Secretion of Genome-Free Hepatitis B Virus – Single Strand Blocking Model for Virion Morphogenesis of Para-retrovirus

As a para-retrovirus, hepatitis B virus (HBV) is an enveloped virus with a double-stranded (DS) DNA genome that is replicated by reverse transcription of an RNA intermediate, the pregenomic RNA or pgRNA. HBV assembly begins with the formation of an “immature” nucleocapsid (NC) incorporating pgRNA, which is converted via reverse transcription within the maturing NC to the DS DNA genome. Only the mature, DS DNA-containing NCs are enveloped and secreted as virions whereas immature NCs containing RNA or single-stranded (SS) DNA are not enveloped. The current model for selective virion morphogenesis postulates that accumulation of DS DNA within the NC induces a “maturation signal” that, in turn, triggers its envelopment and secretion. However, we have found, by careful quantification of viral DNA and NCs in HBV virions secreted in vitro and in vivo, that the vast majority of HBV virions (over 90%) contained no DNA at all, indicating that NCs with no genome were enveloped and secreted as empty virions (i.e., enveloped NCs with no DNA). Furthermore, viral mutants bearing mutations precluding any DNA synthesis secreted exclusively empty virions. Thus, viral DNA synthesis is not required for HBV virion morphogenesis. On the other hand, NCs containing RNA or SS DNA were excluded from virion formation. The secretion of DS DNA-containing as well as empty virions on one hand, and the lack of secretion of virions containing single-stranded (SS) DNA or RNA on the other, prompted us to propose an alternative, “Single Strand Blocking” model to explain selective HBV morphogenesis whereby SS nucleic acid within the NC negatively regulates NC envelopment, which is relieved upon second strand DNA synthesis

Enhanced NFκB and AP-1 transcriptional activity associated with antiestrogen resistant breast cancer

BACKGROUND: Signaling pathways that converge on two different transcription factor complexes, NFκB and AP-1, have been identified in estrogen receptor (ER)-positive breast cancers resistant to the antiestrogen, tamoxifen. METHODS: Two cell line models of tamoxifen-resistant ER-positive breast cancer, MCF7/HER2 and BT474, showing increased AP-1 and NFκB DNA-binding and transcriptional activities, were studied to compare tamoxifen effects on NFκB and AP-1 regulated reporter genes relative to tamoxifen-sensitive MCF7 cells. The model cell lines were treated with the IKK inhibitor parthenolide (PA) or the proteasome inhibitor bortezomib (PS341), alone and in combination with tamoxifen. Expression microarray data available from 54 UCSF node-negative ER-positive breast cancer cases with known clinical outcome were used to search for potential genes signifying upregulated NFκB and AP-1 transcriptional activity in association with tamoxifen resistance. The association of these genes with patient outcome was further evaluated using node-negative ER-positive breast cancer cases identified from three other published data sets (Rotterdam, n = 209; Amsterdam, n = 68; Basel, n = 108), each having different patient age and adjuvant tamoxifen treatment characteristics. RESULTS: Doses of parthenolide and bortezomib capable of sensitizing the two endocrine resistant breast cancer models to tamoxifen were capable of suppressing NFκB and AP-1 regulated gene expression in combination with tamoxifen and also increased ER recruitment of the transcriptional co-repressor, NCoR. Transcript profiles from the UCSF breast cancer cases revealed three NFκB and AP-1 upregulated genes – cyclin D1, uPA and VEGF – capable of dichotomizing node-negative ER-positive cases into early and late relapsing subsets despite adjuvant tamoxfien therapy and most prognostic for younger age cases. Across the four independent sets of node-negative ER-positive breast cancer cases (UCSF, Rotterdam, Amsterdam, Basel), high expression of all three NFκB and AP-1 upregulated genes was associated with earliest metastatic relapse. CONCLUSION: Altogether, these findings implicate increased NFκB and AP-1 transcriptional responses with tamoxifen resistant breast cancer and early metastatic relapse, especially in younger patients. These findings also suggest that agents capable of preventing NFκB and AP-1 gene activation may prove useful in restoring the endocrine responsiveness of such high-risk ER-positive breast cancers

