Development of a novel polyamide-based agent to inhibit EVI1 function

The EVI1 gene at chromosome 3q26 is associated with acute myeloid leukemogenesis, due to both chromosomal rearrangement and to overexpression in the absence of rearrangement. Some rearrangements such as t(3;3) and inv(3) result in overexpression of EVI1 protein, while translocation t(3;21) yields an AML1-MDS1-EVI1 (AME) fusion protein. EVI1 possesses two zinc finger domains, an N-terminal domain with fingers 1–7, which binds to GACAAGATA, and a C-terminal domain (fingers 8–10) which binds GAAGATGAG. Inhibition of EVI1 function with a small molecule compound may provide a targeted therapy for EVI1-expressing leukemias. As a first step towards inhibiting the leukemogenic function of EVI1, we performed structure-function studies on both EVI1 and AME protein to determine what domains are critical for malignant transformation activity. Assays were Rat1 fibroblasts in a soft agar colony forming assay for EVI1; primary bone marrow cells in a serial replating assay for AME. Both assays revealed that mutation of arginine 205 in zinc finger 6 of EVI1, which completely abrogates sequencespecific DNA binding via the N-terminal zinc finger domain, resulted in complete loss of transforming activity; mutations in other domains, such as the C-terminal zinc finger domain, CtBP binding domain, and the domains of AML1 had less of an effect or no effect on transforming activity. In an effort to inhibit EVI1 leukemogenic function, we developed a polyamide, DH-IV-298, designed to block zinc fingers 1–7 binding to the GACAAGATA motif. DNAseI footprinting revealed a specific interaction between DH-IV-298 and the GACAAGATA motif; no significant interaction was observed elsewhere; a mismatch polyamide failed to footprint at equivalent concentrations; and DH-IV-298 failed to bind to a control DNA lacking the GACAAGATA motif. Electromobility shift assay showed that, at a 1:1 polyamide:DNA ratio, DH-IV-298 lowered EVI1:DNA affinity by over 98%, while mismatch was significantly less effective (74% reduction). To assess the effect of DH-IV-298 on EVI1 binding to DNA in vivo, we performed CAT reporter assays in a NIH-3T3-derived cell line with a chromosome-embedded tet-inducible EVI1-VP16 as well as a EVI1-responsive CAT reporter. Removal of tetracycline resulted in a four-fold increase in CAT activity that was completely blocked by DH-IV-298. The mismatch polyamide was significantly less effective than DH-IV-298. Further studies are being performed to assess the effect on endogenous gene expression, and on growth of leukemic cells that express EVI1. These studies provide evidence that a cell permeable small molecule compound may effectively block the activity of a leukemogenic transcription factor

X-ray Observations of XSS J12270-4859 in a New Low State: A Transformation to a Disk-Free Rotation-Powered Pulsar Binary

We present XMM-Newton and Chandra observations of the low-mass X-ray binary XSS J12270--4859, which experienced a dramatic decline in optical/X-ray brightness at the end of 2012, indicative of the disappearance of its accretion disk. In this new state, the system exhibits previously absent orbital-phase-dependent, large-amplitude X-ray modulations with a decline in flux at superior conjunction. The X-ray emission remains predominantly non-thermal but with an order of magnitude lower mean luminosity and significantly harder spectrum relative to the previous high flux state. This phenomenology is identical to the behavior of the radio millisecond pulsar binary PSR J1023+0038 in the absence of an accretion disk, where the X-ray emission is produced in an intra-binary shock driven by the pulsar wind. This further demonstrates that XSS J12270-4859 no longer has an accretion disk and has transformed to a full-fledged eclipsing "redback" system that hosts an active rotation-powered millisecond pulsar. There is no evidence for diffuse X-ray emission associated with the binary that may arise due to outflows or a wind nebula. An extended source situated 1.5' from XSS J12270--4859 is unlikely to be associated, and is probably a previously uncatalogued galaxy cluster.Comment: 8 pages, 6 figures; accepted for publication in the Astrophysical Journa

Plans for a Neutron EDM Experiment at SNS

The electric dipole moment of the neutron, leptons, and atoms provide a unique window to Physics Beyond the Standard Model. We are currently developing a new neutron EDM experiment (the nEDM Experiment). This experiment, which will be run at the 8.9 A Neutron Line at the Fundamental Neutron Physics Beamline (FNPB) at the Spallation Neutron Source (SNS) at the Oak Ridge National Laboratory, will search for the neutron EDM with a sensitivity two orders of magnitude better than the present limit. In this paper, the motivation for the experiment, the experimental method, and the present status of the experiment are discussed.Comment: 9 Pages, 4 Figures, submitted to the proceedings of the Second Meeting of the APS Topical Group on Hadronic Physics, Nashville, TN, October 22-24, 200

