HST Observations of the Serendipitous X-ray Companion to Mrk 273: Cluster at z=0.46?

We have used HST I-band images to identify Mrk 273X, the very unusual high-redshift X-ray-luminous Seyfert 2 galaxy found by ROSAT in the same field-of-view as Mrk 273. We have measured the photometric properties of Mrk 273X and have also analyzed the luminosity distribution of the faint galaxy population seen in the HST image. The luminosity of the galaxy and the properties of the surrounding environment suggest that Mrk 273X is the brightest galaxy in a relatively poor cluster at a redshift near 0.46. Its off-center location in the cluster and the presence of other galaxy groupings in the HST image may indicate that this is a dynamically young cluster on the verge of merging with its neighboring clusters. We find that Mrk 273X is a bright featureless elliptical galaxy with no evidence for a disk. It follows the de Vaucouleurs (r^{1/4}) surface brightness law very well over a range of 8 magnitudes. Though the surface brightness profile does not appear to be dominated by the AGN, the galaxy has very blue colors that do appear to be produced by the AGN. Mrk 273X is most similar to the IC 5063 class of active galaxies --- a hybrid Sy 2 / powerful radio galaxy.Comment: Accepted for publication in the Astrophysical Journal. 8 pages, including 4 postscript figures. Uses emulateapj.sty and psfig.sty. Higher quality version of Figure 1 is available at http://rings.gsfc.nasa.gov/~borne/fig1-markgals.gi

Next Generation CD44v6-Specific CAR-NK Cells Effective against Triple Negative Breast Cancer

There is a medical need to develop new and effective therapies against triple-negative breast cancer (TNBC). Chimeric antigen receptor (CAR) natural killer (NK) cells are a promising alternative to CAR-T cell therapy for cancer. A search for a suitable target in TNBC identified CD44v6, an adhesion molecule expressed in lymphomas, leukemias and solid tumors that is implicated in tumorigenesis and metastases. We have developed a next-generation CAR targeting CD44v6 that incorporates IL-15 superagonist and checkpoint inhibitor molecules. We could show that CD44v6 CAR-NK cells demonstrated effective cytotoxicity against TNBC in 3D spheroid models. The IL-15 superagonist was specifically released upon recognition of CD44v6 on TNBC and contributed to the cytotoxic attack. PD1 ligands are upregulated in TNBC and contribute to the immunosuppressive tumor microenvironment (TME). Competitive inhibition of PD1 neutralized inhibition by PD1 ligands expressed on TNBC. In total, CD44v6 CAR-NK cells are resistant to TME immunosuppression and offer a new therapeutic option for the treatment of BC, including TNBC

A Catalog of Candidate Intermediate-luminosity X-ray Objects

ROSAT, and now Chandra, X-ray images allow studies of extranuclear X-ray point sources in galaxies other than our own. X-ray observations of normal galaxies with ROSAT and Chandra have revealed that off-nuclear, compact, Intermediate-luminosity (Lx[2-10 keV] >= 1e39 erg/s) X-ray Objects (IXOs, a.k.a. ULXs [Ultraluminous X-ray sources]) are quite common. Here we present a catalog and finding charts for 87 IXOs in 54 galaxies, derived from all of the ROSAT HRI imaging data for galaxies with cz <= 5000 km/s from the Third Reference Catalog of Bright Galaxies (RC3). We have defined the cutoff Lx for IXOs so that it is well above the Eddington luminosity of a 1.4 Msun black hole (10^38.3 erg/s), so as not to confuse IXOs with ``normal'' black hole X-ray binaries. This catalog is intended to provide a baseline for follow-up work with Chandra and XMM, and with space- and ground-based survey work at wavelengths other than X-ray. We demonstrate that elliptical galaxies with IXOs have a larger number of IXOs per galaxy than non-elliptical galaxies with IXOs, and note that they are not likely to be merely high-mass X-ray binaries with beamed X-ray emission, as may be the case for IXOs in starburst galaxies. Approximately half of the IXOs with multiple observations show X-ray variability, and many (19) of the IXOs have faint optical counterparts in DSS optical B-band images. Follow-up observations of these objects should be helpful in identifying their nature.Comment: 29 pages, ApJS, accepted (catalog v2.0) (full resolution version of paper and future releases of catalog at http://www.xassist.org/ixocat_hri

ROSAT Blank Field Sources I: Sample Selection and Archival Data

We have identified a population of blank field sources (or `blanks') among the ROSAT bright unidentified X-ray sources with faint optical counterparts. The extreme X-ray over optical flux ratio of blanks is not compatible with the main classes of X-ray emitters except for extreme BL Lacertae objects. From the analysis of ROSAT archival data we found no indication of variability and evidence for only three sources, out of 16, needing absorption in excess of the Galactic value. We also found evidence for an extended nature for only one of the 5 blanks with a serendipitous HRI detection; this source (1WGAJ1226.9+3332) was confirmed as a z=0.89 cluster of galaxies. Palomar images reveal the presence of a red (O-E~2) counterpart in the X-ray error circle for 6 blanks. The identification process brought to the discovery of another high z cluster of galaxies, one (possibly extreme) BL Lac, two ultraluminous X-ray sources in nearby galaxies and two apparently normal type1 AGNs. These AGNs, together with 4 more AGN-like objects seem to form a well defined group: they present unabsorbed X-ray spectra but red Palomar counterparts. We discuss the possible explanations for the discrepancy between the X-ray and optical data, among which: a suppressed big blue bump emission, an extreme dust to gas (~40-60 the Galactic ratio), a high redshift (z>3.5) QSO nature, an atypical dust grain size distribution and a dusty warm absorber. These AGN-like blanks seem to be the bright (and easier to study) analogs of the sources which are found in deep Chandra observations. Three more blanks have a still unknown nature.Comment: 23 pages, 8 figures, accepted by ApJ main journa

