Indicators of replicative damage in equine tendon fibroblast monolayers

<p>Background: Superficial digital flexor tendon (SDFT) injuries of horses usually follow cumulative matrix microdamage; it is not known why the reparative abilities of tendon fibroblasts are overwhelmed or subverted. Relevant in vitro studies of this process require fibroblasts not already responding to stresses caused by the cell culture protocols. We investigated indicators of replicative damage in SDFT fibroblast monolayers, effects of this on their reparative ability, and measures that can be taken to reduce it.</p> <p>Results: We found significant evidence of replicative stress, initially observing consistently large numbers of binucleate (BN) cells. A more variable but prominent feature was the presence of numerous gammaH2AX (γH2AX) puncta in nuclei, this being a histone protein that is phosphorylated in response to DNA double-stranded breaks (DSBs). Enrichment for injury detection and cell cycle arrest factors (p53 (ser15) and p21) occurred most frequently in BN cells; however, their numbers did not correlate with DNA damage levels and it is likely that the two processes have different causative mechanisms. Such remarkable levels of injury and binucleation are usually associated with irradiation, or treatment with cytoskeletal-disrupting agents.</p> <p>Both DSBs and BN cells were greatest in subconfluent (replicating) monolayers. The DNA-damaged cells co-expressed the replication markers TPX2/repp86 and centromere protein F. Once damaged in the early stages of culture establishment, fibroblasts continued to express DNA breaks with each replicative cycle. However, significant levels of cell death were not measured, suggesting that DNA repair was occurring. Comet assays showed that DNA repair was delayed in proportion to levels of genotoxic stress.</p> <p>Conclusions: Researchers using tendon fibroblast monolayers should assess their “health” using γH2AX labelling. Continued use of early passage cultures expressing initially high levels of γH2AX puncta should be avoided for mechanistic studies and ex-vivo therapeutic applications, as this will not be resolved with further replicative cycling. Low density cell culture should be avoided as it enriches for both DNA damage and mitotic defects (polyploidy). As monolayers differing only slightly in baseline DNA damage levels showed markedly variable responses to a further injury, studies of effects of various stressors on tendon cells must be very carefully controlled.</p&gt

Detailed Abundances for the Old Population near the Galactic Center: I. Metallicity distribution of the Nuclear Star Cluster

We report the first high spectral resolution study of 17 M giants kinematically confirmed to lie within a few parsecs of the Galactic Center, using R=24,000 spectroscopy from Keck/NIRSPEC and a new linelist for the infrared K band. We consider their luminosities and kinematics, which classify these stars as members of the older stellar population and the central cluster. We find a median metallicity of =-0.16 and a large spread from approximately -0.3 to +0.3 (quartiles). We find that the highest metallicities are [Fe/H]<+0.6, with most of the stars being at or below the Solar iron abundance. The abundances and the abundance distribution strongly resembles that of the Galactic bulge rather than disk or halo; in our small sample we find no statistical evidence for a dependence of velocity dispersion on metallicity.Comment: 18 pages, 14 figures, accepted for publication in A

Solution of Orthopositronium lifetime Puzzle

The intrinsic decay rate of orthopositronium formed in ${\rm SiO_2}$ powder is measured using the direct $2\gamma$ correction method such that the time dependence of the pick-off annihilation rate is precisely determined. The decay rate of orthopositronium is found to be $7.0396\pm0.0012 (stat.)\pm0.0011 (sys.)\mu s^{-1}$, which is consistent with our previous measurements with about twice the accuracy. Results agree well with the $O(\alpha^2)$ QED prediction, and also with a result reported very recently using nanoporous film

Optical Studies of InGaAs/GaAs Multiple Quantum Wells (MQW\u27s) and GaAs/Si Using Novel Scanning Electron Microscopy Based Techniques

The influence of defects on electron-hole plasma transport in nipi-doped in 0.2Ga0.8As/GaAs multiple quantum wells (MQWs) has been studied using a novel technique called electron beam induced absorption modulation (EBIA) imaging. Modulation doped MQW structures exhibit large optical nonlinearities and are important for the development of all-optical spatial light modulators used in optical computing and communication. The electron-hole plasma is generated by a high-energy electron beam in a scanning electron microscope and is used as a probe to study the MQW absorption modulation induced locally by the electron beam. The influence of structural defects on the diffusive transport of carriers is imaged with a μm-scale resolution. A strong spatial correlation between dark line defects observed in cathodoluminescence (CL) and absorption modulation steps in EBIA reveals the existence of strain-induced band edge fluctuations that are caused by misfit dislocations. The spatial variation of stress in metalorganic chemical vapor deposition grown GaAs/Si has been studied with linearly polarized cathodoluminescence (LPCL). GaAs grown on Si wafers in selective areas (10 to 1000 μm mesas) were studied. The large difference in thermal expansion coefficient between GaAs and Si results in thermal stress-induced microcracks and dislocations which can occur upon cooling from growth temperatures. Using LPCL, we have determined the spatial distribution of the stress tensor from the polarized CL strain-split peak positions and solutions to the orbital-strain Hamiltonian

