a useful indicator of their evolutionary status. We combine these results with projected rotational velocity measurements obtained in a previous paper on these same open clusters. We find that the more massive B-stars experience a spin down as predicted by the theories for the evolution of rotating stars. Furthermore, we find that the members of binary stars also experience a marked spin down with advanced evolutionary state due to tidal interactions. We also derive non-LTE-corrected helium abundances for most of the sample by fitting the He I 4026, 4387, 4471 lines. A large number of helium peculiar stars are found among cooler stars with Teff < 23000 K. The analysis of the high mass stars (8.5 solar masses < M < 16 solar masses) shows that the helium enrichment process progresses through the main sequence (MS) phase and is greater among the faster rotators. This discovery supports the theoretical claim that rotationally induced internal mixing is the main cause of surface chemical anomalies that appear during the MS phase. The lower mass stars appear to have slower rotation rates among the low gravity objects, and they have a large proportion of helium peculiar stars. We suggest that both properties are due to their youth. The low gravity stars are probably pre-main sequence objects that will spin up as they contract. These young objects very likely host a remnant magnetic field from their natal cloud, and these strong fields sculpt out surface regions with unusual chemical abundances.Comment: 50 pages 18 figures, accepted by Ap

A GLIMPSE into the Nature of Galactic Mid-IR Excesses

We investigate the nature of the mid-IR excess for 31 intermediate-mass stars that exhibit an 8 micron excess in either the Galactic Legacy Infrared Mid-Plane Survey Extraordinaire or the Mid-Course Space Experiment using high resolution optical spectra to identify stars surrounded by warm circumstellar dust. From these data we determine projected stellar rotational velocities and estimate stellar effective temperatures for the sample. We estimate stellar ages from these temperatures, parallactic distances, and evolutionary models. Using MIPS [24] measurements and stellar parameters we determine the nature of the infrared excess for 19 GLIMPSE stars. We find that 15 stars exhibit Halpha emission and four exhibit Halpha absorption. Assuming that the mid-IR excesses arise in circumstellar disks, we use the Halpha fluxes to model and estimate the relative contributions of dust and free-free emission. Six stars exhibit Halpha fluxes that imply free-free emission can plausibly explain the infrared excess at [24]. These stars are candidate classical Be stars. Nine stars exhibit Halpha emission, but their Halpha fluxes are insufficient to explain the infrared excesses at [24], suggesting the presence of a circumstellar dust component. After the removal of the free-free component in these sources, we determine probable disk dust temperatures of Tdisk~300-800 K and fractional infrared luminosities of L(IR)/L(*)~10^-3. These nine stars may be pre-main-sequence stars with transitional disks undergoing disk clearing. Three of the four sources showing Halpha absorption exhibit circumstellar disk temperatures ~300-400 K, L(IR)/L(*)~10^-3, IR colors K-[24]< 3.3, and are warm debris disk candidates. One of the four Halpha absorption sources has K-[24]> 3.3 implying an optically thick outer disk and is a transition disk candidate.Comment: 17 figures. Accepted for publication in Ap

What are the hot R Coronae Borealis stars?

We investigate the evolutionary status of four stars: V348 Sgr, DY Cen, and MV Sgr in the Galaxy and HV 2671 in the LMC. These stars have in common random deep declines in visual brightness, which are characteristic of R Coronae Borealis (RCB) stars. RCB stars are typically cool hydrogen-deficient supergiants. The four stars studied in this paper are hotter (Teff = 15–20 kK) than the majority of RCB stars (Teff = 5000–7000 K). Although these are commonly grouped together as the hot RCB stars they do not necessarily share a common evolutionary history. We present new observational data and an extensive collection of archival and previously published data that is reassessed to ensure internal consistency. We find temporal variations of various properties on different timescales that will eventually help us to uncover the evolutionary history of these objects. DY Cen and MV Sgr have typical RCB helium abundances, which exclude any currently known post–asymptotic giant branch (post-AGB) evolutionary models. Moreover, their carbon and nitrogen abundances present us with further problems for their interpretation. V348 Sgr and HV 2671 are in general agreement with a born-again post-AGB evolution, and their abundances are similar to Wolf-Rayet central stars of planetary nebulae (PNs). The three Galactic stars in the sample have circumstellar nebulae, which produce forbidden line radiation (for HV 2671 we have no information). V348 Sgr and DY Cen have low-density, low-expansion velocity nebulae (resolved in the case of V348 Sgr), while MV Sgr has a higher density, higher expansion velocity nebula. All three stars, on the other hand, have split emission lines, which indicate the presence of an equatorial bulge but not of a Keplerian disk. In addition, the historical light curves for the three Galactic hot RCB stars show evidence for a significant fading in their maximum-light brightnesses of ~1 mag over the last 70 yr. From this we deduce that their effective temperatures increased by a few thousand degrees. If V348 Sgr is a born-again star, as we presume, this means that the star is returning from the born-again AGB phase to the phase of a central star of PN. Spectroscopically, no dramatic change is observed over the last 50 years for V348 Sgr and MV Sgr. However, there is some evidence that the winds of V348 Sgr and DY Cen have increased in strength in the last decade. HV 2671, located in the LMC, has not been analyzed in detail but at 5 Å… resolution is almost identical to V348 Sgr. Through the bolometric correction derived for V348 Sgr and the known distance, we can estimate the absolute ν magnitude of HV 2671 (Mν = -3.0 mag) and its bolometric luminosity (~6000 L⊙)

