lncRNA-dependent mechanisms of androgen-receptor-regulated gene activation programs.

Although recent studies have indicated roles of long non-coding RNAs (lncRNAs) in physiological aspects of cell-type determination and tissue homeostasis, their potential involvement in regulated gene transcription programs remains rather poorly understood. The androgen receptor regulates a large repertoire of genes central to the identity and behaviour of prostate cancer cells, and functions in a ligand-independent fashion in many prostate cancers when they become hormone refractory after initial androgen deprivation therapy. Here we report that two lncRNAs highly overexpressed in aggressive prostate cancer, PRNCR1 (also known as PCAT8) and PCGEM1, bind successively to the androgen receptor and strongly enhance both ligand-dependent and ligand-independent androgen-receptor-mediated gene activation programs and proliferation in prostate cancer cells. Binding of PRNCR1 to the carboxy-terminally acetylated androgen receptor on enhancers and its association with DOT1L appear to be required for recruitment of the second lncRNA, PCGEM1, to the androgen receptor amino terminus that is methylated by DOT1L. Unexpectedly, recognition of specific protein marks by PCGEM1-recruited pygopus 2 PHD domain enhances selective looping of androgen-receptor-bound enhancers to target gene promoters in these cells. In 'resistant' prostate cancer cells, these overexpressed lncRNAs can interact with, and are required for, the robust activation of both truncated and full-length androgen receptor, causing ligand-independent activation of the androgen receptor transcriptional program and cell proliferation. Conditionally expressed short hairpin RNA targeting these lncRNAs in castration-resistant prostate cancer cell lines strongly suppressed tumour xenograft growth in vivo. Together, these results indicate that these overexpressed lncRNAs can potentially serve as a required component of castration-resistance in prostatic tumours

Local electronic structure and magnetic properties of LaMn0.5Co0.5O3 studied by x-ray absorption and magnetic circular dichroism spectroscopy

We have studied the local electronic structure of LaMn0.5Co0.5O3 using soft-x-ray absorption spectroscopy at the Co-L_3,2 and Mn-L_3,2 edges. We found a high-spin Co^{2+}--Mn^{4+} valence state for samples with the optimal Curie temperature. We discovered that samples with lower Curie temperatures contain low-spin nonmagnetic Co^{3+} ions. Using soft-x-ray magnetic circular dichroism we established that the Co^{2+} and Mn^{4+} ions are ferromagnetically aligned. We revealed also that the Co^{2+} ions have a large orbital moment: m_orb/m_spin ~ 0.47. Together with model calculations, this suggests the presence of a large magnetocrystalline anisotropy in the material and predicts a non-trivial temperature dependence for the magnetic susceptibility.Comment: 8 pages, 7 figure

A novel promoter controls Cyp19a1 gene expression in mouse adipose tissue

<p>Abstract</p> <p>Background</p> <p>Aromatase, the key enzyme in estrogen biosynthesis, is encoded by the Cyp19a1 gene. Thus far, 3 unique untranslated first exons associated with distinct promoters in the mouse Cyp19a1 gene have been described (brain, ovary, and testis-specific). It remains unknown whether aromatase is expressed in other mouse tissues via novel and tissue-specific promoters.</p> <p>Methods</p> <p>Real-time PCR was used to examine the aromatase expression levels in various C57BL/6 mouse tissues. 5'-rapid amplification of cDNA ends (5'-RACE) was used to determine the transcriptional start sites of Cyp19a1 transcripts. Promoter activity was measured using serial deletion mutants of DNA fused to the luciferase reporter gene. Primary mouse adipose fibroblasts were isolated and cultured from 16-week-old mouse gonadal fat pads.</p> <p>Results</p> <p>We systematically analyzed Cyp19a1 expression in a large number of mouse tissues, and demonstrated for the first time that aromatase was expressed in the male but not female gonadal fat pad. Subcutaneous and brown adipose tissue did not contain detectable Cyp19a1 mRNA. We used 5'-RACE to clone a novel gonadal fat-specific untranslated first exon, which is spliced onto a common junction 15 bp upstream of the translation start site. This adipose-specific first exon was mapped to approximately 75 kb upstream of the translation start site. Transfection of luciferase reporter gene plasmids containing the promoter region upstream of the adipose-specific first exon into murine 3T3-L1 adipose fibroblasts demonstrated significant basal promoter activity conferred primarily by the sequence located at -343/-1 bp. Dexamethasone significantly induced activity of this adipose-specific promoter region. Adipose-specific Cyp19a1 mRNA was expressed in primary mouse adipose fibroblasts and significantly induced by dexamethasone alone or serum plus dexamethasone.</p> <p>Conclusion</p> <p>Taken together, this research identified a novel, adipose-specific first exon of Cyp19a1 and its hormonally regulated promoter region in male murine gonadal fat. These results expand the known 5'-regulatory region of the murine Cyp19a1 gene to 75 kb upstream of the translation start site. Cyp19a1 expression in mouse adipose tissue may play an important role in reproductive biology and lipid metabolism.</p

