Science.gov

Sample records for 10-12 erg cm-2

  1. ERG — EDRN Public Portal

    Cancer.gov

    The ERG gene codes for the ERG protein, a transcription factor that is a member of the erythroblast transformation-specific (ETS) family of transcriptions factors. Members of this family are involved in embryonic development, cell proliferation, differentiation, angiogenesis, inflammation, and apoptosis. ERG is mainly expressed in the nucleus. ERG is also involved in chromosomal translocations, resulting in different fusion gene products, such as TMPSSR2-ERG and NDRG1-ERG in prostate cancer, EWS-ERG in Ewing's sarcoma and FUS-ERG in acute myeloid leukemia. Different isoforms arising from multiple alternatively spliced transcripts have been identified.

  2. Sedimentary record of erg migration

    SciTech Connect

    Porter, M.L.

    1986-06-01

    The sedimentary record of erg (eolian sand sea) migration consists of an idealized threefold division of sand-sea facies sequences. The basal division, here termed the fore-erg, is composed of a hierarchy of eolian sand bodies contained within sediments of the flanking depositional environment. The fore-erg represents the downwind, leading edge of the erg and records the onset of eolian sedimentation. Basin subsidence coupled with erg migration places the medial division, termed the central erg, over the fore-erg strata. Eolian influence on regional sedimentation patterns is greatest in the central erg, and most of the sand transported and deposited in the erg is contained within this region. Reduction in sand supply and continued erg migration will cover the central-erg deposits with a veneer of back-erg deposits. This upper division of the erg facies sequence resembles closely the fore-erg region. Back-erg deposits may be thin due to limited eolian influence on sedimentation or incomplete erg migration, or they may be completely absent because of great susceptibility to postdepositional erosion. Tectonic, climatic, and eustatic influences on sand-sea deposition will produce distinctive variations or modifications of the idealized erg facies sequence. The resulting variants in the sedimentary record of erg migration are illustrated with ancient examples from western North America, Europe, southern Africa, and South America. 38 references, 2 figures, 2 tables.

  3. North Polar Erg

    NASA Technical Reports Server (NTRS)

    2005-01-01

    [figure removed for brevity, see original site]

    Our topic for the weeks of April 4 and April 11 is dunes on Mars. We will look at the north polar sand sea and at isolated dune fields at lower latitudes. Sand seas on Earth are often called 'ergs,' an Arabic name for dune field. A sand sea differs from a dune field in two ways: 1) a sand sea has a large regional extent, and 2) the individual dunes are large in size and complex in form.

    This VIS image was taken at 82 degrees North latitude during Northern spring. As with yesterday's image, the dunes are still partially frost covered. This region is part of the north polar erg (sand sea), note the complexity and regional coverage of the dunes.

    Image information: VIS instrument. Latitude 81.2, Longitude 118.2 East (241.8 West). 19 meter/pixel resolution.

    Note: this THEMIS visual image has not been radiometrically nor geometrically calibrated for this preliminary release. An empirical correction has been performed to remove instrumental effects. A linear shift has been applied in the cross-track and down-track direction to approximate spacecraft and planetary motion. Fully calibrated and geometrically projected images will be released through the Planetary Data System in accordance with Project policies at a later time.

    NASA's Jet Propulsion Laboratory manages the 2001 Mars Odyssey mission for NASA's Office of Space Science, Washington, D.C. The Thermal Emission Imaging System (THEMIS) was developed by Arizona State University, Tempe, in collaboration with Raytheon Santa Barbara Remote Sensing. The THEMIS investigation is led by Dr. Philip Christensen at Arizona State University. Lockheed Martin Astronautics, Denver, is the prime contractor for the Odyssey project, and developed and built the orbiter. Mission operations are conducted jointly from Lockheed Martin and from JPL, a division of the California Institute of Technology in Pasadena.

  4. North Polar Erg

    NASA Technical Reports Server (NTRS)

    2005-01-01

    [figure removed for brevity, see original site]

    Our topic for the weeks of April 4 and April 11 is dunes on Mars. We will look at the north polar sand sea and at isolated dune fields at lower latitudes. Sand seas on Earth are often called 'ergs,' an Arabic name for dune field. A sand sea differs from a dune field in two ways: 1) a sand sea has a large regional extent, and 2) the individual dunes are large in size and complex in form.

    This VIS image was taken at 81 degrees North latitude during Northern spring. This region of the north polar erg is dominated by a different form of dunes than yesterday's image.

    Image information: VIS instrument. Latitude 81.4, Longitude 121.9 East (238.1 West). 19 meter/pixel resolution.

    Note: this THEMIS visual image has not been radiometrically nor geometrically calibrated for this preliminary release. An empirical correction has been performed to remove instrumental effects. A linear shift has been applied in the cross-track and down-track direction to approximate spacecraft and planetary motion. Fully calibrated and geometrically projected images will be released through the Planetary Data System in accordance with Project policies at a later time.

    NASA's Jet Propulsion Laboratory manages the 2001 Mars Odyssey mission for NASA's Office of Space Science, Washington, D.C. The Thermal Emission Imaging System (THEMIS) was developed by Arizona State University, Tempe, in collaboration with Raytheon Santa Barbara Remote Sensing. The THEMIS investigation is led by Dr. Philip Christensen at Arizona State University. Lockheed Martin Astronautics, Denver, is the prime contractor for the Odyssey project, and developed and built the orbiter. Mission operations are conducted jointly from Lockheed Martin and from JPL, a division of the California Institute of Technology in Pasadena.

  5. Sayama CM2 Chondrite: Fresh but Heavily Altered

    NASA Technical Reports Server (NTRS)

    Takaoka, N.; Nakamura, T.; Noguchi, T.; Tonui, E.; Gounelle, M.; Zolensky, M. E.; Ebisawa, N.; Osawa, T.; Okazaki, R.; Nagao, K.; Yoneda, S.

    2001-01-01

    Noble gas composition and mineralogy of Sayama meteorite, that fell in Japan and recently identified as a CM2 chondrite, revealed many unique features, indicating that it experienced extensive aqueous alteration under highly oxidized condition compared with typical CMs. Additional information is contained in the original extended abstract.

  6. Organic Matter Inclusions in CM2 Chondrite Murchison

    NASA Astrophysics Data System (ADS)

    Peeters, Z.; Liebig, B.; Lee, T.

    2015-07-01

    Large (~10 μm) inclusions of pure organic carbon exist in carbonaceous chondrites. We extracted organic inclusions from Murchison, a CM2, and analyzed the sections using XANES, TEM, and nanoSIMS. The results are compared to previous results of CRs.

  7. Benchmarking and performance analysis of the CM-2. [SIMD computer

    NASA Technical Reports Server (NTRS)

    Myers, David W.; Adams, George B., II

    1988-01-01

    A suite of benchmarking routines testing communication, basic arithmetic operations, and selected kernel algorithms written in LISP and PARIS was developed for the CM-2. Experiment runs are automated via a software framework that sequences individual tests, allowing for unattended overnight operation. Multiple measurements are made and treated statistically to generate well-characterized results from the noisy values given by cm:time. The results obtained provide a comparison with similar, but less extensive, testing done on a CM-1. Tests were chosen to aid the algorithmist in constructing fast, efficient, and correct code on the CM-2, as well as gain insight into what performance criteria are needed when evaluating parallel processing machines.

  8. hERG me out.

    PubMed

    Czodrowski, Paul

    2013-09-23

    A detailed analysis of the hERG content inside the ChEMBL database is performed. The correlation between the outcome from binding assays and functional assays is probed. On the basis of descriptor distributions, design paradigms with respect to structural and physicochemical properties of hERG active and hERG inactive compounds are challenged. Finally, classification models with different data sets are trained. All source code is provided, which is based on the Python open source packages RDKit and scikit-learn to enable the community to rerun the experiments. The code is stored on github ( https://github.com/pzc/herg_chembl_jcim).

  9. 50 CFR 10.12 - Definitions.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... Definitions § 10.12 Definitions. Aircraft means any contrivance used for flight in the air. Amphibians means a... dead, including without limitation any wild mammal, bird, reptile, amphibian, fish, mollusk,...

  10. 50 CFR 10.12 - Definitions.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... Definitions § 10.12 Definitions. Aircraft means any contrivance used for flight in the air. Amphibians means a... dead, including without limitation any wild mammal, bird, reptile, amphibian, fish, mollusk,...

  11. 50 CFR 10.12 - Definitions.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... Definitions § 10.12 Definitions. Aircraft means any contrivance used for flight in the air. Amphibians means a... dead, including without limitation any wild mammal, bird, reptile, amphibian, fish, mollusk,...

  12. 50 CFR 10.12 - Definitions.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... Definitions § 10.12 Definitions. Aircraft means any contrivance used for flight in the air. Amphibians means a... dead, including without limitation any wild mammal, bird, reptile, amphibian, fish, mollusk,...

  13. 50 CFR 10.12 - Definitions.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... Definitions § 10.12 Definitions. Aircraft means any contrivance used for flight in the air. Amphibians means a... dead, including without limitation any wild mammal, bird, reptile, amphibian, fish, mollusk,...

  14. Direct particle simulation on the Connection Machine CM-2

    NASA Technical Reports Server (NTRS)

    Dagum, L.

    1992-01-01

    Particle simulation is a useful technique for analyzing low density flows. The Connection Machine CM-2 is a useful test bed for studying the fine-grain data objects decomposition and the coarse-grain domain decomposition single instruction multiple datastream (SIMD) approaches to particle simulation. Both approaches are investigated for the model problem of uniform flow through a channel and the algorithms required for the SIMD domain decomposition approach are presented. An unresolved issue with the domain decomposition approach is the effect of a poor partitioning on flows with real geometries. Initial results with the channel flow problem indicate that a poor partitioning has only a small detrimental effect on the overall performance.

  15. English Literature Grades 10-12.

    ERIC Educational Resources Information Center

    Instructional Objectives Exchange, Los Angeles, CA.

    This collection is composed of objectives and evaluation items in poetry, novel, short story, drama, and essay for Senior High, Grades 10-12. Each objective contains four elements: (1) the objective, (2) measurement items, (3) means for judging the adequacy of the student response, and (4) an IOX (Instructional Objectives Exchange) acceptability…

  16. 43 CFR 10.12 - Civil penalties.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... REPATRIATION REGULATIONS Human Remains, Funerary Objects, Sacred Objects, or Objects of Cultural Patrimony in Museums and Federal Collections § 10.12 Civil penalties. (a) The Secretary's authority to assess civil penalties. The Secretary is authorized by section 9 of the Act to assess civil penalties on any museum...

  17. 43 CFR 10.12 - Civil penalties.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... REPATRIATION REGULATIONS Human Remains, Funerary Objects, Sacred Objects, or Objects of Cultural Patrimony in Museums and Federal Collections § 10.12 Civil penalties. (a) The Secretary's authority to assess civil penalties. The Secretary is authorized by section 9 of the Act to assess civil penalties on any museum...

  18. 43 CFR 10.12 - Civil penalties.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... REPATRIATION REGULATIONS Human Remains, Funerary Objects, Sacred Objects, or Objects of Cultural Patrimony in Museums and Federal Collections § 10.12 Civil penalties. (a) The Secretary's authority to assess civil penalties. The Secretary is authorized by section 9 of the Act to assess civil penalties on any museum...

  19. 43 CFR 10.12 - Civil penalties.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... REPATRIATION REGULATIONS Human Remains, Funerary Objects, Sacred Objects, or Objects of Cultural Patrimony in Museums and Federal Collections § 10.12 Civil penalties. (a) The Secretary's authority to assess civil penalties. The Secretary is authorized by section 9 of the Act to assess civil penalties on any museum...

  20. 43 CFR 10.12 - Civil penalties.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... REPATRIATION REGULATIONS Human Remains, Funerary Objects, Sacred Objects, or Objects of Cultural Patrimony in Museums and Federal Collections § 10.12 Civil penalties. (a) The Secretary's authority to assess civil penalties. The Secretary is authorized by section 9 of the Act to assess civil penalties on any museum...

  1. Presolar grains in the CM2 chondrite Sutter's Mill

    NASA Astrophysics Data System (ADS)

    Zhao, Xuchao; Lin, Yangting; Yin, Qing-Zhu; Zhang, Jianchao; Hao, Jialong; Zolensky, Michael; Jenniskens, Peter

    2014-11-01

    The Sutter's Mill (SM) carbonaceous chondrite is a regolith breccia, composed predominantly of CM2 clasts with varying degrees of aqueous alteration and thermal metamorphism. An investigation of presolar grains in four Sutter's Mill sections, SM43, SM51, SM2-4, and SM18, was carried out using NanoSIMS ion mapping technique. A total of 37 C-anomalous grains and one O-anomalous grain have been identified, indicating an abundance of 63 ppm for presolar C-anomalous grains and 2 ppm for presolar oxides. Thirty-one silicon carbide (SiC), five carbonaceous grains, and one Al-oxide (Al2O3) were confirmed based on their elemental compositions determined by C-N-Si and O-Si-Mg-Al isotopic measurements. The overall abundance of SiC grains in Sutter's Mill (55 ppm) is consistent with those in other CM chondrites. The absence of presolar silicates in Sutter's Mill suggests that they were destroyed by aqueous alteration on the parent asteroid. Furthermore, SM2-4 shows heterogeneous distributions of presolar SiC grains (12-54 ppm) in different matrix areas, indicating that the fine-grained matrix clasts come from different sources, with various thermal histories, in the solar nebula.

  2. Microbiological study of the Murchison CM2 meteorite

    NASA Astrophysics Data System (ADS)

    Pikuta, Elena V.; Hoover, Richard B.

    2012-10-01

    In 1864, Louis Pasteur attempted to cultivate living microorganisms from pristine samples of the Orgueil CI1 carbonaceous meteorite. His results were negative and never published, but recorded it in his laboratory notebooks. At that time, only aerobic liquid or agar-based organic reach media were used, as his research on anaerobes had just started. In our laboratory the Murchison CM2 carbonaceous meteorite was selected to expand on these studies for microbiological study by cultivation on anaerobic mineral media. Since the surface could have been more easily contaminated, interior fragments of a sample of the Murchison meteorite were extracted and crushed under sterile conditions. The resulting powder was then mixed in anoxic medium and injected into Hungate tubes containing anaerobic media with various growth substrates at different pH and salinity and incubated at different temperatures. The goal of the experiments was to determine if living cells would grow from the material of freshly fractured interior fragments of the stone. If any growth occurred, work could then be carried out to assess the nature of the environmental contamination by observations of the culture growth (rates of speed and biodiversity); live/dead fluorescent staining to determine contamination level and DNA analysis to establish the microbial species present. In this paper we report the results of that study.

  3. Targeting TMPRSS2-ERG in Prostate Cancer

    DTIC Science & Technology

    2015-09-01

    ABSTRACT About half of all prostate cancers are known to harbor a genetic mutation that fuses a gene known as ERG to the regulatory region of the gene...activity. We applied this technique to screen genetic and chemical libraries to study ERG mediated tumorigenesis and identify novel therapeutic...agents targeting ERG activity. 15. SUBJECT TERMS Prostate cancer, ERG, gene expression, high throughput screening, small molecule microarray, genetic

  4. Intragenic ERG Deletions Do Not Explain the Biology of ERG-Related Acute Lymphoblastic Leukemia

    PubMed Central

    Potuckova, Eliska; Zuna, Jan; Hovorkova, Lenka; Starkova, Julia; Stary, Jan; Trka, Jan; Zaliova, Marketa

    2016-01-01

    Intragenic ERG deletions occur in 3–5% of B-cell precursor acute lymphoblastic leukemia, specifically in B-other subtype lacking the classifying genetic lesions. They represent the only genetic lesion described so far present in the majority of cases clustering into a subgroup of B-other subtype characterized by a unique gene expression profile, probably sharing a common, however, not yet fully described, biological background. We aimed to elucidate whether ERG deletions could drive the specific biology of this ERG-related leukemia subgroup through expression of aberrant or decreased expression of wild type ERG isoforms. We showed that leukemic cells with endogenous ERG deletion express an aberrant transcript translated into two proteins in transfected cell lines and that one of these proteins colocalizes with wild type ERG. However, we did not confirm expression of the proteins in acute lymphoblastic leukemia cases with endogenous ERG deletion. ERG deletions resulted in significantly lower expression of wild type ERG transcripts compared to B-other cases without ERG deletion. However, cases with subclonal ERG deletion, clustering to the same ERG deletion associated subgroup, presented similar levels of wild type ERG as cases without ERG deletion. In conclusion, our data suggest that neither the expression of aberrant proteins from internally deleted allele nor the reduced expression of wild type ERG seem to provide a plausible explanation of the specific biology of ERG -related leukemia subgroup. PMID:27494621

  5. 24 CFR 10.12 - Additional rulemaking proceedings.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 24 Housing and Urban Development 1 2010-04-01 2010-04-01 false Additional rulemaking proceedings. 10.12 Section 10.12 Housing and Urban Development Office of the Secretary, Department of Housing and Urban Development RULEMAKING: POLICY AND PROCEDURES Procedures § 10.12 Additional rulemaking...

  6. 1 CFR 10.12 - Format, indexes, and ancillaries.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 1 General Provisions 1 2010-01-01 2010-01-01 false Format, indexes, and ancillaries. 10.12 Section 10.12 General Provisions ADMINISTRATIVE COMMITTEE OF THE FEDERAL REGISTER SPECIAL EDITIONS OF THE FEDERAL REGISTER PRESIDENTIAL PAPERS Annual Publication § 10.12 Format, indexes, and ancillaries. (a)...

  7. 1 CFR 10.12 - Format, indexes, and ancillaries.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 1 General Provisions 1 2011-01-01 2011-01-01 false Format, indexes, and ancillaries. 10.12 Section 10.12 General Provisions ADMINISTRATIVE COMMITTEE OF THE FEDERAL REGISTER SPECIAL EDITIONS OF THE FEDERAL REGISTER PRESIDENTIAL PAPERS Annual Publication § 10.12 Format, indexes, and ancillaries. (a)...

  8. 46 CFR 90.10-12 - Gas free.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 4 2012-10-01 2012-10-01 false Gas free. 90.10-12 Section 90.10-12 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CARGO AND MISCELLANEOUS VESSELS GENERAL PROVISIONS Definition of Terms Used in This Subchapter § 90.10-12 Gas free. This term means free from dangerous concentrations...

  9. 46 CFR 90.10-12 - Gas free.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 4 2013-10-01 2013-10-01 false Gas free. 90.10-12 Section 90.10-12 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CARGO AND MISCELLANEOUS VESSELS GENERAL PROVISIONS Definition of Terms Used in This Subchapter § 90.10-12 Gas free. This term means free from dangerous concentrations...

  10. 46 CFR 90.10-12 - Gas free.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 4 2014-10-01 2014-10-01 false Gas free. 90.10-12 Section 90.10-12 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CARGO AND MISCELLANEOUS VESSELS GENERAL PROVISIONS Definition of Terms Used in This Subchapter § 90.10-12 Gas free. This term means free from dangerous concentrations...

  11. 46 CFR 90.10-12 - Gas free.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 4 2010-10-01 2010-10-01 false Gas free. 90.10-12 Section 90.10-12 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CARGO AND MISCELLANEOUS VESSELS GENERAL PROVISIONS Definition of Terms Used in This Subchapter § 90.10-12 Gas free. This term means free from dangerous concentrations...

  12. 46 CFR 90.10-12 - Gas free.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 4 2011-10-01 2011-10-01 false Gas free. 90.10-12 Section 90.10-12 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) CARGO AND MISCELLANEOUS VESSELS GENERAL PROVISIONS Definition of Terms Used in This Subchapter § 90.10-12 Gas free. This term means free from dangerous concentrations...

  13. 10 CFR 10.12 - Interview and other investigation.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 10 Energy 1 2014-01-01 2014-01-01 false Interview and other investigation. 10.12 Section 10.12... Eligibility for Access to Restricted Data or National Security Information or an Employment Clearance § 10.12... pursuant to § 10.11(a) (5) and (12). (c) If the Director, Division of Facilities and Security, cannot...

  14. 10 CFR 10.12 - Interview and other investigation.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 10 Energy 1 2010-01-01 2010-01-01 false Interview and other investigation. 10.12 Section 10.12... Eligibility for Access to Restricted Data or National Security Information or an Employment Clearance § 10.12... pursuant to § 10.11(a) (5) and (12). (c) If the Director, Division of Facilities and Security, cannot...

  15. 10 CFR 10.12 - Interview and other investigation.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 10 Energy 1 2013-01-01 2013-01-01 false Interview and other investigation. 10.12 Section 10.12... Eligibility for Access to Restricted Data or National Security Information or an Employment Clearance § 10.12... pursuant to § 10.11(a) (5) and (12). (c) If the Director, Division of Facilities and Security, cannot...

  16. 10 CFR 10.12 - Interview and other investigation.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 10 Energy 1 2012-01-01 2012-01-01 false Interview and other investigation. 10.12 Section 10.12... Eligibility for Access to Restricted Data or National Security Information or an Employment Clearance § 10.12... pursuant to § 10.11(a) (5) and (12). (c) If the Director, Division of Facilities and Security, cannot...

  17. 10 CFR 10.12 - Interview and other investigation.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 10 Energy 1 2011-01-01 2011-01-01 false Interview and other investigation. 10.12 Section 10.12... Eligibility for Access to Restricted Data or National Security Information or an Employment Clearance § 10.12... pursuant to § 10.11(a) (5) and (12). (c) If the Director, Division of Facilities and Security, cannot...

  18. Generation and diagnostics of pulsed intense ion beams with an energy density of 10 J/cm2

    NASA Astrophysics Data System (ADS)

    Isakova, Yu.; Pushkarev, A.; Khailov, I.; Zhong, H.

    2015-07-01

    The paper presents the results of a study on transportation and focusing of a pulsed ion beam at gigawatt power level, generated by a diode with explosive-emission cathode. The experiments were carried out with the TEMP-4M accelerator operating in double-pulse mode: the first pulse is of negative polarity (500 ns, 100-150 kV), and this is followed by a second pulse of positive polarity (120 ns, 200-250 kV). To reduce the beam divergence, we modified the construction of the diode. The width of the anode was increased compared to that of the cathode. We studied different configurations of planar and focusing strip diodes. It was found that the divergence of the ion beam formed by a planar strip diode, after construction modification, does not exceed 3° (half-angle). Modification to the construction of a focusing diode made it possible to reduce the beam divergence from 8° to 4°-5°, as well as to increase the energy density at the focus up to 10-12 J/cm2, and decrease the shot to shot variation in the energy density from 10%-15% to 5%-6%. When measuring the ion beam energy density above the ablation threshold of the target material (3.5-4 J/cm2), we used a metal mesh with 50% transparency to lower the energy density. The influence of the metal mesh on beam transport has been studied.

  19. Photopic ERG components in retinitis pigmentosa.

    PubMed

    Anastasi, M; Lauricella, M; Ponte, F

    1992-04-01

    The persistence of a residual flicker electroretinogram 20 Hz response in many cases of retinitis pigmentosa, when the Oscillatory Potentials (OPs) were no longer recordable, led the authors to an investigation by Fourier analysis. The study was carried out in 33 patients affected by different hereditary forms of retinitis pigmentosa revealing recordable 20 Hz flash ERG responses. We applied the Fourier analysis to this ERG response and compared the weight percentage of the first two components to the OP added amplitude. The analysis showed that the 20 Hz flash ERG response contains only the first harmonic in patients with no recordable OPs and both harmonics in patients with recordable OPs. This relationship between OPs and 20 Hz second component, that is possibly related to the activity of inner retina as well as the OPs, can demonstrate an alteration of the inner retina which evolves with distinct electrophysiological features from the ERG photoreceptor impairment.

  20. Multielectron ultrastrong laser field ionization of Arn+, Krm+ and Xel+ (n <= 9, m <= 9, l <= 12) at intensities from 1015 W cm-2 to 1018 W cm-2

    NASA Astrophysics Data System (ADS)

    Palaniyappan, S.; Di Chiara, A.; Ghebregziabher, I.; Huskins, E. L.; Falkowski, A.; Pajerowski, D.; Walker, B. C.

    2006-07-01

    Ionization yields are reported for Ar, Kr and Xe in ultrastrong fields from 1015 W cm-2 to 1018 W cm-2. Non-sequential ionization (NSI) is shown to be a robust and general feature in ultrahigh field ionization. NSI yields measured are consistent with the trends predicted by a rescattering model, but as one proceeds to higher Z atoms more NSI is observed than predicted theoretically. Additional recollision mechanisms that may need to be considered in future theories of ultrastrong field-atom interactions include 'chain' NSI, NSI from excited states of the atom (e.g. Rydberg states or inner-shell holes) and the possibility of ultrastrong field enhanced recollision/impact processes.

  1. Compositions of Partly Altered Olivine and Replacement Serpentine in the CM2 Chondrite QUE 93005

    NASA Astrophysics Data System (ADS)

    Velbel, M. A.; Tonui, E. K.; Zolensky, M. E.

    2013-09-01

    Serpentine replacing olivine in QUE 93005 has a narrow range of compositions, regardless of the reactant olivine’s composition. Homogeneity of replacement serpentines in QUE 93005 and other CM2s favors homogeneity of aqueous solutions on >cm scales.

  2. Magnesium isotopic fractionation in chondrules from the Murchison and Murray CM2 carbonaceous chondrites

    NASA Astrophysics Data System (ADS)

    Bouvier, Audrey; Wadhwa, Meenakshi; Simon, Steven B.; Grossman, Lawrence

    2013-03-01

    We present high-precision measurements of the Mg isotopic compositions of a suite of types I and II chondrules separated from the Murchison and Murray CM2 carbonaceous chondrites. These chondrules are olivine- and pyroxene-rich and have low 27Al/24Mg ratios (0.012-0.316). The Mg isotopic compositions of Murray chondrules are on average lighter (δ26Mg ranging from -0.95‰ to -0.15‰ relative to the DSM-3 standard) than those of Murchison (δ26Mg ranging from -1.27‰ to +0.77‰). Taken together, the CM2 chondrules exhibit a narrower range of Mg isotopic compositions than those from CV and CB chondrites studied previously. The least-altered CM2 chondrules are on average lighter (average δ26Mg = -0.39 ± 0.30‰, 2SE) than the moderately to heavily altered CM2 chondrules (average δ26Mg = -0.11 ± 0.21‰, 2SE). The compositions of CM2 chondrules are consistent with isotopic fractionation toward heavy Mg being associated with the formation of secondary silicate phases on the CM2 parent body, but were also probably affected by volatilization and recondensation processes involved in their original formation. The low-Al CM2 chondrules analyzed here do not exhibit any mass-independent variations in 26Mg from the decay of 26Al, with the exception of two chondrules that show only small variations just outside of the analytical error. In the case of the chondrule with the highest Al/Mg ratio (a type IAB chondrule from Murchison), the lack of resolvable 26Mg excess suggests that it either formed >1 Ma after calcium-aluminum-rich inclusions, or that its Al-Mg isotope systematics were reset by secondary alteration processes on the CM2 chondrite parent body after the decay of 26Al.

  3. GFDL's CM2 global coupled climate models. Part I: Formulation and simulation characteristics

    USGS Publications Warehouse

    Delworth, T.L.; Broccoli, A.J.; Rosati, A.; Stouffer, R.J.; Balaji, V.; Beesley, J.A.; Cooke, W.F.; Dixon, K.W.; Dunne, J.; Dunne, K.A.; Durachta, J.W.; Findell, K.L.; Ginoux, P.; Gnanadesikan, A.; Gordon, C.T.; Griffies, S.M.; Gudgel, R.; Harrison, M.J.; Held, I.M.; Hemler, R.S.; Horowitz, L.W.; Klein, S.A.; Knutson, T.R.; Kushner, P.J.; Langenhorst, A.R.; Lee, H.-C.; Lin, S.-J.; Lu, J.; Malyshev, S.L.; Milly, P.C.D.; Ramaswamy, V.; Russell, J.; Schwarzkopf, M.D.; Shevliakova, E.; Sirutis, J.J.; Spelman, M.J.; Stern, W.F.; Winton, M.; Wittenberg, A.T.; Wyman, B.; Zeng, F.; Zhang, R.

    2006-01-01

    The formulation and simulation characteristics of two new global coupled climate models developed at NOAA's Geophysical Fluid Dynamics Laboratory (GFDL) are described. The models were designed to simulate atmospheric and oceanic climate and variability from the diurnal time scale through multicentury climate change, given our computational constraints. In particular, an important goal was to use the same model for both experimental seasonal to interannual forecasting and the study of multicentury global climate change, and this goal has been achieved. Tw o versions of the coupled model are described, called CM2.0 and CM2.1. The versions differ primarily in the dynamical core used in the atmospheric component, along with the cloud tuning and some details of the land and ocean components. For both coupled models, the resolution of the land and atmospheric components is 2?? latitude ?? 2.5?? longitude; the atmospheric model has 24 vertical levels. The ocean resolution is 1?? in latitude and longitude, with meridional resolution equatorward of 30?? becoming progressively finer, such that the meridional resolution is 1/3?? at the equator. There are 50 vertical levels in the ocean, with 22 evenly spaced levels within the top 220 m. The ocean component has poles over North America and Eurasia to avoid polar filtering. Neither coupled model employs flux adjustments. The co ntrol simulations have stable, realistic climates when integrated over multiple centuries. Both models have simulations of ENSO that are substantially improved relative to previous GFDL coupled models. The CM2.0 model has been further evaluated as an ENSO forecast model and has good skill (CM2.1 has not been evaluated as an ENSO forecast model). Generally reduced temperature and salinity biases exist in CM2.1 relative to CM2.0. These reductions are associated with 1) improved simulations of surface wind stress in CM2.1 and associated changes in oceanic gyre circulations; 2) changes in cloud tuning and

  4. Drugs, hERG and sudden death.

    PubMed

    Brown, A M

    2004-06-01

    Early recognition of potential QT/TdP liability is now an essential component of the drug discovery/drug development program. The hERG assay is an indispensable step and a high-quality assay must accompany any investigational new drug (IND) application. While it is the gold standard at present, the hERG assay is too labor-intensive and too low throughput to be used as a screen early in the discovery/development process. A variety of indirect high throughput screens have been used.

  5. Targeting TMPRSS2-ERG in Prostate Cancer

    DTIC Science & Technology

    2014-09-01

    in prostate cancer cells. We will complete validation candidate kinases that modulate the ERG signature using shRNA and new CRISPR technology as an...will plan to validate hits using CRISPR -Cas9 technology as an orthogonal system. The CRISPR -Cas9 system was not available at the time of the

  6. Aliphatic Amines in Antarctic CR2, CM2, and CM1/2 Carbonaceous Chondrites

    NASA Technical Reports Server (NTRS)

    Aponte, Jose C.; McLain, Hannah L.; Dworkin, Jason P.; Elsila, Jamie E.

    2016-01-01

    Meteoritic water-soluble organic compounds provide a unique record of the processes that occurred during the formation of the solar system and the chemistry preceding the origins of life on Earth. We have investigated the molecular distribution, compound-specific delta13C isotopic ratios and enantiomeric compositions of aliphatic monoamines present in the hot acid-water extracts of the carbonaceous chondrites LAP 02342 (CR2), GRA 95229 (CR2), LON 94101 (CM2), LEW 90500 (CM2), and ALH 83100 (CM1/2). Analyses of the concentration of monoamines in these meteorites revealed: (a) the CR2 chondrites studied here contain higher concentrations of monoamines relative to the analyzed CM2 chondrites; (b) the concentration of monoamines decreases with increasing carbon number; and (c) isopropylamine is the most abundant monoamine in these CR2 chondrites, while methylamine is the most abundant amine species in these CM2 and CM1/2 chondrites. The delta13C values of monoamines in CR2 chondrite do not correlate with the number of carbon atoms; however, in CM2 and CM1/2 chondrites, the 13C enrichment decreases with increasing monoamine carbon number. The delta13C values of methylamine in CR2 chondrites ranged from -1 to +10per mille, while in CM2 and CM1/2 chondrites the delta13C values of methylamine ranged from +41 to +59per mille. We also observed racemic compositions of sec-butylamine, 3-methyl-2-butylamine, and sec-pentylamine in the studied carbonaceous chondrites. Additionally, we compared the abundance and delta13C isotopic composition of monoamines to those of their structurally related amino acids. We found that monoamines are less abundant than amino acids in CR2 chondrites, with the opposite being true in CM2 and CM1/2 chondrites. We used these collective data to evaluate different primordial synthetic pathways for monoamines in carbonaceous chondrites and to understand the potential common origins these molecules may share with meteoritic amino acids.

  7. Aliphatic amines in Antarctic CR2, CM2, and CM1/2 carbonaceous chondrites

    NASA Astrophysics Data System (ADS)

    Aponte, José C.; McLain, Hannah L.; Dworkin, Jason P.; Elsila, Jamie E.

    2016-09-01

    Meteoritic water-soluble organic compounds provide a unique record of the processes that occurred during the formation of the solar system and the chemistry preceding the origins of life on Earth. We have investigated the molecular distribution, compound-specific δ13C isotopic ratios and enantiomeric compositions of aliphatic monoamines present in the hot acid-water extracts of the carbonaceous chondrites LAP 02342 (CR2), GRA 95229 (CR2), LON 94101 (CM2), LEW 90500 (CM2), and ALH 83100 (CM1/2). Analyses of the concentration of monoamines in these meteorites revealed: (a) the CR2 chondrites studied here contain higher concentrations of monoamines relative to the analyzed CM2 chondrites; (b) the concentration of monoamines decreases with increasing carbon number; and (c) isopropylamine is the most abundant monoamine in these CR2 chondrites, while methylamine is the most abundant amine species in these CM2 and CM1/2 chondrites. The δ13C values of monoamines in CR2 chondrite do not correlate with the number of carbon atoms; however, in CM2 and CM1/2 chondrites, the 13C enrichment decreases with increasing monoamine carbon number. The δ13C values of methylamine in CR2 chondrites ranged from -1 to +10‰, while in CM2 and CM1/2 chondrites the δ13C values of methylamine ranged from +41 to +59‰. We also observed racemic compositions of sec-butylamine, 3-methyl-2-butylamine, and sec-pentylamine in the studied carbonaceous chondrites. Additionally, we compared the abundance and δ13C isotopic composition of monoamines to those of their structurally related amino acids. We found that monoamines are less abundant than amino acids in CR2 chondrites, with the opposite being true in CM2 and CM1/2 chondrites. We used these collective data to evaluate different primordial synthetic pathways for monoamines in carbonaceous chondrites and to understand the potential common origins these molecules may share with meteoritic amino acids.

  8. Music, Grades 10-12. Secondary Schools Curriculum Guide.

    ERIC Educational Resources Information Center

    Cranston School Dept., RI.

    Nine courses are included in this music guide for grades 10-12: Music Theory; Humanities: Renaissance, Baroque, and Classical; Humanities: Idealism vs. Realism; Humanities: 20th Century Man and His World; A Capella Choir; Chorale; Band; and Basic Keyboard Study. Major objectives, numbered objectives, and activities indicate level of learning,…

  9. Social Studies, Grades 10-12. Secondary Schools Curriculum Guide.

    ERIC Educational Resources Information Center

    Cranston School Dept., RI.

    This curriculum guide provides a one-year course in social studies for grades 10-12. The guide is intended to serve as a resource to teachers, students, department chairmen, guidance personnel, curriculum planners, and anyone else involved in present or future curriculum planning. The course is divided into 22 section areas covering the topics of…

  10. Newspapers and Law-Related Education. Grades 10-12.

    ERIC Educational Resources Information Center

    Diamond, Sandra; Riekes, Linda

    Designed to assist teachers of students in grades 10-12 who wish to use the newspaper as a supplemental tool in law-related education, this guide provides model lessons demonstrating ways in which the daily newspaper can enhance textbook material. Although the guide is based on ongoing law-related education programs in St. Louis Public Schools, it…

  11. Black American Literature, Grades 10-12. Experimental.

    ERIC Educational Resources Information Center

    Gunn, Evelyn, Ed.

    This black American literature course for grades 10-12 is designed to introduce students to the unique contribution that the black American has made to American life. The course guide is divided into four units--Slave Narrative and Autobiography, Poetry, Short Story, Drama--with suggested activites, including class discussions, small group…

  12. Ewing Sarcoma With ERG Gene Rearrangements: A Molecular Study Focusing on the Prevalence of FUS-ERG and Common Pitfalls in Detecting EWSR1-ERG Fusions by FISH

    PubMed Central

    Chen, Sonja; Deniz, Kemal; Sung, Yun-Shao; Zhang, Lei; Dry, Sarah; Antonescu, Cristina R.

    2016-01-01

    The genetics of Ewing sarcoma (ES) are characterized by a canonical fusion involving EWSR1 gene and a member of the ETS family of transcription factors, such as FLI1 and ERG. In fact, ERG gene rearrangements represent the second most common molecular alteration, with EWSR1-ERG being identified in 5–10% of cases, while only a handful of reports document a FUS-ERG fusion. In this study, we focus on ES with ERG gene abnormalities, specifically to investigate the prevalence and clinicopathologic features of FUS-ERG fusions in a large cohort of small blue round cell tumors (SBRCTs) and compare to the eight reported FUS-positive ES. Among the 85 SBRCTs tested, seven (8.2%) cases harbored FUS gene rearrangements; six fused to ERG and one with FEV. During this investigation we came across a number of ERG-rearranged ES lacking both EWSR1 and FUS abnormalities by FISH. In one case, RNA sequencing identified an EWSR1-ERG transcript despite the negative EWSR1 rearrangements by FISH. Additional 3-color FISH fusion assay demonstrated the fusion of EWSR1 and ERG signals in all four cases negative for break-apart EWSR1 FISH. These results emphasize a potential pitfall of relying on EWSR1 FISH assay alone for diagnosis of ES. In cases with classic morphology and/or strong CD99 and ERG immunoreactivity, additional molecular testing should be applied, such as ERG FISH or RT-PCR/next generation sequencing, for a more definitive diagnosis. Although our study group is small, there were no differences noted between the clinical, morphologic features and immunoprofile of the different subsets of ERG-rearranged SBRCTs. PMID:26690869

  13. Muscarinic modulation of erg potassium current

    PubMed Central

    Hirdes, Wiebke; Horowitz, Lisa F; Hille, Bertil

    2004-01-01

    We studied modulation of current in human embryonic kidney tsA-201 cells coexpressing rat erg1 channels with M1 muscarinic receptors. Maximal current was inhibited 30% during muscarinic receptor stimulation, with a small positive shift of the midpoint of activation. Inhibition was attenuated by coexpression of the regulator of G-protein signalling RGS2 or of a dominant-negative protein, Gq, but not by N-ethylmaleimide or C3 toxin. Overexpression of a constitutively active form of Gq (but not of G13 or of Gs) abolished the erg current. Hence it is likely that Gq/11, and not Gi/o or G13, mediates muscarinic inhibition. Muscarinic suppression of erg was attenuated by chelating intracellular Ca2+ to < 1 nm free Ca2+ with 20 mm BAPTA in the pipette, but suppression was normal if internal Ca2+ was strongly clamped to a 129 nm free Ca2+ level with a BAPTA buffer and this was combined with numerous other measures to prevent intracellular Ca2+ transients (pentosan polysulphate, preincubation with thapsigargin, and removal of extracellular Ca2+). Hence a minimum amount of Ca2+ was necessary for the inhibition, but a Ca2+ elevation was not. The ATP analogue AMP-PCP did not prevent inhibition. The protein kinase C (PKC) blockers staurosporine and bisindolylmaleimide I did not prevent inhibition, and the PKC-activating phorbol ester PMA did not mimic it. Neither the tyrosine kinase inhibitor genistein nor the tyrosine phosphatase inhibitor dephostatin prevented inhibition by oxotremorine-M. Hence protein kinases are not needed. Experiments with a high concentration of wortmannin were consistent with recovery being partially dependent on PIP2 resynthesis. Wortmannin did not prevent muscarinic inhibition. Our studies of muscarinic inhibition of erg current suggest a role for phospholipase C, but not the classical downstream messengers, such as PKC or a calcium transient. PMID:15235086

  14. Organic Power Electronics: Transistor Operation in the kA/cm(2) Regime.

    PubMed

    Klinger, Markus P; Fischer, Axel; Kaschura, Felix; Widmer, Johannes; Kheradmand-Boroujeni, Bahman; Ellinger, Frank; Leo, Karl

    2017-03-17

    In spite of interesting features as flexibility, organic thin-film transistors have commercially lagged behind due to the low mobilities of organic semiconductors associated with hopping transport. Furthermore, organic transistors usually have much larger channel lengths than their inorganic counterparts since high-resolution structuring is not available in low-cost production schemes. Here, we present an organic permeable-base transistor (OPBT) which, despite extremely simple processing without any high-resolution structuring, achieve a performance beyond what has so far been possible using organic semiconductors. With current densities above 1 kA cm(-2) and switching speeds towards 100 MHz, they open the field of organic power electronics. Finding the physical limits and an effective mobility of only 0.06 cm(2) V(-1) s(-1), this OPBT device architecture has much more potential if new materials optimized for its geometry will be developed.

  15. The EET87513 clast N: A CM2 fragment in an HED polymict breccia

    NASA Technical Reports Server (NTRS)

    Buchanan, P. C.; Zolensky, M. E.; Reid, A. M.; Barrett, R. A.

    1993-01-01

    Xenoliths of material resembling carbonaceous chondrites have been found in several HED polymict breccias. Most workers concluded that these clasts are related to CM2 meteorites on the basis of texture, bulk composition, and mineralogy. Data on clast N, a carbonaceous chondrite fragment from the howardite EET87513 large enough (approximately 4x5mm on the surface of the slab from which it was separated) to extract bulk samples for INAA and oxygen isotope analysis and to provide a thin section for electron microprobe, SEM, and TEM analysis is reported. Preliminary data for this clast were previously reported. INAA was performed at Oregon State University and bulk oxygen isotopic composition was determined at the University of Chicago. These data confirm that EET87513 clast N is a fragment of CM2 material.

  16. Relativistic Electron Accleration by a Laser of Intensity in Excess of 1020 W cm-2

    NASA Astrophysics Data System (ADS)

    Mangles, S. P. D.; Walton, B.; Wei, M. S.; Clarke, R. J.; Fritzler, S.; Gopal, A.; Hernandez-Gomez, C.; Krushelnick, K.; Najmudin, Z.; Dangor, A. E.

    Relativistic electrons with energies in excess of 300 MeV have been observed resulting from the interaction of a 0.3 PW laser beam focused to intensities of around 3 × 1020 W cm-2 interacting with an underdense plasma. Two dimensional particle in cell simulation of the interaction show that an interaction directly between preheated electrons and the intense laser field is responsible for the maximum acceleration.

  17. Investigation of pyridine carboxylic acids in CM2 carbonaceous chondrites: Potential precursor molecules for ancient coenzymes

    NASA Astrophysics Data System (ADS)

    Smith, Karen E.; Callahan, Michael P.; Gerakines, Perry A.; Dworkin, Jason P.; House, Christopher H.

    2014-07-01

    The distribution and abundances of pyridine carboxylic acids (including nicotinic acid) in eight CM2 carbonaceous chondrites (ALH 85013, DOM 03183, DOM 08003, EET 96016, LAP 02333, LAP 02336, LEW 85311, and WIS 91600) were investigated by liquid chromatography coupled to UV detection and high resolution Orbitrap mass spectrometry. We find that pyridine monocarboxylic acids are prevalent in CM2-type chondrites and their abundance negatively correlates with the degree of pre-terrestrial aqueous alteration that the meteorite parent body experienced. We also report the first detection of pyridine dicarboxylic acids in carbonaceous chondrites. Additionally, we carried out laboratory studies of proton-irradiated pyridine in carbon dioxide-rich ices (a 1:1 mixture) to serve as a model of the interstellar ice chemistry that may have led to the synthesis of pyridine carboxylic acids. Analysis of the irradiated ice residue shows that a comparable suite of pyridine mono- and dicarboxylic acids was produced, although aqueous alteration may still play a role in the synthesis (and ultimate yield) of these compounds in carbonaceous meteorites. Nicotinic acid is a precursor to nicotinamide adenine dinucleotide, a likely ancient molecule used in cellular metabolism in all of life, and its common occurrence in CM2 chondrites may indicate that meteorites may have been a source of molecules for the emergence of more complex coenzymes on the early Earth.

  18. Investigation of Pyridine Carboxylic Acids in CM2 Carbonaceous Chondrites: Potential Precursor Molecules for Ancient Coenzymes

    NASA Technical Reports Server (NTRS)

    Smith, Karen E.; Callahan, Michael P.; Gerakines, Perry A.; Dworkin, Jason P.; House, Christopher H.

    2014-01-01

    The distribution and abundances of pyridine carboxylic acids (including nicotinic acid) in eight CM2 carbonaceous chondrites (ALH 85013, DOM 03183, DOM 08003, EET 96016, LAP 02333, LAP 02336, LEW 85311, and WIS 91600) were investigated by liquid chromatography coupled to UV detection and high resolution Orbitrap mass spectrometry. We find that pyridine monocarboxylic acids are prevalent in CM2-type chondrites and their abundance negatively correlates with the degree of pre-terrestrial aqueous alteration that the meteorite parent body experienced. We also report the first detection of pyridine dicarboxylic acids in carbonaceous chondrites. Additionally, we carried out laboratory studies of proton-irradiated pyridine in carbon dioxide-rich ices (a 1:1 mixture) to serve as a model of the interstellar ice chemistry that may have led to the synthesis of pyridine carboxylic acids. Analysis of the irradiated ice residue shows that a comparable suite of pyridine mono- and dicarboxylic acids was produced, although aqueous alteration may still play a role in the synthesis (and ultimate yield) of these compounds in carbonaceous meteorites. Nicotinic acid is a precursor to nicotinamide adenine dinucleotide, a likely ancient molecule used in cellular metabolism in all of life, and its common occurrence in CM2 chondrites may indicate that meteorites may have been a source of molecules for the emergence of more complex coenzymes on the early Earth.

  19. Investigation of Pyridine Carboxylic Acids in CM2 Carbonaceous Chondrites: Potential Precursor Molecules for Ancient Coenzymes

    NASA Technical Reports Server (NTRS)

    Smith, Karen E.; Callahan, Michael P.; Gerakines, Perry A.; Dworkin, Jason P.; House, Christopher H.

    2014-01-01

    The distribution and abundances of pyridine carboxylic acids (including nicotinic acid) in eight CM2 carbonaceous chondrites (ALH 85013, DOM 03183, DOM 08003, EET 96016, LAP 02333, LAP 02336, LEW 85311, and WIS 91600) were investigated by liquid chromatography coupled to UV detection and high resolution Orbitrap mass spectrometry. We find that pyridine monocarboxylic acids are prevalent in CM2-type chondrites and their abundance negatively correlates with the degree of pre-terrestrial aqueous alteration that the meteorite parent body experienced. We lso report the first detection of pyridine dicarboxylic acids in carbonaceous chondrites. Additionally, we carried out laboratory studies of proton-irradiated pyridine in carbon dioxide-rich ices (a 1:1 mixture) to serve as a model of the interstellar ice chemistry that may have led to the synthesis of pyridine carboxylic acids. Analysis of the irradiated ice residue shows that a comparable suite of pyridine mono- and dicarboxylic acids was produced, although aqueous alteration may still play a role in the synthesis (and ultimate yield) of these compounds in carbonaceous meteorites. Nicotinic acid is a precursor to nicotinamide adenine dinucleotide, a likely ancient molecule used in cellular metabolism in all of life, and its common occurrence in CM2 chondrites may indicate that meteorites may have been a source of molecules for the emergence of more complex coenzymes on the early Earth.