Direct visualization of electroporation-assisted in vivo gene delivery to tumors using intravital microscopy – spatial and time dependent distribution

BACKGROUND: Electroporation is currently receiving much attention as a way to increase drug and DNA delivery. Recent studies demonstrated the feasibility of electrogene therapy using a range of therapeutic genes for the treatment of experimental tumors. However, the transfection efficiency of electroporation-assisted DNA delivery is still low compared to viral methods and there is a clear need to optimize this approach. In order to optimize treatment, knowledge about spatial and time dependency of gene expression following delivery is of utmost importance in order to improve gene delivery. Intravital microscopy of tumors growing in dorsal skin fold window chambers is a useful method for monitoring gene transfection, since it allows non-invasive dynamic monitoring of gene expression in tumors in a live animal. METHODS: Intravital microscopy was used to monitor real time spatial distribution of the green fluorescent protein (GFP) and time dependence of transfection efficiency in syngeneic P22 rat tumor model. DNA alone, liposome-DNA complexes and electroporation-assisted DNA delivery using two different sets of electric pulse parameters were compared. RESULTS: Electroporation-assisted DNA delivery using 8 pulses, 600 V/cm, 5 ms, 1 Hz was superior to other methods and resulted in 22% increase in fluorescence intensity in the tumors up to 6 days post-transfection, compared to the non-transfected area in granulation tissue. Functional GFP was detected within 5 h after transfection. Cells expressing GFP were detected throughout the tumor, but not in the surrounding tissue that was not exposed to electric pulses. CONCLUSIONS: Intravital microscopy was demonstrated to be a suitable method for monitoring time and spatial distribution of gene expression in experimental tumors and provided evidence that electroporation-assisted gene delivery using 8 pulses, 600 V/cm, 5 ms, 1 Hz is an effective method, resulting in early onset and homogenous distribution of gene expression in the syngeneic P22 rat tumor model

Search for new phenomena in final states with an energetic jet and large missing transverse momentum in pp collisions at √ s = 8 TeV with the ATLAS detector

Results of a search for new phenomena in final states with an energetic jet and large missing transverse momentum are reported. The search uses 20.3 fb−1 of √ s = 8 TeV data collected in 2012 with the ATLAS detector at the LHC. Events are required to have at least one jet with pT > 120 GeV and no leptons. Nine signal regions are considered with increasing missing transverse momentum requirements between Emiss T > 150 GeV and Emiss T > 700 GeV. Good agreement is observed between the number of events in data and Standard Model expectations. The results are translated into exclusion limits on models with either large extra spatial dimensions, pair production of weakly interacting dark matter candidates, or production of very light gravitinos in a gauge-mediated supersymmetric model. In addition, limits on the production of an invisibly decaying Higgs-like boson leading to similar topologies in the final state are presente

Thermodynamics of mixing in diopside-jadeite, CaMgSi2O6-NaAlSi2O6, solid solution from staticlattice energy calculations

Static lattice energy calculations (SLEC), based on empirical interatomic potentials, have beenperformed for a set of 800 different structures in a 2 2 4 supercell of C2/c diopside with compositionsbetween diopside and jadeite, and with different states of order of the exchangeable Na/Ca and Mg/Al cations. Excess static energies of these structures have been cluster expanded in a basis set of 37 pair-interaction parameters. These parameters have been used to constrain Monte Carlo simulations of temperature-dependent properties in the range of 273?2,023 K and to calculate a temperature?composition phase diagram. The simulations predict the order?disorder transition in omphacite at1,150 20C in good agreement with the experimental data of Carpenter (Mineral Petrol 78:433?440, 1981). The stronger ordering of Mg/Al within the M1 site than of Ca/Na in the M2 site is attributed to the shorter M1?M1 nearest-neighbor distance, and, consequently, the stronger ordering force. The comparison of the simulated relationship between the order parameters corresponding to M1 and M2 sites with the X-ray refinement data on natural omphacites (Boffa Ballaran et al. in Am Mineral83:419?433, 1998) suggests that the cation ordering becomes kinetically ineffective at about 600C

Identification of novel vascular targets in lung cancer

Background: Lung cancer remains the leading cause of cancer-related death, largely owing to the lack of effective treatments. A tumour vascular targeting strategy presents an attractive alternative; however, the molecular signature of the vasculature in lung cancer is poorly explored. This work aimed to identify novel tumour vascular targets in lung cancer. Methods: Enzymatic digestion of fresh tissue followed by endothelial capture with Ulex lectin-coated magnetic beads was used to isolate the endothelium from fresh tumour specimens of lung cancer patients. Endothelial isolates from the healthy and tumour lung tissue were subjected to whole human genome expression profiling using microarray technology. Results: Bioinformatics analysis identified tumour endothelial expression of angiogenic factors, matrix metalloproteases and cellsurface transmembrane proteins. Predicted novel tumour vascular targets were verified by RNA-seq, quantitative real-time PCR analysis and immunohistochemistry. Further detailed expression profiling of STEAP1 on 82 lung cancer patients confirmed STEAP1 as a novel target in the tumour vasculature. Functional analysis of STEAP1 using siRNA silencing implicates a role in endothelial cell migration and tube formation. Conclusions: The identification of cell-surface tumour endothelial markers in lung is of interest in therapeutic antibody and vaccine development