Fabrication and characterization of a lithium-glass-based composite neutron detector

A novel composite, scintillating material intended for neutron detection and composed of small (1.5 mm) cubes of KG2-type lithium glass embedded in a matrix of scintillating plastic has been developed in the form of a 2.2 in.-diameter, 3.1 in.-tall cylindrical prototype loaded with $\left( 5.82 \pm 0.02 \right)\%$ lithium glass by mass. The response of the material when exposed to ${}^{252}$Cf fission neutrons and various $\gamma$-ray sources has been studied; using the charge-integration method for pulse shape discrimination, good separation between neutron and $\gamma$-ray events is observed and intrinsic efficiencies of $\left( 1.15 \pm 0.16 \right)\times 10^{-2}$ and $\left( 2.28 \pm 0.21 \right)\times 10^{-4}$ for ${}^{252}$Cf fission neutrons and ${}^{60}$Co $\gamma$ rays are obtained; an upper limit for the sensitivity to ${}^{137}$Cs $\gamma$ rays is determined to be $< 3.70 \times 10^{-8}$. The neutron/$\gamma$ discrimination capabilities are improved in circumstances when a neutron capture signal in the lithium glass can be detected in coincidence with a preceding elastic scattering event in the plastic scintillator; with this coincidence requirement, the intrinsic efficiency of the prototype detector for ${}^{60}$Co $\gamma$ rays is $\left( 2.42 \pm 0.61 \right)\times 10^{-6}$ while its intrinsic efficiency for unmoderated ${}^{252}$Cf fission neutrons is $\left( 4.31 \pm 0.59 \right)\times 10^{-3}$. Through use of subregion-integration ratios in addition to the coincidence requirement, the efficiency for $\gamma$ rays from ${}^{60}$Co is reduced to $\left( 7.15 \pm 4.10 \right) \times 10^{-7}$ while the ${}^{252}$Cf fission neutron efficiency becomes $\left( 2.78 \pm 0.38 \right) \times 10^{-3}$.Comment: Final results, figures, and text; published in Nuclear Instruments and Methods in Physics Research Section

Halos of Spiral Galaxies. III. Metallicity Distributions

(Abriged) We report results of a campaign to image the stellar populations in the halos of highly inclined spiral galaxies, with the fields roughly 10 kpc (projected) from the nuclei. We use the F814W (I) and F606W (V) filters in the Wide Field Planetary Camera 2, on board the Hubble Space telescope. Extended halo populations are detected in all galaxies. The color-magnitude diagrams appear to be completely dominated by giant-branch stars, with no evidence for the presence of young stellar populations in any of the fields. We find that the metallicity distribution functions are dominated by metal-rich populations, with a tail extending toward the metal poor end. To first order, the overall shapes of the metallicity distribution functions are similar to what is predicted by simple, single-component model of chemical evolution with the effective yields increasing with galaxy luminosity. However, metallicity distributions significantly narrower than the simple model are observed for a few of the most luminous galaxies in the sample. It appears clear that more luminous spiral galaxies also have more metal-rich stellar halos. The increasingly significant departures from the closed-box model for the more luminous galaxies indicate that a parameter in addition to a single yield is required to describe chemical evolution. This parameter, which could be related to gas infall or outflow either in situ or in progenitor dwarf galaxies that later merge to form the stellar halo, tends to act to make the metallicity distributions narrower at high metallicity.Comment: 20 pages, 8 figures (ApJ, in press

Massive Star Cluster Formation and Destruction in Luminous Infrared Galaxies in GOALS

We present the results of a {\it Hubble Space Telescope} ACS/HRC FUV, ACS/WFC optical study into the cluster populations of a sample of 22 Luminous Infrared Galaxies in the Great Observatories All-Sky LIRG Survey. Through integrated broadband photometry we have derived ages and masses for a total of 484 star clusters contained within these systems. This allows us to examine the properties of star clusters found in the extreme environments of LIRGs relative to lower luminosity star-forming galaxies in the local Universe. We find that by adopting a Bruzual \& Charlot simple stellar population (SSP) model and Salpeter initial mass function, the age distribution of clusters declines as $dN/d\tau = \tau^{-0.9 +/- 0.3}$, consistent with the age distribution derived for the Antennae Galaxies, and interpreted as evidence for rapid cluster disruption occuring in the strong tidal fields of merging galaxies. The large number of $10^{6} M_{\odot}$ young clusters identified in the sample also suggests that LIRGs are capable of producing more high-mass clusters than what is observed to date in any lower luminosity star-forming galaxy in the local Universe. The observed cluster mass distribution of $dN/dM = M^{-1.95 +/- 0.11}$ is consistent with the canonical -2 power law used to describe the underlying initial cluster mass function (ICMF) for a wide range of galactic environments. We interpret this as evidence against mass-dependent cluster disruption, which would flatten the observed CMF relative to the underlying ICMF distribution.Comment: 63 pages, 58 Figures, 56 Tables, Accepted for publication in Ap