  20. Wide-Pulse Evaluation of 0.5 CM2 Silicon Carbide SGTO

    DTIC Science & Technology

    2009-06-01

    the surface of the chip. Figure 3. SGTO packaged at ARL using power package, AuSn die attach, 5-mil wire bonds, and silicone -based potting. Figure 5...WIDE-PULSE EVALUATION OF 0.5 CM2 SILICON CARBIDE SGTO Heather O’Brien, Aderinto Ogunniyi, Charles J. Scozzie U.S. Army Research Laboratory, 2800...3026 E. Cornwallis Rd Research Triangle Park, NC 27709 USA Victor Temple Silicon Power Corp., 958 Main St., Suite A Clifton Park, NY 12065 USA

  1. Organic Power Electronics: Transistor Operation in the kA/cm2 Regime

    NASA Astrophysics Data System (ADS)

    Klinger, Markus P.; Fischer, Axel; Kaschura, Felix; Widmer, Johannes; Kheradmand-Boroujeni, Bahman; Ellinger, Frank; Leo, Karl

    2017-03-01

    In spite of interesting features as flexibility, organic thin-film transistors have commercially lagged behind due to the low mobilities of organic semiconductors associated with hopping transport. Furthermore, organic transistors usually have much larger channel lengths than their inorganic counterparts since high-resolution structuring is not available in low-cost production schemes. Here, we present an organic permeable-base transistor (OPBT) which, despite extremely simple processing without any high-resolution structuring, achieve a performance beyond what has so far been possible using organic semiconductors. With current densities above 1 kA cm‑2 and switching speeds towards 100 MHz, they open the field of organic power electronics. Finding the physical limits and an effective mobility of only 0.06 cm2 V‑1 s‑1, this OPBT device architecture has much more potential if new materials optimized for its geometry will be developed.

  2. Organic Power Electronics: Transistor Operation in the kA/cm2 Regime

    PubMed Central

    Klinger, Markus P.; Fischer, Axel; Kaschura, Felix; Widmer, Johannes; Kheradmand-Boroujeni, Bahman; Ellinger, Frank; Leo, Karl

    2017-01-01

    In spite of interesting features as flexibility, organic thin-film transistors have commercially lagged behind due to the low mobilities of organic semiconductors associated with hopping transport. Furthermore, organic transistors usually have much larger channel lengths than their inorganic counterparts since high-resolution structuring is not available in low-cost production schemes. Here, we present an organic permeable-base transistor (OPBT) which, despite extremely simple processing without any high-resolution structuring, achieve a performance beyond what has so far been possible using organic semiconductors. With current densities above 1 kA cm−2 and switching speeds towards 100 MHz, they open the field of organic power electronics. Finding the physical limits and an effective mobility of only 0.06 cm2 V−1 s−1, this OPBT device architecture has much more potential if new materials optimized for its geometry will be developed. PMID:28303924

  3. Compositions of Partly Altered Olivine and Replacement Serpentine in the CM2 Chondrite QUE93005

    NASA Technical Reports Server (NTRS)

    Velbel, M. A.; Tonui, E. K.; Zolensky, M. E.

    2005-01-01

    Some phyllosilicates in CM carbonaceous chondrites formed by aqueous alteration of anhydrous precursor phases. Although broad trends in the compositions of hydrous phyllosilicates are recognized and believed to be related to trends in degree of aqueous alteration, details of the reactions that formed specific secondary minerals remain obscure. This paper reports compositional relationships between remnants of partially pseudomorphically (or alteromorphically) replaced silicates and their alteration products (serpentine) in the CM2 chondrite QUE93005 and compares it with previously published results for ALH81002. Reactants and products were characterized by optical petrography, backscattered scanning electron microscopy (BSEM), and electron microprobe. By focusing on serpentine formed from known reactants (olivines), and on only those instances in which some of the reactant silicate remains, direct compositional relationships between reactants and products, and the elemental mobility required by the reactions, can be established. Additional information is included in the original extended abstract.

  4. Spectral content of buried Ag foils at 1016 W/cm2 laser illuminationa)

    NASA Astrophysics Data System (ADS)

    Huntington, C. M.; Maddox, B. R.; Park, H.-S.; Prisbrey, S.; Remington, B. A.

    2014-11-01

    Sources of 5-12 keV thermal Heα x-rays are readily generated by laser irradiation of mid-Z foils at intensities >1014 W/cm2, and are widely used as probes for inertial confinement fusion and high-energy-density experiments. Higher energy 17-50 keV x-ray sources are efficiently produced from "cold" Kα emission using short pulse, petawatt lasers at intensities >1018 W/cm2 [H.-S. Park, B. R. Maddox et al., "High-resolution 17-75 keV backlighters for high energy density experiments," Phys. Plasmas 15(7), 072705 (2008); B. R. Maddox, H. S. Park, B. A. Remington et al., "Absolute measurements of x-ray backlighter sources at energies above 10 keV," Phys. Plasmas 18(5), 056709 (2011)]. However, when long pulse (>1 ns) lasers are used with Z > 30 elements, the spectrum contains contributions from both K shell transitions and from ionized atomic states. Here we show that by sandwiching a silver foil between layers of high-density carbon, the ratio of Kα:Heα in the x-ray spectrum is significant increased over directly illuminated Ag foils, with narrower lines from K-shell transitions. Additionally, the emission volume is more localized for the sandwiched target, producing a more planar x-ray sheet. This technique may be useful for generating probes requiring spectral purity and a limited spatial extent, for example, in incoherent x-ray Thomson scattering experiments.

  5. The amino acid composition of the Sutter's Mill CM2 carbonaceous chondrite

    NASA Astrophysics Data System (ADS)

    Burton, Aaron S.; Glavin, Daniel P.; Elsila, Jamie E.; Dworkin, Jason P.; Jenniskens, Peter; Yin, Qing-Zhu

    2014-11-01

    We determined the abundances and enantiomeric compositions of amino acids in Sutter's Mill fragment #2 (designated SM2) recovered prior to heavy rains that fell April 25-26, 2012, and two other meteorite fragments, SM12 and SM51, that were recovered postrain. We also determined the abundance, enantiomeric, and isotopic compositions of amino acids in soil from the recovery site of fragment SM51. The three meteorite stones experienced terrestrial amino acid contamination, as evidenced by the low D/L ratios of several proteinogenic amino acids. The D/L ratios were higher in SM2 than in SM12 and SM51, consistent with rain introducing additional L-amino acid contaminants to SM12 and SM51. Higher percentages of glycine, β-alanine, and γ-amino-n-butyric acid were observed in free form in SM2 and SM51 compared with the soil, suggesting that these free amino acids may be indigenous. Trace levels of D+L-β-aminoisobutyric acid (β-AIB) observed in all three meteorites are not easily explained as terrestrial contamination, as β-AIB is rare on Earth and was not detected in the soil. Bulk carbon and nitrogen and isotopic ratios of the SM samples and the soil also indicate terrestrial contamination, as does compound-specific isotopic analysis of the amino acids in the soil. The amino acid abundances in SM2, the most pristine SM meteorite analyzed here, are approximately 20-fold lower than in the Murchison CM2 carbonaceous chondrite. This may be due to thermal metamorphism in the Sutter's Mill parent body at temperatures greater than observed for other aqueously altered CM2 meteorites.

  6. Organic blend semiconductors and transistors with hole mobility exceeding 10 cm2/Vs (Presentation Recording)

    NASA Astrophysics Data System (ADS)

    Paterson, Alexandra F.; Anthopoulos, Thomas D.

    2015-10-01

    Plastic electronics that can be manufactured using solution-based methods are the subject of great research interest due to their potential for low-cost, large-area electronic applications. The interest in this field has led to considerable research and subsequent advances in device performance. To this end solution-processed organic thin-film transistors (OTFTs) have shown impressive improvements in recent years through the increasing values of charge carrier mobility. Here we report the development of next generation organic blend materials for OTFTs with hole mobilities of 10 cm2/Vs. These high performance devices have been achieved using a novel semiconducting blend system comprising of an amorphous-like conjugated polymer and a high mobility small molecule. The combination of a highly crystalline small molecule with the polymer binder aids the formation of uniform films as well as enables an element of control over the nucleation and growth of the small molecule. The polymer binders investigated belongs to the family of indacenodithiophene-based copolymers which are renowned for their high carrier mobilities regardless of their apparent structural disorder. The addition of the polymer with carefully chosen small molecules is found to further increase the hole mobility of the resulting blend OTFT to over 10 cm2/Vs. These organic devices provide an interesting insight into this rather complex blend system, highlighting the correlation between the morphology developed following solution processing and device performance, as well as exploring the role of each of the two components in the blend in terms of their contribution to charge transport.

  7. Erg cooperates with TGF-β to control mesenchymal differentiation.

    PubMed

    Cox, Megan K; Appelboom, Brittany L; Ban, Ga I; Serra, Rosa

    2014-11-01

    Transforming growth factor β (TGF-β) signaling plays an integral role in skeletal development. Conditional deletion of the TGF-β type II receptor (Tgfbr2) from type II Collagen (Col2a) expressing cells results in defects in development of the annulus fibrosus (AF) of the intervertebral disc (IVD). We previously used microarray analysis to search for marker genes of AF as well as transcription factors regulated by TGF-β during AF development. The transcription factor avian erythroblastosis virus E-26 (v-ets) oncogene related (Erg) was identified in the microarray screen as a candidate regulator of AF development. To study the effects of TGF-β on AF differentiation and the role of Erg in this process, we used mouse sclerotome grown in micromass cultures. At 0.5ng TGF-β/ml, sclerotome cells started to express markers of AF. Regulation of Erg by TGF-β was confirmed in these cells. In addition, TGF-β soaked Affi-gel beads implanted into the axial skeleton of stage HH 25 chick embryos showed that TGF-β could induce expression of Erg mRNA in vivo. Next, an adenovirus to over-express Erg in primary sclerotome micromass cultures was generated. Over-expression of Erg led to a change in cell morphology and inhibition of differentiation into hyaline cartilage as seen by reduced Alcian blue staining and decreased Sox9 and c-Maf expression. Erg was not sufficient to induce expression of AF markers and expression of Sca1, a marker of pluripotent progenitor cells, was up-regulated in Erg expressing cells. When cells that ectopically expressed Erg were treated with TGF-β, enhanced expression of specific differentiation markers was observed suggesting Erg can cooperate with TGF-β to regulate differentiation of the sclerotome. Furthermore, we showed using co-immunopreciptiation that Erg and Smad3 bind to each other suggesting a mechanism for their functional interaction.

  8. Evaporation of metals by high-density (107 A · cm-2) electrical currents

    NASA Astrophysics Data System (ADS)

    Rakhel, A. D.

    1996-09-01

    In the present work, the problem of time evolution of pressure and temperature profiles across a wire through which an electrical current with a density of the order of 107 A · cm-2 flows is solved. The correct boundary conditions for a metal surface are obtained for the case when this metal is rapidly evaporated as a result of high-power Joule heating. The pressure profile appears under these conditions due to pinch-effect and inertia of thermal expansion of the metal; the temperature profile arises because of intensive evaporation from the surface of the wire. The conditions under which a liquid metal is superheated are formulated. On the basis of the analysis of the experimental results on exploding wires, the conclusion is drawn that decay of the metastable state takes place near the binodal. It is shown that the distribution of fine dispersed vapor bubbles is strongly nonuniform across the wire and the process of expansion of the two-phase mixture is very similar to the motion of a wave.

  9. On the Behavior of Phosphorus During the Aqueous Alteration of CM2 Carbonaceous Chondrites

    NASA Technical Reports Server (NTRS)

    Brearley, Adrian J.; Chizmadia, Lysa J.

    2005-01-01

    During the earliest period of solar system formation, water played an important role in the evolution of primitive dust, both after accretion of planetesimals and possible before accretion within the protoplanetary disk. Many chondrites show evidence of variable degrees of aqueous alteration, the CM2 chondrites being among the most studied [1]. This group of chondrites is characterized by mineral assemblages of both primary and secondary alteration phases. Hence, these meteorites retain a particularly important record of the reactions that occurred between primary high temperature nebular phases and water. Studies of these chondrites can provide information on the conditions and environments of aqueous alteration and the mobility of elements during alteration. This latter question is at the core of a debate concerning the location of aqueous alteration, i.e. whether alteration occurred predominantly within a closed system after accretion (parent body alteration) or whether some degree of alteration occurred within the solar nebula or on ephemeral protoplanetary bodies prior to accretion. At the core of the parent body alteration model is the hypothesis that elemental exchange between different components, principally chondrules and matrix, must have occurred. chondrules and matrix, must have occurred. In this study, we focus on the behavior of the minor element, phosphorus. This study was stimulated by observations of the behavior of P during the earliest stages of alteration in glassy mesostasis in type II chondrules in CR chondrites and extends the preliminary observations of on Y791198 to other CM chondrites.

  10. Murchison CM2 chondrite at nanoscale: evidence for hydrated minerals in the protoplanetary disk

    NASA Astrophysics Data System (ADS)

    Trigo, J. M.; Vila-Ruaix, A.; Alonso-Azcárate, J.; Abad, M. M.

    2017-03-01

    The most pristine chondrites are undifferentiated meteorites with highly unequilibrated mineral grains that accreted from the protoplanetary disk about 4.6 Gyrs ago. Here we focus our attention in the study of Murchison, one of the most primitive carbonaceous chondrites belonging to the CM2 group. Despite of being aqueously altered, Murchison matrix is extraordinarily complex at nanoscale, and its study can hold clues to understand the origin of the water incorporated in the parent bodies of carbonaceous chondrites. Murchison comes from an undifferentiated carbon-rich asteroid which formed from the accretion of solid particles formed in the outer protoplanetary disk. Their rock-forming materials felt into the plane of the system where they mixed with organics, and probably with hydrated minerals. Our UHRTEM (ultra-high resolution transmission electron microscopy) data demonstrate that Murchison fine-grained matrix consists of a complex mixture of many ingredients, including chondrule and CAI fragments, stellar grains, phyllosilicates and organic compounds. We describe here some mineral and textural features that exemplify how pristine, and diverse is Murchison matrix. Our results indicate that the study of carbonaceous chondrites at nanoscale can provide a significant progress in our understanding of the accretion of materials and the preservation of presolar grains in the outer regions of the protoplanetary disk.

  11. A minimum column density of 1 g cm(-2) for massive star formation.

    PubMed

    Krumholz, Mark R; McKee, Christopher F

    2008-02-28

    Massive stars are very rare, but their extreme luminosities make them both the only type of young star we can observe in distant galaxies and the dominant energy sources in the Universe today. They form rarely because efficient radiative cooling keeps most star--forming gas clouds close to isothermal as they collapse, and this favours fragmentation into stars of one solar mass or lower. Heating of a cloud by accreting low-mass stars within it can prevent fragmentation and allow formation of massive stars, but the necessary properties for a cloud to form massive stars-and therefore where massive stars form in a galaxy--have not yet been determined. Here we show that only clouds with column densities of at least 1 g cm(-2) can avoid fragmentation and form massive stars. This threshold, and the environmental variation of the stellar initial mass function that it implies, naturally explain the characteristic column densities associated with massive star clusters and the difference between the radial profiles of Halpha and ultraviolet emission in galactic disks. The existence of a threshold also implies that the initial mass function should show detectable variation with environment within the Galaxy, that the characteristic column densities of clusters containing massive stars should vary between galaxies, and that star formation rates in some galactic environments may have been systematically underestimated.

  12. N-15-Rich Organic Globules in a Cluster IDP and the Bells CM2 Chondrite

    NASA Technical Reports Server (NTRS)

    Messenger, S.; Nakamura-Messenger, K.; Keller, Lindsay P.

    2008-01-01

    Organic matter in primitive meteorites and chondritic porous interplanetary dust particles (CP IDPs) is commonly enriched in D/H and 15N/14N relative to terrestrial values [1-3]. These anomalies are ascribed to the partial preservation of presolar cold molecular cloud material [1]. Some meteorites and IDPs contain m-size inclusions with extreme H and N isotopic anomalies [2-4], possibly due to preserved pristine primordial organic grains. We recently showed that the in the Tagish Lake meteorite, the principle carriers of these anomalies are sub- m, hollow organic globules [5]. The globules likely formed by photochemical processing of organic ices in a cold molecular cloud or the outermost regions of the protosolar disk [5]. We proposed that similar materials should be common among primitive meteorites, IDPs, and comets. Similar objects have been observed in organic extracts of carbonaceous chondrites [6-8], however their N and H isotopic compositions are generally unknown. Bulk H and N isotopic compositions may indicate which meteorites best preserve interstellar organic compounds. Thus, we selected the Bells CM2 carbonaceous chondrites for study based on its large bulk 15N (+335 %) and D (+990 %) [9].

  13. Deregulation of DUX4 and ERG in acute lymphoblastic leukemia.

    PubMed

    Zhang, Jinghui; McCastlain, Kelly; Yoshihara, Hiroki; Xu, Beisi; Chang, Yunchao; Churchman, Michelle L; Wu, Gang; Li, Yongjin; Wei, Lei; Iacobucci, Ilaria; Liu, Yu; Qu, Chunxu; Wen, Ji; Edmonson, Michael; Payne-Turner, Debbie; Kaufmann, Kerstin B; Takayanagi, Shin-Ichiro; Wienholds, Erno; Waanders, Esmé; Ntziachristos, Panagiotis; Bakogianni, Sofia; Wang, Jingjing; Aifantis, Iannis; Roberts, Kathryn G; Ma, Jing; Song, Guangchun; Easton, John; Mulder, Heather L; Chen, Xiang; Newman, Scott; Ma, Xiaotu; Rusch, Michael; Gupta, Pankaj; Boggs, Kristy; Vadodaria, Bhavin; Dalton, James; Liu, Yanling; Valentine, Marcus L; Ding, Li; Lu, Charles; Fulton, Robert S; Fulton, Lucinda; Tabib, Yashodhan; Ochoa, Kerri; Devidas, Meenakshi; Pei, Deqing; Cheng, Cheng; Yang, Jun; Evans, William E; Relling, Mary V; Pui, Ching-Hon; Jeha, Sima; Harvey, Richard C; Chen, I-Ming L; Willman, Cheryl L; Marcucci, Guido; Bloomfield, Clara D; Kohlschmidt, Jessica; Mrózek, Krzysztof; Paietta, Elisabeth; Tallman, Martin S; Stock, Wendy; Foster, Matthew C; Racevskis, Janis; Rowe, Jacob M; Luger, Selina; Kornblau, Steven M; Shurtleff, Sheila A; Raimondi, Susana C; Mardis, Elaine R; Wilson, Richard K; Dick, John E; Hunger, Stephen P; Loh, Mignon L; Downing, James R; Mullighan, Charles G

    2016-12-01

    Chromosomal rearrangements deregulating hematopoietic transcription factors are common in acute lymphoblastic leukemia (ALL). Here we show that deregulation of the homeobox transcription factor gene DUX4 and the ETS transcription factor gene ERG is a hallmark of a subtype of B-progenitor ALL that comprises up to 7% of B-ALL. DUX4 rearrangement and overexpression was present in all cases and was accompanied by transcriptional deregulation of ERG, expression of a novel ERG isoform, ERGalt, and frequent ERG deletion. ERGalt uses a non-canonical first exon whose transcription was initiated by DUX4 binding. ERGalt retains the DNA-binding and transactivation domains of ERG, but it inhibits wild-type ERG transcriptional activity and is transforming. These results illustrate a unique paradigm of transcription factor deregulation in leukemia in which DUX4 deregulation results in loss of function of ERG, either by deletion or induced expression of an isoform that is a dominant-negative inhibitor of wild-type ERG function.

  14. A Comparison Between Silicon Carbide from Indarch (EH4) and CM2 Meteorites

    NASA Astrophysics Data System (ADS)

    Russell, S. S.; Alexander, C. M. O'd.; Ott, U.; Zinner, E. K.; Arden, J. W.; Pillinger, C. T.

    1993-07-01

    A light-element stepped-combustion, noble-gas, ion-probe, and SEM study of interstellar SiC from Indarch has been undertaken in order to compare SiC isolated from enstatite chondrites with SiC from the more extensively studied CM2 carbonaceous chondrites. Eighty-five grams of Indarch were etched in HF/HCl, crushed and treated with 9MHF/1MHCl + 1M HCl, Cr(sub)2O(sub)7^2- in H(sub)2SO(sub)4, and then HClO(sub)4, leaving an acid-resistant residue equivalent to 42 ppm of the whole rock. Carbon, nitrogen, and noble-gas data were acquired by stepped combustion and pyrolysis after precombusting the samples to 600 degrees C to oxidize nanometer-sized diamond. The presence of isotopically anomalous SiC in the Indarch residue is indicated by the isotopically heavy CO2 released at high temperature during stepped combustion, with a maximum delta ^13C value of +1420 per mil (^12C/^13C = 36.2), identical to results obtained for typical CM2 samples [1]. In contrast to CM meteorites, however, the peak release of heavy carbon occurs at 1200 degrees C, some 200 degrees C higher than the peak release temperatures of Murchison and Cold Bokkeveld. A similarly high release temperature was seen in the stepped-combustion analysis of the noble gas in the Indarch residue. This suggests a morphological and/or size difference between the SiC present in the two types of meteorites. The nitrogen stepped-combustion profile of the Indarch residue is dominated by the presence of Si(sub)3N(sub)4 of unremarkable isotopic composition (delta ^15N = -56 per mil) that could not be resolved from nitrogen released from SiC. The abundance of SiC in Indarch is estimated, from the stepped combustion data, to be about 1.4 ppm (or 14 ppm SiC in the matrix, not dissimilar to values obtained for CM2 meteorites). A comparison of the noble-gas data from grain-size fractions of Murchison [2] and the bulk Indarch residue data, particularly the Ne-E/Xe-s ratio, suggests that Indarch is enriched in fine-grained Si

  15. Characterizing unforced multi-decadal variability of ENSO: a case study with the GFDL CM2.1 coupled GCM

    NASA Astrophysics Data System (ADS)

    Atwood, A. R.; Battisti, D. S.; Wittenberg, A. T.; Roberts, W. H. G.; Vimont, D. J.

    2016-12-01

    Large multi-decadal fluctuations of El Niño-Southern Oscillation (ENSO) variability simulated in a 4000-year pre-industrial control run of GFDL CM2.1 have received considerable attention due to implications for constraining the causes of past and future changes in ENSO. We evaluated the mechanisms of this low-frequency ENSO modulation through analysis of the extreme epochs of CM2.1 as well as through the use of a linearized intermediate-complexity model of the tropical Pacific, which produces reasonable emulations of observed ENSO variability. We demonstrate that the low-frequency ENSO modulation can be represented by the simplest model of a linear, stationary process, even in the highly nonlinear CM2.1. These results indicate that CM2.1's ENSO modulation is driven by transient processes that operate at interannual or shorter time scales. Nonlinearities and/or multiplicative noise in CM2.1 likely exaggerate the ENSO modulation by contributing to the overly active ENSO variability. In contrast, simulations with the linear model suggest that intrinsically-generated tropical Pacific decadal mean state changes do not contribute to the extreme-ENSO epochs in CM2.1. Rather, these decadal mean state changes actually serve to damp the intrinsically-generated ENSO modulation, primarily by stabilizing the ENSO mode during strong-ENSO epochs. Like most coupled General Circulation Models, CM2.1 suffers from large biases in its ENSO simulation, including ENSO variance that is nearly twice that seen in the last 50 years of observations. We find that CM2.1's overly strong ENSO variance directly contributes to its strong multi-decadal modulation through broadening the distribution of epochal variance, which increases like the square of the long-term variance. These results suggest that the true spectrum of unforced ENSO modulation is likely substantially narrower than that in CM2.1. However, relative changes in ENSO modulation are similar between CM2.1, the linear model tuned to

  16. Fe and O EELS Studies of Ion Irradiated Murchison CM2 Carbonaceous Chondrite Matrix

    NASA Technical Reports Server (NTRS)

    Keller, L. P.; Christofferson, R.; Dukes, C. A.; Baragiola, R. A.; Rahman, Z.

    2015-01-01

    Introduction: The physical and chemical response of hydrated carbonaceous chondrite materials to space weathering processes is poorly understood. Improving this understanding is a key part of establishing how regoliths on primitive carbonaceous asteroids respond to space weathering processes, knowledge that supports future sample return missions (Hayabusa 2 and OSIRISREx) that are targeting objects of this type. We previously reported on He+ irradiation of Murchison matrix and showed that the irradiation resulted in amorphization of the matrix phyllosilicates, loss of OH, and surface vesiculation. Here, we report electron energy-loss spectroscopy (EELS) measurements of the irradiated material with emphasis on the Fe and O speciation. Sample and Methods: A polished thin section of the Murchison CM2 carbonaceous chondrite was irradiated with 4 kilovolts He(+) (normal incidence) to a total dose of 1 x 10(exp 18) He(+) per square centimeter. We extracted thin sections from both irradiated and unirradiated regions in matrix using focused ion beam (FIB) techniques with electron beam deposition for the protective carbon strap to minimize surface damage artifacts from the FIB milling. The FIB sections were analyzed using a JEOL 2500SE scanning and transmission electron microscope (STEM) equipped with a Gatan Tridiem imaging filter. EELS spectra were collected from 50 nanometer diameter regions with an energy resolution of 0.7 electronvolts FWHM at the zero loss. EELS spectra were collected at low electron doses to minimize possible artifacts from electron-beam irradiation damage. Results and Discussion: Fe L (sub 2,3) EELS spectra from matrix phyllosilicates in CM chondrites show mixed Fe(2+)/Fe(3+) oxidation states with Fe(3+)/Sigma Fe approximately 0.5. Fe L(sub 2,3) spectra from the irradiated/ amorphized matrix phyllosilicates show higher Fe(2+)/Fe(3+) ratios compared to spectra obtained from pristine material at depths beyond the implantation/amorphization layer. We

  17. Impact-Induced Chondrule Deformation and Aqueous Alteration of CM2 Murchison

    NASA Technical Reports Server (NTRS)

    Hanna, R. D.; Zolensky, M.; Ketcham, R. A.; Behr, W. M.; Martinez, J. E.

    2014-01-01

    Deformed chondrules in CM2 Murchison have been found to define a prominent foliation [1,2] and lineation [3] in 3D using X-ray computed tomography (XCT). It has been hypothesized that chondrules in foliated chondrites deform by "squeezing" into surrounding pore space [4,5], a process that also likely removes primary porosity [6]. However, shock stage classification based on olivine extinction in Murchison is consistently low (S1-S2) [4-5,7] implying that significant intracrystalline plastic deformation of olivine has not occurred. One objective of our study is therefore to determine the microstructural mechanisms and phases that are accommodating the impact stress and resulting in relative displacements within the chondrules. Another question regarding impact deformation in Murchison is whether it facilitated aqueous alteration as has been proposed for the CMs which generally show a positive correlation between degree of alteration and petrofabric strength [7,2]. As pointed out by [2], CM Murchison represents a unique counterpoint to this correlation: it has a strong petrofabric but a relatively low degree of aqueous alteration. However, Murchison may not represent an inconsistency to the proposed causal relationship between impact and alteration, if it can be established that the incipient aqueous alteration post-dated chondrule deformation. Methods: Two thin sections from Murchison sample USNM 5487 were cut approximately perpendicular to the foliation and parallel to lineation determined by XCT [1,3] and one section was additionally polished for EBSD. Using a combination of optical petrography, SEM, EDS, and EBSD several chondrules were characterized in detail to: determine phases, find microstructures indicative of strain, document the geometric relationships between grain-scale microstructures and the foliation and lineation direction, and look for textural relationships of alteration minerals (tochilinite and Mg-Fe serpentine) that indicate timing of their

  18. Formation of spinel-, hibonite-rich inclusions found in CM2 carbonaceous chrondrites

    SciTech Connect

    Simon, S B; Grossman, L; Hutcheon, I D; Phinney, D L; Weber, P K; Fallon, S J

    2005-11-03

    We report petrography, mineral chemistry, bulk chemistry, and bulk isotopic compositions of a suite of 40 spinel-rich inclusions from the Murchison (CM2) carbonaceous chondrite. Seven types of inclusions are identified based on mineralogy: spinel-hibonite-perovskite; spinel-perovskite-pyroxene; spinel-perovskite-melilite; spinel-hibonite-perovskite-melilite; spinel-hibonite; spinel-pyroxene; and spinel-melilite-anorthite. Hibonite-bearing inclusions have Ti-poor spinel compared to the hibonite-free ones, and spinel-hibonite-perovskite inclusions have the highest average bulk TiO{sub 2} contents (7.8 wt%). The bulk CaO/Al{sub 2}O{sub 3} ratios of the inclusions range from 0.005-0.21, well below the solar value of 0.79. Hibonite-, spinel-rich inclusions consist of phases that are not predicted by condensation calculations to coexist; in the equilibrium sequence, hibonite is followed by melilite, which is followed by spinel. Therefore, hibonite-melilite or melilite-spinel inclusions should be dominant instead. One explanation for the 'missing melilite' is that it condensed as expected but was lost due to evaporation of Mg and Ca during heating and melting of spherule precursors. If this theory were correct, melilite-poor spherules would have isotopically heavy Mg and Ca. Except for one inclusion with F{sub Mg} = 4.3 {+-} 2.6{per_thousand}/amu and another with isotopically light Ca (F{sub Ca} = 3.4 {+-} 2.0{per_thousand}/amu), however, all the inclusions we analyzed have normal isotopic compositions within their 2{sigma} uncertainties. Thus, we found no evidence for significant mass-dependent fractionation. Our preferred explanation for the general lack of melilite among hibonite-, spinel-bearing inclusions is kinetic inhibition of melilite condensation relative to spinel. Because of similarities between the crystal structures of hibonite and spinel, it should be easier for spinel to form from hibonite than for melilite to do so.

  19. Escitalopram block of hERG potassium channels.

    PubMed

    Chae, Yun Ju; Jeon, Ji Hyun; Lee, Hong Joon; Kim, In-Beom; Choi, Jin-Sung; Sung, Ki-Wug; Hahn, Sang June

    2014-01-01

    Escitalopram, a selective serotonin reuptake inhibitor, is the pharmacologically active S-enantiomer of the racemic mixture of RS-citalopram and is widely used in the treatment of depression. The effects of escitalopram and citalopram on the human ether-a-go-go-related gene (hERG) channels expressed in human embryonic kidney cells were investigated using voltage-clamp and Western blot analyses. Both drugs blocked hERG currents in a concentration-dependent manner with an IC50 value of 2.6 μM for escitalopram and an IC50 value of 3.2 μM for citalopram. The blocking of hERG by escitalopram was voltage-dependent, with a steep increase across the voltage range of channel activation. However, voltage independence was observed over the full range of activation. The blocking by escitalopram was frequency dependent. A rapid application of escitalopram induced a rapid and reversible blocking of the tail current of hERG. The extent of the blocking by escitalopram during the depolarizing pulse was less than that during the repolarizing pulse, suggesting that escitalopram has a high affinity for the open state of the hERG channel, with a relatively lower affinity for the inactivated state. Both escitalopram and citalopram produced a reduction of hERG channel protein trafficking to the plasma membrane but did not affect the short-term internalization of the hERG channel. These results suggest that escitalopram blocked hERG currents at a supratherapeutic concentration and that it did so by preferentially binding to both the open and the inactivated states of the channels and by inhibiting the trafficking of hERG channel protein to the plasma membrane.

  20. Indexing molecules for their hERG liability.

    PubMed

    Rayan, Anwar; Falah, Mizied; Raiyn, Jamal; Da'adoosh, Beny; Kadan, Sleman; Zaid, Hilal; Goldblum, Amiram

    2013-07-01

    The human Ether-a-go-go-Related-Gene (hERG) potassium (K(+)) channel is liable to drug-inducing blockage that prolongs the QT interval of the cardiac action potential, triggers arrhythmia and possibly causes sudden cardiac death. Early prediction of drug liability to hERG K(+) channel is therefore highly important and preferably obligatory at earlier stages of any drug discovery process. In vitro assessment of drug binding affinity to hERG K(+) channel involves substantial expenses, time, and labor; and therefore computational models for predicting liabilities of drug candidates for hERG toxicity is of much importance. In the present study, we apply the Iterative Stochastic Elimination (ISE) algorithm to construct a large number of rule-based models (filters) and exploit their combination for developing the concept of hERG Toxicity Index (ETI). ETI estimates the molecular risk to be a blocker of hERG potassium channel. The area under the curve (AUC) of the attained model is 0.94. The averaged ETI of hERG binders, drugs from CMC, clinical-MDDR, endogenous molecules, ACD and ZINC, were found to be 9.17, 2.53, 3.3, -1.98, -2.49 and -3.86 respectively. Applying the proposed hERG Toxicity Index Model on external test set composed of more than 1300 hERG blockers picked from chEMBL shows excellent performance (Matthews Correlation Coefficient of 0.89). The proposed strategy could be implemented for the evaluation of chemicals in the hit/lead optimization stages of the drug discovery process, improve the selection of drug candidates as well as the development of safe pharmaceutical products.

  1. Marginal erg facies: A trial approach toward a descriptive classification

    SciTech Connect

    Caputo, M.V. ); Langford, R.P. )

    1991-03-01

    During the late 1970s and early 1980s, sedimentologists began recognizing the margins of eolian sand seas as separate, components which differed from interior sand seas in geometry, extent, and facies. Stratigraphers have now observed these differences in eolian rocks. Erg margins may be grouped in five ways: (1) by associations with extradunal environments-coastal plain, lacustrine, periglacial, marine (tidal flat, coastal sabkha, beach, and lagoon), and arid alluvial (alluvial fan, fluvial, playa, inland sabkha); (2) by allocyclic controls-eustasy, plate tectonism, and climate; (3) by autocyclic controls-local tectonism, topography, vegetation, hydrology, structure, sediment source and supply, and wind regime; (4) by geographic position-upwind, downwind, and along-wind margins; and (5) by sedimentary facies-texture and architecture. In contrast with erg interiors, erg margins are characterized by smaller, less complex dune-forms related to thinner sand accumulation; elementary dune architecture; more vegetation and bioturbation; high occurrence of sand sheet, zibar, and serir facies; expansive, low-relief interdunes with widely distributed dunes; and a greater proportion of interbedded extradunal deposits. Some of the published studies on ancient eolian systems have identified erg margin facies that have been influences by marine and arid alluvial processes. Few reports have described lacustrine-eolian and periglacial-eolian interactions. This study is an attempt to organize known features of modern and ancient erg margins into a scheme based on erg margin controls.

  2. Characterization of hERG1a and hERG1b potassium channels-a possible role for hERG1b in the I (Kr) current.

    PubMed

    Larsen, Anders Peter; Olesen, Søren-Peter; Grunnet, Morten; Jespersen, Thomas

    2008-09-01

    I (Kr) is the fast component of the delayed rectifier potassium currents responsible for the repolarization of the cardiac muscle. The molecular correlate underlying the I (Kr) current has been identified as the hERG1 channel. Recently, two splice variants of the hERG1 alpha-subunit, hERG1a and hERG1b, have been shown to be co-expressed in human cardiomyocytes. In this paper, we present the electrophysiological characterization of hERG1a, hERG1b, and co-expressed hERG1a/b channels in a mammalian expression system using the whole-cell patch clamp technique. We also quantified the messenger RNA (mRNA) levels of hERG1a and hERG1b in human cardiac tissue, and based on the expressed ratios, we evaluated the resulting currents in Xenopus laevis oocytes. Compared to hERG1a channels, activation was faster for both hERG1b and hERG1a/b channels. The deactivation kinetics was greatly accelerated in the presence of hERG1b, whereas no difference in the time constant of inactivation was observed. The voltage-dependent recovery from inactivation was also similar. However, the time constant of recovery from inactivation was significantly faster for hERG1b channels compared to hERG1a and hERG1a/b. Quantification of hERG1a and hERG1b mRNA in the human heart showed that hERG1b mRNA constitutes, on average, 19% in the right atrium and 12% in the left ventricle of the total hERG1 mRNA. Expression of the observed ratios of hERG1a to hERG1b in X. laevis oocytes showed that these ratios are indeed sufficient to change the deactivation phenotype markedly. The present work suggests that hERG1b is likely to play a role in the formation of the native I (Kr) current.

  3. Nanoimprinted polymer lasers with threshold below 100 W/cm2 using mixed-order distributed feedback resonators.

    PubMed

    Wang, Yue; Tsiminis, Georgios; Kanibolotsky, Alexander L; Skabara, Peter J; Samuel, Ifor D W; Turnbull, Graham A

    2013-06-17

    Organic semiconductor lasers were fabricated by UV-nanoimprint lithography with thresholds as low as 57 W/cm(2) under 4 ns pulsed operation. The nanoimprinted lasers employed mixed-order distributed feedback resonators, with second-order gratings surrounded by first-order gratings, combined with a light-emitting conjugated polymer. They were pumped by InGaN LEDs to produce green-emitting lasers, with thresholds of 208 W/cm(2) (102 nJ/pulse). These hybrid lasers incorporate a scalable UV-nanoimprint lithography process, compatible with high-performance LEDs, therefore we have demonstrated a coherent, compact, low-cost light source.

  4. THE TMPRSS2:ERG REARRANGEMENT, ERG EXPRESSION, AND PROSTATE CANCER OUTCOMES: A COHORT STUDY AND META-ANALYSIS

    PubMed Central

    Pettersson, Andreas; Graff, Rebecca E.; Bauer, Scott R.; Pitt, Michael; Lis, Rosina T.; Stack, Edward C.; Martin, Neil E.; Kunz, Lauren; Penney, Kathryn L.; Ligon, Azra H.; Suppan, Catherine; Flavin, Richard; Sesso, Howard D.; Rider, Jennifer R.; Sweeney, Christopher; Stampfer, Meir; Fiorentino, Michelangelo; Kantoff, Philip W.; Sanda, Martin; Giovannucci, Edward; Ding, Eric L.; Loda, Massimo; Mucci, Lorelei A.

    2013-01-01

    Background Whether the genomic rearrangement TMPRSS2:ERG has prognostic value in prostate cancer is unclear. Methods Among men with prostate cancer in the prospective Physicians’ Health and Health Professionals Follow-Up Studies, we identified rearrangement status by immunohistochemical assessment of ERG protein expression. We used Cox models to examine associations of ERG overexpression with biochemical recurrence and lethal disease (distant metastases or cancer-specific mortality). In a meta-analysis including 47 additional studies, we used random effects models to estimate associations between rearrangement status and outcomes. Results The cohort consisted of 1,180 men treated with radical prostatectomy between 1983 and 2005. During a median follow-up of 12.6 years, 266 men experienced recurrence, and 85 men developed lethal disease. We found no significant association between ERG overexpression and biochemical recurrence (HR: 0.99; 95% CI: 0.78-1.26) or lethal disease (HR: 0.93; 95% CI: 0.61-1.43). The meta-analysis of prostatectomy series included 5,074 men followed for biochemical recurrence (1,623 events), and 2,049 men followed for lethal disease (131 events). TMPRSS2:ERG was associated with stage at diagnosis (RR≥T3 vs. T2: 1.23; 95% CI: 1.16-1.30) but not with biochemical recurrence (RR: 1.00; 95% CI: 0.86-1.17) or lethal disease (RR: 0.99; 95% CI: 0.47-2.09). Conclusions These results suggest that TMPRSS2:ERG, or ERG overexpression, is associated with tumor stage but does not strongly predict recurrence or mortality among men treated with radical prostatectomy. Impact This is the largest prospective cohort study to examine associations of ERG overexpression and lethal prostate cancer among men treated with radical prostatectomy. PMID:22736790

  5. ERG transcriptional networks in primary acute leukemia cells implicate a role for ERG in deregulated kinase signaling.

    PubMed

    Bock, Juliane; Mochmann, Liliana H; Schlee, Cornelia; Farhadi-Sartangi, Nasrin; Göllner, Stefanie; Müller-Tidow, Carsten; Baldus, Claudia D

    2013-01-01

    High expression of the E26 transforming sequence related gene (ERG) is associated with poor prognosis in a subgroup of leukemia patients with acute myeloid (AML) and acute T-lymphoblastic leukemia (T-ALL). In a previous study we proposed that ERG overexpression may deregulate several signaling cascades in acute leukemia. Herein, we further expand those studies by identifying a consensus of biological targets in primary blasts of newly diagnosed acute leukemia patients. Our findings of chromatin immunoprecipitation-on-chip of primary samples revealed 48 significantly enriched single genes including DAAM1 and NUMB. Significantly enriched signaling pathways included WNT/β-catenin, p53, and PI3K/AKT with ERG overexpression inducing dephosphorylation of AKT(Ser473) relative to non ERG expressing K562 cells. Cell based ERG overexpression studies also revealed drug resistance to multi-kinase inhibitor, BAY 43-9006 (Sorafenib) and to the tyrosine kinase inhibitor TKI258. Thus in primary leukemic cells, ERG may contribute to the dysregulation of kinase signaling, which results in resistance to kinase inhibitors.

  6. Successful Capture, Extraction and Identification of Hypervelocity CM2 Meteorite Fragments Shot by Light-Gas Gun

    NASA Technical Reports Server (NTRS)

    Snead, C.; Westphal, A. J.; Dominguez, G.; Zolensky, M. E.

    2003-01-01

    Here we report the successful capture, extraction and identification of two fragments of a CM2 meteorite (ALH83100) into lowdensity aerogel. The shot was carried out at the AVGR at NASAARC. A mixture of powdered ALH83100 and borosilicate glass microspheres was shot at 4.55.0 km/sec into 50 mg cm silicate aerogel.

  7. Comparison of visible and near-infrared reflectance spectra of CM2 carbonaceous chondrites and primitive asteroids

    NASA Technical Reports Server (NTRS)

    Vilas, F.; Hiroi, T.; Zolensky, M. E.

    1993-01-01

    Spectra of primitive asteroids (defined as C, P, and D classes and associated subclasses) were compared to the limited number of spectra of CM2 carbonaceous chondrites. An absorption feature located at 0.7 microns attributed to an Fe(+2) - Fe(+3) charge transfer absorption in iron oxides in phyllosilicates is apparent in some of the CM2 carbonaceous chondrite spectra and many of the asteroid spectra. Sawyer found a correlation between the area of the 0.7 micron feature and the mean semimajor axis of the asteroids. Spectra of a larger sample of carbonaceous chondrites, including 7 CM2 chondrites, covering a spectral interval of 0.30-2.5 microns were recently obtained using the Relab instrument at Brown University. These spectra were compared with spectrophotometric asteroid observations in a separate abstract. Those spectra of CM2 chondrites were isolated into the UV, visible and near-infrared spectral regions in order to compare them with high-quality narrowband reflectance spectra.

  8. Autoinhibitory mechanisms of ERG studied by molecular dynamics simulations

    NASA Astrophysics Data System (ADS)

    Lu, Yan; Salsbury, Freddie R.

    2015-01-01

    ERG, an ETS-family transcription factor, acts as a regulator of differentiation of early hematopoietic cells. It contains an autoinhibitory domain, which negatively regulates DNA-binding. The mechanism of autoinhibitory is still illusive. To understand the mechanism, we study the dynamical properties of ERG protein by molecular dynamics simulations. These simulations suggest that DNA binding autoinhibition associates with the internal dynamics of ERG. Specifically, we find that (1), The N-C terminal correlation in the inhibited ERG is larger than that in uninhibited ERG that contributes to the autoinhibition of DNA-binding. (2), DNA-binding changes the property of the N-C terminal correlation from being anti-correlated to correlated, that is, changing the relative direction of the correlated motions and (3), For the Ets-domain specifically, the inhibited and uninhibited forms exhibit essentially the same dynamics, but the binding of the DNA decreases the fluctuation of the Ets-domain. We also find from PCA analysis that the three systems, even with quite different dynamics, do have highly similar free energy surfaces, indicating that they share similar conformations.

  9. Clonal Evaluation of Prostate Cancer by ERG/SPINK1 Status to Improve Prognosis Prediction

    DTIC Science & Technology

    2015-10-01

    ERG-/SPINK1- ERG+/SPINK1- RPA 80% RRP or RP or ERG+/SPINK1- ERG/SPINK1 IHC C H &E H &E ERG/SPINK1 IHC D Figure 1 B Figure 1. Clonal evaluation of...Examples of Hematoxylin and eosin ( H &E) stained and dual ERG/SPINK1 IHC discontinuously involved prostate biopsy cores from two cases are shown. In C , both...SUBJECT TERMS Multiclonality, ERG, SPINK1, immunohistochemistry, active surveillance, prostate biopsy, prostatectomy 16. SECURITY CLASSIFICATION OF: 17

  10. Endothelial Erg expression is required for embryogenesis and vascular integrity

    PubMed Central

    Han, Rong; Pacifici, Maurizio; Iwamoto, Masahiro; Trojanowska, Maria

    2015-01-01

    abstract Members of the ETS family of transcription factors are involved in several developmental processes including endothelial cell specification and blood vessel formation, but their exact roles remain unclear. The family member Erg is highly expressed in endothelial cells as compared to other developing cell types including chondrocytes, hematopoietic cells and mesodermal cells. To study the specific roles ERG plays in endothelial cell specification and function during early embryogenesis, we conditionally ablated it by mating ErgloxP/loxP and Tie2-Cre mice. We found that mutant embryos died by mid-gestation and that angiogenesis and vascular integrity were highly compromised. Our study reveals that ERG has essential and cell autonomous roles in endothelial cell development and blood vessel maintenance. PMID:26061019

  11. Adiabatic quantum-flux-parametron cell library designed using a 10 kA cm-2 niobium fabrication process

    NASA Astrophysics Data System (ADS)

    Takeuchi, Naoki; Nagasawa, Shuichi; China, Fumihiro; Ando, Takumi; Hidaka, Mutsuo; Yamanashi, Yuki; Yoshikawa, Nobuyuki

    2017-03-01

    Adiabatic quantum-flux-parametron (AQFP) logic is an energy-efficient superconductor logic with zero static power consumption and very small switching energy. In this paper, we report a new AQFP cell library designed using the AIST 10 kA cm-2 Nb high-speed standard process (HSTP), which is a high-critical-current-density version of the AIST 2.5 kA cm-2 Nb standard process (STP2). Since the intrinsic damping of the Josephson junction (JJ) of HSTP is relatively strong, shunt resistors for JJs were removed and the energy efficiency improved significantly. Also, excitation transformers in the new cells were redesigned so that the cells can operate in a four-phase excitation mode. We described the detail of HSTP and the AQFP cell library designed using HSTP, and showed experimental results of cell test circuits.

  12. Genetic analyses involving interactions between the ergosterol biosynthetic enzymes, lanosterol synthase (Erg7p) and 3-ketoreductase (Erg27p), in the yeast Saccharomyces cerevisiae

    PubMed Central

    Teske, B.; Taramino, S.; Bhuiyan, M. S. A.; Kumaraswami, N. S.; Randall, S. K.; Barbuch, R.; Eckstein, J.; Balliano, G.; Bard, M.