Phosphodiesterase 10A Upregulation Contributes to Pulmonary Vascular Remodeling

Phosphodiesterases (PDEs) modulate the cellular proliferation involved in the pathophysiology of pulmonary hypertension (PH) by hydrolyzing cAMP and cGMP. The present study was designed to determine whether any of the recently identified PDEs (PDE7-PDE11) contribute to progressive pulmonary vascular remodeling in PH. All in vitro experiments were performed with lung tissue or pulmonary arterial smooth muscle cells (PASMCs) obtained from control rats or monocrotaline (MCT)-induced pulmonary hypertensive (MCT-PH) rats, and we examined the effects of the PDE10 inhibitor papaverine (Pap) and specific small interfering RNA (siRNA). In addition, papaverine was administrated to MCT-induced PH rats from day 21 to day 35 by continuous intravenous infusion to examine the in vivo effects of PDE10A inhibition. We found that PDE10A was predominantly present in the lung vasculature, and the mRNA, protein, and activity levels of PDE10A were all significantly increased in MCT PASMCs compared with control PASMCs. Papaverine and PDE10A siRNA induced an accumulation of intracellular cAMP, activated cAMP response element binding protein and attenuated PASMC proliferation. Intravenous infusion of papaverine in MCT-PH rats resulted in a 40%–50% attenuation of the effects on pulmonary hypertensive hemodynamic parameters and pulmonary vascular remodeling. The present study is the first to demonstrate a central role of PDE10A in progressive pulmonary vascular remodeling, and the results suggest a novel therapeutic approach for the treatment of PH

Haze in Pluto's atmosphere: Results from SOFIA and ground-based observations of the 2015 June 29 Pluto occultation

On UT 29 June 2015, the occultation by Pluto of a bright star (r′ = 11.9) was observed from the Stratospheric Observatory for Infrared Astronomy (SOFIA) and several ground-based stations in New Zealand and Australia. Pre-event astrometry allowed for an in-flight update to the SOFIA team with the result that SOFIA was deep within the central flash zone (~22 km from center). Analysis of the combined data leads to the result that Pluto's middle atmosphere is essentially unchanged from 2011 and 2013 (Person et al. 2013; Bosh et al. 2015); there has been no significant expansion or contraction of the atmosphere. Additionally, our multi-wavelength observations allow us to conclude that a haze component in the atmosphere is required to reproduce the light curves obtained. This haze scenario has implications for understanding the photochemistry of Pluto's atmosphere

Manganese-Iron Phosphate Nodules at the Groken Site, Gale Crater, Mars

The MSL Curiosity rover investigated dark, Mn-P-enriched nodules in shallow lacustrine/fluvial sediments at the Groken site in Glen Torridon, Gale Crater, Mars. Applying all relevant information from the rover, the nodules are interpreted as pseudomorphs after original crystals of vivianite, (Fe2+,Mn2+)3(PO4)2·8H2O, that cemented the sediment soon after deposition. The nodules appear to have flat faces and linear boundaries and stand above the surrounding siltstone. ChemCam LIBS (laser-induced breakdown spectrometry) shows that the nodules have MnO abundances approximately twenty times those of the surrounding siltstone matrix, contain little CaO, and have SiO2 and Al2O3 abundances similar to those of the siltstone. A deconvolution of APXS analyses of nodule-bearing targets, interpreted here as representing the nodules’ non-silicate components, shows high concentrations of MnO, P2O5, and FeO and a molar ratio P/Mn = 2. Visible to near-infrared reflectance of the nodules (by ChemCam passive and Mastcam multispectral) is dark and relatively flat, consistent with a mixture of host siltstone, hematite, and a dark spectrally bland material (like pyrolusite, MnO2). A drill sample at the site is shown to contain minimal nodule material, implying that analyses by the CheMin and SAM instruments do not constrain the nodules’ mineralogy or composition. The fact that the nodules contain P and Mn in a small molar integer ratio, P/Mn = 2, suggests that the nodules contained a stoichiometric Mn-phosphate mineral, in which Fe did (i.e., could) not substitute for Mn. The most likely such minerals are laueite and strunzite, (Fe2+,Mn2+)3(PO4)2·8H2O and –6H2O, respectively, which occur on Earth as alteration products of other Mn-bearing phosphates including vivianite. Vivianite is a common primary and diagenetic precipitate from low-oxygen, P-enriched waters. Calculated phase equilibria show Mn-bearing vivianite could be replaced by laueite or strunzite and then by hematite plus pyrolusite as the system became more oxidizing and acidic. These data suggest that the nodules originated as vivianite, forming as euhedral crystals in the sediment, enclosing sediment grains as they grew. After formation, the nodules were oxidized—first to laueite/strunzite yielding the diagnostic P/Mn ratio, and then to hematite plus an undefined Mn oxy-hydroxide (like pyrolusite). The limited occurrence of these Mn-Fe-P nodules, both in space and time (i.e., stratigraphic position), suggests a local control on their origin. By terrestrial analogies, it is possible that the nodules precipitated near a spring or seep of Mn-rich water, generated during alteration of olivine in the underlying sediments