The Distribution of Redshifts in New Samples of Quasi-stellar Objects

Two new samples of QSOs have been constructed from recent surveys to test the hypothesis that the redshift distribution of bright QSOs is periodic in $\log(1+z)$. The first of these comprises 57 different redshifts among all known close pairs or multiple QSOs, with image separations $\leq$ 10\arcsec, and the second consists of 39 QSOs selected through their X-ray emission and their proximity to bright comparatively nearby active galaxies. The redshift distributions of the samples are found to exhibit distinct peaks with a periodic separation of $\sim 0.089$ in $\log(1+z)$ identical to that claimed in earlier samples but now extended out to higher redshift peaks $z = 2.63, 3.45$ and 4.47, predicted by the formula but never seen before. The periodicity is also seen in a third sample, the 78 QSOs of the 3C and 3CR catalogues. It is present in these three datasets at an overall significance level $10^{-5}$ - $10^{-6}$, and appears not to be explicable by spectroscopic or similar selection effects. Possible interpretations are briefly discussed.Comment: submitted for publication in the Astronomical Journal, 15 figure

Modified bases enable high-efficiency oligonucleotide-mediated allelic replacement via mismatch repair evasion

Genome engineering using single-stranded oligonucleotides is an efficient method for generating small chromosomal and episomal modifications in a variety of host organisms. The efficiency of this allelic replacement strategy is highly dependent on avoidance of the endogenous mismatch repair (MMR) machinery. However, global MMR inactivation generally results in significant accumulation of undesired background mutations. Here, we present a novel strategy using oligos containing chemically modified bases (2′-Fluoro-Uridine, 5-Methyl-deoxyCytidine, 2,6-Diaminopurine or Iso-deoxyGuanosine) in place of the standard T, C, A or G to avoid mismatch detection and repair, which we tested in Escherichia coli. This strategy increases transient allelic-replacement efficiencies by up to 20-fold, while maintaining a 100-fold lower background mutation level. We further show that the mismatched bases between the full length oligo and the chromosome are often not incorporated at the target site, probably due to nuclease activity at the 5′ and 3′ termini of the oligo. These results further elucidate the mechanism of oligo-mediated allelic replacement (OMAR) and enable improved methodologies for efficient, large-scale engineering of genomes.Synthetic Biology Engineering Research CenterNational Science Foundation (U.S.) (Grant #SA5283-11210)United States. Dept. of Energy (Genomes to Life Center) (Grant #DE-FG02-03ER6344)Wyss Institute for Biologically Inspired Engineerin

Selection-Independent Generation of Gene Knockout Mouse Embryonic Stem Cells Using Zinc-Finger Nucleases

Gene knockout in murine embryonic stem cells (ESCs) has been an invaluable tool to study gene function in vitro or to generate animal models with altered phenotypes. Gene targeting using standard techniques, however, is rather inefficient and typically does not exceed frequencies of 10−6. In consequence, the usage of complex positive/negative selection strategies to isolate targeted clones has been necessary. Here, we present a rapid single-step approach to generate a gene knockout in mouse ESCs using engineered zinc-finger nucleases (ZFNs). Upon transient expression of ZFNs, the target gene is cleaved by the designer nucleases and then repaired by non-homologous end-joining, an error-prone DNA repair process that introduces insertions/deletions at the break site and therefore leads to functional null mutations. To explore and quantify the potential of ZFNs to generate a gene knockout in pluripotent stem cells, we generated a mouse ESC line containing an X-chromosomally integrated EGFP marker gene. Applying optimized conditions, the EGFP locus was disrupted in up to 8% of ESCs after transfection of the ZFN expression vectors, thus obviating the need of selection markers to identify targeted cells, which may impede or complicate downstream applications. Both activity and ZFN-associated cytotoxicity was dependent on vector dose and the architecture of the nuclease domain. Importantly, teratoma formation assays of selected ESC clones confirmed that ZFN-treated ESCs maintained pluripotency. In conclusion, the described ZFN-based approach represents a fast strategy for generating gene knockouts in ESCs in a selection-independent fashion that should be easily transferrable to other pluripotent stem cells

Chandra X-ray Observations of the Spiral Galaxy M81

A Chandra X-Ray Observatory ACIS-S imaging observation is used to study the population of X-ray sources in the nearby Sab galaxy M81 (NGC 3031). A total of 177 sources are detected with 124 located within the D25 isophote to a limiting X-ray luminosity of 3e36 ergs/cm2/s. Source positions, count rates, luminosities in the 0.3-8.0 keV band, limiting optical magnitudes, and potential counterpart identifications are tabulated. Spectral and timing analysis of the 36 brightest sources are reported including the low-luminosity active galactic nucleus, SN 1993J, and the Einstein-discovered ultra-luminous X-ray source X6.Comment: 27 pages, 17 figures, 2 tables, submitted to Ap