    2008-01-01

    Summary Protein-protein interaction studies in the S. cerevisiae ergosterol biosynthetic pathway suggest that enzymes in this pathway may act as an integrated multienzyme complex. The yeast sterol 3-ketoreductase (Erg27p) required for C-4 demethylation of sterols has previously been shown to also be required for the function of the upstream oxidosqualene cyclase/lanosterol synthase (Erg7p); thus, erg27 mutants accumulate oxidosqualenes as precursors rather than 3-ketosterones. In the present study, we have created various mutations in the ERG27 gene. These mutations include 5 C-terminal truncations, 6 internal deletions, and 32 point mutants of which 14 were obtained by site directed mutagenesis and 18 by random mutagenesis. We have characterized these ERG27 mutations by determining the following: Erg27 and Erg7 enzyme activities, presence of Erg27p as determined by western immunoblots, ability to grow on various sterol substrates and GC sterol profiles. Mutations of the predicted catalytic residues, Y202F and K206A, resulted in the endogenous accumulation of 3-ketosterones rather than oxidosqualenes suggesting retention of Erg7 enzyme activity. This novel phenotype demonstrated that the catalytic function of Erg27p can be separated from its Erg7p chaperone ability. Other erg27 mutations resulted in proteins that were present, as determined by western immunoblotting, but unable to interact with the Erg7 protein. We also classify Erg27p as belonging to the SDR (short chain dehydrogenase/reductase) family of enzymes and demonstrate the possibility of homo -or hetero-dimerization of the protein. This study provides new insights into the role of Erg27p in sterol biosynthesis. PMID:18555807

  13. A comparison of ERG abnormalities in XLRS and XLCSNB.

    PubMed

    Bradshaw, Keith; Allen, Louise; Trump, Dorothy; Hardcastle, Alison; George, Nicolas; Moore, Anthony

    2004-03-01

    Dark and light adapted ERGs were recorded in 19 patients with XLRS and in 15 patients with CSNB. Patients were assigned to clinical groups after identification of mutations in the RS1 (16 patients), NYX (11 patients) and CACNA1F (4 patients) genes causing XLRS, 'complete' CSNB and 'incomplete' CSNB, respectively. ERG responses were compared with those of 26 healthy volunteers. Rod responses were most severely affected in patients in the NYX group but a rod-generated b-wave could be identified in the majority of patients in this group. Rod responses were less severely affected in the CACNA1F and RS1 groups and ERGs did not differ significantly between these two groups. Cone reponses were largely unaffected in the NYX group but were abnormal in the RS1 and especially CACNA1F groups. The ERG results suggest that the RS1 and CACNA1F gene products have comparable functional consequences and that all three genes may affect multiple retinal sites.

  14. ERG deregulation induces PIM1 over-expression and aneuploidy in prostate epithelial cells.

    PubMed

    Magistroni, Vera; Mologni, Luca; Sanselicio, Stefano; Reid, James Frances; Redaelli, Sara; Piazza, Rocco; Viltadi, Michela; Bovo, Giorgio; Strada, Guido; Grasso, Marco; Gariboldi, Manuela; Gambacorti-Passerini, Carlo

    2011-01-01

    The ERG gene belongs to the ETS family of transcription factors and has been found to be involved in atypical chromosomal rearrangements in several cancers. To gain insight into the oncogenic activity of ERG, we compared the gene expression profile of NIH-3T3 cells stably expressing the coding regions of the three main ERG oncogenic fusions: TMPRSS2/ERG (tERG), EWS/ERG and FUS/ERG. We found that all three ERG fusions significantly up-regulate PIM1 expression in the NIH-3T3 cell line. PIM1 is a serine/threonine kinase frequently over-expressed in cancers of haematological and epithelial origin. We show here that tERG expression induces PIM1 in the non-malignant prostate cell line RWPE-1, strengthening the relation between tERG and PIM1 up-regulation in the initial stages of prostate carcinogenesis. Silencing of tERG reversed PIM1 induction. A significant association between ERG and PIM1 expression in clinical prostate carcinoma specimens was found, suggesting that such a mechanism may be relevant in vivo. Chromatin Immunoprecipitation experiments showed that tERG directly binds to PIM1 promoter in the RWPE-1 prostate cell line, suggesting that tERG could be a direct regulator of PIM1 expression. The up-regulation of PIM1 induced by tERG over-expression significantly modified Cyclin B1 levels and increased the percentage of aneuploid cells in the RWPE-1 cell line after taxane-based treatment. Here we provide the first evidence for an ERG-mediated PIM1 up-regulation in prostate cells in vitro and in vivo, suggesting a direct effect of ERG transcriptional activity in the alteration of genetic stability.

  15. The ubiquitin ligase TRIM25 targets ERG for degradation in prostate cancer

    PubMed Central

    Wang, Shan; Kollipara, Rahul K.; Humphries, Caroline G.; Ma, Shi-Hong; Hutchinson, Ryan; Li, Rui; Siddiqui, Javed; Tomlins, Scott A.; Raj, Ganesh V.; Kittler, Ralf

    2016-01-01

    Ets related gene (ERG) is a transcription factor that is overexpressed in 40% of prostate tumors due to a gene fusion between ERG and TMPRSS2. Because ERG functions as a driver of prostate carcinogenesis, understanding the mechanisms that influence its turnover may provide new molecular handles to target the protein. Previously, we found that ERG undergoes ubiquitination and then is deubiquitinated by USP9X in prostate cancer cells to prevent its proteasomal degradation. Here, we identify Tripartite motif-containing protein 25 (TRIM25) as the E3 ubiquitin ligase that ubiquitinates the protein prior to its degradation. TRIM25 binds full-length ERG, and it also binds the N-terminally truncated variants of ERG that are expressed in tumors with TMPRSS2-ERG fusions. We demonstrate that TRIM25 polyubiquitinates ERG in vitro and that inactivation of TRIM25 resulted in reduced polyubiquitination and stabilization of ERG. TRIM25 mRNA and protein expression was increased in ERG rearrangement-positive prostate cancer specimens, and we provide evidence that ERG upregulates TRIM25 expression. Thus, overexpression of ERG in prostate cancer may cause an increase in TRIM25 activity, which is mitigated by the expression of the deubiquitinase USP9X, which is required to stabilize ERG. PMID:27626314

  16. New potential binding determinant for hERG channel inhibitors

    NASA Astrophysics Data System (ADS)

    Saxena, P.; Zangerl-Plessl, E.-M.; Linder, T.; Windisch, A.; Hohaus, A.; Timin, E.; Hering, S.; Stary-Weinzinger, A.

    2016-04-01

    Human ether-à-go-go related gene (hERG) 1 channels conduct the rapid delayed rectifier K+ current (IKr) and are essential for the repolarization of the cardiac action potential. hERG1 inhibition by structurally diverse drugs may lead to life threatening arrhythmia. Putative binding determinants of hERG1 channel blockers include T623, S624 and V625 on the pore helix, and residues G648, Y652 and F656, located on segment S6. We and others have previously hypothesized that additional binding determinants may be located on helix S5, which is in close contact with the S6 segments. In order to test this hypothesis, we performed a detailed investigation combining ionic current measurements with two-microelectrode voltage clamp and molecular modeling techniques. We identified a novel aromatic high affinity binding determinant for blockers located in helix S5, F557, which is equally potent as Y652. Modeling supports a direct interaction with the outer pore helix.

  17. Advanced polymer targets for TNSA regime producing 6 MeV protons at 1016 W/cm2 laser intensity

    NASA Astrophysics Data System (ADS)

    Torrisi, L.

    2017-02-01

    High intensity laser pulses, at an intensity of the order of 1016 W/cm2, are employed to irradiate in vacuum polyethylene terephthalate thin foils in the target normal sheath acceleration (TNSA) regime. The plasma obtained in the forward emission is investigated using ion collectors and semiconductor detectors connected in a time-of-flight configuration, Thomson parabola spectrometer, and X-ray streak camera. The results indicate that the foil thickness of 1 micron is optimal to accelerate protons of up to 6.5 MeV. The high ion acceleration can be due to different effects such as the high absorption in the advanced semicrystalline polymer containing spherulite centers, the high resonant absorption in gold nanoparticles embedded in the polymer, the optimal thickness of the used polymer to enhance the electron density in the forward plasma, and the self-focusing effect induced by preplasma created in front of the irradiated target.

  18. Xenoliths in the CM2 Carbonaceous Chondrite LON 94101: Implications for Complex Mixing on the Asteroidal Parent Body

    NASA Technical Reports Server (NTRS)

    Lindgren, P.; Lee, M. R.; Sofe, M.; Zolensky, M. E.

    2011-01-01

    Xenoliths are foreign clasts that oc-cur in various classes of meteorites, e.g. [1,2,3]. A re-cent study reveals the presence of several distinct classes of xenoliths in regolith-bearing meteorites, in-cluding in over 20 different carbonaceous chondrites [4]. The most common types of xenoliths are fine-grained hydrous clasts, often referred to as C1 or CI clasts in the literature, although their mineralogy is actually more similar to hydrous micrometeorites [5,6]. Xenoliths in meteorites present an opportunity to study material not yet classified or available as separate meteorites, and can provide additional information on processes in the dynamic early history of the Solar Sys-tem. Here we have performed chemical and mineralogi-cal analyses of xenoliths in the CM2 carbonaceous chondrite LON 94101, using scanning electron micro-scopy (SEM) and transmission electron microscopy (TEM).

  19. TMPRSS2:ERG blocks neuroendocrine and luminal cell differentiation to maintain prostate cancer proliferation.

    PubMed

    Mounir, Z; Lin, F; Lin, V G; Korn, J M; Yu, Y; Valdez, R; Aina, O H; Buchwalter, G; Jaffe, A B; Korpal, M; Zhu, P; Brown, M; Cardiff, R D; Rocnik, J L; Yang, Y; Pagliarini, R

    2015-07-01

    The biological outcome of TMPRSS2:ERG chromosomal translocations in prostate cancer (PC) remains poorly understood. To address this, we compared the transcriptional effects of TMPRSS2:ERG expression in a transgenic mouse model with those of ERG knockdown in a TMPRSS2:ERG-positive PC cell line. This reveals that ERG represses the expression of a previously unreported set of androgen receptor (AR)-independent neuronal genes that are indicative of neuroendocrine (NE) cell differentiation-in addition to previously reported AR-regulated luminal genes. Cell sorting and proliferation assays performed after sustained ERG knockdown indicate that ERG drives proliferation and blocks the differentiation of prostate cells to both NE and luminal cell types. Inhibition of ERG expression in TMPRSS2:ERG-positive PC cells through blockade of AR signaling is tracked with increased NE gene expression. We also provide evidence that these NE cells are resistant to pharmacological AR inhibition and can revert to the phenotype of parental cells upon restoration of AR/ERG signaling. Our findings highlight an ERG-regulated mechanism capable of repopulating the parent tumor through the transient generation of an anti-androgen therapy-resistant cell population, suggesting that ERG may have a direct role in preventing resistance to anti-androgen therapy.

  20. Stereoselective Blockage of Quinidine and Quinine in the hERG Channel and the Effect of Their Rescue Potency on Drug-Induced hERG Trafficking Defect.

    PubMed

    Yan, Meng; Fan, Pan; Shi, Yanhui; Feng, Lifang; Wang, Junnan; Zhan, Ge; Li, Baoxin

    2016-09-28

    Diastereoisomers of quinidine and quinine are used to treat arrhythmia and malaria, respectively. It has been reported that both drugs block the hERG (human ether-a-go-go-related gene) potassium channel which is essential for myocardium repolarization. Abnormality of repolarization increases risk of arrhythmia. The aim of our research is to study and compare the impacts of quinidine and quinine on hERG. Results show that both drugs block the hERG channel, with quinine 14-fold less potent than quinidine. In addition, they presented distinct impacts on channel dynamics. The results imply their stereospecific block effect on the hERG channel. However, F656C-hERG reversed this stereoselectivity. The mutation decreases affinity of the two drugs with hERG, and quinine was more potent than quinidine in F656C-hERG blockage. These data suggest that F656 residue contributes to the stereoselective pocket for quinidine and quinine. Further study demonstrates that both drugs do not change hERG protein levels. In rescue experiments, we found that they exert no reverse effect on pentamidine- or desipramine-induced hERG trafficking defect, although quinidine has been reported to rescue trafficking-deficient pore mutation hERG G601S based on the interaction with F656. Our research demonstrated stereoselective effects of quinidine and quinine on the hERG channel, and this is the first study to explore their reversal potency on drug-induced hERG deficiency.

  1. Two paths for stabilization of ERG in prostate carcinogenesis: TMPRSS2-ERG fusions and speckle-type pox virus and zinc finger protein mutations

    PubMed Central

    Pascal, Laura E; Wang, Zhou

    2016-01-01

    Speckle-type POZ (pox virus and zinc finger protein) protein (SPOP) is an E3 ubiquitin ligase adaptor protein that specifically promotes the ubiquitination and proteasome degradation of proteins. SPOP mutations are frequent in prostate cancer, and in a previous study, An et al. demonstrated that SPOP induced the degradation of the androgen receptor (AR) suggesting that SPOP is important in maintaining prostate homeostasis. In this current highlighted report, An and colleagues showed that ERG, which has been implicated as an oncoprotein in prostate cancer, contains putative SPOP-binding consensus (SBC) motifs 42ASSSS46 and 423VTSSS427 in the N- and C-terminal of ERG, respectively. The authors went on to demonstrate that SPOP promotes the ubiquitination and degradation of ERG through binding to the degron/SBC motif at the ERG N-terminus. SPOP mutations in the MATH domain prevented recognition and targeting of ERG for ubiquitination and degradation. In addition, N-terminal truncated ERG proteins encoded by the most frequently identified TMPRSS2-ERG rearrangements in prostate cancer (T1-E4 and T1-E5) were resistant to SPOP-mediated degradation, resulting in the stabilization of truncated ERG proteins. Stabilization of ERG protein through either SPOP mutation or TMPRSS2-ERG fusions induced proliferation and invasion in prostate cancer cells. This study along with a recently published similar report provides two previously unrecognized mechanisms for the upregulation of ERG proteins frequently observed in prostate cancers. These findings generate great enthusiasm for the development of targeted therapeutic strategies designed to eliminate ERG protein in prostate cancer cells. PMID:26763545

  2. Stereoselective Blockage of Quinidine and Quinine in the hERG Channel and the Effect of Their Rescue Potency on Drug-Induced hERG Trafficking Defect

    PubMed Central

    Yan, Meng; Fan, Pan; Shi, Yanhui; Feng, Lifang; Wang, Junnan; Zhan, Ge; Li, Baoxin

    2016-01-01

    Diastereoisomers of quinidine and quinine are used to treat arrhythmia and malaria, respectively. It has been reported that both drugs block the hERG (human ether-a-go-go-related gene) potassium channel which is essential for myocardium repolarization. Abnormality of repolarization increases risk of arrhythmia. The aim of our research is to study and compare the impacts of quinidine and quinine on hERG. Results show that both drugs block the hERG channel, with quinine 14-fold less potent than quinidine. In addition, they presented distinct impacts on channel dynamics. The results imply their stereospecific block effect on the hERG channel. However, F656C-hERG reversed this stereoselectivity. The mutation decreases affinity of the two drugs with hERG, and quinine was more potent than quinidine in F656C-hERG blockage. These data suggest that F656 residue contributes to the stereoselective pocket for quinidine and quinine. Further study demonstrates that both drugs do not change hERG protein levels. In rescue experiments, we found that they exert no reverse effect on pentamidine- or desipramine-induced hERG trafficking defect, although quinidine has been reported to rescue trafficking-deficient pore mutation hERG G601S based on the interaction with F656. Our research demonstrated stereoselective effects of quinidine and quinine on the hERG channel, and this is the first study to explore their reversal potency on drug-induced hERG deficiency. PMID:27690007

  3. Antibody-independent Targeted Quantification of TMPRSS2-ERG Fusion Protein Products in Prostate Cancer

    SciTech Connect

    He, Jintang; Sun, Xuefei; Shi, Tujin; Schepmoes, Athena A.; Fillmore, Thomas L.; Petyuk, Vladislav A.; Xie, Fang; Zhao, Rui; Gritsenko, Marina A.; Yang, Feng; Kitabayashi, Naoki; Chae, Sung Suk; Rubin, Mark; Siddiqui, Javed; Wei, John; Chinnaiyan, Arul M.; Qian, Weijun; Smith, Richard D.; Kagan, Jacob; Srivastava, Sudhir; Rodland, Karin D.; Liu, Tao; Camp, David G.

    2014-10-01

    Fusions between the transmembrane protease serine 2 (TMPRSS2) and ETS related gene (ERG) represent one of the most specific biomarkers that define a distinct molecular subtype of prostate cancer. The studies on TMPRSS2-ERG gene fusions have seldom been performed at the protein level, primarily due to the lack of high-quality antibodies or an antibody-independent method that is sufficiently sensitive for detecting the truncated ERG protein products resulting from TMPRSS2-ERG gene fusions and alternative splicing. Herein, we applied a recently developed PRISM (high-pressure high-resolution separations with intelligent selection and multiplexing)-SRM (selected reaction monitoring) strategy for quantifying ERG protein in prostate cancer cell lines and tumors. The highly sensitive PRISM-SRM assays led to confident detection of 6 unique ERG peptides in either the TMPRSS2-ERG positive cell lines or tissues but not in the negative controls, indicating that ERG protein expression is highly correlated with TMPRSS2-ERG gene rearrangements. Significantly, our results demonstrated for the first time that at least two groups of ERG protein isoforms were simultaneously expressed at variable levels in TMPRSS2-ERG positive samples as evidenced by concomitant detection of two mutually exclusive peptides. Three peptides shared across almost all fusion protein products were determined to be the most abundant peptides, and hence can be used as “signature” peptides for detecting ERG overexpression resulting from TMPRSS2-ERG gene fusion. These PRISM-SRM assays provide valuable tools for studying TMPRSS2-ERG gene fusion protein products, thus improving our understanding of the role of TMPRSS2-ERG gene fusion in the biology of prostate cancer.

  4. Small Molecule Inhibitors of ERG and ETV1 in Prostate Cancer

    DTIC Science & Technology

    2015-09-01

    expression of TMPRSS2-ERG protein slows down prostate cancer growth (12, 13). Therefore, ETS proteins emerge as potential novel targets for treatment of...TMPRSS2-ERG, ERG, patient derived xenografts, YK-4-279, Tumor growth, Proliferation, ERG Inhibitor, and treatment resistance. Overall Project...lines LuCaP 23.1 responded to treatment , LuCaP 86.2 had a limited response, and LuCaP 35 did not respond to treatment . The ERG negative line LuCaP

  5. Gas-Phase Oxidation of Cm+ and Cm2+ -- Thermodynamics of neutral and ionized CmO

    SciTech Connect

    Gibson, John K; Haire, Richard G.; Santos, Marta; Pires de Matos, Antonio; Marcalo, Joaquim

    2008-12-08

    Fourier transform ion cyclotron resonance mass spectrometry was employed to study the products and kinetics of gas-phase reactions of Cm+ and Cm2+; parallel studies were carried out with La+/2+, Gd+/2+ and Lu+/2+. Reactions with oxygen-donor molecules provided estimates for the bond dissociation energies, D[M+-O](M = Cm, Gd, Lu). The first ionization energy, IE[CmO], was obtained from the reactivity of CmO+ with dienes, and the second ionization energies, IE[MO+](M = Cm, La, Gd, Lu), from the rates of electron-transfer reactions from neutrals to the MO2+ ions. The following thermodynamic quantities for curium oxide molecules were obtained: IE[CmO]= 6.4+-0.2 eV; IE[CmO+]= 15.8+-0.4 eV; D[Cm-O]= 710+-45 kJ mol-1; D[Cm+-O]= 670+-40 kJ mol-1; and D[Cm2+-O]= 342+-55 kJ mol-1. Estimates for the M2+-O bond energies for M = Cm, La, Gd and Lu are all intermediate between D[N2-O]and D[OC-O]--i.e., 167 kJ mol-1< D[M2+-O]< 532 kJ mol-1 -- such that the four MO2+ ions fulfill the thermodynamic requirement for catalytic O-atom transport from N2O to CO. It was demonstrated that the kinetics are also favorable and that the CmO2+, LaO2+, GdO2+ and LuO2+ dipositive ions each catalyze the gas-phase oxidation of CO to CO2 by N2O. The CmO2+ ion appeared during the reaction of Cm+ with O2 when the intermediate, CmO+, was not collisionally cooled -- although its formation is kinetically and/or thermodynamically unfavorable, CmO2+ is a stable species.

  6. Increased gene copy number of ERG on chromosome 21 but not TMPRSS2–ERG fusion predicts outcome in prostatic adenocarcinomas

    PubMed Central

    Toubaji, Antoun; Albadine, Roula; Meeker, Alan K; Isaacs, William B; Lotan, Tamara; Haffner, Michael C; Chaux, Alcides; Epstein, Jonathan I; Han, Misop; Walsh, Patrick C; Partin, Alan W; De Marzo, Angelo M; Platz, Elizabeth A; Netto, George J

    2012-01-01

    The role of TMPRSS2–ERG gene fusion in prostate cancer prognostication remains controversial. We evaluated the prognostic role of TMPRSS2–ERG fusion using fluorescence in situ hybridization analysis in a case–control study nested in The Johns Hopkins retropubic radical prostatectomy cohort. In all, 10 tissue microarrays containing paired tumors and normal tissues obtained from 172 cases (recurrence) and 172 controls (non-recurrence) matched on pathological grade, stage, race/ethnicity, and age at the time of surgery were analyzed. All radical prostatectomies were performed at our institution between 1993 and 2004. Recurrence was defined as biochemical recurrence, development of clinical evidence of metastasis, or death from prostate carcinoma. Each tissue microarray spot was scored for the presence of TMPRSS2–ERG gene fusion and for ERG gene copy number gains. The odds ratio of recurrence and 95% confidence intervals were estimated from conditional logistic regression. Although the percentage of cases with fusion was slightly lower in cases than in controls (50 vs 57%), the difference was not statistically significant (P=0.20). The presence of fusion due to either deletion or split event was not associated with recurrence. Similarly, the presence of duplicated ERG deletion, duplicated ERG split, or ERG gene copy number gain with a single ERG fusion was not associated with recurrence. ERG gene polysomy without fusion was significantly associated with recurrence (odds ratio 2.0, 95% confidence interval 1.17–3.42). In summary, TMPRSS2–ERG fusion was not prognostic for recurrence after retropubic radical prostatectomy for clinically localized prostate cancer, although men with ERG gene copy number gain without fusion were twice more likely to recur. PMID:21743434

  7. Ablation of the oncogenic transcription factor ERG by deubiquitinase inhibition in prostate cancer.

    PubMed

    Wang, Shan; Kollipara, Rahul K; Srivastava, Nishi; Li, Rui; Ravindranathan, Preethi; Hernandez, Elizabeth; Freeman, Eva; Humphries, Caroline G; Kapur, Payal; Lotan, Yair; Fazli, Ladan; Gleave, Martin E; Plymate, Stephen R; Raj, Ganesh V; Hsieh, Jer-Tsong; Kittler, Ralf

    2014-03-18

    The transcription factor E-twenty-six related gene (ERG), which is overexpressed through gene fusion with the androgen-responsive gene transmembrane protease, serine 2 (TMPRSS2) in ∼40% of prostate tumors, is a key driver of prostate carcinogenesis. Ablation of ERG would disrupt a key oncogenic transcriptional circuit and could be a promising therapeutic strategy for prostate cancer treatment. Here, we show that ubiquitin-specific peptidase 9, X-linked (USP9X), a deubiquitinase enzyme, binds ERG in VCaP prostate cancer cells expressing TMPRSS2-ERG and deubiquitinates ERG in vitro. USP9X knockdown resulted in increased levels of ubiquitinated ERG and was coupled with depletion of ERG. Treatment with the USP9X inhibitor WP1130 resulted in ERG degradation both in vivo and in vitro, impaired the expression of genes enriched in ERG and prostate cancer relevant gene signatures in microarray analyses, and inhibited growth of ERG-positive tumors in three mouse xenograft models. Thus, we identified USP9X as a potential therapeutic target in prostate cancer cells and established WP1130 as a lead compound for the development of ERG-depleting drugs.

  8. Ablation of the oncogenic transcription factor ERG by deubiquitinase inhibition in prostate cancer

    PubMed Central

    Wang, Shan; Kollipara, Rahul K.; Srivastava, Nishi; Li, Rui; Ravindranathan, Preethi; Hernandez, Elizabeth; Freeman, Eva; Humphries, Caroline G.; Kapur, Payal; Lotan, Yair; Fazli, Ladan; Gleave, Martin E.; Plymate, Stephen R.; Raj, Ganesh V.; Hsieh, Jer-Tsong; Kittler, Ralf

    2014-01-01

    The transcription factor E-twenty-six related gene (ERG), which is overexpressed through gene fusion with the androgen-responsive gene transmembrane protease, serine 2 (TMPRSS2) in ∼40% of prostate tumors, is a key driver of prostate carcinogenesis. Ablation of ERG would disrupt a key oncogenic transcriptional circuit and could be a promising therapeutic strategy for prostate cancer treatment. Here, we show that ubiquitin-specific peptidase 9, X-linked (USP9X), a deubiquitinase enzyme, binds ERG in VCaP prostate cancer cells expressing TMPRSS2-ERG and deubiquitinates ERG in vitro. USP9X knockdown resulted in increased levels of ubiquitinated ERG and was coupled with depletion of ERG. Treatment with the USP9X inhibitor WP1130 resulted in ERG degradation both in vivo and in vitro, impaired the expression of genes enriched in ERG and prostate cancer relevant gene signatures in microarray analyses, and inhibited growth of ERG-positive tumors in three mouse xenograft models. Thus, we identified USP9X as a potential therapeutic target in prostate cancer cells and established WP1130 as a lead compound for the development of ERG-depleting drugs. PMID:24591637

  9. TMPRSS2-ERG gene fusion in small cell carcinoma of the prostate.

    PubMed

    Guo, Charles C; Dancer, Jane Y; Wang, Yan; Aparicio, Ana; Navone, Nora M; Troncoso, Patricia; Czerniak, Bogdan A

    2011-01-01

    Recent studies have shown that most prostate cancers carry the TMPRSS2-ERG gene fusion. Here we evaluated the TMPRSS2-ERG gene fusion in small cell carcinoma of the prostate (n = 12) in comparison with small cell carcinoma of the urinary bladder (n = 12) and lung (n = 11). Fluorescence in situ hybridization demonstrated rearrangement of the ERG gene in 8 cases of prostatic small cell carcinoma (67%), and the rearrangement was associated with deletion of the 5' ERG gene in 7 cases, but rearrangement of the ERG gene was not present in any small cell carcinoma of the urinary bladder or lung. Next we evaluated the TMPRSS2-ERG gene fusion in nude mouse xenografts that were derived from 2 prostatic small cell carcinomas carrying the TMPRSS2-ERG gene fusion. Two transcripts encoded by the TMPRSS2-ERG gene fusion were detected by reverse transcriptase polymerase chain reaction, and DNA sequencing demonstrated that the 2 transcripts were composed of fusions of exon 1 of the TMPRSS2 gene to exon 4 or 5 of the ERG gene. Our study demonstrates the specific presence of TMPRSS2-ERG gene fusion in prostatic small cell carcinoma, which may be helpful in distinguishing small cell carcinoma of prostatic origin from nonprostatic origins. The high prevalence of the TMPRSS2-ERG gene fusion in prostatic small cell carcinoma as well as adenocarcinoma implies that small cell carcinoma may share a common pathogenic pathway with adenocarcinoma in the prostate.

  10. Antineoplastic Effects of siRNA against TMPRSS2-ERG Junction Oncogene in Prostate Cancer

    PubMed Central

    Urbinati, Giorgia; Ali, Hafiz Muhammad; Rousseau, Quentin; Chapuis, Hubert; Desmaële, Didier; Couvreur, Patrick; Massaad-Massade, Liliane

    2015-01-01

    TMPRSS2-ERG junction oncogene is present in more than 50% of patients with prostate cancer and its expression is frequently associated with poor prognosis. Our aim is to achieve gene knockdown by siRNA TMPRSS2-ERG and then to assess the biological consequences of this inhibition. First, we designed siRNAs against the two TMPRSS2-ERG fusion variants (III and IV), most frequently identified in patients’ biopsies. Two of the five siRNAs tested were found to efficiently inhibit mRNA of both TMPRSS2-ERG variants and to decrease ERG protein expression. Microarray analysis further confirmed ERG inhibition by both siRNAs TMPRSS2-ERG and revealed one common down-regulated gene, ADRA2A, involved in cell proliferation and migration. The siRNA against TMPRSS2-ERG fusion variant IV showed the highest anti-proliferative effects: Significantly decreased cell viability, increased cleaved caspase-3 and inhibited a cluster of anti-apoptotic proteins. To propose a concrete therapeutic approach, siRNA TMPRSS2-ERG IV was conjugated to squalene, which can self-organize as nanoparticles in water. The nanoparticles of siRNA TMPRSS2-ERG-squalene injected intravenously in SCID mice reduced growth of VCaP xenografted tumours, inhibited oncoprotein expression and partially restored differentiation (decrease in Ki67). In conclusion, this study offers a new prospect of treatment for prostate cancer based on siRNA-squalene nanoparticles targeting TMPRSS2-ERG junction oncogene. PMID:25933120

  11. Structural and dynamic studies of the transcription factor ERG reveal DNA binding is allosterically autoinhibited.

    PubMed

    Regan, Michael C; Horanyi, Peter S; Pryor, Edward E; Sarver, Jessica L; Cafiso, David S; Bushweller, John H

    2013-08-13

    The Ets-Related Gene (ERG) belongs to the Ets family of transcription factors and is critically important for maintenance of the hematopoietic stem cell population. A chromosomal translocation observed in the majority of human prostate cancers leads to the aberrant overexpression of ERG. We have identified regions flanking the ERG Ets domain responsible for autoinhibition of DNA binding and solved crystal structures of uninhibited, autoinhibited, and DNA-bound ERG. NMR-based measurements of backbone dynamics show that uninhibited ERG undergoes substantial dynamics on the millisecond-to-microsecond timescale but autoinhibited and DNA-bound ERG do not. We propose a mechanism whereby the allosteric basis of ERG autoinhibition is mediated predominantly by the regulation of Ets-domain dynamics with only modest structural changes.

  12. Perovskite solar cells with 18.21% efficiency and area over 1 cm2 fabricated by heterojunction engineering

    NASA Astrophysics Data System (ADS)

    Wu, Yongzhen; Yang, Xudong; Chen, Wei; Yue, Youfeng; Cai, Molang; Xie, Fengxian; Bi, Enbing; Islam, Ashraful; Han, Liyuan

    2016-11-01

    Perovskite solar cells (PSCs) are promising low-cost photovoltaic technologies with high solar-to-electric power conversion efficiency (PCE). The heterojunction structure between perovskite and charge extraction layers is crucial to the photovoltaic performance of PSCs. Here, we report efficient inverted-structured PSCs with a perovskite-fullerene graded heterojunction (GHJ), in which the electron-accepting material is distributed in the perovskite layer with a gradient. This structure can enhance the PCE as it improves the photoelectron collection and reduces recombination loss, especially for the formamidinium cation-based perovskite. The conformal fullerene coating on perovskite during the GHJ deposition achieves a full coverage with reduced layer thickness, thus minimizing the resistive loss in larger sized devices. Our strategy enables the fabrication of centimetre-scale PSCs showing high efficiency with small hysteresis and good stability. A PCE of 18.21% was certified by an independent institution for cells with an aperture area of 1.022 cm2.

  13. Epitaxial SrTiO3 films with electron mobilities exceeding 30,000cm2V-1s-1

    NASA Astrophysics Data System (ADS)

    Son, Junwoo; Moetakef, Pouya; Jalan, Bharat; Bierwagen, Oliver; Wright, Nicholas J.; Engel-Herbert, Roman; Stemmer, Susanne

    2010-06-01

    The study of quantum phenomena in semiconductors requires epitaxial structures with exceptionally high charge-carrier mobilities. Furthermore, low-temperature mobilities are highly sensitive probes of the quality of epitaxial layers, because they are limited by impurity and defect scattering. Unlike many other complex oxides, electron-doped SrTiO3 single crystals show high (~104cm2V-1s-1) electron mobilities at low temperatures. High-mobility, epitaxial heterostructures with SrTiO3 have recently attracted attention for thermoelectric applications, field-induced superconductivity and two-dimensional (2D) interface conductivity. Epitaxial SrTiO3 thin films are often deposited by energetic techniques, such as pulsed laser deposition. Electron mobilities in such films are lower than those of single crystals. In semiconductor physics, molecular beam epitaxy (MBE) is widely established as the deposition method that produces the highest mobility structures. It is a low-energetic, high-purity technique that allows for low defect densities and precise control over doping concentrations and location. Here, we demonstrate controlled doping of epitaxial SrTiO3 layers grown by MBE. Electron mobilities in these films exceed those of single crystals. At low temperatures, the films show Shubnikov-de Haas oscillations. These high-mobility SrTiO3 films allow for the study of the intrinsic physics of SrTiO3 and can serve as building blocks for high-mobility oxide heterostructures.

  14. Clasts in the CM2 carbonaceous chondrite Lonewolf Nunataks 94101: Evidence for aqueous alteration prior to complex mixing

    NASA Astrophysics Data System (ADS)

    Lindgren, Paula; Lee, Martin R.; Sofe, Mahmood R.; Zolensky, Michael E.

    2013-06-01

    Clasts in the CM2 carbonaceous chondrite Lonewolf Nunataks (LON) 94101 have been characterized using scanning and transmission electron microscopy and electron microprobe analysis to determine their degrees of aqueous alteration, and the timing of alteration relative to incorporation of clasts into the host. The provenance of the clasts, and the mechanism by which they were incorporated and mixed with their host material are also considered. Results show that at least five distinct types of clasts occur in LON 94101, of which four have been aqueously altered to various degrees and one is largely anhydrous. The fact that they have had different alteration histories implies that the main part of aqueous activity occurred prior to the mixing and assimilation of the clasts with their host. Further, the presence of such a variety of clasts suggests complex mixing in a dynamic environment involving material from various sources. Two of the clasts, one containing approximately 46 vol% carbonate and the other featuring crystals of pyrrhotite up to approximately 1 mm in size, are examples of unusual lithologies and indicate concentration of chemical elements in discrete areas of the parent body(ies), possibly by flow of aqueous solutions.

  15. Spin-Hall-Effect-Assisted Electroresistance in Antiferromagnets via 105 A/cm2 dc Current

    PubMed Central

    Han, Jiahao; Wang, Yuyan; Pan, Feng; Song, Cheng

    2016-01-01

    Antiferromagnet (AFM) spintronics with reduced electrical current is greatly expected to process information with high integration and low power consumption. In Pt/FeMn and Ta/FeMn hybrids, we observe significant resistance variation (up to 7% of the total resistance) manipulated by 105 A/cm2 dc current. We have excluded the contribution of isotropic structural effects, and confirmed the critical role of the spin Hall injection from Pt (or Ta) to FeMn. This electrical current-manipulated resistance (i.e. electroresistance) is proposed to be attributed to the spin-Hall-effect-induced spin-orbit torque in FeMn. Similar results have also been detected in plain IrMn films, where the charge current generates spin current via the spin Hall effect with the existence of Ir atoms. All the measurements are free from external magnetic fields and ferromagnets. Our findings present an interesting step towards high-efficiency spintronic devices. PMID:27546199

  16. Improved Reproducibility for Perovskite Solar Cells with 1 cm(2) Active Area by a Modified Two-Step Process.

    PubMed

    Shen, Heping; Wu, Yiliang; Peng, Jun; Duong, The; Fu, Xiao; Barugkin, Chog; White, Thomas P; Weber, Klaus; Catchpole, Kylie R

    2017-02-22

    With rapid progress in recent years, organohalide perovskite solar cells (PSC) are promising candidates for a new generation of highly efficient thin-film photovoltaic technologies, for which up-scaling is an essential step toward commercialization. In this work, we propose a modified two-step method to deposit the CH3NH3PbI3 (MAPbI3) perovskite film that improves the uniformity, photovoltaic performance, and repeatability of large-area perovskite solar cells. This method is based on the commonly used two-step method, with one additional process involving treating the perovskite film with concentrated methylammonium iodide (MAI) solution. This additional treatment is proved to be helpful for tailoring the residual PbI2 level to an optimal range that is favorable for both optical absorption and inhibition of recombination. Scanning electron microscopy and photoluminescence image analysis further reveal that, compared to the standard two-step and one-step methods, this method is very robust for achieving uniform and pinhole-free large-area films. This is validated by the photovoltaic performance of the prototype devices with an active area of 1 cm(2), where we achieved the champion efficiency of ∼14.5% and an average efficiency of ∼13.5%, with excellent reproducibility.

  17. Spin-Hall-Effect-Assisted Electroresistance in Antiferromagnets via 10(5) A/cm(2) dc Current.

    PubMed

    Han, Jiahao; Wang, Yuyan; Pan, Feng; Song, Cheng

    2016-08-22

    Antiferromagnet (AFM) spintronics with reduced electrical current is greatly expected to process information with high integration and low power consumption. In Pt/FeMn and Ta/FeMn hybrids, we observe significant resistance variation (up to 7% of the total resistance) manipulated by 10(5) A/cm(2) dc current. We have excluded the contribution of isotropic structural effects, and confirmed the critical role of the spin Hall injection from Pt (or Ta) to FeMn. This electrical current-manipulated resistance (i.e. electroresistance) is proposed to be attributed to the spin-Hall-effect-induced spin-orbit torque in FeMn. Similar results have also been detected in plain IrMn films, where the charge current generates spin current via the spin Hall effect with the existence of Ir atoms. All the measurements are free from external magnetic fields and ferromagnets. Our findings present an interesting step towards high-efficiency spintronic devices.

  18. Characteristics of Late Negative ERG Responses Elicited By Sawtooth Flicker

    PubMed Central

    Gowrisankaran, Sowjanya; Genead, Mohamed A.; Anastasakis, Anastasios; Alexander, Kenneth R.

    2012-01-01

    Background This study aimed to determine whether the properties of the late negative responses (LNRs) of the electroretinogram (ERG) elicited by sawtooth flicker are consistent with the characteristics of the photopic negative response generated by a light pulse (PhNRpulse). Methods ERG recordings were obtained from 10 visually normal individuals and from 6 patients with optic atrophy (OA) in response to 8-Hz rapid-on and rapid-off sawtooth flicker and to brief (4 ms) light pulses. All stimuli were either long-wavelength (R), middle-wavelength (G), or a combination of equal luminances of long and middle-wavelengths (Y) presented on a short-wavelength, rod-saturating adapting field. Amplitudes of LNRs were obtained in response to rapid-on (LNRon) and rapid-off (LNRoff) sawtooth flicker, and were also derived from the sum of the ERG waveforms to the two sawtooth phases (LNRadd). Results For the control subjects, PhNRpulse amplitude varied with stimulus wavelength, being largest in response to a long-wavelength pulse, as expected. However, the amplitudes of LNRon, LNRoff, and LNRadd were not significantly different for R, Y, and G sawtooth flicker. Despite the absence of a chromatic effect, LNRoff and LNRadd amplitudes were significantly smaller in the OA patients than in the controls, similar to the results for the PhNRpulse, implying an inner retinal origin for the LNRoff and LNRadd. However, LNRon amplitudes did not differ significantly between the OA patients and controls, although there was a significant correlation between the LNRon and PhNRpulse for R stimuli. Conclusion We conclude that LNRoff and LNRadd but not LNRon can be useful measures to assess the integrity of the inner retina that can complement the PhNRpulse. PMID:23054159

  19. Acepromazine inhibits hERG potassium ion channels expressed in human embryonic kidney 293 cells

    PubMed Central

    Joo, Young Shin; Lee, Hong Joon; Choi, Jin-Sung

    2017-01-01

    The effects of acepromazine on human ether-à-go-go-related gene (hERG) potassium channels were investigated using whole-cell voltage-clamp technique in human embryonic kidney (HEK293) cells transfected with hERG. The hERG currents were recorded with or without acepromazine, and the steady-state and peak tail currents were analyzed for the evaluating the drug effects. Acepromazine inhibited the hERG currents in a concentration-dependent manner with an IC50 value of 1.5 µM and Hill coefficient of 1.1. Acepromazine blocked hERG currents in a voltage-dependent manner between –40 and +10 mV. Before and after application of acepromazine, the half activation potentials of hERG currents changed to hyperpolarizing direction. Acepromazine blocked both the steady-state hERG currents by depolarizing pulse and the peak tail currents by repolarizing pulse; however, the extent of blocking by acepromazine in the repolarizing pulse was more profound than that in the depolarizing pulse, indicating that acepromazine has a high affinity for the open state of the channels, with a relatively lower affinity for the closed state of hERG channels. A fast application of acepromazine during the tail currents inhibited the open state of hERG channels in a concentration-dependent. The steady-state inactivation of hERG currents shifted to the hyperpolarized direction by acepromazine. These results suggest that acepromazine inhibits the hERG channels probably by an open- and inactivated-channel blocking mechanism. Regarding to the fact that the hERG channels are the potential target of drug-induced long QT syndrome, our results suggest that acepromazine can possibly induce a cardiac arrhythmia through the inhibition of hERG channels. PMID:28066143

  20. Secondary School Curriculum Guide. Music. Grades 10-12. Draft Copy.

    ERIC Educational Resources Information Center

    Rogers, Arnold R., Ed.; And Others

    This draft curriculum guide provides curriculum materials that are organized into behavioral objectives with a scope and sequence. Divided by grade and levels, each section includes level objectives and suggested activities. To be covered in grades 10-12 are 5 levels of musical content. They are: 1) why man creates and how he perceives; 2) the…

  1. Steps towards Healthy Growth; Health Education Curriculum Guide. Grades 10-12.

    ERIC Educational Resources Information Center

    Dippo, Jeanette, Ed.

    GRADES OR AGES: Grades 10-12. SUBJECT MATTER: Health Education--includes chapters on a) nutrition; b) disease prevention and control; c) tobacco; d) alcohol; e) drugs; f) personality development; g) family life and sexuality; h) environmental, public, and world health; i) consumer health; and j) safety, first aid, and survival education.…

  2. A Resource File for Social Studies in Utah. Level 10-12: U.S. Studies.

    ERIC Educational Resources Information Center

    Utah State Office of Education, Salt Lake City.

    This resource file contains information for Utah secondary school teachers to help their students at level 10-12 to meet the state's instructional objectives in the social studies. This particular student level emphasizes U.S. studies. The following disciplines are covered in the resource file: psychology, anthropology, sociology, geography,…

  3. Extended chronologies of aqueous alteration in the CM2 carbonaceous chondrites: Evidence from carbonates in Queen Alexandra Range 93005

    NASA Astrophysics Data System (ADS)

    Lee, M. R.; Lindgren, P.; Sofe, M. R.; O'D Alexander, C. M.; Wang, J.

    2012-09-01

    The Antarctic CM2 carbonaceous chondrite QUE 93005 contains four compositionally distinct carbonates, namely breunnerite, calcite, dolomite and a Ca-poor dolomite. These carbonates can form monomineralic grains, or may be intergrown as bimineralic grains consisting of dolomite plus breunnerite and dolomite plus calcite, or polymineralic grains containing an intergrowth of breunnerite, Ca-poor dolomite and calcite. Carbonates in all grain types have inclusions of Fe-Ni sulphides and/or Mg-Fe phyllosilicates. In the bimineralic grains, dolomite crystallised first to be overgrown by breunnerite or partially replaced by calcite. Polymineralic grains are concentrically layered, with breunnerite crystallising first on pore margins to be later etched, then overgrown and partially replaced by Ca-poor dolomite that was itself partly dissolved prior to being overgrown by calcite. Calcite and dolomite have also cemented fractures that cross-cut the fine-grained rims to aqueously altered chondrules and were formed by expansion of the chondrules during their hydration. Overall, the sequence of mineralisation in QUE 93005 was: (1) dolomite, (2) breunnerite, (3) Ca-poor dolomite then (4) calcite. This secular change in carbonate composition and mineralogy reflects changing solution composition and probably also provenance. Mg-Fe phyllosilicates replaced dolomite, breunnerite and Ca-poor dolomite prior to calcite crystallisation, and most or all of the sulphides formed after both the phyllosilicates and calcite. Following sulphide crystallisation, the edges of carbonate grains were abraded, either by impact 'gardening' or as a consequence of fluidisation of the matrix during rapid loss of gas or vapour. Determination of the crystallisation age of dolomite via the Mn-Cr system indicates that aqueous alteration of QUE 93005 began on or before 3.93 ± 0.23 Ma after the formation of the oldest solar system solids. Overall, the water/rock ratio and fO2 during alteration of QUE 93005

  4. Changes in storm tracks and energy transports in a warmer climate simulated by the GFDL CM2.1 model

    NASA Astrophysics Data System (ADS)

    Wu, Yutian; Ting, Mingfang; Seager, Richard; Huang, Huei-Ping; Cane, Mark A.

    2011-07-01

    Storm tracks play a major role in regulating the precipitation and hydrological cycle in midlatitudes. The changes in the location and amplitude of the storm tracks in response to global warming will have significant impacts on the poleward transport of heat, momentum and moisture and on the hydrological cycle. Recent studies have indicated a poleward shift of the storm tracks and the midlatitude precipitation zone in the warming world that will lead to subtropical drying and higher latitude moistening. This study agrees with this key feature for not only the annual mean but also different seasons and for the zonal mean as well as horizontal structures based on the analysis of Geophysical Fluid Dynamics Laboratory (GFDL) CM2.1 model simulations. Further analyses show that the meridional sensible and latent heat fluxes associated with the storm tracks shift poleward and intensify in both boreal summer and winter in the late twenty-first century (years 2081-2100) relative to the latter half of the twentieth century (years 1961-2000). The maximum dry Eady growth rate is examined to determine the effect of global warming on the time mean state and associated available potential energy for transient growth. The trend in maximum Eady growth rate is generally consistent with the poleward shift and intensification of the storm tracks in the middle latitudes of both hemispheres in both seasons. However, in the lower troposphere in northern winter, increased meridional eddy transfer within the storm tracks is more associated with increased eddy velocity, stronger correlation between eddy velocity and eddy moist static energy, and longer eddy length scale. The changing characteristics of baroclinic instability are, therefore, needed to explain the storm track response as climate warms. Diagnosis of the latitude-by-latitude energy budget for the current and future climate demonstrates how the coupling between radiative and surface heat fluxes and eddy heat and moisture

  5. Duplication of the fusion of TMPRSS2 to ERG sequences identifies fatal human prostate cancer

    PubMed Central

    Attard, G; Clark, J; Ambroisine, L; Fisher, G; Kovacs, G; Flohr, P; Berney, D; Foster, CS; Fletcher, A; Gerald, WL; Moller, H; Reuter, V; De Bono, JS; Scardino, P; Cuzick, J; Cooper, CS

    2009-01-01

    New predictive markers for managing prostate cancer are urgently required because of the highly variable natural history of this disease. At the time of diagnosis, Gleason score provides the gold standard for assessing the aggressiveness of prostate cancer. However, the recent discovery of TMPRSS2 fusions to the ERG gene in prostate cancer raises the possibility of using alterations at the ERG locus as additional mechanism-based prognostic indicators. Fluorescence in situ hybridization (FISH) assays were used to assess ERG gene status in a cohort of 445 prostate cancers from patients who had been conservatively managed. The FISH assays detected separation of 5′ (labelled green) and 3′ (labelled red) ERG sequences, which is a consequence of the TMPRSS2–ERG fusion, and additionally identify interstitial deletion of genomic sequences between the tandemly located TMPRSS2 and ERG gene sequences on chromosome 21. Cancers lacking ERG alterations exhibited favourable cause-specific survival (90% survival at 8 years). We identify a novel category of prostate cancers, characterized by duplication of the fusion of TMPRSS2 to ERG sequences together with interstitial deletion of sequences 5′ to ERG (called ‘2+Edel’), which by comparison exhibited extremely poor cause-specific survival (hazard ratio = 6.10, 95% confidence ratio = 3.33–11.15, P < 0.001, 25% survival at 8 years). In multivariate analysis, ‘2+Edel’ provided significant prognostic information (P = 0.003) in addition to that provided by Gleason score and prostate-specific antigen level at diagnosis. Other individual categories of ERG alteration were associated with intermediate or good prognosis. We conclude that determination of ERG gene status, including duplication of the fusion of TMPRSS2 to ERG sequences in 2+Edel, allows stratification of prostate cancer into distinct survival categories. PMID:17637754

  6. Mitochondrial reactive oxygen species mediate hypoxic down-regulation of hERG channel protein.

    PubMed

    Nanduri, Jayasri; Wang, Ning; Bergson, Pamela; Yuan, Guoxiang; Ficker, Eckhard; Prabhakar, Nanduri R

    2008-08-22

    Previous studies suggest that reactive oxygen species (ROS) play an important role in physiological responses to hypoxia. In the present study, we examined the effects of hypoxia on human ether-a-go-go related gene (hERG) channel protein expression and assessed the role of ROS. Hypoxia, in a stimulus- and time-dependent manner, decreased hERG protein with marked reduction in hERG K+ conductance in human embryonic kidney cells stably expressing the hERG alpha subunit. Down-regulation of hERG by hypoxia was not due to increased proteasomal degradation or decreased transcription but due to decreased synthesis of the protein. Hypoxia increased ROS in a time-dependent manner. Antioxidants prevented hypoxia-evoked down-regulation of hERG protein and exogenous oxidants mimicked the effects of hypoxia. Hypoxia-evoked down-regulation of hERG protein and elevation in ROS were absent in p(O) cells, which are devoid of mitochondrial DNA. Inhibitors of NADPH oxidase failed to prevent the effects of hypoxia. These results demonstrate that hypoxia enhances the production of ROS in the mitochondria, resulting in down-regulation of hERG translation and decreased hERG-mediated K+ conductance.