Sonar-induced pressure fields in a post-mortem common dolphin

Author Posting. © Acoustical Society of America, 2012. This article is posted here by permission of Acoustical Society of America for personal use, not for redistribution. The definitive version was published in Journal of the Acoustical Society of America 131 (2012): 1595-1604, doi:10.1121/1.3675005.Potential physical effects of sonar transmissions on marine mammals were investigated by measuring pressure fields induced in a 119-kg, 211-cm-long, young adult male common dolphin (Delphinus delphis) cadaver. The specimen was instrumented with tourmaline acoustic pressure gauges used as receiving sensors. Gauge implantation near critical tissues was guided by intraoperative, high-resolution, computerized tomography (CT) scanning. Instrumented structures included the melon, nares, ear, thoracic wall, lungs, epaxial muscle, and lower abdomen. The specimen was suspended from a frame equipped with a standard 50.8-mm-diameter spherical transducer used as the acoustic source and additional receiving sensors to monitor the transmitted and external, scattered field. Following immersion, the transducer transmitted pulsed sinusoidal signals at 5, 7, and 10 kHz. Quantitative internal pressure fields are reported for all cases except those in which the gauge failed or no received signal was detected. A full necropsy was performed immediately after the experiment to examine instrumented areas and all major organs. No lesions attributable to acoustic transmissions were found, consistent with the low source level and source-receiver distances.Work supported by NOPP through ONR Grant No. N000140710992. Work at CSI additionally supported by ONR Grant No. N000140811231

Measurements of Secondary Cosmic Microwave Background Anisotropies with the South Pole Telescope

We report cosmic microwave background (CMB) power spectrum measurements from the first 100 sq. deg. field observed by the South Pole Telescope (SPT) at 150 and 220 GHz. On angular scales where the primary CMB anisotropy is dominant, ell ~< 3000, the SPT power spectrum is consistent with the standard LambdaCDM cosmology. On smaller scales, we see strong evidence for a point source contribution, consistent with a population of dusty, star-forming galaxies. After we mask bright point sources, anisotropy power on angular scales of 3000 50 at both frequencies. We combine the 150 and 220 GHz data to remove the majority of the point source power, and use the point source subtracted spectrum to detect Sunyaev-Zel'dovich (SZ) power at 2.6 sigma. At ell=3000, the SZ power in the subtracted bandpowers is 4.2 +/- 1.5 uK^2, which is significantly lower than the power predicted by a fiducial model using WMAP5 cosmological parameters. This discrepancy may suggest that contemporary galaxy cluster models overestimate the thermal pressure of intracluster gas. Alternatively, this result can be interpreted as evidence for lower values of sigma8. When combined with an estimate of the kinetic SZ contribution, the measured SZ amplitude shifts sigma8 from the primary CMB anisotropy derived constraint of 0.794 +/- 0.028 down to 0.773 +/- 0.025. The uncertainty in the constraint on sigma8 from this analysis is dominated by uncertainties in the theoretical modeling required to predict the amplitude of the SZ power spectrum for a given set of cosmological parameters.Comment: 28 pages, 11 figures, submitted to Ap

Angular Power Spectra of the Millimeter Wavelength Background Light from Dusty Star-forming Galaxies with the South Pole Telescope

We use data from the first 100 square-degree field observed by the South Pole Telescope (SPT) in 2008 to measure the angular power spectrum of temperature anisotropies contributed by the background of dusty star-forming galaxies (DSFGs) at millimeter wavelengths. From the auto and cross-correlation of 150 and 220 GHz SPT maps, we significantly detect both Poisson distributed and, for the first time at millimeter wavelengths, clustered components of power from a background of DSFGs. The spectral indices between 150 and 220 GHz of the Poisson and clustered components are found to be 3.86 +- 0.23 and 3.8 +- 1.3 respectively, implying a steep scaling of the dust emissivity index beta ~ 2. The Poisson and clustered power detected in SPT, BLAST (at 600, 860, and 1200 GHz), and Spitzer (1900 GHz) data can be understood in the context of a simple model in which all galaxies have the same graybody spectrum with dust emissivity index of beta = 2 and dust temperature T_d = 34 K. In this model, half of the 150 GHz background light comes from redshifts greater than 3.2. We also use the SPT data to place an upper limit on the amplitude of the kinetic Sunyaev-Zel'dovich power spectrum at l = 3000 of 13 uK^2 at 95% confidence.Comment: 18 pages, 9 figure

Galaxy Clusters Selected with the Sunyaev-Zel'dovich Effect from 2008 South Pole Telescope Observations