The redshifts of bright sub-mm sources

One of the key goals in observational cosmology over the next few years will be to establish the redshift distribution of the recently-discovered sub-mm source population. In this brief review I discuss and summarize the redshift information which has been gleaned to date for the ~ 50 bright sub-mm sources which have been uncovered via the six main classes of survey performed with SCUBA on the JCMT over the last 2-3 years. Despite the biases inherent in some of these surveys, and the crudeness of the redshift information available in others, I conclude that all current information suggests that only 10-15 % of luminous sub-mm sources lie at z < 2, and that the median redshift of this population is z ~ 3. I suggest that such a high median redshift is arguably not unexpected given current theories designed to explain the correlation between black-hole mass and spheroid mass found at low redshift. In such scenarios, peak AGN emission is expected to correspond to, or even to cause termination of major star-formation activity in the host spheroid. In contrast, maximum dust emission is expected to occur roughly half-way through the star-formation process. Given that optical emission from bright quasars peaks at z = 2.5, dust-emission from massive ellipticals might be reasonably expected to peak at some point in the preceding ~ 1 Gyr, at z ~ 3. Confirmation or refutation of this picture requires significantly-improved redshift information on bright samples of SCUBA sources.Comment: 11 pages, 5 figures, FIRSED2000 conference proceedings, eds. I.M. van Bemmel, B. Wilkes, & P. Barthel Elsevier New Astronomy Review

Chelator free gallium-68 radiolabelling of silica coated iron oxide nanorods via surface interactions

The commercial availability of combined magnetic resonance imaging (MRI)/positron emission tomography (PET) scanners for clinical use has increased demand for easily prepared agents which offer signal or contrast in both modalities. Herein we describe a new class of silica coated iron–oxide nanorods (NRs) coated with polyethylene glycol (PEG) and/or a tetraazamacrocyclic chelator (DO3A). Studies of the coated NRs validate their composition and confirm their properties as in vivo T₂ MRI contrast agents. Radiolabelling studies with the positron emitting radioisotope gallium-68 (t1/2 = 68 min) demonstrate that, in the presence of the silica coating, the macrocyclic chelator was not required for preparation of highly stable radiometal-NR constructs. In vivo PET-CT and MR imaging studies show the expected high liver uptake of gallium-68 radiolabelled nanorods with no significant release of gallium-68 metal ions, validating our innovation to provide a novel simple method for labelling of iron oxide NRs with a radiometal in the absence of a chelating unit that can be used for high sensitivity liver imaging

Inter-model comparison of global hydroxyl radical (OH) distributions and their impact on atmospheric methane over the 2000–2016 period

The modeling study presented here aims to estimate how uncertainties in global hydroxyl radical (OH) distributions, variability, and trends may contribute to resolving discrepancies between simulated and observed methane (CH4) changes since 2000. A multi-model ensemble of 14 OH fields was analyzed and aggregated into 64 scenarios to force the offline atmospheric chemistry transport model LMDz (Laboratoire de Meteorologie Dynamique) with a standard CH4 emission scenario over the period 2000–2016. The multi-model simulated global volume-weighted tropospheric mean OH concentration ([OH]) averaged over 2000–2010 ranges between 8:7*10^5 and 12:8*10^5 molec cm-3. The inter-model differences in tropospheric OH burden and vertical distributions are mainly determined by the differences in the nitrogen oxide (NO) distributions, while the spatial discrepancies between OH fields are mostly due to differences in natural emissions and volatile organic compound (VOC) chemistry. From 2000 to 2010, most simulated OH fields show an increase of 0.1–0:3*10^5 molec cm-3 in the tropospheric mean [OH], with year-to-year variations much smaller than during the historical period 1960–2000. Once ingested into the LMDz model, these OH changes translated into a 5 to 15 ppbv reduction in the CH4 mixing ratio in 2010, which represents 7%–20% of the model-simulated CH4 increase due to surface emissions. Between 2010 and 2016, the ensemble of simulations showed that OH changes could lead to a CH4 mixing ratio uncertainty of > 30 ppbv. Over the full 2000–2016 time period, using a common stateof- the-art but nonoptimized emission scenario, the impact of [OH] changes tested here can explain up to 54% of the gap between model simulations and observations. This result emphasizes the importance of better representing OH abundance and variations in CH4 forward simulations and emission optimizations performed by atmospheric inversions