  7. Noninvasive Prenatal Diagnosis Significance of ERG Methylation as a Biomarker in Down’s Syndrome

    PubMed Central

    Liu, Xiangju; Xue, Ming

    2017-01-01

    Background Down’s syndrome (DS) is a genetic disease with chromosome abnormality due to the increasing chromosome 21. This study focused on the clinical application value of ERG methylation level in blood of pregnant women as a biomarker in Down’s syndrome. Material/Methods The sham group consisted of 210 nonpregnant women, the positive control group consisted of 33 women with a delivery history of DS fetus, and the negative control group consisted of 60 women with eutocia history. A combination of restriction enzyme digestion experiment and PCR was performed to examine ERG methylation levels, methylation sites, and distribution in blood of pregnant women and in chorion tissues from abortion samples. Gene sequencing was performed to determine the ERG sequence in chromosome 21. Homology between normal tissues and chorion tissues from abortion samples was analyzed with bioinformatics technology. Results ERG methylation in chorion tissues from 210 abortion samples at 8, 9, and 10 weeks gestational age were determined; however, no ERG methylation was determined in blood of pregnant women. Gene sequencing indicated that normal ERG sequence in chromosome 21 was in fetus chorion tissues, and these ERG sequences were aberrantly methylated. Bioinformatics result showed that homology and DNA methylation level was discrepancy in normal tissues and chorion tissues from abortion samples. Conclusions It was worthwhile to use ERG methylation as biomarker in noninvasive prenatal diagnosis, and ERG methylation should be applied with consent of pregnancy and her relatives. PMID:28111453

  8. Fluconazole inhibits hERG K(+) channel by direct block and disruption of protein trafficking.

    PubMed

    Han, Shengna; Zhang, Yu; Chen, Qiu; Duan, Yanyan; Zheng, Tenghao; Hu, Xiangjie; Zhang, Zhao; Zhang, Lirong

    2011-01-10

    Fluconazole, a commonly used azole antifungal drug, can induce QT prolongation, which may lead to Torsades de Pointes and sudden death. To investigate the arrhythmogenic side effects of fluconazole, we studied the effect of fluconazole on human ether-a-go-go-related gene (hERG) K(+) channels (wild type, Y652A and F656C) expressed in human embryonic kidney (HEK293) cells using a whole-cell patch clamp technique, Western blot analysis and confocal microscopy. Fluconazole inhibited wild type hERG currents in a concentration-dependent manner, with a half-maximum block concentration (IC(50)) of 48.2±9.4μM. Fluconazole did not change other channel kinetics (activation and steady-state inactivation) of hERG channel. Mutations in drug- binding sites (Y652A or F656C) of the hERG channel significantly attenuated the hERG current blockade by fluconazole. In addition, fluconazole inhibited the trafficking of hERG protein by Western blot analysis and confocal microscopy, respectively. These findings indicate that fluconazole may cause acquired long QT syndrome (LQTS) via a direct inhibition of hERG current and by disrupting hERG protein trafficking, and the mutations Y652 and F656 may be obligatory determinants in inhibition of hERG current for fluconazole.

  9. ISCEV standard for clinical multifocal electroretinography (mfERG) (2011 edition)

    PubMed Central

    Hood, Donald C.; Bach, Michael; Brigell, Mitchell; Keating, David; Kondo, Mineo; Lyons, Jonathan S.; Marmor, Michael F.; McCulloch, Daphne L.; Palmowski-Wolfe, Anja M.

    2015-01-01

    The clinical multifocal electroretinogram (mfERG) is an electrophysiological test of local retinal function. With this technique, many local ERG responses are recorded quasi-simultaneously from the cone-driven retina under light-adapted conditions. This document, from the International Society for Clinical Electrophysiology of Vision (ISCEV: www.iscev.org), replaces the ISCEV guidelines for the mfERG published in 2007. Standards for performance of the basic clinical mfERG test with a stimulus array of 61 or 103 hexagons, as well as for reporting the results, are specified. PMID:22038576

  10. Blockage of hERG current and the disruption of trafficking as induced by roxithromycin.

    PubMed

    Han, Sheng-Na; Yang, Song-Hua; Zhang, Yu; Duan, Yan-Yan; Sun, Xiao-Yan; Chen, Qiu; Fan, Tian-Li; Ye, Zhen-Kun; Huang, Chen-Zheng; Hu, Xiang-Jie; Zhang, Zhao; Zhang, Li-Rong

    2013-12-01

    Roxithromycin is an oral macrolide antibiotic agent that has been repeatedly reported to provoke excessive prolongation of the Q-T interval and torsades de pointes in clinical settings. To investigate the mechanisms underlying the arrhythmogenic side effects of roxithromycin, we studied the molecular mechanisms of roxithromycin on human ether-à-go-go-related gene (hERG) K(+) channels expressed in human embryonic kidney (HEK293) cells. Roxithromycin was found to inhibit wild-type (WT) hERG currents in a concentration-dependent manner with a half-maximum block concentration (IC50) of 55.8 ± 9.1 μmol/L. S6 residue hERG mutants (Y652A and F656C) showed reduced levels of hERG current blockage attributable to roxithromycin. Roxithromycin also inhibited the trafficking of hERG protein to the cell membrane, as confirmed by Western blot analysis and confocal microscopy. These findings indicate that roxithromycin may cause acquired long-QT syndrome via direct inhibition of hERG current and by disruption of hERG protein trafficking. Mutations in drug-binding sites (Y652A or F656C) of the hERG channel were found to attenuate hERG current blockage by roxithromycin, but did not significantly alter the disruption of trafficking.

  11. Biomimetic membrane platform containing hERG potassium channel and its application to drug screening.

    PubMed

    Arslan Yildiz, Ahu; Kang, CongBao; Sinner, Eva-Kathrin

    2013-04-07

    The hERG (human ether-à-go-go-related gene) potassium channel has been extensively studied by both academia and industry because of its relation to inherited or drug-induced long QT syndrome (LQTS). Unpredicted hERG and drug interaction affecting channel activity is of main concern for drug discovery. Although there are several methods to test hERG and drug interaction, it is still necessary to develop some efficient and economic ways to probe hERG and drug interactions. To contribute this aim, we have developed a biomimetic lipid membrane platform into which the hERG channel can be folded. Expression and integration of the hERG channel was achieved using a cell-free (CF) expression system. The folding of hERG in the biomimetic membrane system was investigated using Surface Plasmon Enhanced Fluorescence Spectroscopy (SPFS) and Imaging Surface Plasmon Resonance (iSPR). In addition, the hERG channel folded into our biomimetic membrane platform was used for probing the channel and drug interactions through fluorescence polarization (FP) assay. Our results suggest that the biomimetic system employed is capable of detecting the interaction between hERG and different channel blockers at varied concentrations. We believe that our current approach could be applied to other membrane proteins for drug screening or other protein-related interactions.

  12. An optical fiber spool for laser stabilization with reduced acceleration sensitivity to 10-12/g

    NASA Astrophysics Data System (ADS)

    Hu, Yong-Qi; Dong, Jing; Huang, Jun-Chao; Li, Tang; Liu, Liang

    2015-10-01

    Environmental vibration causes mechanical deformation in optical fibers, which induces excess frequency noise in fiber-stabilized lasers. In order to solve such a problem, we propose an ultralow acceleration sensitivity fiber spool with symmetrically mounted structure. By numerical analysis with the finite element method, we obtain the optimal geometry parameters of the spool with which the horizontal and vertical acceleration sensitivity can be reduced to 3.25 × 10-12/g and 5.38 × 10-12/g respectively. Moreover, the structure features the insensitivity to the variation of geometry parameters, which will minimize the influence from numerical simulation error and manufacture tolerance. Project supported by the National Natural Science Foundation of China (Grant Nos. 11034008 and 11274324) and the Key Research Program of the Chinese Academy of Sciences (Grant No. KJZD-EW-W02).

  13. Development of a peptide-based vaccine targeting TMPRSS2:ERG fusion positive prostate cancer

    PubMed Central

    Kissick, Haydn Thomas; Sanda, Martin George; Dunn, Laura Kathleen; Arredouani, Mohamed Simo

    2013-01-01

    Identification of novel vaccine targets is critical for the design and advancement of prostate cancer (PCa) immunotherapy. Ideal targets are proteins that are abundant in prostate tumors while absent in extra-prostatic tissues. The fusion of the androgen-regulated TMPRSS2 gene with the ETS transcription factor ERG occurs in approximately 50% of prostate cancer cases and results in aberrant ERG expression. Because expression of ERG is very low in peripheral tissue, we evaluated the suitability of this protein as an antigen target in PCa vaccines. ERG-derived HLA-A*0201-restricted immunogenic epitopes were identified through a 3-step strategy that included in silico, in vitro, and in vivo validation. Algorithms were used to predict potential HLA-A*0201-binding epitopes. High scoring epitopes were tested for binding to HLA-A*0201 using the T2-based stabilization assay in vitro. Five peptides were found to bind HLA-A*0201 and were subsequently tested for immunogenicity in humanized HLA-A*0201 transgenic mice. The in vivo screening identified three immunogenic peptides. One of these peptides, ERG295, overcame peripheral tolerance in HLA-A*0201 mice that expressed prostate restricted ERG. Also, this peptide induced an antigen specific response against ERG-expressing human prostate tumor cells. Finally, tetramer assay showed detectable and responsive ERG295-specific cytotoxic lymphocytes in peripheral blood of HLA-A*0201+ prostate cancer patients. Detection of ERG-specific CTLs in both mice and the blood of prostate cancer patients indicates that ERG-specific tolerance can be overcome. Additionally, these data suggest that ERG is a suitable target antigen for PCa immunotherapy. PMID:24149465

  14. Development of a peptide-based vaccine targeting TMPRSS2:ERG fusion-positive prostate cancer.

    PubMed

    Kissick, Haydn Thomas; Sanda, Martin George; Dunn, Laura Kathleen; Arredouani, Mohamed Simo

    2013-12-01

    Identification of novel vaccine targets is critical for the design and advancement of prostate cancer (PCa) immunotherapy. Ideal targets are proteins that are abundant in prostate tumors while absent in extra-prostatic tissues. The fusion of the androgen-regulated TMPRSS2 gene with the ETS transcription factor ERG occurs in approximately 50 % of prostate cancer cases and results in aberrant ERG expression. Because expression of ERG is very low in peripheral tissue, we evaluated the suitability of this protein as an antigen target in PCa vaccines. ERG-derived HLA-A*0201-restricted immunogenic epitopes were identified through a 3-step strategy that included in silico, in vitro, and in vivo validation. Algorithms were used to predict potential HLA-A*0201-binding epitopes. High-scoring epitopes were tested for binding to HLA-A*0201 using the T2-based stabilization assay in vitro. Five peptides were found to bind HLA-A*0201 and were subsequently tested for immunogenicity in humanized, HLA-A*0201 transgenic mice. The in vivo screening identified three immunogenic peptides. One of these peptides, ERG295, overcame peripheral tolerance in HLA-A*0201 mice that expressed prostate-restricted ERG. Also, this peptide induced an antigen-specific response against ERG-expressing human prostate tumor cells. Finally, tetramer assay showed detectable and responsive ERG295-specific cytotoxic lymphocytes in peripheral blood of HLA-A*0201(+) prostate cancer patients. Detection of ERG-specific CTLs in both mice and the blood of prostate cancer patients indicates that ERG-specific tolerance can be overcome. Additionally, these data suggest that ERG is a suitable target antigen for PCa immunotherapy.

  15. Identification of quaternary ammonium compounds as potent inhibitors of hERG potassium channels

    PubMed Central

    Xia, Menghang; Shahane, Sampada; Huang, Ruili; Titus, Steven A.; Shum, Enoch; Zhao, Yong; Southall, Noel; Zheng, Wei; Witt, Kristine L.; Tice, Raymond R.; Austin, Christopher P.

    2011-01-01

    The human ether-a-go-go-related gene (hERG) channel, a member of a family of voltage-gated potassium (K+) channels, plays a critical role in the repolarization of the cardiac action potential. The reduction of hERG channel activity as a result of adverse drug effects or genetic mutations may cause QT interval prolongation and potentially lead to acquired long QT syndrome. Thus, screening for hERG channel activity is important in drug development. Cardiotoxicity associated with the inhibition of hERG channels by environmental chemicals is also a public health concern. To assess the inhibitory effects of environmental chemicals on hERG channel function, we screened the National Toxicology Program (NTP) collection of 1408 compounds by measuring thallium influx into cells through hERG channels. Seventeen compounds with hERG channel inhibition were identified with IC50 potencies ranging from 0.26 to 22 μM. Twelve of these compounds were confirmed as hERG channel blockers in an automated whole cell patch clamp experiment. In addition, we investigated the structure-activity relationship of seven compounds belonging to the quaternary ammonium compound (QAC) series on hERG channel inhibition. Among four active QAC compounds, tetra-n-octylammonium bromide was the most potent with an IC50 value of 260 nM in the thallium influx assay and 80 nM in the patch clamp assay. The potency of this class of hERG channel inhibitors appears to depend on the number and length of their aliphatic side-chains surrounding the charged nitrogen. Profiling environmental compound libraries for hERG channel inhibition provides information useful in prioritizing these compounds for cardiotoxicity assessment in vivo. PMID:21362439

  16. Identification of quaternary ammonium compounds as potent inhibitors of hERG potassium channels

    SciTech Connect

    Xia Menghang; Shahane, Sampada A.; Huang, Ruili; Titus, Steven A.; Shum, Enoch; Zhao Yong; Southall, Noel; Zheng, Wei; Witt, Kristine L.; Tice, Raymond R.; Austin, Christopher P.

    2011-05-01

    The human ether-a-go-go-related gene (hERG) channel, a member of a family of voltage-gated potassium (K{sup +}) channels, plays a critical role in the repolarization of the cardiac action potential. The reduction of hERG channel activity as a result of adverse drug effects or genetic mutations may cause QT interval prolongation and potentially leads to acquired long QT syndrome. Thus, screening for hERG channel activity is important in drug development. Cardiotoxicity associated with the inhibition of hERG channels by environmental chemicals is also a public health concern. To assess the inhibitory effects of environmental chemicals on hERG channel function, we screened the National Toxicology Program (NTP) collection of 1408 compounds by measuring thallium influx into cells through hERG channels. Seventeen compounds with hERG channel inhibition were identified with IC{sub 50} potencies ranging from 0.26 to 22 {mu}M. Twelve of these compounds were confirmed as hERG channel blockers in an automated whole cell patch clamp experiment. In addition, we investigated the structure-activity relationship of seven compounds belonging to the quaternary ammonium compound (QAC) series on hERG channel inhibition. Among four active QAC compounds, tetra-n-octylammonium bromide was the most potent with an IC{sub 50} value of 260 nM in the thallium influx assay and 80 nM in the patch clamp assay. The potency of this class of hERG channel inhibitors appears to depend on the number and length of their aliphatic side-chains surrounding the charged nitrogen. Profiling environmental compound libraries for hERG channel inhibition provides information useful in prioritizing these compounds for cardiotoxicity assessment in vivo.

  17. BET bromodomain-mediated interaction between ERG and BRD4 promotes prostate cancer cell invasion

    PubMed Central

    Blee, Alexandra M.; Liu, Shujun; Wang, Liguo; Huang, Haojie

    2016-01-01

    Prostate cancer (PCa) that becomes resistant to hormone castration and next-generation androgen receptor (AR)-targeted therapies, called castration-resistant prostate cancer (CRPC), poses a significant clinical challenge. A better understanding of PCa progression and key molecular mechanisms could bring novel therapies to light. One potential therapeutic target is ERG, a transcription factor aberrantly up-regulated in PCa due to chromosomal rearrangements between androgen-regulated gene TMPRSS2 and ERG. Here we show that the most common PCa-associated truncated ERG T1–E4 (ERGΔ39), encoded by fusion between TMPRSS2 exon 1 and ERG exon 4, binds to bromodomain-1 (BD1) of bromodomain containing protein 4 (BRD4), a member of the bromodomain and extraterminal domain (BET) family. This interaction is partially abrogated by BET inhibitors JQ1 and iBET762. Meta-analysis of published ERG (T1–E4) and BRD4 chromatin immunoprecipitation-sequencing (ChIP-seq) data demonstrates overlap in a substantial portion of their binding sites. Gene expression profile analysis shows some ERG-BRD4 co-target genes are upregulated in CRPC compared to hormone-naïve counterparts. We provide further evidence that ERG-mediated invasion of PCa cells was significantly enhanced by an acetylation-mimicking mutation in ERG that augments the ERG-BRD4 interaction. Our findings reveal that PCa-associated ERG can interact and co-occupy with BRD4 in the genome, and suggest this druggable interaction is critical for ERG-mediated cell invasion and PCa progression. PMID:27223260

  18. IL-6 Overexpression in ERG-Positive Prostate Cancer Is Mediated by Prostaglandin Receptor EP2.

    PubMed

    Merz, Constanze; von Mässenhausen, Anne; Queisser, Angela; Vogel, Wenzel; Andrén, Ove; Kirfel, Jutta; Duensing, Stefan; Perner, Sven; Nowak, Michael

    2016-04-01

    Prostate cancer is the most diagnosed cancer in men and multiple risk factors and genetic alterations have been described. The TMPRSS2-ERG fusion event and the overexpression of the transcription factor ERG are present in approximately 50% of all prostate cancer patients, however, the clinical outcome is still controversial. Prostate tumors produce various soluble factors, including the pleiotropic cytokine IL-6, regulating cellular processes such as proliferation and metastatic segregation. Here, we used prostatectomy samples in a tissue microarray format and analyzed the co-expression and the clinicopathologic data of ERG and IL-6 using immunohistochemical double staining and correlated the read-out with clinicopathologic data. Expression of ERG and IL-6 correlated strongly in prostate tissue samples. Forced expression of ERG in prostate tumor cell lines resulted in significantly increased secretion of IL-6, whereas the down-regulation of ERG decreased IL-6 secretion. By dissecting the underlying mechanism in prostate tumor cell lines we show the ERG-mediated up-regulation of the prostanoid receptors EP2 and EP3. The prostanoid receptor EP2 was overexpressed in human prostate cancer tissue. Furthermore, the proliferation rate and IL-6 secretion in DU145 cells was reduced after treatment with EP2-receptor antagonist. Collectively, our study shows that the expression of ERG in prostate cancer is linked to the expression of IL-6 mediated by the prostanoid receptor EP2.

  19. Nucleotide resolution analysis of TMPRSS2 and ERG rearrangements in prostate cancer

    PubMed Central

    Weier, Christopher; Haffner, Michael C.; Mosbruger, Timothy; Esopi, David M.; Hicks, Jessica; Zheng, Qizhi; Fedor, Helen; Isaacs, William B.; De Marzo, Angelo M.; Nelson, William G.; Yegnasubramanian, Srinivasan

    2013-01-01

    TMPRSS2-ERG rearrangements occur in approximately 50% of prostate cancers and therefore represent one of the most frequently observed structural rearrangements in all cancers. However, little is known about the genomic architecture of such rearrangements. We therefore designed and optimized a pipeline involving target-capture of TMPRSS2 and ERG genomic sequences coupled with paired-end next generation sequencing to resolve genomic rearrangement breakpoints in TMPRSS2 and ERG at nucleotide resolution in a large series of primary prostate cancer specimens (n = 83). This strategy showed >90% sensitivity and specificity in identifying TMPRSS2-ERG rearrangements, and allowed identification of intra- and inter-chromosomal rearrangements involving TMPRSS2 and ERG with known and novel fusion partners. Our results indicate that rearrangement breakpoints show strong clustering in specific intronic regions of TMPRSS2 and ERG. The observed TMPRSS2-ERG rearrangements often exhibited complex chromosomal architecture associated with several intra- and inter-chromosomal rearrangements. Nucleotide resolution analysis of breakpoint junctions revealed that the majority of TMPRSS2 and ERG rearrangements (~88%) occurred at or near regions of microhomology or involved insertions of one or more base pairs. This architecture implicates nonhomologous end joining (NHEJ) and microhomology mediated end joining (MMEJ) pathways in the generation of such rearrangements. These analyses have provided important insights into the molecular mechanisms involved in generating prostate cancer-specific recurrent rearrangements. PMID:23447416

  20. Biological Effects of TMPRSS2/ERG Fusion Isoforms in Human Prostate Cancer

    DTIC Science & Technology

    2009-02-01

    40–5. 11. Lapointe J, Kim YH, Miller MA, et al. A variant TMPRSS2 isoform and ERG fusion product in prostate cancer with implications for molecular ... diagnosis . Mod Pathol 2007;20:467–73. 12. Demichelis F, Fall K, Perner S, et al. TMPRSS2:ERG gene fusion associated with lethal prostate cancer in a

  1. Effects of an hERG activator, ICA-105574, on electrophysiological properties of canine hearts.

    PubMed

    Asayama, Mahoko; Kurokawa, Junko; Shirakawa, Kiyoshi; Okuyama, Hisashi; Kagawa, Toshiki; Okada, Jun-ichi; Sugiura, Seiryo; Hisada, Toshiaki; Furukawa, Tetsushi

    2013-01-01

    In short QT syndrome, inherited gain-of-function mutations in the human ether a-gogo-related gene (hERG) K(+) channel have been associated with development of fatal arrhythmias. This implies that drugs that activate hERG as a side effect may likewise pose significant arrhythmia risk. hERG activators have been found to have diverse mechanisms of activation, which may reflect their distinct binding sites. Recently, the new hERG activator ICA-105574 was introduced, which disables inactivation of the hERG channel with very high potency. We explored characteristics of this new drug in several experimental models. Patch clamp experiments were used to verify activation of hERG channels by ICA-105574 in human embryonic kidney cells stably-expressing hERG channels. ICA-105574 significantly shortened QT and QTc intervals and monophasic action potential duration (MAP(90)) in Langendorff-perfused guinea-pig hearts. We also administered ICA-105574 to anesthetized dogs while recording ECG and drug plasma concentrations. ICA-105574 (10 mg/kg) significantly shortened QT and QTc intervals, with a free plasma concentration of approximately 1.7 µM at the point of maximal effect. Our data showed that unbound ICA-105574 caused QT shortening in dogs at concentrations comparable to the half maximal effective concentration (EC(50), 0.42 µM) of hERG activation in the patch clamp studies.

  2. Ageing studies on CuInS2/In2S3 junction (2.5×2cm2) deposited using automated spray machine

    NASA Astrophysics Data System (ADS)

    Santhosh, M. V.; Kartha, C. Sudha; Vijayakumar, K. P.

    2014-04-01

    CuInS2/In2S3 heterojunction photovoltaic device was realized in an area of 2.5 × 2 cm2 using automated spray pyrolysis machine which shows an open-circuit voltage of 432mV, short circuit current density of 6.33mA/cm2, fill factor of 34% and efficiency of 0.94%. Performance of the device was monitored up to 100 days and it was working quite well without the application of any protective coatings. The device maintains a fill factor of around 32% up to 80 days but other photovoltaic parameters had slight decrease.

  3. The relationship of TMPRSS2-ERG gene fusion between primary and metastatic prostate cancers

    PubMed Central

    Guo, Charles C.; Wang, Yan; Xiao, Li; Troncoso, Patricia; Czerniak, Bogdan A.

    2013-01-01

    Recent studies have revealed the presence of TMPRSS2-ERG gene fusion in both primary and metastatic prostatic cancers (PCAs). However, the relationship between primary and corresponding metastatic PCAs with respect to the status of this gene fusion remains unclear. Using fluorescence in situ hybridization, we evaluated the rearrangement of the ERG gene in the radical prostatectomy (RP) specimens and corresponding lymph node metastases from 19 patients with PCA. The mean age of the patients was 61 years and the median Gleason score in the RP specimens was 7 (4+3). PCA was unifocal in 6 cases and multifocal in 13 cases, including 10 with 2 foci and 3 with 3 foci. In the primary PCAs, rearrangement of the ERG gene was observed in 13 cases and associated with deletion of the 5’ ERG gene in 8 cases. In the metastases, the ERG rearrangement was present in 10 cases and associated with deletion of the 5’ ERG gene in 6 cases. In unifocal PCAs, the status of the ERG rearrangement was concordant between the primary PCA and metastasis in 5 of 6 cases. In multifocal PCA, despite a significant interfocal discordance, the status of the ERG rearrangement was concordant between the index (largest) primary tumor focus and metastasis in all 13 cases. Our study demonstrates a close relationship of the TMPRSS2-ERG gene fusion status between primary and metastatic PCA. The concordance of the ERG gene rearrangement status between the index primary tumor focus and metastasis suggests that metastasis most likely arises from the index tumor focus in multifocal PCA. PMID:21937078

  4. The relationship of TMPRSS2-ERG gene fusion between primary and metastatic prostate cancers.

    PubMed

    Guo, Charles C; Wang, Yan; Xiao, Li; Troncoso, Patricia; Czerniak, Bogdan A

    2012-05-01

    Recent studies have revealed the presence of TMPRSS2-ERG gene fusion in both primary and metastatic prostate cancers. However, the relationship between primary and corresponding metastatic prostate cancers with respect to the status of this gene fusion remains unclear. Using fluorescence in situ hybridization, we evaluated the rearrangement of the ERG gene in the radical prostatectomy specimens and corresponding lymph node metastases from 19 patients with prostate cancer. The mean age of the patients was 61 years, and the median Gleason score in the radical prostatectomy specimens was 7 (4 + 3). Prostate cancer was unifocal in 6 cases and multifocal in 13 cases, including 10 with 2 foci and 3 with 3 foci. In the primary prostate cancers, rearrangement of the ERG gene was observed in 13 cases and associated with deletion of the 5' ERG gene in 8 cases. In the metastases, the ERG rearrangement was present in 10 cases and associated with deletion of the 5' ERG gene in 6 cases. In unifocal prostate cancers, the status of the ERG rearrangement was concordant between the primary prostate cancer and metastasis in 5 of 6 cases. In multifocal prostate cancer, despite a significant interfocal discordance, the status of the ERG rearrangement was concordant between the index (largest) primary tumor focus and metastasis in all 13 cases. Our study demonstrates a close relationship of the TMPRSS2-ERG gene fusion status between primary and metastatic prostate cancer. The concordance of the ERG gene rearrangement status between the index primary tumor focus and metastasis suggests that metastasis most likely arises from the index tumor focus in multifocal prostate cancer.

  5. Heterogeneity and chronology of 6q15 deletion and ERG-fusion in prostate cancer

    PubMed Central

    Krohn, Antje; Freudenthaler, Fabian; Bauer, Melanie; Salomon, Georg; Heinzer, Hans; Michl, Uwe; Steurer, Stefan; Simon, Ronald; Sauter, Guido; Schlomm, Thorsten; Minner, Sarah

    2016-01-01

    Prostate cancer is notorious for its heterogeneity, which poses a problem for the applicability of diagnostic molecular markers. However, heterogeneity analysis can provide valuable information on the chronology in which molecular alterations arise. Here, we constructed a heterogeneity tissue microarray (TMA) comprising samples from 10 different tumor areas of 189 prostate cancers each in order to study the sequence of two frequent molecular alterations, i.e. 6q15 deletion and TMPRSS2:ERG fusion. Previous work shows a marked inverse relationship between these alterations, suggesting that presence of one of these alterations might impact development of the other. 6q15 deletion was analyzed by fluorescence in situ hybridization and ERG-expression by immunohistochemistry. Only 6.6% of 334 ERG-positive but 28.4% of 440 ERG-negative TMA spots showed 6q15 deletions (p < 0.0001). A breakdown of these data to the level of tumor foci revealed 6q deletions in 138 tumor foci that were large enough to have at least 3 analyzable TMA spots. These included 42 tumor foci with homogeneous ERG positivity and 16 with homogeneous 6q15 deletions. Remarkably, six of the 42 homogeneously ERG-positive tumor foci (14.3%) harbored small 6q15-deleted areas, but none of the 34 6q15-deleted foci showed areas of ERG positivity (p = 0.022). In conclusion, our data suggest that ERG-fusion can precede 6q15 deletion, but not vice versa. The complete absence of ERG-positive tumor areas in 6q15-deleted tumor foci further suggest that the functional consequences of 6q15 deletions may prevent the development of TMPRSS2:ERG fusions. PMID:26684029

  6. Endoxifen, the active metabolite of tamoxifen, inhibits cloned hERG potassium channels.

    PubMed

    Chae, Yun Ju; Lee, Keon Jin; Lee, Hong Joon; Sung, Ki-Wug; Choi, Jin-Sung; Lee, Eun Hui; Hahn, Sang June

    2015-04-05

    The effects of tamoxifen, and its active metabolite endoxifen (4-hydroxy-N-desmethyl-tamoxifen), on hERG currents stably expressed in HEK cells were investigated using the whole-cell patch-clamp technique and an immunoblot assay. Tamoxifen and endoxifen inhibited hERG tail currents at -50mV in a concentration-dependent manner with IC50 values of 1.2 and 1.6μM, respectively. The steady-state activation curve of the hERG currents was shifted to the hyperpolarizing direction in the presence of endoxifen. The voltage-dependent inhibition of hERG currents by endoxifen increased steeply in the voltage range of channel activation. The inhibition by endoxifen displayed a shallow voltage dependence (δ=0.18) in the full activation voltage range. A fast application of endoxifen induced a reversible block of hERG tail currents during repolarization in a concentration-dependent manner, which suggested an interaction with the open state of the channel. Endoxifen also decreased the hERG current elicited by a 5s depolarizing pulse to +60mV to inactivate the hERG currents, suggesting an interaction with the activated (open and/or inactivated) states of the channels. Tamoxifen and endoxifen inhibited the hERG channel protein trafficking to the plasma membrane in a concentration-dependent manner with endoxifen being more potent than tamoxifen. These results indicated that tamoxifen and endoxifen inhibited the hERG current by direct channel blockage and by the disruption of channel trafficking to the plasma membrane in a concentration-dependent manner. A therapeutic concentration of endoxifen inhibited the hERG current by preferentially interacting with the activated (open and/or inactivated) states of the channel.

  7. Quantifying the ON and OFF Contributions to the Flash ERG with the Discrete Wavelet Transform

    PubMed Central

    Gauvin, Mathieu; Sustar, Maja; Little, John M.; Brecelj, Jelka; Lina, Jean-Marc; Lachapelle, Pierre

    2017-01-01

    Purpose Discrete wavelet transform (DWT) analyses suggest that the 20- and 40-Hz components of the short-flash photopic electroretinogram (ERG) are closely related to the ON and OFF pathways, respectively. With the DWT, we examined how the ERG ON and OFF components are modulated by the stimulus intensity and/or duration. Methods Discrete wavelet transform descriptors (20, 40 Hz and 40:20-Hz ratio) were extracted from ERGs evoked to 25 combinations of flash durations (150–5 ms) and strengths (0.8–2.8 log cd.m−2). Results In ERGs evoked to the 150-ms stimulus (to separate the ON and OFF ERGs), the 40:20-Hz ratio of ON ERGs (mean ± SD: 0.49 ± 0.04) was significantly smaller (P < 0.05) than that of OFF ERGs (1.71 ± 0.18) owing to a significantly (P < 0.05) higher contribution of the 20 and 40 Hz components to the ON and OFF ERGs, respectively. With brighter stimuli, the ON and OFF components increased similarly (P < 0.05). While progressively shorter flashes had no impact (P > 0.05) on the ON component, it exponentially enhanced (P < 0.05) the OFF component. Conclusions Discrete wavelet transform allows for an accurate determination of ON and OFF retinal pathways even in ERGs evoked to a short flash. To our knowledge, the significant OFF facilitatory effect evidenced with shorter stimuli has not previously been reported. Translational Relevance The DWT approach should offer a rapid, easy, and reproducible approach to retrospectively and prospectively evaluate the function of the retinal ON and OFF pathways using the standard (short-flash duration) clinical ERG stimulus. PMID:28097047

  8. Tropical Cyclone Paka's Initial Explosive Development (10-12 December, 1997)

    NASA Technical Reports Server (NTRS)

    Rodgers, Edward B.; Halverson, Jeff; Simpson, Joanne; Olson, William; Pierce, Harold

    1999-01-01

    Convection associated with an equatorial westerly wind burst was first observed late November during the strong El Nino of 1997 at approximately 2000 km southwest of the Hawaiian Islands. This region of convection lead to the formation of twin tropical cyclones, one in the southern hemisphere named Pam and the other in the northern hemisphere named Paka. During the first week in December, tropical cyclone Paka, the system of concern, reached tropical storm stage as it moved rapidly westward at relatively low latitudes. During the 10-12 of December, Paka rapidly developed into a typhoon.

  9. a Theoretical Investigation on 10-12 Potential of Hydrogen-Hydrogen Covalent Bond

    NASA Astrophysics Data System (ADS)

    Taneri, Sencer

    2013-05-01

    This is an analytical investigation of well-known 10-12 potential of hydrogen-hydrogen covalent bond. In this research, we will make an elaboration of the well-known 6-12 Lennard-Jones potential in case of this type of bond. Though the results are illustrated in many text books and literature, an analytical analysis for these potentials is missing almost everywhere. The power laws are valid for small radial distances, which are calculated to some extent. The internuclear separation as well as the binding energy of the hydrogen molecule are evaluated with success.

  10. Dietary lycopene intake and risk of prostate cancer defined by ERG protein expression1

    PubMed Central

    Graff, Rebecca E; Pettersson, Andreas; Lis, Rosina T; Ahearn, Thomas U; Markt, Sarah C; Wilson, Kathryn M; Rider, Jennifer R; Fiorentino, Michelangelo; Finn, Stephen; Kenfield, Stacey A; Loda, Massimo; Giovannucci, Edward L; Rosner, Bernard; Mucci, Lorelei A

    2016-01-01

    Background: There is limited evidence that supports etiologically distinct molecular subtypes of prostate cancer, the identification of which may improve prevention. Given their antioxidant properties, we hypothesized that lycopene and tomato sauce may be especially protective against diseases harboring the common gene fusion transmembrane protease, serine 2 (TMPRSS2):v-ets avian erythroblastosis virus E26 oncogene homolog (ERG). Objective: We aimed to examine associations between estimated lycopene and tomato sauce intake and the risk of prostate cancer defined by ERG protein expression subtype. Design: Our study population consisted of a prospective cohort of 46,719 men from the Health Professionals Follow-Up Study. TMPRSS2:ERG was assessed by ERG immunohistochemistry on tumor tissue microarrays constructed from radical prostatectomy specimens. We used multivariable competing risk models to calculate HRs and 95% CIs for the risk of ERG-positive and, separately, ERG-negative disease. We implemented inverse probability weighting to account for evaluating ERG status only in surgically treated cases. Results: During 23 y of follow-up, 5543 men were diagnosed with prostate cancer, among whom 884 were assayed for ERG (426 ERG-positive). With inclusion of only the latter cases, increasing cumulative average tomato sauce intake was associated with a decreased risk of prostate cancer overall (≥2 servings/wk compared with <1 serving/mo; multivariable HR: 0.70; 95% CI: 0.52, 0.95; P-trend = 0.002). With respect to molecular subtypes, cumulative average tomato sauce intake was associated with a decreased risk of ERG-positive disease (HR: 0.54; 95% CI: 0.37, 0.81; P-trend = 0.004) but not with ERG-negative disease (HR: 0.96; 95% CI: 0.62, 1.50; P-trend = 0.10) (P-heterogeneity = 0.04). Increasing quintiles of lycopene intake were associated with a decreased risk of both subtypes (P-heterogeneity = 0.79). Inverse probability weighting did not materially change the results

  11. Compilation and physicochemical classification analysis of a diverse hERG inhibition database

    NASA Astrophysics Data System (ADS)

    Didziapetris, Remigijus; Lanevskij, Kiril

    2016-12-01

    A large and chemically diverse hERG inhibition data set comprised of 6690 compounds was constructed on the basis of ChEMBL bioactivity database and original publications dealing with experimental determination of hERG activities using patch-clamp and competitive displacement assays. The collected data were converted to binary format at 10 µM activity threshold and subjected to gradient boosting machine classification analysis using a minimal set of physicochemical and topological descriptors. The tested parameters involved lipophilicity (log P), ionization (p K a ), polar surface area, aromaticity, molecular size and flexibility. The employed approach allowed classifying the compounds with an overall 75-80 % accuracy, even though it only accounted for non-specific interactions between hERG and ligand molecules. The observed descriptor-response profiles were consistent with common knowledge about hERG ligand binding site, but also revealed several important quantitative trends, as well as slight inter-assay variability in hERG inhibition data. The results suggest that even weakly basic groups (p K a < 6) might substantially contribute to hERG inhibition potential, whereas the role of lipophilicity depends on the compound's ionization state, and the influence of log P decreases in the order of bases > zwitterions > neutrals > acids. Given its robust performance and clear physicochemical interpretation, the proposed model may provide valuable information to direct drug discovery efforts towards compounds with reduced risk of hERG-related cardiotoxicity.

  12. hERG1 Potassium Channels: Novel Biomarkers in Human Solid Cancers

    PubMed Central

    Lastraioli, Elena; Lottini, Tiziano; Bencini, Lapo; Bernini, Marco; Arcangeli, Annarosa

    2015-01-01

    Because of their high incidence and mortality solid cancers are a major health problem worldwide. Although several new biomarkers and potential targets for therapy have been identified through biomolecular research in the last years, the effects on patients' outcome are still unsatisfactory. Increasing evidence indicates that hERG1 potassium channels are overexpressed in human primary cancers of different origin and several associations between hERG1 expression and clinicopathological features and/or outcome are emerging. Aberrant hERG1 expression may be exploited either for early diagnosis (especially in those cancers where it is expressed in the initial steps of tumor progression) or for therapy purposes. Indeed, hERG1 blockage impairs tumor cell growth both in vitro and in vivo in preclinical mouse model. hERG1-based tumor therapy in humans, however, encounters the major hindrance of the potential cardiotoxicity that many hERG1 blockers exert. In this review we focus on recent advances in translational research in some of the most frequent human solid cancers (breast, endometrium, ovary, pancreas, esophagus, stomach, and colorectum) that have been shown to express hERG1 and that are a major health problem. PMID:26339650

  13. ERG signaling in prostate cancer is driven through PRMT5-dependent methylation of the Androgen Receptor

    PubMed Central

    Mounir, Zineb; Korn, Joshua M; Westerling, Thomas; Lin, Fallon; Kirby, Christina A; Schirle, Markus; McAllister, Gregg; Hoffman, Greg; Ramadan, Nadire; Hartung, Anke; Feng, Yan; Kipp, David Randal; Quinn, Christopher; Fodor, Michelle; Baird, Jason; Schoumacher, Marie; Meyer, Ronald; Deeds, James; Buchwalter, Gilles; Stams, Travis; Keen, Nicholas; Sellers, William R; Brown, Myles; Pagliarini, Raymond A

    2016-01-01

    The TMPRSS2:ERG gene fusion is common in androgen receptor (AR) positive prostate cancers, yet its function remains poorly understood. From a screen for functionally relevant ERG interactors, we identify the arginine methyltransferase PRMT5. ERG recruits PRMT5 to AR-target genes, where PRMT5 methylates AR on arginine 761. This attenuates AR recruitment and transcription of genes expressed in differentiated prostate epithelium. The AR-inhibitory function of PRMT5 is restricted to TMPRSS2:ERG-positive prostate cancer cells. Mutation of this methylation site on AR results in a transcriptionally hyperactive AR, suggesting that the proliferative effects of ERG and PRMT5 are mediated through attenuating AR’s ability to induce genes normally involved in lineage differentiation. This provides a rationale for targeting PRMT5 in TMPRSS2:ERG positive prostate cancers. Moreover, methylation of AR at arginine 761 highlights a mechanism for how the ERG oncogene may coax AR towards inducing proliferation versus differentiation. DOI: http://dx.doi.org/10.7554/eLife.13964.001 PMID:27183006

  14. ERG is a novel and reliable marker for endothelial cells in central nervous system tumors.

    PubMed

    Haber, Matthew A; Iranmahboob, Amir; Thomas, Cheddhi; Liu, Mengling; Najjar, Amanda; Zagzag, David

    2015-01-01

    ETS-related gene (ERG) is a transcription factor that has been linked to angiogenesis. Very little research has been done to assess ERG expression in central nervous system (CNS) tumors. We evaluated 57 CNS tumors, including glioblastomas (GBMs) and hemangioblastomas (HBs), as well as two arteriovenous malformations and four samples of normal brain tissue with immunohistochemistry using a specific ERG rabbit monoclonal antibody. In addition, immunostains for CD31, CD34, and α-smooth muscle actin (α-SMA) were performed on all samples. CD31 demonstrated variable and sometimes weak immunoreactivity for endothelial cells. Furthermore, in 1 case of a GBM, CD34 stained not only endothelial cells, but also tumor cells. In contrast, we observed that ERG was only expressed in the nuclei of endothelial cells, for example, in the hyperplastic vascular complexes that comprise the glomeruloid microvascular proliferation seen in GBMs. Conversely, α-SMA immunoreactivity was identified in the abluminal cells of these hyperplastic vessels. Quantitative evaluation with automated methodology and custom Matlab 2008b software was used to calculate percent staining of ERG in each case. We observed significantly higher quantitative expression of ERG in HBs than in other CNS tumors. Our results show that ERG is a novel, reliable, and specific marker for endothelial cells within CNS tumors that can be used to better study the process of neovascularization.

  15. Improved throughput of PatchXpress hERG assay using intracellular potassium fluoride.

    PubMed

    Zeng, Haoyu; Penniman, Jacob R; Kinose, Fumi; Kim, David; Trepakova, Elena S; Malik, Manish G; Dech, Spencer J; Balasubramanian, Bharathi; Salata, Joseph J

    2008-04-01

    Blockade of the human ether-a-go-go-related gene (hERG) potassium channel, with a consequent possibility of QT prolongation and increased susceptibility to a characteristic polymorphic ventricular arrhythmia, torsade de pointes, is an important cause of withdrawal of drugs from the market. In the aftermath of recent drug withdrawals, regulatory agencies now require in vitro hERG screening of all pharmaceutical compounds that are targeted for human use. To minimize the potential for failure in later-stage drug development, many pharmaceutical and biotechnology companies have begun to use automated patch clamp systems with higher throughput than conventional manual patch-clamp techniques to conduct routine functional hERG screening during drug discovery and early development. We have optimized an automated patch-clamp hERG screening method for the PatchXpress 7000A system (Molecular Devices, Sunnyvale, CA) using potassium fluoride (KF) in the internal recording solution. In this study we show that (1) the biophysical and pharmacological properties of hERG current recorded with KF are similar to those with standard potassium chloride solutions, (2) use of KF significantly improves the success rate of hERG screening using PatchXpress without compromising data quality, and (3) utilization of KF can significantly increase the throughput of hERG screening with PatchXpress.

  16. Global Analysis Reveals Families of Chemical Motifs Enriched for hERG Inhibitors

    PubMed Central

    Du, Fang; Babcock, Joseph J.; Yu, Haibo; Zou, Beiyan; Li, Min

    2015-01-01

    Promiscuous inhibition of the human ether-à-go-go-related gene (hERG) potassium channel by drugs poses a major risk for life threatening arrhythmia and costly drug withdrawals. Current knowledge of this phenomenon is derived from a limited number of known drugs and tool compounds. However, in a diverse, naïve chemical library, it remains unclear which and to what degree chemical motifs or scaffolds might be enriched for hERG inhibition. Here we report electrophysiology measurements of hERG inhibition and computational analyses of >300,000 diverse small molecules. We identify chemical ‘communities’ with high hERG liability, containing both canonical scaffolds and structurally distinctive molecules. These data enable the development of more effective classifiers to computationally assess hERG risk. The resultant predictive models now accurately classify naïve compound libraries for tendency of hERG inhibition. Together these results provide a more complete reference map of characteristic chemical motifs for hERG liability and advance a systematic approach to rank chemical collections for cardiotoxicity risk. PMID:25700001

  17. FR171456 is a specific inhibitor of mammalian NSDHL and yeast Erg26p

    PubMed Central

    Helliwell, Stephen B.; Karkare, Shantanu; Bergdoll, Marc; Rahier, Alain; Leighton-Davis, Juliet R.; Fioretto, Celine; Aust, Thomas; Filipuzzi, Ireos; Frederiksen, Mathias; Gounarides, John; Hoepfner, Dominic; Hofmann, Andreas; Imbert, Pierre-Eloi; Jeker, Rolf; Knochenmuss, Richard; Krastel, Philipp; Margerit, Anais; Memmert, Klaus; Miault, Charlotte V.; Rao Movva, N.; Muller, Alban; Naegeli, Hans-Ulrich; Oberer, Lukas; Prindle, Vivian; Riedl, Ralph; Schuierer, Sven; Sexton, Jessica A.; Tao, Jianshi; Wagner, Trixie; Yin, Hong; Zhang, Juan; Roggo, Silvio; Reinker, Stefan; Parker, Christian N.