A simple echocardiographic estimate of right ventricular-arterial coupling to assess severity and outcome in pulmonary hypertension on chronic lung disease

SCOPUS: le.jDecretOANoAutActifinfo:eu-repo/semantics/publishe

TRIO OF STELLAR OCCULTATIONS BY PLUTO ONE YEAR PRIOR TO NEW HORIZONS' ARRIVAL

We observed occultations by Pluto during a predicted series of events in 2014 July with the 1 m telescope of the Mt. John Observatory in New Zealand. The predictions were based on updated astrometry obtained in the previous months at the USNO, CTIO, and Lowell Observatories. We successfully detected occultations by Pluto of an R = 18 mag star on July 23 (14:23:32 ± 00:00:04 UTC to 14:25:30 ± 00:00:04 UTC), with a drop of 75% of the unocculted stellar signal, and of an R = 17 star on July 24 (11:41:30 ± 00:00:08 UTC to 11:43:28 ± 00:00:08 UTC), with a drop of 80% of the unocculted stellar signal, both with 20 s exposures with our frame-transfer Portable Occultation, Eclipse, and Transit System. Since Pluto had a geocentric velocity of 22.51 km s[superscript −1] on July 23 and 22.35 km s[superscript −1] on July 24, these intervals yield limits on the chord lengths (surface and lower atmosphere) of 2700 ± 130 km and 2640 ± 250 km, respectively, indicating that the events were near central, and therefore provide astrometric constraints on the prediction method. Our coordinated observations with the 4 m AAT in Australia on July 23 and the 6.5 m Magellan/Clay on Las Campanas, the 4.1 m Southern Astrophysical Research Telescope on Cerro Pachön, the 2.5 m DuPont on Las Campanas (LCO), the 0.6 m SARA-South on Cerro Tololo of the Southeastern Association for Research in Astronomy (SARA), the MPI/ESO 2.2 m on La Silla, and the 0.45 m Cerro Calán telescope and 0.36 telescope in Constitución in Chile on July 27 and 31, which would have provided higher-cadence observations for studies of Pluto's atmosphere, were largely foiled by clouds, but led to detection with the LCO Magellan/Clay and DuPont Telescopes on July 31 of the grazing occultation of a previously unknown 15th-magnitude star, completing the trio of occultations successfully observed and reported in this paper.National Research Foundation (South Africa

Pluto Occultation on 2015 June 29 UTC With Central Flash and Atmospheric Spikes Just Before the New Horizons Flyby

We observed the occultation by Pluto of a 12th magnitude star, one of the two brightest occultation stars ever in our dozen years of continual monitoring of Pluto's atmosphere through such studies, on 2015 June 29 UTC. At the Univ. of Canterbury Mt. John Observatory (New Zealand), under clear skies throughout, we used a POETS frame-transfer CCD at 10 Hz with GPS timing on the 1-m McLellan telescope as well as an infrared camera on an 0.6-m telescope and three-color photometry at a slower cadence on a second 0.6-m telescope. At the Auckland Observatory, we used a POETS and a PICO on 0.5-m and 0.4-m telescopes, with 0.4 s and 2 s cadences, respectively, obtaining ingress observations before clouds moved in. The Mt. John light curves show a central flash, indicating that we were close to the center of the occultation path. Analysis of our light curves show that Pluto's atmosphere remains robust. The presence of spikes at both sites in the egress and ingress shows atmospheric layering. We coordinated our observations with aircraft observations (Bosh et al., 2017) with the Stratospheric Observatory for Infrared Astronomy (SOFIA). Our chords helped constrain the path across Pluto that SOFIA saw. Our ground-based and airborne stellar-occultation effort came only just over two weeks of Earth days and two Pluto days before the flyby of NASA's New Horizons spacecraft