We present a detection-significance-limited catalog of 21 Sunyaev-Zel'dovich selected galaxy clusters. These clusters, along with 1 unconfirmed candidate, were identified in 178 deg^2 of sky surveyed in 2008 by the South Pole Telescope to a depth of 18 uK-arcmin at 150 GHz. Optical imaging from the Blanco Cosmology Survey (BCS) and Magellan telescopes provided photometric (and in some cases spectroscopic) redshift estimates, with catalog redshifts ranging from z=0.15 to z>1, with a median z = 0.74. Of the 21 confirmed galaxy clusters, three were previously identified as Abell clusters, three were presented as SPT discoveries in Staniszewski et al, 2009, and three were first identified in a recent analysis of BCS data by Menanteau et al, 2010; the remaining 12 clusters are presented for the first time in this work. Simulated observations of the SPT fields predict the sample to be nearly 100% complete above a mass threshold of M_200 ~ 5x10^14 M_sun/h at z = 0.6. This completeness threshold pushes to lower mass with increasing redshift, dropping to ~4x10^14 M_sun/h at z=1. The size and redshift distribution of this catalog are in good agreement with expectations based on our current understanding of galaxy clusters and cosmology. In combination with other cosmological probes, we use the cluster catalog to improve estimates of cosmological parameters. Assuming a standard spatially flat wCDM cosmological model, the addition of our catalog to the WMAP 7-year analysis yields sigma_8 = 0.81 +- 0.09 and w = -1.07 +- 0.29, a ~50% improvement in precision on both parameters over WMAP7 alone.Comment: 19 pages, 9 figures, 4 appendice

Mycolactone-dependent depletion of endothelial cell thrombomodulin is strongly associated with fibrin deposition in Buruli ulcer lesions

A well-known histopathological feature of diseased skin in Buruli ulcer (BU) is coagulative necrosis caused by the Mycobacterium ulcerans macrolide exotoxin mycolactone. Since the underlying mechanism is not known, we have investigated the effect of mycolactone on endothelial cells, focussing on the expression of surface anticoagulant molecules involved in the protein C anticoagulant pathway. Congenital deficiencies in this natural anticoagulant pathway are known to induce thrombotic complications such as purpura fulimans and spontaneous necrosis. Mycolactone profoundly decreased thrombomodulin (TM) expression on the surface of human dermal microvascular endothelial cells (HDMVEC) at doses as low as 2ng/ml and as early as 8hrs after exposure. TM activates protein C by altering thrombin's substrate specificity, and exposure of HDMVEC to mycolactone for 24 hours resulted in an almost complete loss of the cells' ability to produce activated protein C. Loss of TM was shown to be due to a previously described mechanism involving mycolactone-dependent blockade of Sec61 translocation that results in proteasome-dependent degradation of newly synthesised ER-transiting proteins. Indeed, depletion from cells determined by live-cell imaging of cells stably expressing a recombinant TM-GFP fusion protein occurred at the known turnover rate. In order to determine the relevance of these findings to BU disease, immunohistochemistry of punch biopsies from 40 BU lesions (31 ulcers, nine plaques) was performed. TM abundance was profoundly reduced in the subcutis of 78% of biopsies. Furthermore, it was confirmed that fibrin deposition is a common feature of BU lesions, particularly in the necrotic areas. These findings indicate that there is decreased ability to control thrombin generation in BU skin. Mycolactone's effects on normal endothelial cell function, including its ability to activate the protein C anticoagulant pathway are strongly associated with this. Fibrin-driven tisischemia could contribute to the development of the tissue necrosis seen in BU lesions

X-ray Properties of the First SZE-selected Galaxy Cluster Sample from the South Pole Telescope

We present results of X-ray observations of a sample of 15 clusters selected via their imprint on the cosmic microwave background (CMB) from the thermal Sunyaev-Zel'dovich (SZ) effect. These clusters are a subset of the first SZ-selected cluster catalog, obtained from observations of 178 deg^2 of sky surveyed by the South Pole Telescope. Using X-ray observations with Chandra and XMM-Newton, we estimate the temperature, T_X, and mass, M_g, of the intracluster medium (ICM) within r_500 for each cluster. From these, we calculate Y_X=M_g T_X and estimate the total cluster mass using a M_500-Y_X scaling relation measured from previous X-ray studies. The integrated Comptonization, Y_SZ, is derived from the SZ measurements, using additional information from the X-ray measured gas density profiles and a universal temperature profile. We calculate scaling relations between the X-ray and SZ observables, and find results generally consistent with other measurements and the expectations from simple self-similar behavior. Specifically, we fit a Y_SZ-Y_X relation and find a normalization of 0.82 +- 0.07, marginally consistent with the predicted ratio of Y_SZ/Y_X=0.91+-0.01 that would be expected from the density and temperature models used in this work. Using the Y_X derived mass estimates, we fit a Y_SZ-M_500 relation and find a slope consistent with the self-similar expectation of Y_SZ ~ M^5/3 with a normalization consistent with predictions from other X-ray studies. We compare the X-ray mass estimates to previously published SZ mass estimates derived from cosmological simulations of the SPT survey. We find that the SZ mass estimates are lower by a factor of 0.89+-0.06, which is within the ~15% systematic uncertainty quoted for the simulation-based SZ masses.Comment: 28 pages, 19 figures, submitted to Ap

Measurement of Electron Backscattering in the Energy Range of Neutron β\beta-Decay

We report on the first detailed measurements of electron backscattering from low Z targets at energies up to 124 keV. Both energy and angular distributions of the backscattered electrons are measured and compared with electron transport simulations based on the Geant4 and Penelope Monte Carlo simulation codes. Comparisons are also made with previous, less extensive, measurements and with measurements at lower energies.Comment: 9 pages, 8 figures, accepted for publication in Phys. Rev.