    2015-01-01

    FR171456 is a natural product with cholesterol-lowering properties in animal models, but its molecular target is unknown, which hinders further drug development. Here we show that FR171456 specifically targets the sterol-4-alpha-carboxylate-3-dehydrogenase (Saccharomyces cerevisiae—Erg26p, Homo sapiens—NSDHL (NAD(P) dependent steroid dehydrogenase-like)), an essential enzyme in the ergosterol/cholesterol biosynthesis pathway. FR171456 significantly alters the levels of cholesterol pathway intermediates in human and yeast cells. Genome-wide yeast haploinsufficiency profiling experiments highlight the erg26/ERG26 strain, and multiple mutations in ERG26 confer resistance to FR171456 in growth and enzyme assays. Some of these ERG26 mutations likely alter Erg26 binding to FR171456, based on a model of Erg26. Finally, we show that FR171456 inhibits an artificial Hepatitis C viral replicon, and has broad antifungal activity, suggesting potential additional utility as an anti-infective. The discovery of the target and binding site of FR171456 within the target will aid further development of this compound. PMID:26456460

  18. Structure and Function of the Splice Variants of TMPRSS2-ERG, a Prevalent Genomic Alteration in Prostate Cancer

    DTIC Science & Technology

    2012-09-01

    the two main tumor foci in this section, one tumor focus was ERG positive. In ERG positive tumor focus, malignant epithelium or normal endotheials...ERG was not expressed in normal mouse prostate epithelium (Mohamed et al., 2010). Similar findings were noted for human prostate (Furusato et al...2010). These observations highlight the cancer-specific aberrant expression and function of ERG in the prostate epithelium . Figure 10

  19. hERG1 channels drive tumour malignancy and may serve as prognostic factor in pancreatic ductal adenocarcinoma

    PubMed Central

    Lastraioli, E; Perrone, G; Sette, A; Fiore, A; Crociani, O; Manoli, S; D'Amico, M; Masselli, M; Iorio, J; Callea, M; Borzomati, D; Nappo, G; Bartolozzi, F; Santini, D; Bencini, L; Farsi, M; Boni, L; Di Costanzo, F; Schwab, A; Onetti Muda, A; Coppola, R; Arcangeli, A

    2015-01-01

    Background: hERG1 channels are aberrantly expressed in human cancers. The expression, functional role and clinical significance of hERG1 channels in pancreatic ductal adenocarcinoma (PDAC) is lacking. Methods: hERG1 expression was tested in PDAC primary samples assembled as tissue microarray by immunohistochemistry using an anti-hERG1 monoclonal antibody (α-hERG1-MoAb). The functional role of hERG1 was studied in PDAC cell lines and primary cultures. ERG1 expression during PDAC progression was studied in Pdx-1-Cre,LSL-KrasG12D/+,LSL-Trp53R175H/+ transgenic (KPC) mice. ERG1 expression in vivo was determined by optical imaging using Alexa-680-labelled α-hERG1-MoAb. Results: (i) hERG1 was expressed at high levels in 59% of primary PDAC; (ii) hERG1 blockade decreased PDAC cell growth and migration; (iii) hERG1 was physically and functionally linked to the Epidermal Growth Factor-Receptor pathway; (iv) in transgenic mice, ERG1 was expressed in PanIN lesions, reaching high expression levels in PDAC; (v) PDAC patients whose primary tumour showed high hERG1 expression had a worse prognosis; (vi) the α-hERG1-MoAb could detect PDAC in vivo. Conclusions: hERG1 regulates PDAC malignancy and its expression, once validated in a larger cohort also comprising of late-stage, non-surgically resected cases, may be exploited for diagnostic and prognostic purposes in PDAC either ex vivo or in vivo. PMID:25719829

  20. Mechanism of inhibition by olanzapine of cloned hERG potassium channels.

    PubMed

    Lee, Hong Joon; Choi, Jin-Sung; Hahn, Sang June

    2015-11-16

    Olanzapine is widely used in the treatment of schizophrenia and related psychoses. We investigated the effects of olanzapine on human ether-a-go-go related gene (hERG) channels stably expressed in human embryonic kidney (HEK) cells using the whole-cell patch-clamp technique. Olanzapine inhibited hERG tail currents at -50mV in a concentration-dependent manner with an IC50 value of 8.0μM and a Hill coefficient of 0.9. The voltage-dependent inhibition of the hERG currents by olanzapine increased steeply in the voltage range of channel activation. Olanzapine also shifted the steady-state activation curve of the hERG currents in a hyperpolarizing direction. At more depolarized potentials where the channels were fully activated (between 0 and +50mV), the olanzapine inhibition was voltage-independent. The steady-state inactivation curve of the hERG currents was shifted in the hyperpolarizing direction in the presence of olanzapine. A fast application of olanzapine induced a reversible inhibition of the hERG tail currents during repolarization in a concentration-dependent manner with an IC50 value of 11.9μM, suggesting an open-channel block. Olanzapine also decreased the hERG current elicited by a 5s depolarizing pulse to +60mV to inactivate the hERG currents, suggesting an inhibition of the activated (open and/or inactivated) states of the channels. These results indicated that olanzapine inhibited the hERG current by preferentially interacting with the activated states of the channel.

  1. Evaluating the Efficacy of ERG Targeted Therapy In Vivo for Prostate Tumors

    DTIC Science & Technology

    2013-04-01

    single oncogene mice to genetically analyze cooperation in vivo. Investigate using molecular techniques if ERG modulates proliferation, apoptosis and... breeding between our tetO-ERG mice and Dr. Bieberich’s Hoxb13-rtTA mice will be initiated shortly. We will reinitiate our studies on the ability of ERG...If adding animals or procedures to category D or E for the first time. please see "Modify Pain Category"’ below. Number R8Q(Ifl$tfld Pain Cat ~tgory

  2. Evaluation of MG-101 course Machine guarding'' taught in Stanford, California, November 10--12, 1992

    SciTech Connect

    Wright, T.S.

    1993-03-01

    This report summarizes trainee evaluations for the Safety Training Section course, Machine Guarding,'' (MG-101) which was conducted November 10--12, 1992 at Stanford Linear Accelerator, in Stanford, California. This report summarizes the quantitative course evaluations that trainees provided upon completion of the course, including presents examination results, and recommendations for course improvement. Numeric course ratings were generally positive and show that the course material and instruction were very effective. Written comments supported the positive numeric ratings. The course content and knowledge gained by the trainees exceeded most of the students' expectations of the course. This course is now in the format that has been requested by past attendees. It appears to be meeting their needs. Results from the final examination showed that students gained appropriate knowledge from the course.

  3. Solution-processed small molecule-polymer blend organic thin-film transistors with hole mobility greater than 5 cm2/Vs.

    PubMed

    Smith, Jeremy; Zhang, Weimin; Sougrat, Rachid; Zhao, Kui; Li, Ruipeng; Cha, Dongkyu; Amassian, Aram; Heeney, Martin; McCulloch, Iain; Anthopoulos, Thomas D

    2012-05-08

    Using phase-separated organic semiconducting blends containing a small molecule, as the hole transporting material, and a conjugated amorphous polymer, as the binder material, we demonstrate solution-processed organic thin-film transistors with superior performance characteristics that include; hole mobility >5 cm(2) /Vs, current on/off ratio ≥10(6) and narrow transistor parameter spread. These exceptional characteristics are attributed to the electronic properties of the binder polymer and the advantageous nanomorphology of the blend film.

  4. New narrow infrared absorption features in the spectrum of Io between 3600 and 3100 cm (2.8-3.2 micrometers)

    NASA Technical Reports Server (NTRS)

    Sandford, Scott A.; Geballe, Thomas R.; Salama, Farid; Goorvitch, David

    1994-01-01

    We report the discovery of a series of infrared absorption bands between 3600 and 3100/cm (2.8-3.2 micrometers) in the spectrum of Io. Individual narrow bands are detected at 3553, 3514.5, 3438, 3423, 3411.5, and 3401/cm (2.815, 2.845, 2.909, 2.921, 2.931, and 2.940 micrometers, respectively). The positions and relative strengths of these bands, and the difference of their absolute strengths between the leading and trailing faces of Io, indicate that they are due to SO2. The band at 3438/cm (2.909 micrometers) could potentially have a contribution from an additional molecular species. The existence of these bands in the spectrum of Io indicates that a substantial fraction of the SO2 on Io must reside in transparent ices having relatively large crystal sizes. The decrease in the continuum observed at the high frequency ends of the spectra is probably due to the low frequency side of the recently detected, strong 3590/cm (2.79 micrometer) feature. This band is likely due to the combination of a moderately strong SO2 band and an additional absorption from another molecular species, perhaps H2O isolated in SO2 at low concentrations. A broad (FWHM approximately = 40-60/cm), weak band is seen near 3160/cm (3.16 micrometers) and is consistent with the presence of small quantities of H2O isolated in SO2-rich ices. There is no evidence in the spectra for the presence of H2O vapor on Io. Thus, the spectra presented here neither provide unequivocal evidence for the presence of H2O on Io nor preclude it at the low concentrations suggested by past studies.

  5. Feasibility Study for Implementing Magnetic Suspension in the Glenn Research Center 225 cm2 Supersonic Wind Tunnel for Testing the Dynamic Stability of Blunt Bodies

    NASA Technical Reports Server (NTRS)

    Sevier, Abigail; Davis, David O.; Schoenenberger, Mark; Barnhart, Paul

    2016-01-01

    The implementation of a magnetic suspension system in the NASA Glenn Research Center (GRC) 225 cm2 Supersonic Wind Tunnel would be a powerful test technique that could accurately determine the dynamic stability of blunt body entry vehicles with no sting interference. This paper explores initial design challenges to be evaluated before implementation, including defining the lowest possible operating dynamic pressure and corresponding model size, developing a compatible video analysis technique, and incorporating a retractable initial support sting.

  6. A toxin to nervous, cardiac, and endocrine ERG K+ channels isolated from Centruroides noxius scorpion venom.

    PubMed

    Gurrola, G B; Rosati, B; Rocchetti, M; Pimienta, G; Zaza, A; Arcangeli, A; Olivotto, M; Possani, L D; Wanke, E

    1999-05-01

    Toxins isolated from a variety of venoms are tools for probing the physiological function and structure of ion channels. The ether-a-go-go-related genes (erg) codify for the K+ channels (ERG), which are crucial in neurons and are impaired in human long-QT syndrome and Drosophila 'seizure' mutants. We have isolated a peptide from the scorpion Centruroides noxius Hoffmann that has no sequence homologies with other toxins, and demonstrate that it specifically inhibits (IC50=16+/-1 nM) only ERG channels of different species and distinct histogenesis. These results open up the possibility of investigating ERG channel structure-function relationships and novel pharmacological tools with potential therapeutic efficacy.

  7. Azole resistance in Cryptococcus gattii from the Pacific Northwest: Investigation of the role of ERG11.

    PubMed

    Gast, Charles E; Basso, Luiz R; Bruzual, Igor; Wong, Brian

    2013-11-01

    Cryptococcus gattii is responsible for an expanding epidemic of serious infections in Western Canada and the Northwestern United States (Pacific Northwest). Some patients with these infections respond poorly to azole antifungals, and high azole MICs have been reported in Pacific Northwest C. gattii. In this study, multiple azoles (but not amphotericin B) had higher MICs for 25 Pacific Northwest C. gattii than for 34 non-Pacific Northwest C. gattii or 20 Cryptococcus neoformans strains. We therefore examined the roles in azole resistance of overexpression of or mutations in the gene (ERG11) encoding the azole target enzyme. ERG11/ACT1 mRNA ratios were higher in C. gattii than in C. neoformans, but these ratios did not differ in Pacific Northwest and non-Pacific Northwest C. gattii strains, nor did they correlate with fluconazole MICs within any group. Three Pacific Northwest C. gattii strains with low azole MICs and 2 with high azole MICs had deduced Erg11p sequences that differed at one or more positions from that of the fully sequenced Pacific Northwest C. gattii strain R265. However, the azole MICs for conditional Saccharomyces cerevisiae erg11 mutants expressing the 5 variant ERG11s were within 2-fold of the azole MICs for S. cerevisiae expressing the ERG11 gene from C. gattii R265, non-Pacific Northwest C. gattii strain WM276, or C. neoformans strains H99 or JEC21. We conclude that neither ERG11 overexpression nor variations in ERG11 coding sequences was responsible for the high azole MICs observed for the Pacific Northwest C. gattii strains we studied.

  8. Biological and Genomic Differences of ERG Oncoprotein-Stratified Prostate Cancers from African and Caucasian Americans

    DTIC Science & Technology

    2014-10-01

    for AfricanAmerican ( AA ) men as compared to Caucasian Americans (CA). Biological basis for this disparity has not been established. Oncogenic...underlying biological or genetic differences of prostate cancer (CaP) incidence and/or progression between African American ( AA ) and Caucasian American (CA...positive and ERG negative CaP. It is anticipated that molecular determinants of aggressive CaP in AA men include somatic mutations (TMPRSS2-ERG

  9. Tbx20 controls the expression of the KCNH2 gene and of hERG channels.

    PubMed

    Caballero, Ricardo; Utrilla, Raquel G; Amorós, Irene; Matamoros, Marcos; Pérez-Hernández, Marta; Tinaquero, David; Alfayate, Silvia; Nieto-Marín, Paloma; Guerrero-Serna, Guadalupe; Liu, Qing-Hua; Ramos-Mondragón, Roberto; Ponce-Balbuena, Daniela; Herron, Todd; Campbell, Katherine F; Filgueiras-Rama, David; Peinado, Rafael; López-Sendón, José L; Jalife, José; Delpón, Eva; Tamargo, Juan

    2017-01-17

    Long QT syndrome (LQTS) exhibits great phenotype variability among family members carrying the same mutation, which can be partially attributed to genetic factors. We functionally analyzed the KCNH2 (encoding for Kv11.1 or hERG channels) and TBX20 (encoding for the transcription factor Tbx20) variants found by next-generation sequencing in two siblings with LQTS in a Spanish family of African ancestry. Affected relatives harbor a heterozygous mutation in KCNH2 that encodes for p.T152HfsX180 Kv11.1 (hERG). This peptide, by itself, failed to generate any current when transfected into Chinese hamster ovary (CHO) cells but, surprisingly, exerted "chaperone-like" effects over native hERG channels in both CHO cells and mouse atrial-derived HL-1 cells. Therefore, heterozygous transfection of native (WT) and p.T152HfsX180 hERG channels generated a current that was indistinguishable from that generated by WT channels alone. Some affected relatives also harbor the p.R311C mutation in Tbx20. In human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs), Tbx20 enhanced human KCNH2 gene expression and hERG currents (IhERG) and shortened action-potential duration (APD). However, Tbx20 did not modify the expression or activity of any other channel involved in ventricular repolarization. Conversely, p.R311C Tbx20 did not increase KCNH2 expression in hiPSC-CMs, which led to decreased IhERG and increased APD. Our results suggest that Tbx20 controls the expression of hERG channels responsible for the rapid component of the delayed rectifier current. On the contrary, p.R311C Tbx20 specifically disables the Tbx20 protranscriptional activity over KCNH2 Therefore, TBX20 can be considered a KCNH2-modifying gene.

  10. The Central Asian ergs: A study by remote sensing and geographic information systems

    NASA Astrophysics Data System (ADS)

    Maman, Shimrit; Blumberg, Dan G.; Tsoar, Haim; Mamedov, Batyr; Porat, Naomi

    2011-12-01

    Ergs are large, relatively flat areas of deserts covered by wind-swept sand and with varying degrees of vegetation cover. The ergs of Central Asia extend from Turkmenistan to the Syr-Darya River in Kazakhstan. They are crossed by the Amu-Darya River, to the north and south of which lie Kyzyl-Kum and Kara-Kum, respectively. This research focuses on identifying and mapping the ergs of Central Asia and analyzing the climate factors that set the dunes in motion and that later stabilize them. The vast area encompassed by the Central Asian ergs and their inaccessibility make traditional mapping methods virtually impossible. A variety of spaceborne imagery with varying spectral and spatial resolutions was used. These images provided the basis for mapping sand distribution, dune forms, and vegetation cover. Wilson (1973) defined the Central Asian ergs as active based on precipitation. Our results, in contrast, show that these ergs are mostly stabilized, with the estimated sand mantled area for the Kara-Kum desert ˜260,000 km 2, and for the Kyzyl-Kum it is ˜195,500 km 2. Meteorological analysis of wind and precipitation data indicate a low wind power environment (DP < 200) and sufficient rainfall (>100 mm) to support vegetation. The age of the sand samples was determined by optically stimulated luminescence (OSL) (˜5-7 Ka), which provides insight about past climate characteristics. GIS analysis was performed in parallel with field work to obtain validation and verification.

  11. Behavior of the Aurora During 10-12 May, 1999 When The Solar Wind Nearly Disappeared

    NASA Technical Reports Server (NTRS)

    Parks, G.; Brittnacher, M.; Chua, D.; Fillingim, M.; Germany, G.; Spann, J.; Rose, M. Franklin (Technical Monitor)

    2000-01-01

    The aurora was still active with occasional pseudobreakup events when the solar wind density diminished to unusually small densities (0.2 cc -1) during May 10-12, 1999. The aurora was observed at high magnetic latitudes indicating that the electron precipitation source moved northward as the geomagnetic activity decreased. The events we have studied indicate that the solar wind density alone is not the primary parameter that controls the auroral activity. The weak auroral activity was observed with 150 nT magnetic bays and when the interplanetary magnetic field (IMF) Bz was small and positive resulting in small _ parameter. A new auroral feature was observed on May 11, 1999, between 0900-2000 UT. The electron precipitation was energetic, uniform, and covered the region commonly identified as the polar cap. This precipitation lasted for more than 10 hours and was stable over time scales of tens of minutes. On May 12, as the solar wind began to recover, a prolonged period of dayside activity occurred and was followed by a typical aurora at 0500 UT.

  12. Dynamics of cell and tissue growth acquired by means of 25 mm2 to 10 cm2 lens-free imaging

    NASA Astrophysics Data System (ADS)

    Momey, F.; Coutard, J.-G.; Bordy, T.; Navarro, F.; Menneteau, M.; Dinten, J.-M.; Allier, C.

    2015-03-01

    In this paper, we discuss a new methodology based on lens-free imaging to perform wound healing assay with unprecedented statistics. Our video lens-free microscopy setup is a simple optical system featuring only a CMOS sensor and a semi coherent illumination system. Yet it is a powerful means for the real-time monitoring of cultivated cells. It presents several key advantages, e.g., integration into standard incubator, compatibility with standard cell culture protocol, simplicity and ease of use. It can perform the follow-up in a large field of view (25 mm2) of several crucial parameters during the culture of cells i.e. their motility, their proliferation rate or their death. Consequently the setup can gather large statistics both in space and time. But in the case of tissue growth experiments, the field of view of 25 mm2 remains not sufficient and results can be biased depending on the position of the device with respect to the recipient of the cell culture. Hence, to conduct exhaustive wound healing assay, here we propose to enlarge the field of view up to 10 cm2 through two different approaches. The first method consists in performing a scan of the cell culture by moving the source/sensor couple and then stitch the stack of images. The second is to make an acquisition by scanning with a line scan camera. The two approaches are compared in term of resolution, complexity and acquisition time. Next we have performed acquisitions of wound healing assay (keratinocytes HaCaT) both in real-time (25 mm2) and in final point (10 cm2) to assess the combination of these two complementary modalities. In the future, we aim at combining directly super wide field of view acquisitions (>10 cm2) with real time ability inside the incubator.

  13. Absence of ocular effects after either single or repeated exposure to 10 mW/cm(2) from a 60 GHz CW source.

    PubMed

    Kues, H A; D'Anna, S A; Osiander, R; Green, W R; Monahan, J C

    1999-12-01

    This study was designed to examine ocular effects associated with exposure to millimeter waves (60 GHz). Rabbits served as the primary experimental subjects. To confirm the results of the rabbit experiments in a higher species, the second phase of the study used nonhuman primates (Macaca mulatta). First, this study used time-resolved infrared radiometry to assess the field distribution patterns produced by different antennas operating at 60 GHz. These results allowed us to select an antenna that produced a uniform energy distribution and the best distance at which to expose our experimental subjects. The study then examined ocular changes after exposure at an incident power density of 10 mW/cm(2). Acute exposure of both rabbits and nonhuman primates consisted of a single 8 h exposure, and the repeated exposure protocol consisted of five separate 4 h exposures on consecutive days. One eye in each animal was exposed and the contralateral eye served as the sham-exposed control. After postexposure diagnostic examinations, animals were euthanized and the eyes were removed. Ocular tissue was examined by both light and transmission electron microscopy. Neither microscopic examinations nor the diagnostic procedures performed on the eyes of acute and repeatedly exposed rabbits found any ocular changes that could be attributed to millimeter-wave exposure at 10 mW/cm(2). Examination of the primates after comparable exposures also failed to detect any ocular changes due to exposure. On the basis of our results, we conclude that single or repeated exposure to 60 GHz CW radiation at 10 mW/cm(2) does not result in any detectable ocular damage.

  14. Laser Driven Compression Equations of State and Hugoniot Pressure Measurements in Thick Solid Metallic Targets at ˜0.17-13 TW/cm2

    NASA Astrophysics Data System (ADS)

    Remo, John L.

    2010-10-01

    An electro-optic laser probe was developed to obtain parameters for high energy density equations of state (EoS), Hugoniot pressures (PH), and strain rates for high energy density laser irradiation intensity, I, experiments at ˜170 GW/cm2 (λ = 1064 nm) to ˜13 TW/cm2 (λ = 527 nm) on Al, Cu, Ti, Fe, Ni metal targets in a vacuum. At I ˜7 TW/cm2 front surface plasma pressures and temperatures reached 100's GPa and over two million K. Rear surface PH ranged from 7-120 GPa at average shock wave transit velocities 4.2-8.5 km/s, depending on target thickness and I. A surface plasma compression ˜100's GPa generated an impulsive radial expanding shock wave causing compression, rarefactions, and surface elastic and plastic deformations depending on I. A laser/fiber optic system measured rear surface shock wave emergence and particle velocity with ˜3 GHz resolution by monitoring light deflection from diamond polished rear surfaces of malleable metallic targets, analogous to an atomic force microscope. Target thickness, ˜0.5-2.9 mm, prevented front surface laser irradiation penetration, due to low radiation skin depth, from altering rear surface reflectivity (refractive index). At ˜10 TW electromagnetic plasma pulse noise generated from the target chamber overwhelmed detector signals. Pulse frequency analysis using Moebius loop antennae probed transient noise characteristics. Average shock (compression) and particle (rear surface displacement) velocity measurements determined rear surface PH and GPa) EoS that are compared with gas guns.

  15. 4H-SiC Power Bipolar Junction Transistor with a Very Low Specific On-resistance of 2.9 mOmega.cm2

    DTIC Science & Technology

    2006-04-12

    pp1381-1382, 2004. [2] C.-F. Huang and J. A. Cooper, Jr., “High current gain 4H-SiC NPN Bipolar Junction Transistors ,” IEEE Electron Device Lett...4H-SiC Power Bipolar Junction Transistor with a Very Low Specific On-resistance of 2.9 mΩ.cm2 Jianhui Zhang, member, IEEE, Petre Alexandrov...specific on-resistance (Rsp,on) of power 4H-SiC bipolar junction transistors (BJT). A 4H-SiC BJT based on a 12 um drift-layer shows a record low

  16. Giant Field Localization in 2-D Photonic Crystal Cavities with Defect Resonances: Bringing Nonlinear Optics to the W/cm2 Level

    DTIC Science & Technology

    2012-01-01

    5f. WORK UNIT NUMBER 7. PERFORMING ORGANIZATION NAME(S) AND ADDRESS(ES) AEgis Tech,Nanogenesis Division ,410 Jan Davis Dr,Huntsville,AL,35806 8...cm2 level Nadia Mattiucci,1 Mark J. Bloemer,2 and Giuseppe D’Aguanno1,a 1AEgis Tech., Nanogenesis Division 410 Jan Davis Dr, Huntsville, AL 35806, USA...Opt. Comm. 285, 1945 (2012). 24 N. Hô, J. M. Laniel, R. Valée, and A. Villeneuve, “ Photosensitivity of As2S3 chalcogenide thin films at 1.5μm

  17. MgZnO/ZnO heterostructures with electron mobility exceeding 1 × 106 cm2/Vs

    PubMed Central

    Falson, Joseph; Kozuka, Yusuke; Uchida, Masaki; Smet, Jurgen H.; Arima, Taka-hisa; Tsukazaki, Atsushi; Kawasaki, Masashi

    2016-01-01

    The inherently complex chemical and crystallographic nature of oxide materials has suppressed the purities achievable in laboratory environments, obscuring the rich physical degrees of freedom these systems host. In this manuscript we provide a systematic approach to defect identification and management in oxide molecular beam epitaxy grown MgZnO/ZnO heterostructures which host two-dimensional electron systems. We achieve samples displaying electron mobilities in excess of 1 × 106 cm2/Vs. This data set for the MgZnO/ZnO system firmly establishes that the crystalline quality has become comparable to traditional semiconductor materials. PMID:27229479

  18. Tuning hERG out: Antitarget QSAR Models for Drug Development

    PubMed Central

    Braga, Rodolpho C.; Alves, Vinícius M.; Silva, Meryck F. B.; Muratov, Eugene; Fourches, Denis; Tropsha, Alexander; Andrade, Carolina H.

    2015-01-01

    Several non-cardiovascular drugs have been withdrawn from the market due to their inhibition of hERG K+ channels that can potentially lead to severe heart arrhythmia and death. As hERG safety testing is a mandatory FDA-required procedure, there is a considerable interest for developing predictive computational tools to identify and filter out potential hERG blockers early in the drug discovery process. In this study, we aimed to generate predictive and well-characterized quantitative structure–activity relationship (QSAR) models for hERG blockage using the largest publicly available dataset of 11,958 compounds from the ChEMBL database. The models have been developed and validated according to OECD guidelines using four types of descriptors and four different machine-learning techniques. The classification accuracies discriminating blockers from non-blockers were as high as 0.83–0.93 on external set. Model interpretation revealed several SAR rules, which can guide structural optimization of some hERG blockers into non-blockers. We have also applied the generated models for screening the World Drug Index (WDI) database and identify putative hERG blockers and non-blockers among currently marketed drugs. The developed models can reliably identify blockers and non-blockers, which could be useful for the scientific community. A freely accessible web server has been developed allowing users to identify putative hERG blockers and non-blockers in chemical libraries of their interest (http://labmol.farmacia.ufg.br/predherg). PMID:24805060

  19. Molecular basis of hERG potassium channel blockade by the class Ic antiarrhythmic flecainide

    PubMed Central

    Melgari, Dario; Zhang, Yihong; El Harchi, Aziza; Dempsey, Christopher E.; Hancox, Jules C.

    2015-01-01

    The class Ic antiarrhythmic drug flecainide inhibits KCNH2-encoded “hERG” potassium channels at clinically relevant concentrations. The aim of this study was to elucidate the underlying molecular basis of this action. Patch clamp recordings of hERG current (IhERG) were made from hERG expressing cells at 37 °C. Wild-type (WT) IhERG was inhibited with an IC50 of 1.49 μM and this was not significantly altered by reversing the direction of K+ flux or raising external [K+]. The use of charged and uncharged flecainide analogues showed that the charged form of the drug accesses the channel from the cell interior to produce block. Promotion of WT IhERG inactivation slowed recovery from inhibition, whilst the N588K and S631A attenuated-inactivation mutants exhibited IC50 values 4–5 fold that of WT IhERG. The use of pore-helix/selectivity filter (T623A, S624A V625A) and S6 helix (G648A, Y652A, F656A) mutations showed < 10-fold shifts in IC50 for all but V625A and F656A, which respectively exhibited IC50s 27-fold and 142-fold their WT controls. Docking simulations using a MthK-based homology model suggested an allosteric effect of V625A, since in low energy conformations flecainide lay too low in the pore to interact directly with that residue. On the other hand, the molecule could readily form π–π stacking interactions with aromatic residues and particularly with F656. We conclude that flecainide accesses the hERG channel from the cell interior on channel gating, binding low in the inner cavity, with the S6 F656 residue acting as a principal binding determinant. PMID:26159617

  20. Colorimetric TMPRSS2-ERG Gene Fusion Detection in Prostate Cancer Urinary Samples via Recombinase Polymerase Amplification.

    PubMed

    Koo, Kevin M; Wee, Eugene J H; Trau, Matt

    2016-01-01

    TMPRSS2 (Exon 1)-ERG (Exon 4) is the most frequent gene fusion event in prostate cancer (PC), and is highly PC-specific unlike the current serum prostate specific antigen (PSA) biomarker. However, TMPRSS2-ERG levels are currently measured with quantitative reverse-transcription PCR (RT-qPCR) which is time-consuming and requires costly equipment, thus limiting its use in clinical diagnostics. Herein, we report a novel rapid, cost-efficient and minimal-equipment assay named "FusBLU" for detecting TMPRSS2-ERG gene fusions from urine. TMPRSS2-ERG mRNA was amplified by isothermal reverse transcription-recombinase polymerase amplification (RT-RPA), magnetically-isolated, and detected through horseradish peroxidase (HRP)-catalyzed colorimetric reaction. FusBLU was specific for TMPRSS2-ERG mRNA with a low visual detection limit of 10(5) copies. We also demonstrated assay readout versatility on 3 potentially useful platforms. The colorimetric readout was detectable by naked eye for a quick yes/no evaluation of gene fusion presence. On the other hand, a more quantitative TMPRSS2-ERG detection was achievable by absorbance/electrochemical measurements. FusBLU was successfully applied to 12 urinary samples and results were validated by gold-standard RT-qPCR. We also showed that sediment RNA was likely the main source of TMPRSS2-ERG mRNA in urinary samples. We believe that our assay is a potential clinical screening tool for PC and could also have wide applications for other disease-related fusion genes.

  1. Colorimetric TMPRSS2-ERG Gene Fusion Detection in Prostate Cancer Urinary Samples via Recombinase Polymerase Amplification

    PubMed Central

    Koo, Kevin M.; Wee, Eugene J.H.; Trau, Matt

    2016-01-01

    TMPRSS2 (Exon 1)-ERG (Exon 4) is the most frequent gene fusion event in prostate cancer (PC), and is highly PC-specific unlike the current serum prostate specific antigen (PSA) biomarker. However, TMPRSS2-ERG levels are currently measured with quantitative reverse-transcription PCR (RT-qPCR) which is time-consuming and requires costly equipment, thus limiting its use in clinical diagnostics. Herein, we report a novel rapid, cost-efficient and minimal-equipment assay named “FusBLU” for detecting TMPRSS2-ERG gene fusions from urine. TMPRSS2-ERG mRNA was amplified by isothermal reverse transcription-recombinase polymerase amplification (RT-RPA), magnetically-isolated, and detected through horseradish peroxidase (HRP)-catalyzed colorimetric reaction. FusBLU was specific for TMPRSS2-ERG mRNA with a low visual detection limit of 105 copies. We also demonstrated assay readout versatility on 3 potentially useful platforms. The colorimetric readout was detectable by naked eye for a quick yes/no evaluation of gene fusion presence. On the other hand, a more quantitative TMPRSS2-ERG detection was achievable by absorbance/electrochemical measurements. FusBLU was successfully applied to 12 urinary samples and results were validated by gold-standard RT-qPCR. We also showed that sediment RNA was likely the main source of TMPRSS2-ERG mRNA in urinary samples. We believe that our assay is a potential clinical screening tool for PC and could also have wide applications for other disease-related fusion genes. PMID:27375789

  2. Androgen receptor and gene network: Micromechanics reassemble the signaling machinery of TMPRSS2-ERG positive prostate cancer cells

    PubMed Central

    2014-01-01

    Prostate cancer is a gland tumor in the male reproductive system. It is a multifaceted and genomically complex disease. Transmembrane protease, serine 2 and v-ets erythroblastosis virus E26 homolog (TMPRSS2-ERG) gene fusions are the common molecular signature of prostate cancer. Although tremendous advances have been made in unraveling various facets of TMPRSS2-ERG-positive prostate cancer, many research findings must be sequentially collected and re-interpreted. It is important to understand the activation or repression of target genes and proteins in response to various stimuli and the assembly in signal transduction in TMPRSS2-ERG fusion-positive prostate cancer cells. Accordingly, we divide this multi-component review ofprostate cancer cells into several segments: 1) The role of TMPRSS2-ERG fusion in genomic instability and methylated regulation in prostate cancer and normal cells; 2) Signal transduction cascades in TMPRSS2-ERG fusion-positive prostate cancer; 3) Overexpressed genes in TMPRSS2-ERG fusion-positive prostate cancer cells; 4) miRNA mediated regulation of the androgen receptor (AR) and its associated protein network; 5) Quantitative control of ERG in prostate cancer cells; 6) TMPRSS2-ERG encoded protein targeting; In conclusion, we provide a detailed understanding of TMPRSS2-ERG fusion related information in prostate cancer development to provide a rationale for exploring TMPRSS2-ERG fusion-mediated molecular network machinery. PMID:24739220

  3. The A395T mutation in ERG11 gene confers fluconazole resistance in Candida tropicalis causing candidemia.

    PubMed

    Tan, Jingwen; Zhang, Jinqing; Chen, Wei; Sun, Yi; Wan, Zhe; Li, Ruoyu; Liu, Wei

    2015-04-01

    The mechanism of fluconazole resistance in Candida tropicalis is still unclear. Recently, we isolated a fluconazole-resistant strain of C. tropicalis from the blood specimen of a patient with candidemia in China. In vitro antifungal susceptibility of the isolate was determined by using CLSI M27-A3 and E-test methods. The sequence of ERG11 gene was then analyzed, and the three-dimensional model of Erg11p encoded by ERG11 gene was also investigated. The sequencing of ERG11 gene revealed the mutation of A395T in this fluconazole-resistant isolate of C. tropicalis, resulting in the Y132F substitution in Erg11p. Sequence alignment and three-dimensional model comparison of Erg11ps showed high similarity between fluconazole-susceptible isolates of C. tropicalis and Candida albicans. The comparison of the three-dimensional models of Erg11ps demonstrated that the position of the Y132F substitution in this isolate of C. tropicalis is identical to the isolate of C. albicans with fluconazole resistance resulting from Y132F substitution in Erg11p. Hence, we ascertain that the Y132F substitution of Erg11p caused by A395T mutation in ERG11 gene confers the fluconazole resistance in C. tropicalis.

  4. Suppression of the hERG potassium channel response to premature stimulation by reduction in extracellular potassium concentration.

    PubMed

    Melgari, Dario; Du, Chunyun; El Harchi, Aziza; Zhang, Yihong; Hancox, Jules C

    2014-10-01

    Potassium channels encoded by human ether-à-go-go-related gene (hERG) mediate the cardiac rapid delayed rectifier K(+) current (IKr), which participates in ventricular repolarization and has a protective role against unwanted premature stimuli late in repolarization and early in diastole. Ionic current carried by hERG channels (IhERG) is known to exhibit a paradoxical dependence on external potassium concentration ([K(+)]e), but effects of acute [K(+)]e changes on the response of IhERG to premature stimulation have not been characterized. Whole-cell patch-clamp measurements of hERG current were made at 37°C from hERG channels expressed in HEK293 cells. Under conventional voltage-clamp, both wild-type (WT) and S624A pore-mutant IhERG during depolarization to +20 mV and subsequent repolarization to -40 mV were decreased when superfusate [K(+)]e was decreased from 4 to 1 mmol/L. When [K(+)]e was increased from 4 to 10 mmol/L, pulse current was increased and tail IhERG was decreased. Increasing [K(+)]e produced a +10 mV shift in voltage-dependent inactivation of WT IhERG and slowed inactivation time course, while lowering [K(+)]e from 4 to 1 mmol/L produced little change in inactivation voltage dependence, but accelerated inactivation time course. Under action potential (AP) voltage-clamp, lowering [K(+)]e reduced the amplitude of IhERG during the AP and suppressed the maximal IhERG response to premature stimuli. Raising [K(+)]e increased IhERG early during the AP and augmented the IhERG response to premature stimuli. Our results are suggestive that during hypokalemia not only is the contribution of IKr to ventricular repolarization reduced but its ability to protect against unwanted premature stimuli also becomes impaired.

  5. HIV Tat protein inhibits hERG K+ channels: a potential mechanism of HIV infection induced LQTs.

    PubMed

    Bai, Yun-Long; Liu, Hui-Bin; Sun, Bo; Zhang, Ying; Li, Qi; Hu, Chao-Wei; Zhu, Jiu-Xin; Gong, Dong-Mei; Teng, Xue; Zhang, Qin; Yang, Bao-Feng; Dong, De-Li

    2011-11-01

    HIV-infected patients have a high prevalence of long QT syndrome (LQTs). hERG K(+) channel encoded by human ether-a-go-go related gene contributes to IKr K(+) currents responsible for the repolarization of cardiomyocytes. Inhibition of hERG K(+) channels leads to LQTs. HIV Tat protein, the virus transactivator protein, plays a pivotal role in AIDS. The aim of the present study is to examine the effects of HIV Tat protein on hERG K(+) channels stably expressed in HEK293 cells. The hERG K(+) currents were recorded by whole-cell patch-clamp technique and the hERG channel expression was measured by real-time PCR and Western blot techniques. HIV Tat protein at 200 ng/ml concentration showed no acute effect on hERG currents, but HIV Tat protein (200 ng/ml) incubation for 24 h significantly inhibited hERG currents. In HIV Tat incubated cells, the inactivation and the recovery time from inactivation of hERG channels were significantly changed. HIV Tat protein incubation (200 ng/ml) for 24h had no effect on the hERG mRNA expression, but dose-dependently inhibited hERG protein expression. The MTT assay showed that HIV Tat protein at 50 ng/ml and 200 ng/ml had no effect on the cell viability. HIV Tat protein increased reactive oxygen species (ROS) generation and the inhibition of hERG channel protein expression by HIV Tat protein was prevented by antioxidant tempol. HIV Tat protein in vivo treatment reduced IKr currents and prolonged action potential duration of guinea pig cardiomyocytes. We conclude that HIV Tat protein inhibits hERG K(+) currents through the inhibition of hERG protein expression, which might be the potential mechanism of HIV infection induced LQTs.

  6. A novel assessment of nefazodone-induced hERG inhibition by electrophysiological and stereochemical method

    SciTech Connect

    Shin, Dae-Seop; Park, Myoung Joo; Lee, Hyang-Ae; Lee, Joo Yun; Chung, Hee-Chung; Yoo, Dae Seok; Chae, Chong Hak; Park, Sang-Joon; Kim, Ki-Suk; Bae, Myung Ae

    2014-02-01

    Nefazodone was used widely as an antidepressant until it was withdrawn from the U.S. market in 2004 due to hepatotoxicity. We have investigated methods to predict various toxic effects of drug candidates to reduce the failure rate of drug discovery. An electrophysiological method was used to assess the cardiotoxicity of drug candidates. Small molecules, including withdrawn drugs, were evaluated using a patch-clamp method to establish a database of hERG inhibition. Nefazodone inhibited hERG channel activity in our system. However, nefazodone-induced hERG inhibition indicated only a theoretical risk of cardiotoxicity. Nefazodone inhibited the hERG channel in a concentration-dependent manner with an IC{sub 50} of 45.3 nM in HEK-293 cells. Nefazodone accelerated both the recovery from inactivation and its onset. Nefazodone also accelerated steady-state inactivation, although it did not modify the voltage-dependent character. Alanine mutants of hERG S6 and pore region residues were used to identify the nefazodone-binding site on hERG. The hERG S6 point mutants Y652A and F656A largely abolished the inhibition by nefazodone. The pore region mutant S624A mildly reduced the inhibition by nefazodone but T623A had little effect. A docking study showed that the aromatic rings of nefazodone interact with Y652 and F656 via π–π interactions, while an amine interacted with the S624 residue in the pore region. In conclusion, Y652 and F656 in the S6 domain play critical roles in nefazodone binding. - Highlights: • Nefazodone inhibits hERG channels with an IC{sub 50} of 45.3 nM in HEK-293 cells. • Nefazodone blocks hERG channels by binding to the open channels. • Y652 and F656 are important for binding of nefazodone. • The aromatic rings of nefazodone interact with Y652 and F656 via π–π interactions.

  7. Postreceptoral contributions to the light-adapted ERG of mice lacking b-waves.

    PubMed

    Shirato, Suguru; Maeda, Hidetaka; Miura, Gen; Frishman, Laura J

    2008-06-01

    The purpose of this study was to determine the contributions of postreceptoral neurons to the light-adapted ERG of the Nob mouse, a model for complete-type congenital stationary night blindness (CSNB1) that lacks a b-wave from depolarizing bipolar cells. Ganzfeld ERGs were recorded from anesthetized adult control mice, control mice injected intravitreally with L-2-amino-4-phosphonobutyric acid (Control APB mice) to remove On pathway activity, and Nob mice. ERGs also were recorded after PDA (cis-2,3-piperidine-dicarboxylic acid, 3-5mM) was injected to block transmission to hyperpolarizing (Off) bipolar and horizontal cells, and all third-order neurons. Stimuli were brief (<4ms, 0.4-2.5log sc td s) and long (200ms, 2.5-4.6log sc td) LED flashes (lambda(max)=513nm, on a rod suppressing background (2.6log sc td). Sinusoidal modulation of the LEDs (mean, 2.6log sc td; contrast, 100%; 3-36Hz) was used to study flicker ERGs. Brief-flash ERGs of Nob mice presented as long-lasting negative waves with a positive-going intrusion that started about 50ms after the flash and peaked around 120ms. Control APB mice had similar responses, and in both cases, PDA removed the positive-going intrusion. For long flashes, PDA removed a small, slow "d-wave" after light offset. With sinusoidal stimulation, the fundamental (F1) amplitude of control mice ERG peaked at 8Hz ( approximately 70microV). For Nob mice the peak was approximately 20microV at 6Hz before PDA and approximately 10muV at 3Hz or lower after PDA. F1 responses were present up to 21Hz in control and Nob eyes and 15Hz in Nob eyes after PDA. Between 3 and 6Hz, F1 phase was 170-210 degrees more delayed in Nob than control mice; phase was hardly altered by PDA. With vector analysis, a substantial postreceptoral input to the Nob flicker ERG was revealed. In control mice, the second harmonic (F2) response showed peaks of approximately 10mocrpV at 3Hz and 13Hz. Nob mice showed almost no F2. In summary, in this study it was found that

  8. Performance of n-in-p Pixel Detectors Irradiated at Fluences up to 5x1015 neq/cm2 for the Future ATLAS Upgrades

    NASA Astrophysics Data System (ADS)

    Macchiolo, A.; Gallrapp, C.; La Rosa, A.; Nisius, R.; Pernegger, H.; Richter, R. H.; Weigell, P.

    We present the results of the characterization of novel n-in-p planar pixel detectors, designed for the future upgrades of the ATLAS pixel system. N-in-p silicon devices are a promising candidate to replace the n-in-n sensors thanks to their radiation hardness and cost effectiveness, that allow for enlarging the area instrumented with pixel detectors. The n-in-p modules presented here are composed of pixel sensors produced by CiS connected by bump-bonding to the ATLAS readout chip FE-I3. The characterization of these devices has been performed with the ATLAS pixel read-out systems, TurboDAQ and USBPIX, before and after irradiation with 25 MeV protons and neutrons up to a fluence of 5x1015 neq/cm2. The charge collection measurements carried out with radioactive sources have proven the feasibility of employing this kind of detectors up to these particle fluences. The collected charge has been measured to be for any fluence in excess of twice the value of the FE-I3 threshold, tuned to 3200 e. The first results from beam test data with 120 GeV pions at the CERN-SPS are also presented, demonstrating a high tracking efficiency before irradiation and a high collected charge for a device irradiated at 1015 neq/cm2. This work has been performed within the framework of the RD50 Collaboration.

  9. 5 × 5 cm2 silicon photonic crystal slabs on glass and plastic foil exhibiting broadband absorption and high-intensity near-fields

    PubMed Central

    Becker, C.; Wyss, P.; Eisenhauer, D.; Probst, J.; Preidel, V.; Hammerschmidt, M.; Burger, S.

    2014-01-01

    Crystalline silicon photonic crystal slabs are widely used in various photonics applications. So far, the commercial success of such structures is still limited owing to the lack of cost-effective fabrication processes enabling large nanopatterned areas (≫ 1 cm2). We present a simple method for producing crystalline silicon nanohole arrays of up to 5 × 5 cm2 size with lattice pitches between 600 and 1000 nm on glass and flexible plastic substrates. Exclusively up-scalable, fast fabrication processes are applied such as nanoimprint-lithography and silicon evaporation. The broadband light trapping efficiency of the arrays is among the best values reported for large-area experimental crystalline silicon nanostructures. Further, measured photonic crystal resonance modes are in good accordance with light scattering simulations predicting strong near-field intensity enhancements greater than 500. Hence, the large-area silicon nanohole arrays might become a promising platform for ultrathin solar cells on lightweight substrates, high-sensitive optical biosensors, and nonlinear optics. PMID:25073935

  10. Tailoring gadolinium-doped ceria-based solid oxide fuel cells to achieve 2 W cm(-2) at 550 °C.

    PubMed

    Lee, Jin Goo; Park, Jeong Ho; Shul, Yong Gun

    2014-06-04

    Low-temperature operation is necessary for next-generation solid oxide fuel cells due to the wide variety of their applications. However, significant increases in the fuel cell losses appear in the low-temperature solid oxide fuel cells, which reduce the cell performance. To overcome this problem, here we report Gd0.1Ce0.9O1.95-based low-temperature solid oxide fuel cells with nanocomposite anode functional layers, thin electrolytes and core/shell fibre-structured Ba0.5Sr0.5Co0.8Fe0.2O3-δ-Gd0.1Ce0.9O1.95 cathodes. In particular, the report describes the use of the advanced electrospinning and Pechini process in the preparation of the core/shell-fibre-structured cathodes. The fuel cells show a very high performance of 2 W cm(-2) at 550 °C in hydrogen, and are stable for 300 h even under the high current density of 1 A cm(-2). Hence, the results suggest that stable and high-performance solid oxide fuel cells at low temperatures can be achieved by modifying the microstructures of solid oxide fuel cell components.

  11. AMINO ACID ANALYSES OF THE ANTARCTIC CM2 METEORITES ALH 83100 AND LEW 90500 USING LIQUID CHROMATOGRAPHY-TIME OF FLIGHT-MASS SPECTROMETRY

    NASA Technical Reports Server (NTRS)

    Glavin, D. P.; Dworkin, J. P.; Aubrey, A.; Botta, O.; Doty, J. H., III; Bada, J. L.

    2001-01-01

    The investigation of organic compounds in primitive carbonaceous meteorites provides a record of the chemical processes that occurred in the early solar system. In particular, amino acids have been shown to be potential indicators in tracing the nature of carbonaceous chondrite parent bodies [ 13. The delivery of amino acids by carbonaceous chondrites to the early Earth could have been any important source of the Earth's prebiotic organic inventory [2]. Over 80 different amino acids have been detected in the Murchison CM2 meteorite, most of them completely non-existent in the terrestrial biosphere [3]. We have optimized a new liquid chromatography-time-of-flight-mass spectrometry (LC-ToF-MS) technique coupled with OPAMAC derivatization in order to detect amino acids in meteorite extracts by UV fluorescence and exact mass simultaneously. The detection limit of the LC-ToF-MS instrument for amino acids is at least 3 orders of magnitude lower than traditional GC-MS techniques. Here we report on the first analyses of amino acids and their enantiomeric abundances in the CM2 carbonaceous meteorites ALH 83100, LEW 90500, and Murchison using this new LC-ToF-MS instrument configuration. Amino acid analyses of any kind for the CM meteorite ALH 83100 have not previously been reported.

  12. Current density and catalyst-coated membrane resistance distribution of hydro-formed metallic bipolar plate fuel cell short stack with 250 cm2 active area

    NASA Astrophysics Data System (ADS)

    Haase, S.; Moser, M.; Hirschfeld, J. A.; Jozwiak, K.

    2016-01-01

    An automotive fuel cell with an active area of 250 cm2 is investigated in a 4-cell short stack with a current and temperature distribution device next to the bipolar plate with 560 current and 140 temperature segments. The electrical conductivities of the bipolar plate and gas diffusion layer assembly are determined ex-situ with this current scan shunt module. The applied fuel cell consists of bipolar plates constructed of 75-μm-thick, welded stainless-steel foils and a graphitic coating. The electrical conductivities of the bipolar plate and gas diffusion layer assembly are determined ex-situ with this module with a 6% deviation in in-plane conductivity. The current density distribution is evaluated up to 2.4 A cm-2. The entire cell's investigated volumetric power density is 4.7 kW l-1, and its gravimetric power density is 4.3 kW kg-1 at an average cell voltage of 0.5 V. The current density distribution is determined without influencing the operating cell. In addition, the current density distribution in the catalyst-coated membrane and its effective resistivity distribution with a finite volume discretisation of Ohm's law are evaluated. The deviation between the current density distributions in the catalyst-coated membrane and the bipolar plate is determined.

  13. ERG dependence distinguishes developmental control of hematopoietic stem cell maintenance from hematopoietic specification

    PubMed Central

    Taoudi, Samir; Bee, Thomas; Hilton, Adrienne; Knezevic, Kathy; Scott, Julie; Willson, Tracy A.; Collin, Caitlin; Thomas, Tim; Voss, Anne K.; Kile, Benjamin T.; Alexander, Warren S.; Pimanda, John E.; Hilton, Douglas J.

    2011-01-01

    Although many genes are known to be critical for early hematopoiesis in the embryo, it remains unclear whether distinct regulatory pathways exist to control hematopoietic specification versus hematopoietic stem cell (HSC) emergence and function. Due to their interaction with key regulators of hematopoietic commitment, particular interest has focused on the role of the ETS family of transcription factors; of these, ERG is predicted to play an important role in the initiation of hematopoiesis, yet we do not know if or when ERG is required. Using in vitro and in vivo models of hematopoiesis and HSC development, we provide strong evidence that ERG is at the center of a distinct regulatory program that is not required for hematopoietic specification or differentiation but is critical for HSC maintenance during embryonic development. We show that, from the fetal period, ERG acts as a direct upstream regulator of Gata2 and Runx1 gene activity. Without ERG, physiological HSC maintenance fails, leading to the rapid exhaustion of definitive hematopoiesis. PMID:21245161

  14. Early lineage priming by trisomy of Erg leads to myeloproliferation in a Down syndrome model.

    PubMed

    Ng, Ashley P; Hu, Yifang; Metcalf, Donald; Hyland, Craig D; Ierino, Helen; Phipson, Belinda; Wu, Di; Baldwin, Tracey M; Kauppi, Maria; Kiu, Hiu; Di Rago, Ladina; Hilton, Douglas J; Smyth, Gordon K; Alexander, Warren S

    2015-05-01

    Down syndrome (DS), with trisomy of chromosome 21 (HSA21), is the commonest human aneuploidy. Pre-leukemic myeloproliferative changes in DS foetal livers precede the acquisition of GATA1 mutations, transient myeloproliferative disorder (DS-TMD) and acute megakaryocytic leukemia (DS-AMKL). Trisomy of the Erg gene is required for myeloproliferation in the Ts(1716)65Dn DS mouse model. We demonstrate here that genetic changes specifically attributable to trisomy of Erg lead to lineage priming of primitive and early multipotential progenitor cells in Ts(1716)65Dn mice, excess megakaryocyte-erythroid progenitors, and malignant myeloproliferation. Gene expression changes dependent on trisomy of Erg in Ts(1716)65Dn multilineage progenitor cells were correlated with those associated with trisomy of HSA21 in human DS hematopoietic stem and primitive progenitor cells. These data suggest a role for ERG as a regulator of hematopoietic lineage potential, and that trisomy of ERG in the context of DS foetal liver hemopoiesis drives the pre-leukemic changes that predispose to subsequent DS-TMD and DS-AMKL.

  15. Clinical S-cone ERG recording with a commercial hand-held full-field stimulator.

    PubMed

    Marmor, Michael F; Cabael, Lorella; Shukla, Shefalee; Hwang, John C; Marcus, Mira

    2004-07-01

    Our purpose was to explore S-cone ERG protocols for a commercial full-field hand-held stimulator that contains colored LEDs, and to see whether the test would be useful as a part of routine ERG testing. S-cone responses were elicited by blue flashes over a longer-wavelength background. With the standard stimulator containing blue (461 nm), green (513 nm) and red (652 nm) LEDs, we were unable to obtain satisfactory responses. Reproducible S-cone ERGs were obtained with a stimulator that had been custom-fitted with shorter-wavelength blue (440 nm) LEDs for stimulation, and orange (590 nm) LEDs for background adaptation. S-cone responses took only a few minutes to record, and the typical waveform showed a slow peak at 45-50 ms with amplitude 3-9 microV, but ranging from 0 microV to more than 10 microV. Larger waves appeared in a patient with enhanced S-cone syndrome. S-cone responses could also be obtained with an alternating blue-orange flicker protocol. We added the S-cone response to our regular ERG protocol for a number of months. Although most normal subjects and patients showed recognizable S-cone responses with this stimulator, the amplitudes were small and there was too much variability to make the technique effective for routine clinical testing. In general, the S-cone responses followed the standard cone ERG responses in disease.

  16. ERG Expression Levels in Prostate Tumors Reflect Functional Status of the Androgen Receptor (AR) as a Consequence of Fusion of ERG with AR Regulated Gene Promoters

    DTIC Science & Technology

    2010-01-01

    literature review. Asian J Androl 2008; 10: 855-63. [30] Donovan MJ , Osman I, Khan FM, et al. Androgen receptor expression is associated with...Nat Rev Urol 2009; 6: 429-39. [8] Heinlein CA, Chang C. Androgen receptor in prostate cancer. Endocr Rev 2004; 25: 276-308. [9] Linja MJ , Visakorpi...Klezovitch O , Risk M, Coleman I, et al. A causal role for ERG in neoplastic transformation of prostate epithelium. Proc Natl Acad Sci USA 2008; 105: 2105

  17. Monoallelic expression of TMPRSS2/ERG in prostate cancer stem cells.

    PubMed

    Polson, Euan S; Lewis, John L; Celik, Hamza; Mann, Vincent M; Stower, Michael J; Simms, Matthew S; Rodrigues, Greta; Collins, Anne T; Maitland, Norman J

    2013-01-01

    While chromosomal translocations have a fundamental role in the development of several human leukaemias, their role in solid tumour development has been somewhat more controversial. Recently, it was shown that up to 80% of prostate tumours harbour at least one such gene fusion, and that the most common fusion event, between the prostate-specific TMPRSS2 gene and the ERG oncogene, is a critical, and probably early factor in prostate cancer development. Here we demonstrate the presence and expression of this significant chromosomal rearrangement in prostate cancer stem cells. Moreover, we show that in the prostate epithelial hierarchy from both normal and tumour tissues, TMPRSS2 transcription is subjected to tight monoallelic regulation, which is retained upon asymmetric division and relaxed during epithelial cell differentiation. The presence and expression of TMPRSS2/ERG in prostate stem cells would provide ERG-driven survival advantages, allowing maintenance of this mutated genotype.

  18. Continuous-wavelet-transform analysis of the multifocal ERG waveform in glaucoma diagnosis.

    PubMed

    Miguel-Jiménez, J M; Blanco, R; De-Santiago, L; Fernández, A; Rodríguez-Ascariz, J M; Barea, R; Martín-Sánchez, J L; Amo, C; Sánchez-Morla, E; Boquete, L

    2015-09-01

    The vast majority of multifocal electroretinogram (mfERG) signal analyses to detect glaucoma study the signals' amplitudes and latencies. The purpose of this paper is to investigate application of wavelet analysis of mfERG signals in diagnosis of glaucoma. This analysis method applies the continuous wavelet transform (CWT) to the signals, using the real Morlet wavelet. CWT coefficients resulting from the scale of maximum correlation are used as inputs to a neural network, which acts as a classifier. mfERG recordings are taken from the eyes of 47 subjects diagnosed with chronic open-angle glaucoma and from those of 24 healthy subjects. The high sensitivity in the classification (0.894) provides reliable detection of glaucomatous sectors, while the specificity achieved (0.844) reflects accurate detection of healthy sectors. The results obtained in this paper improve on the previous findings reported by the authors using the same visual stimuli and database.

  19. The role of the ERG in the diagnosis and treatment of Age-Related Macular Degeneration.

    PubMed

    Gerth, Christina

    2009-02-01

    Age-related macular degeneration (AMD) is affecting an increasing number of people, with 2.95 million people estimated to be affected in the USA by 2020. Possible preventive agents, such as vitamins and supplements have been studied and new treatment options for AMD have been developed in recent years. What role does electrophysiology play as a sensitive outcome measure? The most commonly used tests are the full-field electroretinogram (ffERG) and the multifocal ERG (mfERG). Test results from patients with AMD and reduced central vision need special attention in respect to fixation pattern, age-matched control data, and retinal luminance. Advantages, disadvantages and limitations of techniques will be considered, together with a review of published studies.

  20. The oxygen isotope evolution of parent body aqueous solutions as recorded by multiple carbonate generations in the Lonewolf Nunataks 94101 CM2 carbonaceous chondrite

    NASA Astrophysics Data System (ADS)

    Lee, M. R.; Sofe, M. R.; Lindgren, P.; Starkey, N. A.; Franchi, I. A.

    2013-11-01

    The CM2 carbonaceous chondrite LON 94101 contains aragonite and two generations of calcite that provide snapshots of the chemical and isotopic evolution of aqueous solutions during parent body alteration. Aragonite was the first carbonate to crystallize. It is rare, heterogeneously distributed within the meteorite matrix, and its mean oxygen isotope values are δ18O 39.9 ± 0.6‰, Δ17O -0.3 ± 1.0‰ (1σ). Calcite precipitated soon afterwards, and following a fall in solution Mg/Ca ratios, to produce small equant grains with a mean oxygen isotope value of δ18O 37.5 ± 0.7‰, Δ17O 1.4 ± 1.1‰ (1σ). These grains were partially or completely replaced by serpentine and tochilinite prior to precipitation of the second generation of calcite, which occluded an open fracture to form a millimetre-sized vein, and replaced anhydrous silicates within chondrules and the matrix. The vein calcite has a mean composition of δ18O 18.4 ± 0.3‰, Δ17O -0.5 ± 0.5‰ (1σ). Petrographic and isotopic results therefore reveal two discrete episodes of mineralisation that produced calcite generations with contrasting δ18O, and mean Δ17O values. The aragonite and equant calcite crystallized over a relatively brief period early in the aqueous alteration history of the parent body, and from static fluids that were evolving chemically in response to mineral dissolution and precipitation. The second calcite generation crystallized from solutions of a lower Δ17O, and a lower δ18O and/or higher temperature. As two generations of calcite whose petrographic characteristics and oxygen isotopic compositions are similar to those in LON 94101 occur in at least one other CM2, multiphase carbonate mineralisation could be the typical outcome of the sequence of chemical reactions during parent body aqueous alteration. It is equally possible however that the second generation of calcite formed in response to an event such as impact fracturing and concomitant fluid mobilisation that affected

  1. Rab11-dependent Recycling of the Human Ether-a-go-go-related Gene (hERG) Channel*

    PubMed Central

    Chen, Jeffery; Guo, Jun; Yang, Tonghua; Li, Wentao; Lamothe, Shawn M.; Kang, Yudi; Szendrey, John A.; Zhang, Shetuan

    2015-01-01

    The human ether-a-go-go-related gene (hERG) encodes the pore-forming subunit of the rapidly activating delayed rectifier potassium channel (IKr). A reduction in the hERG current causes long QT syndrome, which predisposes affected individuals to ventricular arrhythmias and sudden death. We reported previously that hERG channels in the plasma membrane undergo vigorous internalization under low K+ conditions. In the present study, we addressed whether hERG internalization occurs under normal K+ conditions and whether/how internalized channels are recycled back to the plasma membrane. Using patch clamp, Western blot, and confocal imaging analyses, we demonstrated that internalized hERG channels can effectively recycle back to the plasma membrane. Low K+-enhanced hERG internalization is accompanied by an increased rate of hERG recovery in the plasma membrane upon reculture following proteinase K-mediated clearance of cell-surface proteins. The increased recovery rate is not due to enhanced protein synthesis, as hERG mRNA expression was not altered by low K+ exposure, and the increased recovery was observed in the presence of the protein biosynthesis inhibitor cycloheximide. GTPase Rab11, but not Rab4, is involved in the recycling of hERG channels. Interfering with Rab11 function not only delayed hERG recovery in cells after exposure to low K+ medium but also decreased hERG expression and function in cells under normal culture conditions. We concluded that the recycling pathway plays an important role in the homeostasis of plasma membrane-bound hERG channels. PMID:26152716

  2. Structure and Function of the Splice Variants of TMPRSS2-ERG, a Prevalent Genomic Alteration in Prostate Cancer

    DTIC Science & Technology

    2009-09-01

    analysis of TMPRSS2-ERG expression in prostate tumors. Quantitative analyses of the transcript levels of various TMPRSS2-ETS fusion genes were done in LCM ...variables of the patient cohort are summarized in Supplementary Table S1. The workflow of LCM and quantitative reverse transcription-PCR analysis is...between TMPRSS2 and ERG have been identified and few of these fusions result into deletion of ~33-39 amino acids at the N- terminal end of ERG

  3. Structure and Function of the Splice Variants of TMPRSS2-ERG, a Prevalent Genomic Alteration in Prostate Cancer

    DTIC Science & Technology

    2011-09-01

    Bieberich4, Isabell A. Sesterhenn5, Albert Dobi1, Shiv Srivastava1 and Taduru L. Sreenath1  1. Center For Prostate Disease Research, Department of...Expression of human ERG protein and consequently its functions in normal and disease states needs to be better understood in light of its suggested...functions in normal and disease conditions. Key words: Ets Related Gene, ERG, Expression, ERG MAb, Mouse, Development. Introduction Chromosomal

  4. External protons destabilize the activated voltage sensor in hERG channels.

    PubMed

    Shi, Yu Patrick; Cheng, Yen May; Van Slyke, Aaron C; Claydon, Tom W

    2014-03-01

    Extracellular acidosis shifts hERG channel activation to more depolarized potentials and accelerates channel deactivation; however, the mechanisms underlying these effects are unclear. External divalent cations, e.g., Ca(2+) and Cd(2+), mimic these effects and coordinate within a metal ion binding pocket composed of three acidic residues in hERG: D456 and D460 in S2 and D509 in S3. A common mechanism may underlie divalent cation and proton effects on hERG gating. Using two-electrode voltage clamp, we show proton sensitivity of hERG channel activation (pKa = 5.6), but not deactivation, was greatly reduced in the presence of Cd(2+) (0.1 mM), suggesting a common binding site for the Cd(2+) and proton effect on activation and separable effects of protons on activation and deactivation. Mutational analysis confirmed that D509 plays a critical role in the pH dependence of activation, as shown previously, and that cooperative actions involving D456 and D460 are also required. Importantly, neutralization of all three acidic residues abolished the proton-induced shift of activation, suggesting that the metal ion binding pocket alone accounts for the effects of protons on hERG channel activation. Voltage-clamp fluorimetry measurements demonstrated that protons shifted the voltage dependence of S4 movement to more depolarized potentials. The data indicate a site and mechanism of action for protons on hERG activation gating; protonation of D456, D460 and D509 disrupts interactions between these residues and S4 gating charges to destabilize the activated configuration of S4.

  5. Functional Consequences of Methionine Oxidation of hERG Potassium Channels

    PubMed Central

    Su, Zhi; Limberis, James; Martin, Ruth L.; Xu, Rong; Kolbe, Katrin; Heinemann, Stefan H.; Hoshi, Toshinori; Cox, Bryan F.; Gintant, Gary A.

    2010-01-01

    Reactive species oxidatively modify numerous proteins including ion channels. Oxidative sensitivity of ion channels is often conferred by amino acids containing sulfur atoms, such as cysteine and methionine. Functional consequences of oxidative modification of methionine in hERG1 (human ether à go-go related gene 1), which encodes cardiac IKr channels, are unknown. Here we used chloramine-T (ChT), which preferentially oxidizes methionine, to examine the functional consequences of methionine oxidation of hERG channels stably expressed in a human embryonic kidney cell line (HEK 293) and native hERG channels in a human neuroblastoma cell line (SH-SY5Y). ChT (300 µM) significantly decreased whole-cell hERG current in both HEK 293 and SH-SY5Y cells. In HEK 293 cells, the effects of ChT on hERG current were time- and concentration-dependent, and were markedly attenuated in the presence of enzyme methionine sulfoxide reductase A that specifically repairs oxidized methionine. After treatment with ChT, the channel deactivation upon repolarization to −60 or −100 mV was significantly accelerated. The effect of ChT on channel activation kinetics was voltage-dependent; activation slowed during depolarization to +30 mV but accelerated during depolarization to 0 or −10 mV. In contrast, the reversal potential, inactivation kinetics, and voltage-dependence of steady-state inactivation remained unaltered. Our results demonstrate that the redox status of methionine is an important modulator of hERG channel. PMID:17624316

  6. ERG promotes T-acute lymphoblastic leukemia and is transcriptionally regulated in leukemic cells by a stem cell enhancer.

    PubMed

    Thoms, Julie A I; Birger, Yehudit; Foster, Sam; Knezevic, Kathy; Kirschenbaum, Yael; Chandrakanthan, Vashe; Jonquieres, Georg; Spensberger, Dominik; Wong, Jason W; Oram, S Helen; Kinston, Sarah J; Groner, Yoram; Lock, Richard; MacKenzie, Karen L; Göttgens, Berthold; Izraeli, Shai; Pimanda, John E

    2011-06-30

    The Ets-related gene (ERG) is an Ets-transcription factor required for normal blood stem cell development. ERG expression is down-regulated during early T-lymphopoiesis but maintained in T-acute lymphoblastic leukemia (T-ALL), where it is recognized as an independent risk factor for adverse outcome. However, it is unclear whether ERG is directly involved in the pathogenesis of T-ALL and how its expression is regulated. Here we demonstrate that transgenic expression of ERG causes T-ALL in mice and that its knockdown reduces the proliferation of human MOLT4 T-ALL cells. We further demonstrate that ERG expression in primary human T-ALL cells is mediated by the binding of other T-cell oncogenes SCL/TAL1, LMO2, and LYL1 in concert with ERG, FLI1, and GATA3 to the ERG +85 enhancer. This enhancer is not active in normal T cells but in transgenic mice targets expression to fetal liver c-kit(+) cells, adult bone marrow stem/progenitors and early CD4(-)CD8(-) double-negative thymic progenitors. Taken together, these data illustrate that ERG promotes T-ALL and that failure to extinguish activity of stem cell enhancers associated with regulatory transcription factors such as ERG can contribute to the development of leukemia.

  7. Three-dimensional nanostructured bilayer solid oxide fuel cell with 1.3 W/cm(2) at 450 °C.

    PubMed

    An, Jihwan; Kim, Young-Beom; Park, Joonsuk; Gür, Turgut M; Prinz, Fritz B

    2013-09-11

    Obtaining high power density at low operating temperatures has been an ongoing challenge in solid oxide fuel cells (SOFC), which are efficient engines to generate electrical energy from fuels. Here we report successful demonstration of a thin-film three-dimensional (3-D) SOFC architecture achieving a peak power density of 1.3 W/cm(2) obtained at 450 °C. This is made possible by nanostructuring of the ultrathin (60 nm) electrolyte interposed with a nanogranular catalytic interlayer at the cathode/electrolyte interface. We attribute the superior cell performance to significant reduction in both the ohmic and the polarization losses due to the combined effects of employing an ultrathin film electrolyte, enhancement of effective area by 3-D architecture, and superior catalytic activity by the ceria-based interlayer at the cathode. These insights will help design high-efficiency SOFCs that operate at low temperatures with power densities that are of practical significance.

  8. Compositions of Partly Altered Olivine and Replacement Serpentine in the CM2 Chondrites QUE93005 and Nogoya: Implications for Scales of Elemental Redistribution During Aqueous Alteration

    NASA Technical Reports Server (NTRS)

    Velbel, M. A.; Tonui, E. K.; Zolensky, M. E.

    2003-01-01

    Some phyllosilicates in CM carbonaceous chondrites formed by aqueous alteration of anhydrous precursor phases. Although broad trends in the compositions of hydrous phyllosilicates are recognized and believed to be related to trends in degree of aqueous alteration, details of the reactions that formed specific secondary minerals remain obscure. This paper reports compositional relationships between remnants of partially pseudomorphically replaced silicates and their alteration products (serpentine) in the CM2 chondrites QUE93005 and Nogoya and compares both with previously published results for Allan Hills 81002. By focusing on serpentine formed from known reactants (olivines), and on only those instances in which some of the reactant silicate remains, direct compositional relationships between reactants and products, and the elemental mobility required by the reactions, can be established.

  9. Oncogenicity and Selective Inhibition of ERG Splicing Variants in Prostate Cancer

    DTIC Science & Technology

    2012-03-01

    Genet 41: 619-624. 9. Demichelis F, Fall K, Perner S, Andren O, Schmidt F, et al. (2007) TMPRSS2:ERG gene fusion associated with lethal prostate...cancer in a watchful waiting cohort. Oncogene 26: 4596-4599. 10. Nam RK, Sugar L, Wang Z, Yang W, Kitching R, et al. (2007) Expression of TMPRSS2:ERG...Dotan ZA, Koutcher JA, Di Cristofano A, et al. (2003) Pten dose dictates cancer progression in the prostate. PLoS Biol 1: E59. 15. Squire JA (2009

  10. Strand V: Education for Survival. First Aid and Survival Education. Health Curriculum Materials Grades 10-12.

    ERIC Educational Resources Information Center

    New York State Education Dept., Albany. Bureau of Secondary Curriculum Development.

    GRADES OR AGES: Grades 10-12. SUBJECT MATTER: First aid and survival education. ORGANIZATION AND PHYSICAL APPEARANCE: The guide is divided into six sections: transportation of the injured, automobile accidents, conditions resulting from nuclear explosion, chemical warfare, natural catastrophes, and psychological first aid. The publication format…

  11. Dynamics of Clothing II. Curriculum Guide. A Family and Consumer Sciences Education Course of Study for Grades 10-12.

    ERIC Educational Resources Information Center

    Hunger, Dean-Ellen, Ed.; Hancey, Helen-Louise; Hendrickson, Diane; Hicks, Camille; Munns, Barbara; Price, Barbara

    This document is a nine-unit curriculum guide for a high school (grades 10-12) course in clothing instruction. The units contain one or two lessons on the following topics: (1) psychological aspects of clothing (behavior, image, and dress; self-concept and self-image); (2) wardrobe selections (wardrobe consumerism, wardrobe evaluation and…

  12. Biological and Genomic Differences of ERG Oncoprotein-Stratified Prostate Cancers from African and Caucasian Americans

    DTIC Science & Technology

    2015-10-01

    It is anticipated that molecular determinants of aggressive prostate cancer in African American men, including somatic mutations and SNPs associated...molecular stratification, germline variants (SNPs), admixture mapping, European and African ancestry, somatic mutations, aggressive cancer... aggressive CaP in AA men include somatic mutations (TMPRSS2-ERG) and germline variants (SNPs). The objective will be achieved by the following specific

  13. Role of the pH in state-dependent blockade of hERG currents

    NASA Astrophysics Data System (ADS)

    Wang, Yibo; Guo, Jiqing; Perissinotti, Laura L.; Lees-Miller, James; Teng, Guoqi; Durdagi, Serdar; Duff, Henry J.; Noskov, Sergei Yu.

    2016-10-01

    Mutations that reduce inactivation of the voltage-gated Kv11.1 potassium channel (hERG) reduce binding for a number of blockers. State specific block of the inactivated state of hERG block may increase risks of drug-induced Torsade de pointes. In this study, molecular simulations of dofetilide binding to the previously developed and experimentally validated models of the hERG channel in open and open-inactivated states were combined with voltage-clamp experiments to unravel the mechanism(s) of state-dependent blockade. The computations of the free energy profiles associated with the drug block to its binding pocket in the intra-cavitary site display startling differences in the open and open-inactivated states of the channel. It was also found that drug ionization may play a crucial role in preferential targeting to the open-inactivated state of the pore domain. pH-dependent hERG blockade by dofetilie was studied with patch-clamp recordings. The results show that low pH increases the extent and speed of drug-induced block. Both experimental and computational findings indicate that binding to the open-inactivated state is of key importance to our understanding of the dofetilide’s mode of action.

  14. Hematopoietic overexpression of the transcription factor Erg induces lymphoid and erythro-megakaryocytic leukemia

    PubMed Central

    Carmichael, Catherine L.; Metcalf, Donald; Henley, Katya J.; Kruse, Elizabeth A.; Di Rago, Ladina; Mifsud, Sandra; Alexander, Warren S.; Kile, Benjamin T.

    2012-01-01

    The transcription factor encoded by the E-twenty-six (ETS)-related gene, ERG, is an essential regulator of hematopoietic stem cell function and a potent human oncoprotein. Enforced expression of ERG in murine hematopoietic cells leads to the development of a well-characterized lymphoid leukemia and a less well-defined non lymphoid disease. To clarify the latter, we generated murine bone marrow chimeras with enforced Erg expression in engrafted hematopoietic progenitor cells. As expected, these mice developed lymphoid leukemia. However, the previously reported non lymphoid disease that developed was shown to be a uniform, transplantable leukemia with both erythroid and megakaryocytic characteristics. In vivo, this disease had the overall appearance of an erythroleukemia, with an accumulation of immature erythroblasts that infiltrated the bone marrow, spleen, liver, and lung. However, when stimulated in vitro, leukemic cell clones exhibited both erythroid and megakaryocytic differentiation, suggesting that transformation occurred in a bipotential progenitor. Thus, in mice, Erg overexpression induces the development of not only lymphoid leukemia but also erythro-megakaryocytic leukemia. PMID:22936051

  15. Small Molecule Inhibitors of ERG and ETV1 in Prostate Cancer

    DTIC Science & Technology

    2013-09-01

    potential novel targets for treatment of primary and/or metastatic disease in prostate cancer. We developed small molecule inhibitors that target...3396-400. 6. Demichelis F, Fall K, Perner S, Andren O, Schmidt F, Setlur SR, et al. TMPRSS2:ERG gene fusion associated with lethal prostate cancer in

  16. The oncogenic role of the ETS transcription factors MEF and ERG.

    PubMed

    Sashida, Goro; Bazzoli, Elena; Menendez, Silvia; Liu, Yan; Nimer, Stephen D

    2010-09-01

    Several ETS transcription factors, including MEF/ELF4 and ERG, can function as oncogenes and are overexpressed in human cancer. MEF cooperates in tumorigenesis in retroviral insertional mutagenesis-based mouse models of cancer and MEF is overexpressed in human lymphoma and ovarian cancer tissues via unknown mechanisms. ERG (Ets related gene) overexpression or increased activity has been found in various human cancers, including sarcomas, acute myeloid leukemia and prostate cancer, where the ERG gene is rearranged due to chromosomal translocations. We have been examining how MEF functions as an oncogene and recently showed that MEF can cooperate with H-Ras(G12V) and can inhibit both p53 and p16 expression thereby promoting transformation. In fact, in cells lacking p53, the absence of Mef abrogates H-Ras(G12V)-induced transformation of mouse embryonic fibroblasts, at least in part due to increased p16 expression. We discuss the known mechanisms by which the ETS transcription factors MEF and ERG contribute to the malignant transformation of cells.

  17. Evaluating the Efficacy of ERG Targeted Therapy In Vivo for Prostate Tumors

    DTIC Science & Technology

    2014-04-01

    performed and transcription of ERG confirmed with specimens using qPCR. IHC for cleaved caspase 3 (CC3) and Ki-67. Senescence markers such as p15, p16 ...luciferase, CC3, Ki-67, p15, p16 , p21 and p27. Whole lobe and organ Western blotting using the same antibodies will also be performed and transcription of

  18. The discovery of CCR3/H1 dual antagonists with reduced hERG risk.

    PubMed

    Bahl, Ash; Barton, Patrick; Bowers, Keith; Brough, Steven; Evans, Richard; Luckhurst, Christopher A; Mochel, Tobias; Perry, Matthew W D; Rigby, Aaron; Riley, Robert J; Sanganee, Hitesh; Sisson, Adam; Springthorpe, Brian

    2012-11-01

    A series of dual CCR3/H(1) antagonists based on a bispiperidine scaffold were discovered. Introduction of an acidic group overcame hERG liability. Bioavailability was optimised by modulation of physico-chemical properties and physical form to deliver a compound suitable for clinical evaluation.

  19. Correlations between ERG, OCT, and Anatomical Findings in the rd10 Mouse

    PubMed Central

    Johnen, Sandra; Müller, Frank; Pfarrer, Christiane

    2014-01-01

    Background. To evaluate the correlation between ERG, OCT, and microscopic findings in the rd10 mouse. Methods. C57BL/6J wild type mice and rd10 mice were compared at the age of 2, 3, 5, 7, 9, 12, 24, and 48 weeks (each age group n = 3) using full-field electroretinography (ERG), spectral domain Optical Coherence Tomography (sd-OCT), fluorescein angiography (FA), Hematoxylin & Eosin histology (HE), and immunohistology (IH). Results. While in wild type mice, the amplitude of a- and b-wave increased with light intensity and with the age of the animals, the rd10 mice showed extinction of the ERG beginning with the age of 5 weeks. In OCT recordings, the thickness of the retina decreased up to 9 weeks of age, mainly based on the degradation of the outer nuclear layer (ONL). Afterwards, the ONL was no longer visible in the OCT. HE staining and immunohistological findings confirmed the in vivo data. Conclusion. ERG and OCT are useful methods to evaluate the retinal function and structure in vivo. The retinal changes seen in the OCT closely match those observed in histological staining. PMID:24683495

  20. Quantitative Gene Expression of ERG9 in Model Saccharomyces cerevisiae: Chamomile Extract For Human Cancer Treatment

    PubMed Central

    Hosseinpour, Maryam; Mobini-Dehkordi, Mohsen

    2016-01-01

    Introduction Over expression of squalene synthase gene causes induction of growth tumour and reduction of apoptosis. This gene which is conserved between Saccharomyces cerevisiae yeast and humans, is named (ERG9). Aim In this work, we studied the effect of Matricaria recutita extract on ERG9 gene (squalene synthase) expression in S.cerevisiae which was used as organism model in cancer therapy. Materials and Methods S. cerevisiae was cultured in YPD medium plus 0,250, 1000 and 3000 μg/ml of Matricaria recutita extract and we evaluated the (ERG9) gene expression by Real-time RT-PCR method after 24 hours. Statistical analysis used At least 3 independent experiments were done. Data were analyzed using One-way ANOVA and Dunnett’s test. A p-value of less than 0.01 was considered as significant. Results We found that 250, 1000 and 3000 μg/ml of Matricaria recutita extract could reduce expression of ERG9 gene significantly (p<0.01). Interestingly, the expression of this gene was completely inhibited in 1000 and 3000 μg/ml concentrations. Conclusion This study predicted that Matricaria recutita extract produced anti-cancer effects in humans, because it could inhibit the expression of an analogue key gene in this malignant disease. Further investigations should be made, to study its molecular mechanism of action at the mammal cell level. PMID:27630863

  1. ERG--Energy Resources Game: Simulation Gaming of Regional Energy Management

    ERIC Educational Resources Information Center

    Wolf, Lyle P.; Laessig, Robert E.

    1973-01-01

    ERG--the Energy Resources Game--is a computer based game which explores questions regarding regional energy supply and demand, such as population and economic growth goals; acceptable levels of dependence on imported energy; and acceptable levels of environmental impact. (JA)

  2. Effect of Voriconazole on Candida tropicalis Biofilms: Relation with ERG Genes Expression.

    PubMed

    Fernandes, Tânia; Silva, Sónia; Henriques, Mariana

    2016-10-01

    Candida tropicalis has emerged as the third most prevalent fungal pathogens and its ability to form biofilms has been considered one of the most important virulence factors, since biofilms represent high tolerance to antifungal agents. However, the mechanisms of C. tropicalis biofilm resistance to antifungals remain poorly understood. Thus, the main aim of this work was to infer about the effect of voriconazole on the formation and control of C. tropicalis biofilms and disclose its relationship with ERG genes' expression. Planktonic cells tolerance of several C. tropicalis clinical isolates to voriconazole was determined through of antifungal susceptibility test, and the effect of this azole against C. tropicalis biofilm formation and pre-formed biofilms was evaluated by cultivable cells determination and total biomass quantification. ERG genes expression was analyzed by quantitative real-time polymerase chain reaction. This work showed that C. tropicalis resistance to voriconazole is strain dependent and that voriconazole was able to partially control biofilm formation, but was unable to eradicate C. tropicalis pre-formed biofilms. Moreover, C. tropicalis biofilms resistance to voriconazole seems to be associated with alterations of sterol content in the cell membrane, resulting in ERG genes overexpression. Voriconazole is unable to control C. tropicalis biofilms, and the overexpression of ERG genes is a possible mechanism of biofilm resistance.

  3. Screening for diabetic retinopathy using new mydriasis-free, full-field flicker ERG recording device

    PubMed Central

    Fukuo, Motonobu; Kondo, Mineo; Hirose, Akira; Fukushima, Harumi; Ikesugi, Kengo; Sugimoto, Masahiko; Kato, Kumiko; Uchigata, Yasuko; Kitano, Shigehiko

    2016-01-01

    Diabetic retinopathy (DR) is a leading cause of blindness among working-age adults. Therefore, it is important to detect DR accurately during mass screening. The purpose of this study was to determine whether a small, hand-held, mydriasis-free, full-field flicker electroretinographic (ERGs) device called RETeval can be used to screen for DR. To accomplish this, we recorded full-field flicker ERGs with this device from 48 normal eyes and 118 eyes with different severities of DR in patients with diabetes mellitus (DM). This system delivered a constant flash retinal luminance by adjusting the flash luminance that compensated for changes in the pupil size. Our results showed that there were significant correlations between the severity of DR and the implicit times (P < 0.001; r = 0.55) and the amplitudes (P = 0.001; r = −0.29). When the implicit time was used for the index, the area under the receiver operating characteristic curve was 0.84 for the detection of DR, and was 0.89 for the detection of DR requiring ophthalmic treatments. These results suggest that the implicit times of the flicker ERGs recorded by the small, mydryasis-free ERG system can be used as an adjunctive tool to screen for DR. PMID:27824158

  4. Modeling the Mechanism of GR/c-Jun/Erg Crosstalk in Apoptosis of Acute Lymphoblastic Leukemia.

    PubMed

    Chen, Daphne Wei-Chen; Krstic-Demonacos, Marija; Schwartz, Jean-Marc

    2012-01-01

    Acute lymphoblastic leukemia (ALL) is one of the most common forms of malignancy that occurs in lymphoid progenitor cells, particularly in children. Synthetic steroid hormones glucocorticoids (GCs) are widely used as part of the ALL treatment regimens due to their apoptotic function, but their use also brings about various side effects and drug resistance. The identification of the molecular differences between the GCs responsive and resistant cells therefore are essential to decipher such complexity and can be used to improve therapy. However, the emerging picture is complicated as the activities of genes and proteins involved are controlled by multiple factors. By adopting the systems biology framework to address this issue, we here integrated the available knowledge together with experimental data by building a series of mathematical models. This rationale enabled us to unravel molecular interactions involving c-Jun in GC induced apoptosis and identify Ets-related gene (Erg) as potential biomarker of GC resistance. The results revealed an alternative possible mechanism where c-Jun may be an indirect GR target that is controlled via an upstream repressor protein. The models also highlight the importance of Erg for GR function, particularly in GC sensitive C7 cells where Erg directly regulates GR in agreement with our previous experimental results. Our models describe potential GR-controlled molecular mechanisms of c-Jun/Bim and Erg regulation. We also demonstrate the importance of using a systematic approach to translate human disease processes into computational models in order to derive information-driven new hypotheses.

  5. Role of the pH in state-dependent blockade of hERG currents

    PubMed Central

    Wang, Yibo; Guo, Jiqing; Perissinotti, Laura L.; Lees-Miller, James; Teng, Guoqi; Durdagi, Serdar; Duff, Henry J.; Noskov, Sergei Yu.

    2016-01-01

    Mutations that reduce inactivation of the voltage-gated Kv11.1 potassium channel (hERG) reduce binding for a number of blockers. State specific block of the inactivated state of hERG block may increase risks of drug-induced Torsade de pointes. In this study, molecular simulations of dofetilide binding to the previously developed and experimentally validated models of the hERG channel in open and open-inactivated states were combined with voltage-clamp experiments to unravel the mechanism(s) of state-dependent blockade. The computations of the free energy profiles associated with the drug block to its binding pocket in the intra-cavitary site display startling differences in the open and open-inactivated states of the channel. It was also found that drug ionization may play a crucial role in preferential targeting to the open-inactivated state of the pore domain. pH-dependent hERG blockade by dofetilie was studied with patch-clamp recordings. The results show that low pH increases the extent and speed of drug-induced block. Both experimental and computational findings indicate that binding to the open-inactivated state is of key importance to our understanding of the dofetilide’s mode of action. PMID:27731415

  6. Modified MBE hardware and techniques and role of gallium purity for attainment of two dimensional electron gas mobility >35×106 cm2/V s in AlGaAs/GaAs quantum wells grown by MBE

    NASA Astrophysics Data System (ADS)

    Gardner, Geoffrey C.; Fallahi, Saeed; Watson, John D.; Manfra, Michael J.

    2016-05-01

    We provide evidence that gallium purity is the primary impediment to attainment of ultra-high mobility in a two-dimensional electron gas (2DEG) in AlGaAs/GaAs heterostructures grown by molecular beam epitaxy (MBE). The purity of gallium can be enhanced dramatically by in-situ high temperature outgassing within an operating MBE. Based on analysis of data from an initial growth campaign in a new MBE system and modifications employed for a 2nd growth campaign, we have produced 2DEGs with low temperature mobility μ in excess of 35×106 cm2/V s at density n=3.0×1011/cm2 and μ=18×106 cm2/V s at n=1.1×1011/cm2. Our 2nd campaign data indicate that gallium purity remains the factor currently limiting μ<40×106 cm2/V s. We describe strategies to overcome this limitation.

  7. Hypoxia inhibits maturation and trafficking of hERG K(+) channel protein: Role of Hsp90 and ROS.

    PubMed

    Nanduri, Jayasri; Bergson, Pamela; Wang, Ning; Ficker, Eckhard; Prabhakar, Nanduri R

    2009-10-16

    We previously reported that reactive oxygen species (ROS) generated during hypoxia decrease hERG current density and protein expression in HEK cells stably expressing hERG protein. In the present study, we investigated the molecular mechanisms involved in hypoxia-induced downregulation of hERG protein. Culturing cells at low temperatures and addition of chemical chaperones during hypoxia restored hERG expression and currents to normoxic levels while antiarrhythmic drugs, which selectively block hERG channels, had no effect on hERG protein levels. Pulse chase studies showed that hypoxia blocks maturation of the core glycosylated form in the endoplasmic reticulum (ER) to the fully glycosylated form on the cell surface. Co-immunoprecipitation experiments revealed that hypoxia inhibited interaction of hERG with Hsp90 chaperone required for maturation, which was restored in the presence of ROS scavengers. These results demonstrate that ROS generated during hypoxia prevents maturation of the hERG protein by inhibiting Hsp90 interaction resulting in decreased protein expression and currents.

  8. Targeting the DNA-binding activity of the human ERG transcription factor using new heterocyclic dithiophene diamidines

    PubMed Central

    Nhili, Raja; Peixoto, Paul; Depauw, Sabine; Flajollet, Sébastien; Dezitter, Xavier; Munde, Manoj M.; Ismail, Mohamed A.; Kumar, Arvind; Farahat, Abdelbasset A.; Stephens, Chad E.; Duterque-Coquillaud, Martine; David Wilson, W.; Boykin, David W.; David-Cordonnier, Marie-Hélène

    2013-01-01

    Direct modulation of gene expression by targeting oncogenic transcription factors is a new area of research for cancer treatment. ERG, an ETS-family transcription factor, is commonly over-expressed or translocated in leukaemia and prostate carcinoma. In this work, we selected the di-(thiophene-phenyl-amidine) compound DB1255 as an ERG/DNA binding inhibitor using a screening test of synthetic inhibitors of the ERG/DNA interaction followed by electrophoretic mobility shift assays (EMSA) validation. Spectrometry, footprint and biosensor-surface plasmon resonance analyses of the DB1255/DNA interaction evidenced sequence selectivity and groove binding as dimer. Additional EMSA evidenced the precise DNA-binding sequence required for optimal DB1255/DNA binding and thus for an efficient ERG/DNA complex inhibition. We further highlighted the structure activity relationships from comparison with derivatives. In cellulo luciferase assay confirmed this modulation both with the constructed optimal sequences and the Osteopontin promoter known to be regulated by ERG and which ERG-binding site was protected from DNaseI digestion on binding of DB1255. These data showed for the first time the ERG/DNA complex modulation, both in vitro and in cells, by a heterocyclic diamidine that specifically targets a portion of the ERG DNA recognition site. PMID:23093599

  9. L- and M-cone input to 12Hz and 30Hz flicker ERGs across the human retina.

    PubMed

    Challa, N K; McKeefry, D; Parry, N R A; Kremers, J; Murray, I J; Panorgias, A

    2010-09-01

    We recorded L- and M-cone isolating ERGs from human subjects using a silent substitution technique at temporal rates of 12 and 30 Hz. These frequencies isolate the activity of cone-opponent and non-opponent post-receptoral mechanisms, respectively. ERGs were obtained using a sequence of stimuli with different spatial configurations comprising; (1) circular stimuli of different sizes which increased in 10° steps up to 70°diameter, or (2) annular stimuli with a 70° outer diameter but with different sized central ablations from 10° up to 60°. L- and M-cone isolating ERGs were obtained from five colour normal subjects using a DTL fibre electrode. Fourier analysis of the ERGs was performed and we measured the amplitude of the first harmonic of the response. For 12 Hz ERGs the L:M cone response amplitude ratio (L:M(ERG)) was close to unity and remained stable irrespective of the spatial configuration of the stimulus. The maintenance of this balanced ratio points to the existence of cone selective input across the human retina for the L-M cone opponent mechanism. For 30 Hz the L:M(ERG) ratio was greater than unity but varied depending upon which region of the retina was being stimulated. This variation we consider to be a consequence of the global response properties of M-cone ERGs rather than representing a real variation in L:M cone ratios across the retina.

  10. Two C4-sterol methyl oxidases (Erg25) catalyse ergosterol intermediate demethylation and impact environmental stress adaptation in Aspergillus fumigatus

    PubMed Central

    Blosser, Sara J.; Merriman, Brittney; Grahl, Nora; Chung, Dawoon

    2014-01-01

    The human pathogen Aspergillus fumigatus adapts to stress encountered in the mammalian host as part of its ability to cause disease. The transcription factor SrbA plays a significant role in this process by regulating genes involved in hypoxia and low-iron adaptation, antifungal drug responses and virulence. SrbA is a direct transcriptional regulator of genes encoding key enzymes in the ergosterol biosynthesis pathway, including erg25A and erg25B, and ΔsrbA accumulates C4-methyl sterols, suggesting a loss of Erg25 activity [C4-sterol methyl oxidase (SMO)]. Characterization of the two genes encoding SMOs in Aspergillus fumigatus revealed that both serve as functional C4-demethylases, with Erg25A serving in a primary role, as Δerg25A accumulates more C4-methyl sterol intermediates than Δerg25B. Single deletion of these SMOs revealed alterations in canonical ergosterol biosynthesis, indicating that ergosterol may be produced in an alternative fashion in the absence of SMO activity. A Δerg25A strain displayed moderate susceptibility to hypoxia and the endoplasmic reticulum stress-inducing agent DTT, but was not required for virulence in murine or insect models of invasive aspergillosis. Inducing expression of erg25A partially restored the hypoxia growth defect of ΔsrbA. These findings implicated Aspergillus fumigatus SMOs in the maintenance of canonical ergosterol biosynthesis and indicated an overall involvement in the fungal stress response. PMID:25107308

  11. Fluconazole susceptibility and ERG11 gene expression in vaginal candida species isolated from lagos Nigeria

    PubMed Central

    Pam, Victoria K; Akpan, Juliet U; Oduyebo, Oyinlola O; Nwaokorie, Francisca O; Fowora, Muinah A; Oladele, Rita O; Ogunsola, Folasade T; Smith, Stella I

    2012-01-01

    Fluconazole resistance is an important type of resistance in Candida because in most countries, fluconazole is the drug of choice for vulvovaginal candidiasis. Candida species resist fluconazole by various mechanisms but there is paucity of data on these in our environment. Such mechanisms include among others, over-expression of the ERG11 gene, which codes for synthesis of the target enzymes in the fungus. The aim of this study was to screen Candida spp. resistant to fluconazole for the expression of ERG11 gene. Fluconazole susceptibility test was performed on 28 clinical strains of Candida species previously obtained from students of a School of Nursing in Lagos, Nigeria. They were identified by API Candida, CHROMagar candida and germ tube test. Using 25 mcg discs, fluconazole susceptibility was determined by the disc diffusion method and results were interpreted in accordance with the Clinical Laboratory Standard Institute (CLSI) criteria; sensitive (S), resistant (R) and susceptible dose dependent (SDD). The R and SDD isolates were subsequently evaluated for the presence of ERG11 gene. Of the 28 clinical isolates, 14 were identified as C. albicans and six as C. tropicalis. The remaining isolates were identified as C. glabrata (2), C. famata (2) C. kefyr (2) one each of C. parapsilosis and C. guilliermondii respectively. In this study, 18 were susceptible (S) to fluconazole, eight were SDD and two were resistant to the antifungal agent. Out of the 14 C. albicans isolates, 12 were susceptible, one showed high level resistance and similar number showed susceptible dose dependence. ERG11 was detected in three susceptible dose dependent Candida species. This analysis demonstrates that susceptible dose dependence should not be overlooked as it may be associated with the presence of ERG11 gene and resistance to fluconazole. There is a need to consider routine antifungal susceptibility testing for Candida species causing vulvovaginitis. PMID:22493755

  12. Allosteric modulators of the hERG K{sup +} channel

    SciTech Connect

    Yu, Zhiyi Klaasse, Elisabeth Heitman, Laura H. IJzerman, Adriaan P.

    2014-01-01

    Drugs that block the cardiac K{sup +} channel encoded by the human ether-à-go-go gene (hERG) have been associated with QT interval prolongation leading to proarrhythmia, and in some cases, sudden cardiac death. Because of special structural features of the hERG K{sup +} channel, it has become a promiscuous target that interacts with pharmaceuticals of widely varying chemical structures and a reason for concern in the pharmaceutical industry. The structural diversity suggests that multiple binding sites are available on the channel with possible allosteric interactions between them. In the present study, three reference compounds and nine compounds of a previously disclosed series were evaluated for their allosteric effects on the binding of [{sup 3}H]astemizole and [{sup 3}H]dofetilide to the hERG K{sup +} channel. LUF6200 was identified as an allosteric inhibitor in dissociation assays with both radioligands, yielding similar EC{sub 50} values in the low micromolar range. However, potassium ions increased the binding of the two radioligands in a concentration-dependent manner, and their EC{sub 50} values were not significantly different, indicating that potassium ions behaved as allosteric enhancers. Furthermore, addition of potassium ions resulted in a concentration-dependent leftward shift of the LUF6200 response curve, suggesting positive cooperativity and distinct allosteric sites for them. In conclusion, our investigations provide evidence for allosteric modulation of the hERG K{sup +} channel, which is discussed in the light of findings on other ion channels. - Highlights: • Allosteric modulators on the hERG K{sup +} channel were evaluated in binding assays. • LUF6200 was identified as a potent allosteric inhibitor. • Potassium ions were found to behave as allosteric enhancers. • Positive cooperativity and distinct allosteric sites for them were proposed.

  13. Prostate cancer of transition zone origin lacks TMPRSS2-ERG gene fusion.

    PubMed

    Guo, Charles C; Zuo, Geyan; Cao, Dongdong; Troncoso, Patricia; Czerniak, Bogdan A

    2009-07-01

    Recent studies have shown a unique chromosomal rearrangement that leads to the fusion of 5'-transmembrane protein serine proteinase-2 (TMPRSS2) with the EST-related gene (ERG) in prostate cancer. In this study, we used fluorescence in situ hybridization to evaluate TMPRSS2-ERG gene fusion in prostate cancer of different zonal origins. Radical prostatectomy specimens with multifocal prostate cancer were obtained from 30 patients who were treated at our institution. Two separate tumor foci in each specimen, one in the peripheral zone and the other in the transition zone, were selected for gene fusion analysis. The selected peripheral zone tumor foci had a mean Gleason score of 6.8 (range, 6-7) and a mean tumor volume of 1.2 cm(3) (range, 0.1-4.6 cm(3)). The selected transition zone tumor foci had a mean Gleason score of 6.7 (range, 5-8) and a mean tumor volume of 4.0 cm(3) (range, 0.5-9.0 cm(3)). ERG gene rearrangement was not observed in any transition zone tumors; however, it was found in the peripheral zone tumors in 13 cases (43%). In 10 cases, the rearrangement was associated with the deletion of the 5'-end of ERG. In conclusion, we found that TMPRSS2-ERG gene fusion is associated with the zonal origin of prostate cancer. This gene fusion is prevalent in prostate cancer arising from the peripheral zone, but is lacking in prostate cancer arising from the transition zone.

  14. Inhibition of hERG Potassium Channels by Celecoxib and Its Mechanism

    PubMed Central

    Frolov, Roman V.; Ignatova, Irina I.; Singh, Satpal

    2011-01-01

    Background Celecoxib (Celebrex), a widely prescribed selective inhibitor of cyclooxygenase-2, can modulate ion channels independently of cyclooxygenase inhibition. Clinically relevant concentrations of celecoxib can affect ionic currents and alter functioning of neurons and myocytes. In particular, inhibition of Kv2.1 channels by celecoxib leads to arrhythmic beating of Drosophila heart and of rat heart cells in culture. However, the spectrum of ion channels involved in human cardiac excitability differs from that in animal models, including mammalian models, making it difficult to evaluate the relevance of these observations to humans. Our aim was to examine the effects of celecoxib on hERG and other human channels critically involved in regulating human cardiac rhythm, and to explore the mechanisms of any observed effect on the hERG channels. Methods and Results Celecoxib inhibited the hERG, SCN5A, KCNQ1 and KCNQ1/MinK channels expressed in HEK-293 cells with IC50s of 6.0 µM, 7.5 µM, 3.5 µM and 3.7 µM respectively, and the KCND3/KChiP2 channels expressed in CHO cells with an IC50 of 10.6 µM. Analysis of celecoxib's effects on hERG channels suggested gating modification as the mechanism of drug action. Conclusions The above channels play a significant role in drug-induced long QT syndrome (LQTS) and short QT syndrome (SQTS). Regulatory guidelines require that all new drugs under development be tested for effects on the hERG channel prior to first administration in humans. Our observations raise the question of celecoxib's potential to induce cardiac arrhythmias or other channel related adverse effects, and make a case for examining such possibilities. PMID:22039467

  15. p16 upregulation is linked to poor prognosis in ERG negative prostate cancer.

    PubMed

    Burdelski, Christoph; Dieckmann, Tatsiana; Heumann, Asmus; Hube-Magg, Claudia; Kluth, Martina; Beyer, Burkhard; Steuber, Thomas; Pompe, Raisa; Graefen, Markus; Simon, Ronald; Minner, Sarah; Tsourlakis, Maria Christina; Koop, Christina; Izbicki, Jakob; Sauter, Guido; Krech, Till; Schlomm, Thorsten; Wilczak, Waldemar; Lebok, Patrick

    2016-09-01

    Altered expression of the p16 tumor suppressor is frequently found in prostate cancer, but its role for tumor development and patient prognosis is disputed. In order to clarify the prognostic role of p16 and to draw conclusions on interactions with key molecular features of prostate cancer, we studied p16 expression in a tissue microarray (TMA) with more than 12,400 prostate cancers and attached clinical, pathological, and molecular data such as ERG status and deletions of 3p13, 5q21, 6q15, and PTEN. p16 immunostaining was absent in non-neoplastic prostate cells but was found in 37 % of 9627 interpretable prostate cancers. Finding p16 expression in 58 % of ERG positive but in only 22 % of ERG negative cancers (p < 0.0001), highlights the known androgen-dependence of both genes. Significant associations between p16 upregulation and tumor phenotype or patient prognosis were strictly limited to the subset of ERG negative cancers. For example, p16 positivity increased from 15 % in Gleason ≤3 + 3 to 38 % in Gleason ≥4 + 4 cancers (p < 0.0001) and was associated with early PSA recurrence (p < 0.0001). p16 upregulation was strongly linked to deletions of PTEN (p < 0.0001), highlighting the interaction of both genes in growth control. In conclusion, p16 upregulation is a strong prognostic factor in ERG negative cancers. The strict limitation of its prognostic impact to a molecularly defined subgroup challenges the concept of molecular prognosis testing without considering molecular subtypes.

  16. Ion acceleration in electrostatic field of charged cavity created by ultra-short laser pulses of 1020-1021 W/cm2

    NASA Astrophysics Data System (ADS)

    Bychenkov, V. Yu.; Singh, P. K.; Ahmed, H.; Kakolee, K. F.; Scullion, C.; Jeong, T. W.; Hadjisolomou, P.; Alejo, A.; Kar, S.; Borghesi, M.; Ter-Avetisyan, S.

    2017-01-01

    Ion acceleration resulting from the interaction of ultra-high intensity and ultra-high contrast (˜10-10) laser pulses with thin A l foil targets at 30° angle of laser incidence is studied. Proton maximum energies of 30 and 18 MeV are measured along the target normal rear and front sides, respectively, showing intensity scaling as Ib . For the target front bf r o n t= 0.5-0.6 and for the target rear br e a r= 0.7-0.8 is observed in the intensity range 1020-1021 W/cm2. The fast scaling from the target rear ˜I0.75 can be attributed enhancement of laser energy absorption as already observed at relatively low intensities. The backward acceleration of the front side protons with intensity scaling as ˜I0.5 can be attributed to the to the formation of a positively charged cavity at the target front via ponderomotive displacement of the target electrons at the interaction of relativistic intense laser pulses with a solid target. The experimental results are in a good agreement with theoretical predictions.

  17. Aging characteristics of blue InGaN micro-light emitting diodes at an extremely high current density of 3.5 kA cm-2

    NASA Astrophysics Data System (ADS)

    Tian, Pengfei; Althumali, Ahmad; Gu, Erdan; Watson, Ian M.; Dawson, Martin D.; Liu, Ran

    2016-04-01

    The aging characteristics of blue InGaN micro-light emitting diodes (micro-LEDs) with different sizes have been studied at an extremely high current density 3.5 kA cm-2 for emerging micro-LED applications including visible light communication (VLC), micro-LED pumped organic lasers and optogenetics. The light output power of micro-LEDs first increases and then decreases due to the competition of Mg activation in p-GaN layer and defect generation in the active region. The smaller micro-LEDs show less light output power degradation compared with larger micro-LEDs, which is attributed to the lower junction temperature of smaller micro-LEDs. It is found that the high current density without additional junction temperature cannot induce significant micro-LED degradation at room temperature but the combination of the high current density and high junction temperature leads to strong degradation. Furthermore, the cluster LEDs, composed of a micro-LED array, have been developed with both high light output power and less light output degradation for micro-LED applications in solid state lighting and VLC.

  18. First operation and performance of a 200 lt double phase LAr LEM-TPC with a 40 × 76 cm2 readout

    NASA Astrophysics Data System (ADS)

    Badertscher, A.; Curioni, A.; Degunda, U.; Epprecht, L.; Gendotti, A.; Horikawa, S.; Knecht, L.; Lussi, D.; Natterer, G.; Nguyen, K.; Resnati, F.; Rubbia, A.; Viant, T.

    2013-04-01

    In this paper we describe the design, construction, and operation of a first large area double-phase liquid argon Large Electron Multiplier Time Projection Chamber (LAr LEM-TPC). The detector has a maximum drift length of 60 cm and the readout consists of a 40 × 76 cm2 LEM and 2D projective anode to multiply and collect drifting charges. Scintillation light is detected by means of cryogenic PMTs positioned below the cathode. To record both charge and light signals, we have developed a compact acquisition system, which is scalable up to ton-scale detectors with thousands of charge readout channels. The acquisition system, as well as the design and the performance of custom-made charge sensitive preamplifiers, are described. The complete experimental setup has been operated for a first time during a period of four weeks at CERN in the cryostat of the ArDM experiment, which was equipped with liquid and gas argon purification systems. The detector, exposed to cosmic rays, recorded events with a single-channel signal-to-noise ratio in excess of 30 for minimum ionising particles. Cosmic muon tracks and their δ-rays were used to assess the performance of the detector, and to estimate the liquid argon purity and the gain at different amplification fields.

  19. Nanostructured multielement (TiHfZrNbVTa)N coatings before and after implantation of N+ ions (1018 cm-2): Their structure and mechanical properties

    NASA Astrophysics Data System (ADS)

    Pogrebnjak, A. D.; Bondar, O. V.; Borba, S. O.; Abadias, G.; Konarski, P.; Plotnikov, S. V.; Beresnev, V. M.; Kassenova, L. G.; Drodziel, P.

    2016-10-01

    Multielement high entropy alloy (HEA) nitride (TiHfZrNbVTa)N coatings were deposited by vacuum arc and their structural and mechanical stability after implantation of high doses of N+ ions, 1018 cm-2, were investigated. The crystal structure and phase composition were characterized by X-ray diffraction (XRD) and Transmission Electron Microscopy, while depth-resolved nanoindentation tests were used to determine the evolution of hardness and elastic modulus along the implantation depth. XRD patterns show that coatings exhibit a main phase with fcc structure, which preferred orientation varies from (1 1 1) to (2 0 0), depending on the deposition conditions. First-principles calculations reveal that the presence of Nb atoms could favor the formation of solid solution with fcc structure in multielement HEA nitride. TEM results showed that amorphous and nanostructured phases were formed in the implanted coating sub-surface layer (∼100 nm depth). Concentration of nitrogen reached 90 at% in the near-surface layer after implantation, and decreased at higher depth. Nanohardness of the as-deposited coatings varied from 27 to 38 GPa depending on the deposition conditions. Ion implantation led to a significant decrease of the nanohardness to 12 GPa in the implanted region, while it reaches 24 GPa at larger depths. However, the H/E ratio is ⩾0.1 in the sub-surface layer due to N+ implantation, which is expected to have beneficial effect on the wear properties.

  20. Performance of a 64-channel, 3.2×3.2 cm2 SiPM tile for TOF-PET application

    NASA Astrophysics Data System (ADS)

    Ferri, Alessandro; Acerbi, Fabio; Gola, Alberto; Piemonte, Claudio; Paternoster, Giovanni; Zorzi, Nicola

    2016-07-01

    In this work, we present a new 3.2×3.2 cm2 detector tile, composed of 8×8 single SiPMs, having a regular 4 mm pitch in both the X and Y directions. The tile fill factor is 85%. We produced two versions of the tile with different SiPM technologies: RGB-HD and NUV. The first one features square micro-cells with 25 μm pitch, a PDE peaked at 550 nm and a DCR of 300 kHz/mm2, at 20 °C and at maximum detection efficiency. The second one features micro-cells with 40 μm pitch and a PDE peaked in the blue part of the spectrum. The dark count rate at 20 °C and at maximum PDE is 100 kHz/mm2. In this work, we show the energy and timing resolution measurements at 511 keV obtained coupling the two tiles to an 8×8 LYSO array with a pixel size of 4×4×22 mm3, perfectly matching the photo-detector array.

  1. Predicting the optimal process window for the coating of single-crystalline organic films with mobilities exceeding 7 cm2/Vs.

    NASA Astrophysics Data System (ADS)

    Janneck, Robby; Vercesi, Federico; Heremans, Paul; Genoe, Jan; Rolin, Cedric

    2016-09-01

    Organic thin film transistors (OTFTs) based on single crystalline thin films of organic semiconductors have seen considerable development in the recent years. The most successful method for the fabrication of single crystalline films are solution-based meniscus guided coating techniques such as dip-coating, solution shearing or zone casting. These upscalable methods enable rapid and efficient film formation without additional processing steps. The single-crystalline film quality is strongly dependent on solvent choice, substrate temperature and coating speed. So far, however, process optimization has been conducted by trial and error methods, involving, for example, the variation of coating speeds over several orders of magnitude. Through a systematic study of solvent phase change dynamics in the meniscus region, we develop a theoretical framework that links the optimal coating speed to the solvent choice and the substrate temperature. In this way, we can accurately predict an optimal processing window, enabling fast process optimization. Our approach is verified through systematic OTFT fabrication based on films grown with different semiconductors, solvents and substrate temperatures. The use of best predicted coating speeds delivers state of the art devices. In the case of C8BTBT, OTFTs show well-behaved characteristics with mobilities up to 7 cm2/Vs and onset voltages close to 0 V. Our approach also explains well optimal recipes published in the literature. This route considerably accelerates parameter screening for all meniscus guided coating techniques and unveils the physics of single crystalline film formation.

  2. Adsorption of 2,4,6,8,10,12-hexanitro-2,4,6,8,10,12-hexaazaisowurtzitane (CL-20) on a soil organic matter. A DFT M05 computational study.

    PubMed

    Sviatenko, Liudmyla K; Gorb, Leonid; Shukla, Manoj K; Seiter, Jennifer M; Leszczynska, Danuta; Leszczynski, Jerzy

    2016-04-01

    Adsorption of 2,4,6,8,10,12-hexanitro-2,4,6,8,10,12-hexaazaisowurtzitane (CL-20) by soil organic matter considering the Leonardite Humic Acid (LHA) model at the M05/tzvp level of Density Functional Theory (DFT) applying cluster approximation has been investigated. Different orientations of CL-20 toward LHA surface were examined. It was found that deprotonation of LHA is required to obtain stable complexes with CL-20. Hydrogen bonds between CL-20 and deprotonated LHA were analyzed applying the atoms in molecules (AIM) theory. An attachment or removal of an electron with respect to the complex does not have significant effect on mutual orientation of the adsorbent in complexes. It was shown that adsorbed CL-20 does not undergo redox transformation and, therefore, adsorption on soil organic matter may be responsible for decrease of the degradation rate of CL-20 in soil.

  3. Analytical platform evaluation for quantification of ERG in prostate cancer using protein and mRNA detection methods

    SciTech Connect

    He, Jintang; Schepmoes, Athena A.; Shi, Tujin; Wu, Chaochao; Fillmore, Thomas L.; Gao, Yuqian; Smith, Richard D.; Qian, Weijun; Rodland, Karin D.; Liu, Tao; Camp, David G.; Rastogi, Anshu; Tan, Shyh-Han; Yan, Wusheng; Mohamed, Ahmed A.; Huang, Wei; Banerjee, Sreedatta; Kagan, Jacob; Srivastava, Sudhir; McLeod, David; Srivastava, Shiv; Petrovics, Gyorgy; Dobi, Albert; Srinivasan, Alagarsamy

    2015-01-01

    Background: The established methods for detecting prostate cancer (CaP) are based on tests using PSA (blood), PCA3 (urine), and AMACR (tissue) as biomarkers in patient samples. The demonstration of ERG oncoprotein overexpression due to gene fusion in CaP has thus provided ERG as an additional biomarker. Based on this, we hypothesized that ERG protein quantification methods can be of use in the diagnosis of prostate cancer. Methods: Therefore, an antibody-free assay for ERG3 protein detection was developed based on PRISM (high-pressure high-resolution separations with intelligent selection and multiplexing)-SRM (selected reaction monitoring) mass spectrometry. We utilized TMPRSS2-ERG positive VCaP and TMPRSS2-ERG negative LNCaP cells to simulate three different sample types (cells, tissue, and post-DRE urine sediment). Results: Recombinant ERG3 protein spiked into LNCaP cell lysates could be detected at levels as low as 20 pg by PRISM-SRM analysis. The sensitivity of the PRISM-SRM assay was around approximately 10,000 VCaP cells in a mixed cell population model of VCaP and LNCaP cells. Interestingly, ERG protein could be detected in as few as 600 VCaP cells spiked into female urine. The sensitivity of the in-house enzyme-linked immunosorbent assay (ELISA) was similar to the PRISM-SRM assay, with detection of 30 pg of purified recombinant ERG3 protein and 10,000 VCaP cells. On the other hand, qRT-PCR exhibited a higher sensitivity, as TMPRSS2-ERG transcripts were detected in as few as 100 VCaP cells, in comparison to NanoString methodologies which detected ERG from 10,000 cells. Conclusions: Based on this data, we propose that the detection of both ERG transcriptional products with RNA-based assays, as well as protein products of ERG using PRISM-SRM assays, may be of clinical value in developing diagnostics and prognostics assays for prostate cancer given their sensitivity, specificity, and reproducibility.

  4. Comparative evaluation of hand wrist radiographs with cervical vertebrae for skeletal maturation in 10-12 years old children.

    PubMed

    Kamal, M; Goyal, S

    2006-09-01

    A comparative evaluation of hand wrist and cervical vertebrae was done to know the validity of cervical vertebrae as maturity indicators. A sample of 50 subjects (25 females and 25 males) in the age group of 10-12 years were selected on criteria of normal occlusion and the result showed that cervical vertebrae can be used with the same confidence as hand wrist radiographs to evaluate skeletal maturity, thus avoiding the need for an additional radiograph.

  5. Aqueous Alteration of Carbonaceous Chondrites: New Insights from Comparative Studies of Two Unbrecciated CM2 Chondrites, Y 791198 and ALH 81002

    NASA Technical Reports Server (NTRS)

    Chizmadia, L. J.; Brearley, A. J.

    2004-01-01

    Carbonaceous chondrites are an important resource for understanding the physical and chemical conditions in the early solar system. In particular, a long-standing question concerns the role of water in the cosmochemical evolution of carbonaceous chondrites. It is well established that extensive hydration of primary nebular phases occurred in the CM and CI chondrites, but the location where this alteration occurred remains controversial. In the CM2 chondrites, hydration formed secondary phases such as serpentine, tochilinite, pentlandite, carbonate and PCP. There are several textural observations which suggest that alteration occurred before the accretion of the final CM parent asteroid, i.e. preaccretionary alteration. Conversely, there is a significant body of evidence that supports parent-body alteration. In order to test these two competing hypotheses further, we studied two CM chondrites, Y-791198 and ALH81002, two meteorites that exhibit widely differing degrees of aqueous alteration. In addition, both meteorites have primary accretionary textures, i.e. experienced minimal asteroidal brecciation. Brecciation significantly complicates the task of unraveling alteration histories, mixing components that have been altered to different degrees from different locations on the same asteroidal parent body. Alteration in Y-791198 is mostly confined to chondrule mesostases, FeNi metal and fine-grained matrix and rims. In comparison, the primary chondrule silicates in ALH81002 have undergone extensive replacement by secondary hydrous phases. This study focuses on compositional and textural relationships between chondrule mesostasis and the associated rim materials. Our hypothesis is: both these components are highly susceptible to aqueous alteration and should be sensitive recorders of the alteration process. For parent body alteration, we expect systematic coupled mineralogical and compositional changes in rims and altered mesostasis, as elemental exchange between these

  6. Investigation of rat bone fracture healing using pulsed 1.5 MHz, 30 mW/cm(2) burst ultrasound--axial distance dependency.

    PubMed

    Fung, Chak-Hei; Cheung, Wing-Hoi; Pounder, Neill M; de Ana, F Javier; Harrison, Andrew; Leung, Kwok-Sui

    2014-03-01

    This study investigated the effect of LIPUS on fracture healing when fractures were exposed to ultrasound at three axial distances: z=0 mm, 60 mm, and 130 mm. We applied LIPUS to rat fracture at these three axial distances mimicking the exposure condition of human fractures at different depths under the soft tissue. Measurement of LIPUS shows pressure variations in near field (nearby transducer); uniform profile was found beyond it (far field). We asked whether different positions of the fracture within the ultrasound field cause inconsistent biological effect during the healing process. Closed femoral fractured Sprague-Dawley rats were randomized into control, near-field (0mm), mid-near field (60 mm) or far-field (130 mm) groups. Daily LIPUS treatment (plane, but apodized source, see details in the text; 2.2 cm in diameter; 1.5 MHz sine waves repeating at 1 kHz PRF; spatial average temporal average intensity, ISATA=30 mW/cm(2)) was given to fracture site at the three axial distances. Weekly radiographs and endpoint microCT, histomorphometry, and mechanical tests were performed. The results showed that the 130 mm group had the highest tissue mineral density; and significantly higher mechanical properties than control at week 4. The 60 mm and 0 mm groups had significantly higher (i.e. p<0.05) woven bone percentage than control group in radiological, microCT and histomorphometry measurements. In general, LIPUS at far field augmented callus mineralization and mechanical properties; while near field and mid-near field enhanced woven bone formation. Our results indicated the therapeutic effect of LIPUS is dependent on the axial distance of the ultrasound beam. Therefore, the depth of fracture under the soft tissue affects the biological effect of LIPUS. Clinicians have to be aware of the fracture depth when LIPUS is applied transcutaneously.

  7. Chloromethane Utilization Gene Cluster from Hyphomicrobium chloromethanicum Strain CM2T and Development of Functional Gene Probes To Detect Halomethane-Degrading Bacteria

    PubMed Central

    McAnulla, Craig; Woodall, Claire A.; McDonald, Ian R.; Studer, Alex; Vuilleumier, Stephane; Leisinger, Thomas; Murrell, J. Colin

    2001-01-01

    Hyphomicrobium chloromethanicum CM2T, an aerobic methylotrophic member of the α subclass of the class proteobacteria, can grow with chloromethane as the sole carbon and energy source. H. chloromethanicum possesses an inducible enzyme system for utilization of chloromethane, in which two polypeptides (67-kDa CmuA and 35-kDa CmuB) are expressed. Previously, four genes, cmuA, cmuB, cmuC, and purU, were shown to be essential for growth of Methylobacterium chloromethanicum on chloromethane. The cmuA and cmuB genes were used as probes to identify homologs in H. chloromethanicum. A cmu gene cluster (9.5 kb) in H. chloromethanicum contained 10 open reading frames: folD (partial), pduX, orf153, orf207, orf225, cmuB, cmuC, cmuA, fmdB, and paaE (partial). CmuA from H. chloromethanicum (67 kDa) showed high identity to CmuA from M. chloromethanicum and contains an N-terminal methyltransferase domain and a C-terminal corrinoid-binding domain. CmuB from H. chloromethanicum is related to a family of methyl transfer proteins and to the CmuB methyltransferase from M. chloromethanicum. CmuC from H. chloromethanicum shows identity to CmuC from M. chloromethanicum and is a putative methyltransferase. folD codes for a methylene-tetrahydrofolate cyclohydrolase, which may be involved in the C1 transfer pathway for carbon assimilation and CO2 production, and paaE codes for a putative redox active protein. Molecular analyses and some preliminary biochemical data indicated that the chloromethane utilization pathway in H. chloromethanicum is similar to the corrinoid-dependent methyl transfer system in M. chloromethanicum. PCR primers were developed for successful amplification of cmuA genes from newly isolated chloromethane utilizers and enrichment cultures. PMID:11133460

  8. Hydrogen and carbon isotopic ratios of polycyclic aromatic compounds in two CM2 carbonaceous chondrites and implications for prebiotic organic synthesis

    NASA Astrophysics Data System (ADS)

    Huang, Yongsong; Aponte, José C.; Zhao, Jiaju; Tarozo, Rafael; Hallmann, Christian

    2015-09-01

    Study of meteoritic organic compounds offers a unique opportunity to understand the origins of the organic matter in the early Solar System. Meteoritic polycyclic aromatic hydrocarbons (PAHs) and heteropolycyclic aromatic compounds (HACs) have been studied for over fifty years, however; their hydrogen stable isotopic ratios (δD) have never been reported. Compound-specific δD measurements of PAHs and HACs are important, in part because the carbon isotopic ratios (δ13C) of various meteoritic PAHs cannot be readily distinguished from their terrestrial counterparts and it is difficult to rule out terrestrial contamination based on carbon isotopic ratios alone. In this study, we have extracted and identified more than sixty PAHs and HACs present in two CM2 carbonaceous chondrites Murchison and LON 94101. Their carbon and hydrogen stable isotopic ratios (δ13C and δD) were measured and used to discuss about their synthetic environments and formation mechanisms. The concentration of aromatic compounds is ∼30% higher in Murchison than in the Antarctic meteorite LON 94101, but both samples contained similar suites of PAHs and HACs. All PAHs and HACs found exhibited positive δD values (up to 1100‰) consistent with an extraterrestrial origin, indicating the relatively low δ13C values are indeed an inherent feature of the meteoritic aromatic compounds. The hydrogen isotopic data suggest aromatic compounds in carbonaceous chondrites were mainly formed in the cold interstellar environments. Molecular level variations in hydrogen and carbon isotopic values offer new insights to the formation pathways for the aromatic compounds in carbonaceous chondrites.

  9. One-year Outcomes of Pachymetry and Epithelium Thicknesses after Accelerated (45 mW/cm2) Transepithelial Corneal Collagen Cross-linking for Keratoconus Patients

    PubMed Central

    Zhang, Xiaoyu; Sun, Ling; Chen, Yingjun; Li, Meiyan; Tian, Mi; Zhou, Xingtao

    2016-01-01

    The thickness of corneal pachymetry and the epithelium after accelerated (45 mW/cm2) transepithelial corneal collagen cross-linking (CXL) for keratoconus were assessed in this prospective case series study. Twenty-eight patients were treated for keratoconus. The mean Kmax was 56.18 ± 7.90. The thinnest point, as assessed by optical coherence tomography (OCT), was 443.18 ± 39.75 μm. Accelerated transepithelial CXL was performed, and corrected distance visual acuity (CDVA), corneal topography, and OCT were recorded at 1 week postoperatively as well as at 1, 3, 6, and 12 months. The surgery was uneventful in all eyes. Postoperative epithelial edema was observed and faded in 3 days. The postoperative Kmax was 54.56 ± 8.81, 55.78 ± 8.11, 56.37 ± 8.71, 55.80 ± 7.92, and 55.47 ± 8.24 at 1 week, 1 month, 3 months, 6 months, and 12 months, respectively (all, P > 0.05). The thinnest postoperative corneal point, 439.04 ± 44.99 μm, was observed at 12 months (P = 0.109). The epithelial thickness decreased during the first postoperative week then showed a gradual recovery. Postoperative pachymetry thickness showed no significant changes for up to 12 months. Postoperative epithelial thickness decreased temporarily, then stabilized at month 12. Accelerated transepithelial CXL was shown to be effective and safe for the treatment of keratoconus. PMID:27597655

  10. Effect of Tube-Based X-Ray Microtomography Imaging on the Amino Acid and Amine Content of the Murchison CM2 Chondrite

    NASA Technical Reports Server (NTRS)

    Glavin, D. P.; Friedrich, J. M.; Aponte, J. C.; Dworkin, J. P.; Ebel, D. S.; Elsila, J. E.; Hill, M.; McLain, H. L.; Towbin, W. H.

    2017-01-01

    X-ray and synchrotron X-ray micro-computed tomography (micro-CT) are increasingly being used for three dimensional reconnaissance imaging of chondrites and returned extraterrestrial material prior to detailed chemical and mineralogical analyses. Although micro-CT imaging is generally considered to be a non-destructive technique since silicate and metallic minerals in chondrites are not affected by X-ray exposures at the intensities and wavelengths typically used, there are concerns that the use of micro-CT could be detrimental to the organics in carbonaceous chondrites. We recently conducted a synchrotron micro-CT experiment on a powdered sample of the Murchison CM2 carbonaceous chondrite exposed to a monochromatic high energy (approximately 48 kiloelectronvolts) total X-ray radiation dose of approximately 1 kilogray (kGy) using the Advanced Photon Source beamline 13-BMD (13-Bending Magnet-D Beamline) at Argonne National Laboratory and found that there were no detectable changes in the amino acid abundances or enantiomeric compositions in the chondrite after exposure relative to a Murchison control sample that was not exposed. However, lower energy bremsstrahlung X-rays could interact more with amino acids and other lower molecular weight amines in meteorites. To test for this possibility, three separate micro-CT imaging experiments of the Murchison meteorite using the GE Phoenix v/tome/x s 240 kilovolt microfocus high resolution tungsten target X-ray tube instrument at the American Museum of Natural History (AMNH) were conducted and the amino acid abundances and enantiomeric compositions were determined. We also investigated the abundances of the C1-C5 amines in Murchison which were not analyzed in the first study.

  11. Assessing the Contribution of the Oscillatory Potentials to the Genesis of the Photopic ERG with the Discrete Wavelet Transform

    PubMed Central

    Dorfman, Allison L.; Trang, Nataly; Gauthier, Mercedes; Little, John M.; Lina, Jean-Marc

    2016-01-01

    The electroretinogram (ERG) is composed of slow (i.e., a-, b-waves) and fast (i.e., oscillatory potentials: OPs) components. OPs have been shown to be preferably affected in some diseases (such as diabetic retinopathy), while the a- and b-waves remain relatively intact. The purpose of this study was to determine the contribution of OPs to the building of the ERG and to examine whether a signal mostly composed of OPs could also exist. DWT analyses were performed on photopic ERGs (flash intensities: −2.23 to 2.64 log cd·s·m−2 in 21 steps) obtained from normal subjects (n = 40) and patients (n = 21) affected with a retinopathy. In controls, the %OP value (i.e., OPs energy/ERG energy) is stimulus- and amplitude-independent (range: 56.6–61.6%; CV = 6.3%). In contrast, the %OPs measured from the ERGs of our patients varied significantly more (range: 35.4%–89.2%; p < 0.05) depending on the pathology, some presenting with ERGs that are almost solely composed of OPs. In conclusion, patients may present with a wide range of %OP values. Findings herein also support the hypothesis that, in certain conditions, the photopic ERG can be mostly composed of high-frequency components. PMID:28101507

  12. Differential hERG ion channel activity of ultrasmall gold nanoparticles.

    PubMed

    Leifert, Annika; Pan, Yu; Kinkeldey, Anne; Schiefer, Frank; Setzler, Julia; Scheel, Olaf; Lichtenbeld, Hera; Schmid, Günter; Wenzel, Wolfgang; Jahnen-Dechent, Willi; Simon, Ulrich

    2013-05-14

    Understanding the mechanism of toxicity of nanomaterials remains a challenge with respect to both mechanisms involved and product regulation. Here we show toxicity of ultrasmall gold nanoparticles (AuNPs). Depending on the ligand chemistry, 1.4-nm-diameter AuNPs failed electrophysiology-based safety testing using human embryonic kidney cell line 293 cells expressing human ether-á-go-go-Related gene (hERG), a Food and Drug Administration-established drug safety test. In patch-clamp experiments, phosphine-stabilized AuNPs irreversibly blocked hERG channels, whereas thiol-stabilized AuNPs of similar size had no effect in vitro, and neither particle blocked the channel in vivo. We conclude that safety regulations may need to be reevaluated and adapted to reflect the fact that the binding modality of surface functional groups becomes a relevant parameter for the design of nanoscale bioactive compounds.

  13. Differential hERG ion channel activity of ultrasmall gold nanoparticles

    PubMed Central

    Leifert, Annika; Pan, Yu; Kinkeldey, Anne; Schiefer, Frank; Setzler, Julia; Scheel, Olaf; Lichtenbeld, Hera; Schmid, Günter; Wenzel, Wolfgang; Jahnen-Dechent, Willi; Simon, Ulrich

    2013-01-01

    Understanding the mechanism of toxicity of nanomaterials remains a challenge with respect to both mechanisms involved and product regulation. Here we show toxicity of ultrasmall gold nanoparticles (AuNPs). Depending on the ligand chemistry, 1.4-nm-diameter AuNPs failed electrophysiology-based safety testing using human embryonic kidney cell line 293 cells expressing human ether-á-go-go-Related gene (hERG), a Food and Drug Administration-established drug safety test. In patch-clamp experiments, phosphine-stabilized AuNPs irreversibly blocked hERG channels, whereas thiol-stabilized AuNPs of similar size had no effect in vitro, and neither particle blocked the channel in vivo. We conclude that safety regulations may need to be reevaluated and adapted to reflect the fact that the binding modality of surface functional groups becomes a relevant parameter for the design of nanoscale bioactive compounds. PMID:23630249

  14. hERG1 potassium channel in cancer cells: a tool to reprogram immortality.

    PubMed

    Gentile, Saverio

    2016-10-01

    It has been well established that changes in ion fluxes across cellular membranes as a function of time is fundamental in maintaining cellular homeostasis of every living cell. Consequently, dysregulation of ion channels activity is a critical event in pathological conditions of several tissues, including cancer. Nevertheless, the role of ion channels in cancer biology is still not well understood and very little is known about the possible therapeutic opportunities offered by the use of the vast collection of drugs that target ion channels. In this review, we focus on the recent advances in understanding the role of the voltage-gated hERG1 potassium channel in cancer and on the effects of pharmacologic manipulation of the hERG1 in cancer cells aiming to provide insights into the biochemical signaling and cellular processes that are altered by using these drugs.

  15. ERG/AKR1C3/AR constitutes a feed-forward loop for AR signaling in prostate cancer cells

    PubMed Central

    Powell, Katelyn; Semaan, Louie; Conley-LaComb, M. Katie; Asangani, Irfan; Wu, Yi-Mi; Ginsburg, Kevin B.; Williams, Julia; Squire, Jeremy A.; Maddipati, Krishna R.; Cher, Michael L.; Chinni, Sreenivasa R.

    2015-01-01

    Purpose Intratumoral androgen synthesis in prostate cancer (PCa) contributes to the development of castration-resistant prostate cancer (CRPC). Several enzymes responsible for androgen biosynthesis have been shown to be overexpressed in CRPC, thus contributing to CRPC in a castrated environment. The TMPRSS2-ERG transcription factor has been shown to be present in primary PCa tumors as well as CRPC tumors. We hypothesize that TMPRSS2-ERG fusions regulate androgen biosynthetic enzyme (ABE) gene expression and the production of androgens, which contributes to the development of CRPC. Experimental design We used a panel of assays including lentivirus transduction, gene expression, chromatin immunoprecipitation and sequencing, Liquid chromatography-Mass spectrometric quantitation, immunocytochemistry, immunohistochemistry and bio-informatics analysis of gene microarray data bases to determine ERG regulation of androgen synthesis. Results We found that ERG regulated the expression of the ABE AKR1C3 in PCa cells via direct binding to the AKR1C3 gene. Knockdown of ERG resulted in reduced AKR1C3 expression, which caused a reduction in both DHT synthesis and PSA expression in VCaP PCa cells treated with 5α-androstanedione, a DHT precursor metabolite. Immunohistochemical staining revealed that ERG was co-expressed with AKR1C3 in PCa tissue samples. Conclusions These data suggest that AKR1C3 catalyzes the biochemical reduction of 5α-Androstanedione to DHT in PCa cells, and that ERG regulates this step through upregulation of AKR1C3 expression. Elucidation of ERG regulation of ABEs in CRPC may help to stratify TMPRSS2-ERG fusion-positive PCa patients in the clinic for anti-AR driven therapies; and AKR1C3 may serve as a valuable therapeutic target in the treatment of CRPC. PMID:25754347

  16. Mechanisms of pharmacological rescue of trafficking defective hERG mutant channels in human long QT syndrome

    PubMed Central

    Gong, Qiuming; Jones, Melanie A.; Zhou, Zhengfeng

    2006-01-01

    Long QT syndrome type 2 is caused by mutations in the human ether-a-go-go-related gene (hERG). We previously reported that the N470D mutation is retained in the endoplasmic reticulum (ER) but can be rescued to the plasma membrane by hERG channel blocker E-4031. The mechanisms of ER retention and how E-4031 rescues the N470D mutant are poorly understood. In this study, we investigated the interaction of hERG channels with ER chaperone protein calnexin. Using coimmunoprecipitation, we showed that the immature forms of both wild type hERG and N470D associated with calnexin. The association required N-linked glycosylation of hERG channels. Pulse-chase analysis revealed that N470D had a prolonged association with calnexin compared to wild type hERG, and E-4031 shortened the time course of calnexin association with N470D. To test whether the prolonged association of N470D with calnexin is due to defective folding of mutant channels, we studied hERG channel folding using trypsin digestion method. We found that N470D and the immature form of wild type hERG were more sensitive to trypsin digestion than the mature form of wild type hERG. In the presence of E-4031, N470D became more resistant to trypsin even in the conditions that its ER-to-Golgi transport was blocked by brefeldin A. These results suggest that defective folding of N470D contributes to its prolonged association with calnexin and ER retention, and that E-4031 may restore proper folding of the N470D channel leading to its cell surface expression. PMID:16361248

  17. Highly Specific Targeting of the TMPRSS2/ERG Fusion Gene in Prostate Cancer Using Liposomal Nanotechnology

    DTIC Science & Technology

    2012-06-01

    time due to elimination by reticuloendothelial system. To increase stability and blood circulation half- life coating nanoparticles with polymers such...ERG fusion gene in prostate cancer using liposomal nanotechnology PRINCIPAL INVESTIGATOR: Bulent Ozpolat, M.D., Ph.D...fusion gene in prostate cancer using liposomal nanotechnology 5b. GRANT NUMBER W81XWH-09-1-0385 5c. PROGRAM ELEMENT NUMBER 6. AUTHOR(S) 5d

  18. Movement of the S4 segment in the hERG potassium channel during membrane depolarization.

    PubMed

    Elliott, David J S; Dondas, Naciye Y; Munsey, Tim S; Sivaprasadarao, Asipu

    2009-12-01

    The hERG potassium channel is a member of the voltage gated potassium (Kv) channel family, comprising a pore domain and four voltage sensing domains (VSDs). Like other Kv channels, the VSD senses changes in membrane voltage and transmits the signal to gates located in the pore domain; the gates open at positive potentials (activation) and close at negative potentials, thereby controlling the ion flux. hERG, however, differs from other Kv channels in that it is activated slowly but inactivated rapidly - a property that is crucial for the role it plays in the repolarization of the cardiac action potential. Voltage-gating requires movement of gating charges across the membrane electric field, which is accomplished by the transmembrane movement of the fourth transmembrane segment, S4, of the VSD containing the positively charged arginine or lysine residues. Here we ask if the functional differences between hERG and other Kv channels could arise from differences in the transmembrane movement of S4. To address this, we have introduced single cysteine residues into the S4 region of the VSD, expressed the mutant channels in Xenopus oocytes and examined the effect of membrane impermeable para-chloromercuribenzene sulphonate on function by the two-electrode voltage clamp technique. Our results show that depolarization results in the accessibility of seven consecutive S4 residues, including the first two charged residues, K525 and R528, to extracellularly applied reagent. These data indicate that the extent of S4 movement in hERG is similar to other Kv channels, including the archabacterial KvAP and the Shaker channel of Drosophila.

  19. Erg and AP-1 as determinants of glucocorticoid response in acute lymphoblastic leukemia.

    PubMed

    Chen, D W-C; Saha, V; Liu, J-Z; Schwartz, J-M; Krstic-Demonacos, M

    2013-06-20

    Glucocorticoids (GCs) are among the most widely prescribed medications in clinical practice. The beneficial effects of GCs in acute lymphoblastic leukemia (ALL) are based on their ability to induce apoptosis, but the underlying transcriptional mechanisms remain poorly defined. Computational modeling has enormous potential in the understanding of biological processes such as apoptosis and the discovery of novel regulatory mechanisms. We here present an integrated analysis of gene expression kinetic profiles using microarrays from GC sensitive and resistant ALL cell lines and patients, including newly generated and previously published data sets available from the Gene Expression Omnibus. By applying time-series clustering analysis in the sensitive ALL CEM-C7-14 cells, we identified 358 differentially regulated genes that we classified into 15 kinetic profiles. We identified GC response element (GRE) sequences in 33 of the upregulated known or potential GC receptor (GR) targets. Comparative study of sensitive and resistant ALL showed distinct gene expression patterns and indicated unexpected similarities between sensitivity-restored and resistant ALL. We found that activator protein 1 (AP-1), Ets related gene (Erg) and GR pathways were differentially regulated in sensitive and resistant ALL. Erg protein levels were substantially higher in CEM-C1-15-resistant cells, c-Jun was significantly induced in sensitive cells, whereas c-Fos was expressed at low levels in both. c-Jun was recruited on the AP-1 site on the Bim promoter, whereas a transient Erg occupancy on the GR promoter was detected. Inhibition of Erg and activation of GR lead to increased apoptosis in both sensitive and resistant ALL. These novel findings significantly advance our understanding of GC sensitivity and can be used to improve therapy of leukemia.

  20. Evaluating the Efficacy of ERG-Targeted Therapy in Vivo for Prostate Tumors

    DTIC Science & Technology

    2015-04-01

    from other tissue specific drivers such as those that drive expression ubiquitously (see Figure 2 Appendix) or to the liver (data no shown). Thus...there is the formal possibility that the HE combination is not compatible, but we are still at a preliminary stage. 1b. Collecting tissues from AE...doxycycline (Table 1B). 1c. Collecting tissues from AE mice turned OFF to characterize inducible ERG expression (months 8-14). 12 week old males will be

  1. Small Molecule Inhibitors of ERG and ETV1 in Prostate Cancer

    DTIC Science & Technology

    2014-09-01

    SUPPLEMENTARY NOTES 14. ABSTRACT A group of chromosomal translocations were recently discovered in prostate cancer that fuses the 5` region of TMPRSS2 (a...common ETS member involved in prostate cancer chromosomal translocations is ERG but other members such as ETV1, ETV4 and ETV5 have been also observed...prognostic marker (6-8). Cell culture and transgenic animal models suggest that increased expression of ETS members, as a result of the chromosomal

  2. Understanding hERG inhibition with QSAR models based on a one-dimensional molecular representation

    NASA Astrophysics Data System (ADS)

    Diller, David J.; Hobbs, Doug W.

    2007-07-01

    Blockage of the potassium channel encoded by the human ether-a-go-go related gene (hERG) is well understood to be the root cause of the cardio-toxicity of numerous approved and investigational drugs. As such, a cascade of in vitro and in vivo assays have been developed to filter compounds with hERG inhibitory activity. Quantitative structure activity relationship (QSAR) models are used at the very earliest part of this cascade to eliminate compounds that are likely to have this undesirable activity prior to synthesis. Here a new QSAR technique based on the one-dimensional representation is described in the context of the development of a model to predict hERG inhibition. The model is shown to perform close to the limits of the quality of the data used for model building. In order to make optimal use of the available data, a general robust mathematical scheme was developed and is described to simultaneously incorporate quantitative data, such as IC50 = 50 nM, and qualitative data, such as inactive or IC50 > 30 μM into QSAR models without discarding any experimental information.

  3. BDNF improves the efficacy ERG amplitude maintenance by transplantation of retinal stem cells in RCS rats.

    PubMed

    Tian, Chunyu; Weng, Chuan Chuang; Yin, Zheng Qin

    2010-01-01

    The aim of this study was to evaluate the efficacy of subretinal transplantation of rat retinal stem cell when combined with Brain-derived neurotrophic factor (BDNF) in a rat model of retinal degeneration - Royal College of Surgeons (RCS) rats. Retinal stem cells were derived from embryonic day 17 Long-Evans rats and pre-labeled with fluorescence pigment-DiI prior to transplant procedures. RCS rats received injections of retinal stem cells, stem cells+BDNF, phosphate buffered saline or BNDF alone (n = 3 eyes for each procedure). At 1, 2 and 3 months after transplantation, the electroretinogram (ERG) was assessed and the outer nuclear layer thickness measured. The eyes receiving retinal stem cell and stem cell+BDNF transplants showed better photoreceptor maintenance than the other groups (P < 0.01) at all time points. One month after retina transplantation, the amplitudes of rod-ERG and Max-ERG b waves were significantly higher the eyes with stem cells+BDNF (P < 0.01), however, this difference was not seen at two and three months post transplantation. BDNF treatment alone group (without transplanted cells) had no effect when compared to buffer injections. The present results indicate that BDNF can enhance the short-term efficacy of the retinal stem cell transplantation in treating retinal degenerative disease.

  4. Deletion of Interstitial Genes between TMPRSS2 and ERG Promotes Prostate Cancer Progression.

    PubMed

    Linn, Douglas E; Penney, Kathryn L; Bronson, Roderick T; Mucci, Lorelei A; Li, Zhe

    2016-04-01

    TMPRSS2-ERG gene fusions that occur frequently in human prostate cancers can be generated either through insertional chromosomal rearrangement or by intrachromosomal deletion. Genetically, a key difference between these two mechanisms is that the latter results in deletion of a ∼3-Mb interstitial region containing genes with unexplored roles in prostate cancer. In this study, we characterized two mouse models recapitulating TMPRSS2-ERG insertion or deletion events in the background of prostate-specific PTEN deficiency. We found that only the mice that lacked the interstitial region developed prostate adenocarcinomas marked by poor differentiation and epithelial-to-mesenchymal transition. Mechanistic investigations identified several interstitial genes, including Ets2 and Bace2, whose reduced expression correlated in the gene homologs in human prostate cancer with biochemical relapse and lethal disease. Accordingly, PTEN-deficient mice with prostate-specific knockout of Ets2 exhibited marked progression of prostate adenocarcinomas that was partly attributed to activation of MAPK signaling. Collectively, our findings established that Ets2 is a tumor suppressor gene in prostate cancer, and its loss along with other genes within the TMPRSS2-ERG interstitial region contributes to disease progression. Cancer Res; 76(7); 1869-81. ©2016 AACR.

  5. Novel point mutations in the ERG11 gene in clinical isolates of azole resistant Candida species.

    PubMed

    Silva, Danielly Beraldo dos Santos; Rodrigues, Luana Mireli Carbonera; Almeida, Adriana Araújo de; Oliveira, Kelly Mari Pires de; Grisolia, Alexéia Barufatti

    2016-03-01

    The azoles are the class of medications most commonly used to fight infections caused by Candida sp. Typically, resistance can be attributed to mutations in ERG11 gene (CYP51) which encodes the cytochrome P450 14α-demethylase, the primary target for the activity of azoles. The objective of this study was to identify mutations in the coding region of theERG11 gene in clinical isolates of Candida species known to be resistant to azoles. We identified three new synonymous mutations in the ERG11 gene in the isolates of Candida glabrata (C108G, C423T and A1581G) and two new nonsynonymous mutations in the isolates of Candida krusei--A497C (Y166S) and G1570A (G524R). The functional consequence of these nonsynonymous mutations was predicted using evolutionary conservation scores. The G524R mutation did not have effect on 14α-demethylase functionality, while the Y166S mutation was found to affect the enzyme. This observation suggests a possible link between the mutation and dose-dependent sensitivity to voriconazole in the clinical isolate of C. krusei. Although the presence of the Y166S in phenotype of reduced azole sensitivity observed in isolate C. krusei demands investigation, it might contribute to the search of new therapeutic agents against resistant Candida isolates.

  6. Development, interpretation and temporal evaluation of a global QSAR of hERG electrophysiology screening data

    NASA Astrophysics Data System (ADS)

    Gavaghan, Claire L.; Arnby, Catrin Hasselgren; Blomberg, Niklas; Strandlund, Gert; Boyer, Scott

    2007-04-01

    A `global' model of hERG K+ channel was built to satisfy three basic criteria for QSAR models in drug discovery: (1) assessment of the applicability domain, (2) assuring that model decisions can be interpreted by medicinal chemists and (3) assessment of model performance after the model was built. A combination of D-optimal onion design and hierarchical partial least squares modelling was applied to construct a global model of hERG blockade in order to maximize the applicability domain of the model and to enhance its interpretability. Additionally, easily interpretable hERG specific fragment-based descriptors were developed. Model performance was monitored, throughout a time period of 15 months, after model implementation. It was found that after this time duration a greater proportion of molecules were outside the model's applicability domain and that these compounds had a markedly higher average prediction error than those from molecules within the model's applicability domain. The model's predictive performance deteriorated within 4 months after building, illustrating the necessity of regular updating of global models within a drug discovery environment.

  7. A k-nearest neighbor classification of hERG K+ channel blockers

    NASA Astrophysics Data System (ADS)

    Chavan, Swapnil; Abdelaziz, Ahmed; Wiklander, Jesper G.; Nicholls, Ian A.

    2016-03-01

    A series of 172 molecular structures that block the hERG K+ channel were used to develop a classification model where, initially, eight types of PaDEL fingerprints were used for k-nearest neighbor model development. A consensus model constructed using Extended-CDK, PubChem and Substructure count fingerprint-based models was found to be a robust predictor of hERG activity. This consensus model demonstrated sensitivity and specificity values of 0.78 and 0.61 for the internal dataset compounds and 0.63 and 0.54 for the external (PubChem) dataset compounds, respectively. This model has identified the highest number of true positives (i.e. 140) from the PubChem dataset so far, as compared to other published models, and can potentially serve as a basis for the prediction of hERG active compounds. Validating this model against FDA-withdrawn substances indicated that it may even be useful for differentiating between mechanisms underlying QT prolongation.

  8. Determining temperature distribution in tissue in the focal plane of the high (>100 W/cm(2)) intensity focused ultrasound beam using phase shift of ultrasound echoes.

    PubMed

    Karwat, Piotr; Kujawska, Tamara; Lewin, Peter A; Secomski, Wojciech; Gambin, Barbara; Litniewski, Jerzy

    2016-02-01

    In therapeutic applications of High Intensity Focused Ultrasound (HIFU) the guidance of the HIFU beam and especially its focal plane is of crucial importance. This guidance is needed to appropriately target the focal plane and hence the whole focal volume inside the tumor tissue prior to thermo-ablative treatment and beginning of tissue necrosis. This is currently done using Magnetic Resonance Imaging that is relatively expensive. In this study an ultrasound method, which calculates the variations of speed of sound in the locally heated tissue volume by analyzing the phase shifts of echo-signals received by an ultrasound scanner from this very volume is presented. To improve spatial resolution of B-mode imaging and minimize the uncertainty of temperature estimation the acoustic signals were transmitted and received by 8 MHz linear phased array employing Synthetic Transmit Aperture (STA) technique. Initially, the validity of the algorithm developed was verified experimentally in a tissue-mimicking phantom heated from 20.6 to 48.6 °C. Subsequently, the method was tested using a pork loin sample heated locally by a 2 MHz pulsed HIFU beam with focal intensity ISATA of 129 W/cm(2). The temperature calibration of 2D maps of changes in the sound velocity induced by heating was performed by comparison of the algorithm-determined changes in the sound velocity with the temperatures measured by thermocouples located in the heated tissue volume. The method developed enabled ultrasound temperature imaging of the heated tissue volume from the very inception of heating with the contrast-to-noise ratio of 3.5-12 dB in the temperature range 21-56 °C. Concurrently performed, conventional B-mode imaging revealed CNR close to zero dB until the temperature reached 50 °C causing necrosis. The data presented suggest that the proposed method could offer an alternative to MRI-guided temperature imaging for prediction of the location and extent of the thermal lesion prior to applying the

  9. PCA3 Reference Set Application: T2-Erg-Martin Sanda-Emory (2014) — EDRN Public Portal

    Cancer.gov

    We hypothesize that combining T2:erg (T2:erg) fusion and PCA3 detection in urine collected after digital rectal exam can improve the specificity of identifying clinically significant prostate cancer presence over the standard PSA and DRE. To address this hypothesis we propose to validate the performance of the urinary T2:erg in a multiplex model predicting the diagnosis of clinically significant prostate cancer on subsequent prostate biopsy using post-DRE pre biopsy urine specimens from a cohort of 900 men on the EDRN’s PCA3 trial.

  10. Analytical platform evaluation for quantification of ERG in prostate cancer using protein and mRNA detection methods

    DOE PAGES

    He, Jintang; Schepmoes, Athena A.; Shi, Tujin; ...

    2015-01-01

    Background: The established methods for detecting prostate cancer (CaP) are based on tests using PSA (blood), PCA3 (urine), and AMACR (tissue) as biomarkers in patient samples. The demonstration of ERG oncoprotein overexpression due to gene fusion in CaP has thus provided ERG as an additional biomarker. Based on this, we hypothesized that ERG protein quantification methods can be of use in the diagnosis of prostate cancer. Methods: Therefore, an antibody-free assay for ERG3 protein detection was developed based on PRISM (high-pressure high-resolution separations with intelligent selection and multiplexing)-SRM (selected reaction monitoring) mass spectrometry. We utilized TMPRSS2-ERG positive VCaP and TMPRSS2-ERGmore » negative LNCaP cells to simulate three different sample types (cells, tissue, and post-DRE urine sediment). Results: Recombinant ERG3 protein spiked into LNCaP cell lysates could be detected at levels as low as 20 pg by PRISM-SRM analysis. The sensitivity of the PRISM-SRM assay was around approximately 10,000 VCaP cells in a mixed cell population model of VCaP and LNCaP cells. Interestingly, ERG protein could be detected in as few as 600 VCaP cells spiked into female urine. The sensitivity of the in-house enzyme-linked immunosorbent assay (ELISA) was similar to the PRISM-SRM assay, with detection of 30 pg of purified recombinant ERG3 protein and 10,000 VCaP cells. On the other hand, qRT-PCR exhibited a higher sensitivity, as TMPRSS2-ERG transcripts were detected in as few as 100 VCaP cells, in comparison to NanoString methodologies which detected ERG from 10,000 cells. Conclusions: Based on this data, we propose that the detection of both ERG transcriptional products with RNA-based assays, as well as protein products of ERG using PRISM-SRM assays, may be of clinical value in developing diagnostics and prognostics assays for prostate cancer given their sensitivity, specificity, and reproducibility.« less

  11. Estimates of L:M cone ratio from ERG flicker photometry and genetics.

    PubMed

    Carroll, Joseph; Neitz, Jay; Neitz, Maureen

    2002-01-01

    Estimates of L:M cone ratio for males with normal color vision were derived using the flicker-photometric electroretinogram (ERG). These were obtained by best fitting ERG spectral sensitivity functions to a weighted sum of long (L)- and middle (M)-wavelength-sensitive cone spectral absorption curves. Using the ERG, measurements can be made with extremely high precision, which leaves variation in the wavelength of maximal sensitivity (lambda(max)) of the cone photopigments as the major remaining source of inaccuracy in determining the ratio of cone contributions. Here that source of inaccuracy was largely eliminated through the use of individualized L-cone spectral absorption curves deduced from L-pigment gene sequences. The method was used on 62 normal males as part of an effort to obtain a true picture of how normal variations in L:M cone ratio are distributed. The percentage of L cones in the average eye was 65%L [where %L = 100 X L / (L+M)]. There were huge individual differences ranging from 28%-93%L, corresponding to more than a 30-fold range in L:M ratio (0.4-13). However, the most extreme values were relatively rare; 80% of the subjects fell within +/-15 %L of the mean, corresponding to a 4-fold range in L:M ratio (1-4). The method remedies major weaknesses inherent in earlier applications of flicker photometry to estimate cone ratio; however, it continues to depend on the assumption that the average L cone produces a response with an identical amplitude to that of the average M cone. A comparison of the ERG results with the distribution of cone ratios estimated from cone pigment messenger RNA in cadaver eyes indicates that the assumption generally holds true. However, there may be a small number of exceptions in which individuals have normally occurring (but relatively rare) amino acid substitutions in one of their pigments that significantly affect the physiology of the cone class containing that pigment, so as to reduce the amplitude of its contribution

  12. Prevalence of Malocclusion among 10-12-year-old Schoolchildren in Kozhikode District, Kerala: An Epidemiological Study

    PubMed Central

    Jeseem, MT; Kumar, TV Anupam

    2016-01-01

    ABSTRACT Background: A malocclusion is an irregularity of the teeth or a malrelationship of the dental arches beyond the range of what is accepted as normal. Objectives: To determine the prevalence of malocclusion in children aged 10-12 years in Kozhikode district of Kerala, South India. Materials and methods: A descriptive cross-sectional study was conducted among schoolchildren aged 10-12 years in six schools in Kozhikode district of Kerala, South India. A total of 2,366 children satisfied the inclusion criteria. Occlusal characteristics like crossbite, open bite, deep bite, protrusion of teeth, midline deviations, midline diastema and tooth rotation were recorded. The data were tabulated and analyzed using Chi-square test. Results: The results revealed that the overall prevalence of malocclusion was 83.3%. Of this, 69.8% of the children had Angle’s class I malocclusion, 9.3% had class II malocclusion (division 1 = 8.85%, division 2 = 0.5%) and 4.1% had class III malocclusion; 23.2% showed an increased overjet (>3 mm), 0.4% reverse overjet, 35.6% increased overbite (>3 mm), 0.29% open bite, 7.2% crossbite with 4.6% crossbite of complete anterior teeth, 63.3% deviation of midline, 0.76% midline diastema and 3.25% rotated tooth. No significant differences in gender distributions of malocclusions were noted except for increased overjet and overbite. Conclusion: There is high prevalence of malocclusion among schoolchildren in Kozhikode district of Kerala. Early interception and early correction of these malocclusions will eliminate the potential irregularities and malpositions in the developing dentofacial complex. How to cite this article: Narayanan RK, Jeseem MT, Kumar TVA. Prevalence of Malocclusion among 10-12-year-old Schoolchildren in Kozhikode District, Kerala: An Epidemiological Study. Int J Clin Pediatr Dent 2016;9(1):50-55. PMID:27274156

  13. Characterization of a hERG screen using the IonWorks HT: comparison to a hERG rubidium efflux screen.

    PubMed

    Sorota, Steve; Zhang, Xue-Song; Margulis, Michael; Tucker, Kristal; Priestley, Tony

    2005-02-01

    The introduction of parallel patch clamp instruments offers the promise of moderate-throughput, high-fidelity voltage clamp for drug screening assays. One such device, the IonWorks HT (Molecular Devices, Sunnyvale, CA), was evaluated and compared to conventional human ethera- go-go-related gene (hERG) patch clamp data and an alternative functional screen based on rubidium flux. Data generated by the IonWorks HT and rubidium assays were compared to determine if either offered superior predictive value compared to conventional patch clamp. Concentration-effect curves for a panel of known hERG blockers were shifted to higher concentrations on the IonWorks HT compared to conventional voltage clamp determinations. The magnitude of the potency shifts was compound-specific and ranged from no shift (e.g., quinidine) to over 200-fold (astemizole). When the extreme value for astemizole was disregarded, the potency shift for 13 other known reference standards was 12-fold or less, with an average shift of fivefold. The same subset of compounds in the rubidium efflux assay exhibited an average potency shift of 12-fold. To provide a simulation of how the IonWorks HT assay might perform in a single concentration screening mode, a panel of test compounds was evaluated. The IonWorks HT screen did not outperform the rubidium efflux screen in predicting conventional voltage clamp measurements. The most likely explanation appears to rest with variable and compound-specific potency shifts in the IonWorks HT assay. The variable potency shifts make it difficult to select a screening concentration that meets the criterion of a high positive predictive value while avoiding false-positives.

  14. Effects of Tannic Acid, Green Tea and Red Wine on hERG Channels Expressed in HEK293 Cells.

    PubMed

    Chu, Xi; Guo, Yusong; Xu, Bingyuan; Li, Wenya; Lin, Yue; Sun, Xiaorun; Ding, Chunhua; Zhang, Xuan

    2015-01-01

    Tannic acid presents in varying concentrations in plant foods, and in relatively high concentrations in green teas and red wines. Human ether-à-go-go-related gene (hERG) channels expressed in multiple tissues (e.g. heart, neurons, smooth muscle and cancer cells), and play important roles in modulating cardiac action potential repolarization and tumor cell biology. The present study investigated the effects of tannic acid, green teas and red wines on hERG currents. The effects of tannic acid, teas and red wines on hERG currents stably transfected in HEK293 cells were studied with a perforated patch clamp technique. In this study, we demonstrated that tannic acid inhibited hERG currents with an IC50 of 3.4 μM and ~100% inhibition at higher concentrations, and significantly shifted the voltage dependent activation to more positive potentials (Δ23.2 mV). Remarkably, a 100-fold dilution of multiple types of tea (green tea, oolong tea and black tea) or red wine inhibited hERG currents by ~90%, and significantly shifted the voltage dependent activation to more positive potentials (Δ30.8 mV and Δ26.0 mV, respectively). Green tea Lung Ching and red wine inhibited hERG currents, with IC50 of 0.04% and 0.19%, respectively. The effects of tannic acid, teas and red wine on hERG currents were irreversible. These results suggest tannic acid is a novel hERG channel blocker and consequently provide a new mechanistic evidence for understanding the effects of tannic acid. They also revealed the potential pharmacological basis of tea- and red wine-induced biology activities.

  15. ADMET Evaluation in Drug Discovery. 16. Predicting hERG Blockers by Combining Multiple Pharmacophores and Machine Learning Approaches.

    PubMed

    Wang, Shuangquan; Sun, Huiyong; Liu, Hui; Li, Dan; Li, Youyong; Hou, Tingjun

    2016-08-01

    Blockade of human ether-à-go-go related gene (hERG) channel by compounds may lead to drug-induced QT prolongation, arrhythmia, and Torsades de Pointes (TdP), and therefore reliable prediction of hERG liability in the early stages of drug design is quite important to reduce the risk of cardiotoxicity-related attritions in the later development stages. In this study, pharmacophore modeling and machine learning approaches were combined to construct classification models to distinguish hERG active from inactive compounds based on a diverse data set. First, an optimal ensemble of pharmacophore hypotheses that had good capability to differentiate hERG active from inactive compounds was identified by the recursive partitioning (RP) approach. Then, the naive Bayesian classification (NBC) and support vector machine (SVM) approaches were employed to construct classification models by integrating multiple important pharmacophore hypotheses. The integrated classification models showed improved predictive capability over any single pharmacophore hypothesis, suggesting that the broad binding polyspecificity of hERG can only be well characterized by multiple pharmacophores. The best SVM model achieved the prediction accuracies of 84.7% for the training set and 82.1% for the external test set. Notably, the accuracies for the hERG blockers and nonblockers in the test set reached 83.6% and 78.2%, respectively. Analysis of significant pharmacophores helps to understand the multimechanisms of action of hERG blockers. We believe that the combination of pharmacophore modeling and SVM is a powerful strategy to develop reliable theoretical models for the prediction of potential hERG liability.

  16. Investigation of miscellaneous hERG inhibition in large diverse compound collection using automated patch-clamp assay

    PubMed Central

    Yu, Hai-bo; Zou, Bei-yan; Wang, Xiao-liang; Li, Min

    2016-01-01

    Aim: hERG potassium channels display miscellaneous interactions with diverse chemical scaffolds. In this study we assessed the hERG inhibition in a large compound library of diverse chemical entities and provided data for better understanding of the mechanisms underlying promiscuity of hERG inhibition. Methods: Approximately 300 000 compounds contained in Molecular Library Small Molecular Repository (MLSMR) library were tested. Compound profiling was conducted on hERG-CHO cells using the automated patch-clamp platform–IonWorks Quattro™. Results: The compound library was tested at 1 and 10 μmol/L. IC50 values were predicted using a modified 4-parameter logistic model. Inhibitor hits were binned into three groups based on their potency: high (IC50<1 μmol/L), intermediate (1 μmol/L< IC50<10 μmol/L), and low (IC50>10 μmol/L) with hit rates of 1.64%, 9.17% and 16.63%, respectively. Six physiochemical properties of each compound were acquired and calculated using ACD software to evaluate the correlation between hERG inhibition and the properties: hERG inhibition was positively correlative to the physiochemical properties ALogP, molecular weight and RTB, and negatively correlative to TPSA. Conclusion: Based on a large diverse compound collection, this study provides experimental evidence to understand the promiscuity of hERG inhibition. This study further demonstrates that hERG liability compounds tend to be more hydrophobic, high-molecular, flexible and polarizable. PMID:26725739

  17. Inhibition of hERG potassium channel by the antiarrhythmic agent mexiletine and its metabolite m-hydroxymexiletine

    PubMed Central

    Gualdani, Roberta; Tadini-Buoninsegni, Francesco; Roselli, Mariagrazia; Defrenza, Ivana; Contino, Marialessandra; Colabufo, Nicola Antonio; Lentini, Giovanni

    2015-01-01

    Mexiletine is a sodium channel blocker, primarily used in the treatment of ventricular arrhythmias. Moreover, recent studies have demonstrated its therapeutic value to treat myotonic syndromes and to relieve neuropathic pain. The present study aims at investigating the direct blockade of hERG potassium channel by mexiletine and its metabolite m-hydroxymexiletine (MHM). Our data show that mexiletine inhibits hERG in a time- and voltage-dependent manner, with an IC50 of 3.7 ± 0.7 μmol/L. Analysis of the initial onset of current inhibition during a depolarizing test pulse indicates mexiletine binds preferentially to the open state of the hERG channel. Looking for a possible mexiletine alternative, we show that m-hydroxymexiletine (MHM), a minor mexiletine metabolite recently reported to be as active as the parent compound in an arrhythmia animal model, is a weaker hERG channel blocker, compared to mexiletine (IC50 = 22.4 ± 1.2 μmol/L). The hERG aromatic residues located in the S6 helix (Tyr652 and Phe656) are crucial in the binding of mexiletine and the different affinities of mexiletine and MHM with hERG channel are interpreted by modeling their corresponding binding interactions through ab initio calculations. The simulations demonstrate that the introduction of a hydroxyl group on the meta-position of the aromatic portion of mexiletine weakens the interaction of the drug xylyloxy moiety with Tyr652. These results provide further insights into the molecular basis of drug/hERG interactions and, in agreement with previously reported results on clofilium and ibutilide analogs, support the possibility of reducing hERG potency and related toxicity by modifying the aromatic pattern of substitution of clinically relevant compounds. PMID:26516576

  18. Postsynaptic ERG potassium channels limit muscle excitability to allow distinct egg-laying behavior states in Caenorhabditis elegans.

    PubMed

    Collins, Kevin M; Koelle, Michael R

    2013-01-09

    Caenorhabditis elegans regulates egg laying by alternating between an inactive phase and a serotonin-triggered active phase. We found that the conserved ERG [ether-a-go-go (EAG) related gene] potassium channel UNC-103 enables this two-state behavior by limiting excitability of the egg-laying muscles. Using both high-speed video recording and calcium imaging of egg-laying muscles in behaving animals, we found that the muscles appear to be excited at a particular phase of each locomotor body bend. During the inactive phase, this rhythmic excitation infrequently evokes calcium transients or contraction of the egg-laying muscles. During the serotonin-triggered active phase, however, these muscles are more excitable and each body bend is accompanied by a calcium transient that drives twitching or full contraction of the egg-laying muscles. We found that ERG-null mutants lay eggs too frequently, and that ERG function is necessary and sufficient in the egg-laying muscles to limit egg laying. ERG K(+) channels localize to postsynaptic sites in the egg-laying muscle, and mutants lacking ERG have more frequent calcium transients and contractions of the egg-laying muscles even during the inactive phase. Thus ERG channels set postsynaptic excitability at a threshold so that further adjustments of excitability by serotonin generate two distinct behavioral states.

  19. Dual requirement for the ETS transcription factors Fli-1 and Erg in hematopoietic stem cells and the megakaryocyte lineage

    PubMed Central

    Kruse, Elizabeth A.; Loughran, Stephen J.; Baldwin, Tracey M.; Josefsson, Emma C.; Ellis, Sarah; Watson, Dennis K.; Nurden, Paquita; Metcalf, Donald; Hilton, Douglas J.; Alexander, Warren S.; Kile, Benjamin T.

    2009-01-01

    Fli-1 and Erg are closely related members of the Ets family of transcription factors. Both genes are translocated in human cancers, including Ewing's sarcoma, leukemia, and in the case of Erg, more than half of all prostate cancers. Although evidence from mice and humans suggests that Fli-1 is required for megakaryopoiesis, and that Erg is required for normal adult hematopoietic stem cell (HSC) regulation, their precise physiological roles remain to be defined. To elucidate the relationship between Fli-1 and Erg in hematopoiesis, we conducted an analysis of mice carrying mutations in both genes. Our results demonstrate that there is a profound genetic interaction between Fli-1 and Erg. Double heterozygotes displayed phenotypes more dramatic than single heterozygotes: severe thrombocytopenia, with a significant deficit in megakaryocyte numbers and evidence of megakaryocyte dysmorphogenesis, and loss of HSCs accompanied by a reduction in the number of committed hematopoietic progenitor cells. These results illustrate an indispensable requirement for both Fli-1 and Erg in normal HSC and megakaryocyte homeostasis, and suggest these transcription factors may coregulate common target genes. PMID:19666492

  20. Translational toxicology and rescue strategies of the hERG channel dysfunction: biochemical and molecular mechanistic aspects

    PubMed Central

    Zhang, Kai-ping; Yang, Bao-feng; Li, Bao-xin

    2014-01-01

    The human ether-à-go-go related gene (hERG) potassium channel is an obligatory anti-target for drug development on account of its essential role in cardiac repolarization and its close association with arrhythmia. Diverse drugs have been removed from the market owing to their inhibitory activity on the hERG channel and their contribution to acquired long QT syndrome (LQTS). Moreover, mutations that cause hERG channel dysfunction may induce congenital LQTS. Recently, an increasing number of biochemical and molecular mechanisms underlying hERG-associated LQTS have been reported. In fact, numerous potential biochemical and molecular rescue strategies are hidden within the biogenesis and regulating network. So far, rescue strategies of hERG channel dysfunction and LQTS mainly include activators, blockers, and molecules that interfere with specific links and other mechanisms. The aim of this review is to discuss the rescue strategies based on hERG channel toxicology from the biochemical and molecular perspectives. PMID:25418379

  1. A DNA methylation microarray-based study identifies ERG as a gene commonly methylated in prostate cancer

    PubMed Central

    Schwartzman, Jacob; Mongoue-Tchokote, Solange; Gibbs, Angela; Gao, Lina; Corless, Christopher L; Jin, Jennifer; Zarour, Luai; Higano, Celestia; True, Lawrence D; Vessella, Robert L; Wilmot, Beth; Bottomly, Daniel; McWeeney, Shannon K; Bova, G. Steven; Partin, Alan W; Mori, Motomi

    2011-01-01

    DNA methylation of promoter regions is a common event in prostate cancer, one of the most common cancers in men worldwide. Because prior reports demonstrating that DNA methylation is important in prostate cancer studied a limited number of genes, we systematically quantified the DNA methylation status of 1,505 CpG dinucleotides for 807 genes in 78 paraffin-embedded prostate cancer samples and three normal prostate samples. The ERG gene, commonly repressed in prostate cells in the absence of an oncogenic fusion to the TMPRSS2 gene, was one of the most commonly methylated genes, occurring in 74% of prostate cancer specimens. In an independent group of patient samples, we confirmed that ERG DNA methylation was common, occurring in 57% of specimens, and cancer-specific. The ERG promoter is marked by repressive chromatin marks mediated by polycomb proteins in both normal prostate cells and prostate cancer cells, which may explain ERG's predisposition to DNA methylation and the fact that tumors with ERG DNA methylation were more methylated, in general. These results demonstrate that bead arrays offer a high-throughput method to discover novel genes with promoter DNA methylation such as ERG, whose measurement may improve our ability to more accurately detect prostate cancer. PMID:21946329

  2. Dual requirement for the ETS transcription factors Fli-1 and Erg in hematopoietic stem cells and the megakaryocyte lineage.

    PubMed

    Kruse, Elizabeth A; Loughran, Stephen J; Baldwin, Tracey M; Josefsson, Emma C; Ellis, Sarah; Watson, Dennis K; Nurden, Paquita; Metcalf, Donald; Hilton, Douglas J; Alexander, Warren S; Kile, Benjamin T

    2009-08-18

    Fli-1 and Erg are closely related members of the Ets family of transcription factors. Both genes are translocated in human cancers, including Ewing's sarcoma, leukemia, and in the case of Erg, more than half of all prostate cancers. Although evidence from mice and humans suggests that Fli-1 is required for megakaryopoiesis, and that Erg is required for normal adult hematopoietic stem cell (HSC) regulation, their precise physiological roles remain to be defined. To elucidate the relationship between Fli-1 and Erg in hematopoiesis, we conducted an analysis of mice carrying mutations in both genes. Our results demonstrate that there is a profound genetic interaction between Fli-1 and Erg. Double heterozygotes displayed phenotypes more dramatic than single heterozygotes: severe thrombocytopenia, with a significant deficit in megakaryocyte numbers and evidence of megakaryocyte dysmorphogenesis, and loss of HSCs accompanied by a reduction in the number of committed hematopoietic progenitor cells. These results illustrate an indispensable requirement for both Fli-1 and Erg in normal HSC and megakaryocyte homeostasis, and suggest these transcription factors may coregulate common target genes.

  3. Novel characteristics of a trafficking-defective G572R-hERG channel linked to hereditary long QT syndrome.

    PubMed

    Lian, Jiangfang; Huang, Na; Zhou, Junbo; Ge, Shijun; Huang, Xiaoyan; Huo, Jianhua; Liu, Liying; Xu, Weifeng; Zhang, Shun; Yang, Xi; Zhou, Jianqing; Huang, Chen

    2010-10-01

    HISTORIQUE : Le syndrome du QT long congénital est une maladie génétique hétérogène associée à un retard de la repolarisation cardiaque, à des intervalles QT prolongés, à l'apparition d'arythmies ventriculaires et à une mort subite. Le syndrome du QT long congénital de type 2 (QTL2) est causé par des mutations du gène KCNH2 ou hERG. Le gène hERG code la sous-unité alpha K11.1 du canal K+ redresseur retardé du cœur qui s'active rapidement. Des études des canaux hERG mutants indiquent que la plupart des mutations faux-sens QTL2 produisent des canaux K11.1 au trafic défectueux. OBJECTIF : Dépister les mécanismes sous-jacents du canal G572R-hERG au moyen d'analyses moléculaires et électrophysiologiques. MÉTHODOLOGIE ET RÉSULTATS : Afin d'établir les propriétés électrophysiologiques des canaux mutants G572R-hERG, les sous-unités mutantes hERG étaient exprimées de manière hétérologue dans des cellules HEK293, seules ou en combinaison avec des sous-unités hERG de type sauvage (WT). Les techniques du patch-clamp ont permis d'enregistrer les courants, tandis que le marquage des protéines par double immunofluorescence et le transfert Western ont permis d'examiner le trafic cellulaire des sous-unités mutantes. Exprimées seules, les sous-unités G572R-hERG étaient absentes de la membrane cellulaire et ne produisaient pas de courants décelables. Coexprimé avec les sous-unités WT-hERG, le canal G572R-hERG réduisait la densité du courant et modifiait les propriétés de synchronisation du canal WT-hERG. CONCLUSION : La mutation faux-sens du gène G572R associée au gène hERG, comme la plupart des mutations faux-sens QTL2, produit un phénotype au trafic défectueux. De plus, le canal G572R-hERG entraîne une perte de fonction du gène hERG par son puissant effet négatif dominant sur le canal WT-hERG.

  4. A comparison among different techniques for human ERG signals processing and classification.

    PubMed

    Barraco, R; Persano Adorno, D; Brai, M; Tranchina, L

    2014-02-01

    Feature detection in biomedical signals is crucial for deepening our knowledge about the involved physiological processes. To achieve this aim, many analytic approaches can be applied but only few are able to deal with signals whose time dependent features provide useful clinical information. Among the biomedical signals, the electroretinogram (ERG), that records the retinal response to a light flash, can improve our comprehension of the complex photoreceptoral activities. The present study is focused on the analysis of the early response of the photoreceptoral human system, known as a-wave ERG-component. This wave reflects the functional integrity of the photoreceptors, rods and cones, whose activation dynamics are not yet completely understood. Moreover, since in incipient photoreceptoral pathologies eventual anomalies in a-wave are not always detectable with a "naked eye" analysis of the traces, the possibility to discriminate pathologic from healthy traces, by means of appropriate analytical techniques, could help in clinical diagnosis. In the present paper, we discuss and compare the efficiency of various techniques of signal processing, such as Fourier analysis (FA), Principal Component Analysis (PCA), Wavelet Analysis (WA) in recognising pathological traces from the healthy ones. The investigated retinal pathologies are Achromatopsia, a cone disease and Congenital Stationary Night Blindness, affecting the photoreceptoral signal transmission. Our findings prove that both PCA and FA of conventional ERGs, don't add clinical information useful for the diagnosis of ocular pathologies, whereas the use of a more sophisticated analysis, based on the wavelet transform, provides a powerful tool for routine clinical examinations of patients.

  5. Branch retinal artery occlusion with visual field and multifocal erg in Susac syndrome: a case report.

    PubMed

    van Winden, Marianne; Salu, Paul

    2010-12-01

    A 19-year-old woman presented with subacute encephalopathy and subsequently developed hearing loss and occlusions of branches of the central retinal artery. The triad of microangiopathy of the brain, retina and cochlea is typical for Susac syndrome. The etiology of this syndrome is still unknown, but the prognosis is good in most cases. Spontaneous resolution usually occurs, but early treatment minimizes the risk of sequelae. Multifocal ERG may be useful to assess subclinical retinal dysfunction after recovery of subjective symptoms of BRAO in Susac syndrome.

  6. MicroRNA-224 targets ERG2 and contributes to malignant progressions of meningioma

    SciTech Connect

    Wang, Maomao; Deng, Xiaodong; Ying, Qi; Jin, Tingyan; Li, Ming; Liang, Chong

    2015-05-01

    MicroRNA-224 is overexpressed in various malignant tumors with poor prognosis, which plays a critical role in biological processes including cell proliferation, apoptosis and several developmental and physiological progressions. However, the potential association between miR-224 and clinical outcome in patients with meningiomas remains unknown. Here, we investigate miR-224 expression and biological functions in meningiomas. MiR-224 expression was measured by Northern blot analysis and quantitative reverse transcription-polymerase chain reaction (qRT-PCR) in meningioma and normal brain tissues. Kaplan–Meier analysis and Cox regression analysis were used to exam its correlation with clinicopathological features and prognostic value. The biological effects of miR-224 on the cell proliferation and apoptosis in meningioma cells were examined by MTT assay and apoptosis assay. We found the expression levels of miR-224 were significantly higher in meningioma tissues than that in normal brain, positively correlated with advanced pathological grade. Kaplan–Meier analysis indicated that meningioma patients with low miR-224 expression exhibited significantly prolonged overall and recurrence-free survival. Furthermore, we demonstrated that ERG2 was an identical candidate target gene of MiR-224 in vitro. Our results indicated that downregulation of miR-224 suppressed cell growth and resulted in the enhancement of cell apoptosis through activation of the ERG2-BAK-induced apoptosis pathway. Our findings imply the miR-224 expression could predict the overall survival and recurrence-free survival of patients with meningioma and it might be a promising therapeutic target for treating malignant meningiomas. - Highlights: • MiR-224 expression is correlates with prognosis in meningioma patients. • ERG2 is a novel downstream target of miR-224. • MiR-224 suppressed cell growth and enhanced apoptosis in IOMM-Lee and CH157 cells. • MiR-224 is an upstream regulator of the ERG2

  7. Assessment of temperature-induced hERG channel blockade variation by drugs.

    PubMed

    Kauthale, Rahul R; Dadarkar, Shruta S; Husain, Raghib; Karande, Vikas V; Gatne, Madhumanjiri M

    2015-07-01

    Drug-induced QT prolongation has been reported in humans and animals. This potentially lethal effect can be induced by drugs interacting with a cardiac potassium channel, namely hERG (human ether-a go-go-related gene) leading to arrhythmia or torsade de pointes (TdP). Hence, in vitro evaluation of therapeutics for their effects on the rapid delayed rectifier current (IKr) mediated by the K(+) ion channel encoded by hERG is a valuable tool for identifying potential arrhythmic side effects during drug safety testing. Our objective was to evaluate the temperature-induced hERG channel blockade variation by human and veterinary drugs using the IonFlux 16 system. A panel of eight drugs was tested for IKr inhibition at both ambient (23 °C) and physiological (37 °C) temperatures at various concentrations using IonFlux 16, an automated patch clamp system. Our results established that both amiodarone (IC(50)  = 0.56 μM at 23 °C and 0.30 μM at 37 °C) and β-estradiol (IC(50)  = 24.72 μM at 23 °C and 8.17 μM at 37 °C) showed a dose-dependent IKr blockade with a higher blockade at 37 °C. Whereas, blockade of IKr by both ivermectin (IC(50)  = 12.52 μM at 23 °C and 24.41 μM at 37 °C) and frusemide (IC(50)  = 12.58 μM at 23 °C and 25.55 μM at 37 °C) showed a dose-dependent IKr blockade with a lower blockade at 37 °C. Gentamicin, enrofloxacin, xylazine and albendazole did not block IKr at both the assessed temperatures. Collectively, these results demonstrate that the effect of temperature variation should be taken into consideration during the evaluation of test drugs for their hERG channel blockade potential.

  8. Metabolically distinct weight loss by 10,12 CLA and caloric restriction highlight the importance of subcutaneous white adipose tissue for glucose homeostasis in mice

    PubMed Central

    Wang, Shari; Goodspeed, Leela; Wietecha, Tomasz; Houston, Barbara; Omer, Mohamed; Ogimoto, Kayoko; Subramanian, Savitha; Gowda, G. A. Nagana; O’Brien, Kevin D.; Kaiyala, Karl J.; Morton, Gregory J.; Chait, Alan

    2017-01-01

    Background Widely used as a weight loss supplement, trans-10,cis-12 conjugated linoleic acid (10,12 CLA) promotes fat loss in obese mice and humans, but has also been associated with insulin resistance. Objective We therefore sought to directly compare weight loss by 10,12 CLA versus caloric restriction (CR, 15–25%), an acceptable healthy method of weight loss, to determine how 10,12 CLA-mediated weight loss fails to improve glucose metabolism. Methods Obese mice with characteristics of human metabolic syndrome were either supplemented with 10,12 CLA or subjected to CR to promote weight loss. Metabolic endpoints such as energy expenditure, glucose and insulin tolerance testing, and trunk fat distribution were measured. Results By design, 10,12 CLA and CR caused equivalent weight loss, with greater fat loss by 10,12 CLA accompanied by increased energy expenditure, reduced respiratory quotient, increased fat oxidation, accumulation of alternatively activated macrophages, and browning of subcutaneous white adipose tissue (WAT). Moreover, 10,12 CLA-supplemented mice better defended their body temperature against a cold challenge. However, 10,12 CLA concurrently induced the detrimental loss of subcutaneous WAT without reducing visceral WAT, promoted reduced plasma and WAT adipokine levels, worsened hepatic steatosis, and failed to improve glucose metabolism. Obese mice undergoing CR were protected from subcutaneous-specific fat loss, had improved hepatic steatosis, and subsequently showed the expected improvements in WAT adipokines, glucose metabolism and WAT inflammation. Conclusions These results suggest that 10,12 CLA mediates the preferential loss of subcutaneous fat that likely contributes to hepatic steatosis and maintained insulin resistance, despite significant weight loss and WAT browning in mice. Collectively, we have shown that weight loss due to 10,12 CLA supplementation or CR results in dramatically different metabolic phenotypes, with the latter

  9. Surface complexation studied via combined grazing-incidence EXAFS and surface diffraction: Arsenate on hematite (0001) and (10-12)

    USGS Publications Warehouse

    Waychunas, G.; Trainor, T.; Eng, P.; Catalano, J.; Brown, G.; Davis, J.; Rogers, J.; Bargar, J.

    2005-01-01

    X-ray diffraction [crystal-truncation-rod (CTR)] studies of the surface structure of moisture-equilibrated hematite reveal sites for complexation not present on the bulk oxygen-terminated surface, and impose constraints on the types of inner-sphere sorption topologies. We have used this improved model of the hematite surface to analyze grazing-incidence EXAFS results for arsenate sorption on the c(0001) and r(10-12) surfaces measured in two electric vector polarizations. This work shows that the reconfiguration of the surface under moist conditions is responsible for an increased adsorption density of arsenate complexes on the (0001) surface relative to predicted ideal termination, and an abundance of "edge-sharing" bidentate complexes on both studied surfaces. We consider possible limitations on combining the methods due to differing surface sensitivities, and discuss further analysis possibilities using both methods. ?? Springer-Verlag 2005.

  10. Colorimetric Detection of Some Highly Hydrophobic Flavonoids Using Polydiacetylene Liposomes Containing Pentacosa-10,12-diynoyl Succinoglycan Monomers

    PubMed Central

    Yun, Deokgyu; Jeong, Daham; Cho, Eunae; Jung, Seunho

    2015-01-01

    Flavonoids are a group of plant secondary metabolites including polyphenolic molecules, and they are well known for antioxidant, anti-allergic, anti-inflammatory and anti-viral propertied. In general, flavonoids are detected with various non-colorimetric detection methods such as column liquid chromatography, thin-layer chromatography, and electrochemical analysis. For the first time, we developed a straightforward colorimetric detection system allowing recognition of some highly hydrophobic flavonoids such as alpha-naphthoflavone and beta-naphthoflavone, visually using 10,12-pentacosadiynoic acid (PCDA) derivatized with succinoglycan monomers isolated from Sinorhizobium meliloti. Besides changes in visible spectrum, we also demonstrate fluorescence changes using our detection system in the presence of those flavonoids. The succinoglycan monomers attached to PCDA molecules may function as an unstructured molecular capturer for some highly hydrophobic flavonoids by hydrophobic interactions, and transmit their molecular interactions as a color change throughout the PCDA liposome. PMID:26600071

  11. Evaluation of the TMPRSS2:ERG fusion for the detection of prostate cancer: a systematic review and meta-analysis.

    PubMed

    Yao, Yanhong; Wang, Haitao; Li, BaoGuo; Tang, Yong

    2014-03-01

    The diagnostic value of TMPRSS2:ERG detection in patients with prostate cancer is controversial. We performed a meta-analysis to consolidate current evidence regarding the use of TMPRSS2:ERG detection assays to diagnose prostate cancers. PubMed, Web of knowledge and other databases were searched for relevant original articles published until July 30, 2013. Methodological quality was assessed using the Quality Assessment of Diagnostic Accuracy Studies (QUADAS) tool. Studies that investigated the presence of TMPRSS2:ERG in the body fluid, needle biopsy and prostatectomy tissue of patients with prostate cancer were identified and reviewed. Sensitivities, specificities, and positive likelihood ratios (LR+) and negative likelihood ratios (LR-) of TMPRSS2:ERG detection in individual studies were calculated and meta-analyzed by random effects model. Thirty-two studies met the inclusion criteria for the meta-analysis. Overall sensitivity of TMPRSS2:ERG detection assays was 47.4% (95% CI, 45.5-49.3%); specificity, LR+, and LR- was 92.6% (95% CI, 91.5-93.7%), 8.94 (95% CI, 5.65-14.13) and 0.49 (95% CI, 0.43-0.55). The pooled sensitivity and specificity in the body fluid subgroup was 44.7% (95% CI, 41.5-47.9 %) and 85.8% (95% CI, 83.5-87.8%), respectively. The pooled sensitivity and specificity based on the reverse transcripts PCR was 49.0% (95% CI, 45.9-52.1%) and 90.2% (95% CI, 88.2-92.0%), respectively. TMPRSS2:ERG may not be used as first-line screening test. However, due to the high specificity, TMPRSS2: ERG detection maybe can serve as a quick and noninvasive method for confirming prostate cancer diagnosis.

  12. A functional Kv1.2-hERG chimaeric channel expressed in Pichia pastoris

    NASA Astrophysics Data System (ADS)

    Dhillon, Mandeep S.; Cockcroft, Christopher J.; Munsey, Tim; Smith, Kathrine J.; Powell, Andrew J.; Carter, Paul; Wrighton, David C.; Rong, Hong-Lin; Yusaf, Shahnaz P.; Sivaprasadarao, Asipu

    2014-02-01

    Members of the six-transmembrane segment family of ion channels share a common structural design. However, there are sequence differences between the members that confer distinct biophysical properties on individual channels. Currently, we do not have 3D structures for all members of the family to help explain the molecular basis for the differences in their biophysical properties and pharmacology. This is due to low-level expression of many members in native or heterologous systems. One exception is rat Kv1.2 which has been overexpressed in Pichia pastoris and crystallised. Here, we tested chimaeras of rat Kv1.2 with the hERG channel for function in Xenopus oocytes and for overexpression in Pichia. Chimaera containing the S1-S6 transmembrane region of HERG showed functional and pharmacological properties similar to hERG and could be overexpressed and purified from Pichia. Our results demonstrate that rat Kv1.2 could serve as a surrogate to express difficult-to-overexpress members of the six-transmembrane segment channel family.

  13. Retinal electrophysiology for toxicology studies: applications and limits of ERG in animals and ex vivo recordings.

    PubMed

    Rosolen, Serge Georges; Kolomiets, Bogdan; Varela, Oscar; Picaud, Serge

    2008-06-01

    Assessing retinal drug toxicity is becoming increasingly important as different molecules are now developed for the treatment of neurodegenerative diseases and vascular disorders. In pharmacology and toxicology, the electroretinogram (ERG) and the multielectrode array (MEA) recording techniques can be used to quantify the possible side effects of retino-active xenobiotics. Toxicity testing requires the use of rodent as well as non-rodent models for extrapolation to the human model when determining risk and safety. Animal species differ in their retinal anatomo-physiology: most rodents used in toxicology studies are essentially nocturnal species, whereas the non-rodent laboratory species normally used (e.g. dogs, pigs and monkeys) are diurnal. The ratio between the photoreceptor populations which varies from species to species, should be considered when designing the experiment protocol and the interpretation. The described ERG procedures are designed to comply with all applicable good laboratory practice standards. Use of these procedures should yield an acceptable level of intra- and inter-subject variability for compiling a historical database, and for detecting possible retinal toxicity in animal studies. They could therefore be used as specific and standardized tools for screening of potential retinotoxic molecules in drug discovery and development in order to compare methods and results with those obtained in human electrophysiological assessments. Recording of ganglion cell light responses on ex vivo retina with the MEA technique can further demonstrate how retino-active xenobiotics affect retinal visual information processing by eliminating potential obstacles related to bioavailability and blood barrier permeability.

  14. Loss of miR-449a in ERG-associated prostate cancer promotes the invasive phenotype by inducing SIRT1

    PubMed Central

    Kumar, Parameet; Sharad, Shashwat; Petrovics, Gyorgy; Mohamed, Ahmed; Dobi, Albert; Sreenath, Taduru L.; Srivastava, Shiv; Biswas, Roopa

    2016-01-01

    Epigenetic regulation by SIRT1, a multifaceted NAD+-dependent protein deacetylase, is one of the most common factors modulating cellular processes in a broad range of diseases, including prostate cancer (CaP). SIRT1 is over-expressed in CaP cells, however the associated mechanism is not well understood. To identify whether specific microRNAs might mediate this linkage, we have screened a miRNA library for differential expression in ERG-associated CaP tissues. Of 20 differentially and significantly expressed miRNAs that distinguish ERG-positive tumors from ERG-negative tumors, we find miR-449a is highly suppressed in ERG-positive tumors. We establish that SIRT1 is a direct target of miR-449a and is also induced by ERG in ERG-associated CaP. Our data suggest that attenuation of miR-449a promotes the invasive phenotype of the ERG-positive CaP in part by inducing the expression of SIRT1 in prostate cancer cells. Furthermore, we also find that suppression of SIRT1 results in a significant reduction in ERG expression in ERG-positive CaP cells, indicating a feed-back regulatory loop associated with ERG, miR-449a and SIRT1. We also report that ERG suppresses p53 acetylation perhaps through miR-449a-SIRT1 axis in CaP cells. Our findings provide new insight into the function of miRNAs in regulating ERG-associated CaP. Thus, miR-449a activation or SIRT1 suppression may represent new therapeutic opportunity for ERG-associated CaP. PMID:26988912

  15. Loss of miR-449a in ERG-associated prostate cancer promotes the invasive phenotype by inducing SIRT1.

    PubMed

    Kumar, Parameet; Sharad, Shashwat; Petrovics, Gyorgy; Mohamed, Ahmed; Dobi, Albert; Sreenath, Taduru L; Srivastava, Shiv; Biswas, Roopa

    2016-04-19

    Epigenetic regulation by SIRT1, a multifaceted NAD+-dependent protein deacetylase, is one of the most common factors modulating cellular processes in a broad range of diseases, including prostate cancer (CaP). SIRT1 is over-expressed in CaP cells, however the associated mechanism is not well understood. To identify whether specific microRNAs might mediate this linkage, we have screened a miRNA library for differential expression in ERG-associated CaP tissues. Of 20 differentially and significantly expressed miRNAs that distinguish ERG-positive tumors from ERG-negative tumors, we find miR-449a is highly suppressed in ERG-positive tumors. We establish that SIRT1 is a direct target of miR-449a and is also induced by ERG in ERG-associated CaP. Our data suggest that attenuation of miR-449a promotes the invasive phenotype of the ERG-positive CaP in part by inducing the expression of SIRT1 in prostate cancer cells. Furthermore, we also find that suppression of SIRT1 results in a significant reduction in ERG expression in ERG-positive CaP cells, indicating a feed-back regulatory loop associated with ERG, miR-449a and SIRT1. We also report that ERG suppresses p53 acetylation perhaps through miR-449a-SIRT1 axis in CaP cells. Our findings provide new insight into the function of miRNAs in regulating ERG-associated CaP. Thus, miR-449a activation or SIRT1 suppression may represent new therapeutic opportunity for ERG-associated CaP.

  16. A quantitative proteomics approach identifies ETV6 and IKZF1 as new regulators of an ERG-driven transcriptional network

    PubMed Central

    Unnikrishnan, Ashwin; Guan, Yi F.; Huang, Yizhou; Beck, Dominik; Thoms, Julie A. I.; Peirs, Sofie; Knezevic, Kathy; Ma, Shiyong; de Walle, Inge V.; de Jong, Ineke; Ali, Zara; Zhong, Ling; Raftery, Mark J.; Taghon, Tom; Larsson, Jonas; MacKenzie, Karen L.; Van Vlierberghe, Pieter; Wong, Jason W. H.; Pimanda, John E.

    2016-01-01

    Aberrant stem cell-like gene regulatory networks are a feature of leukaemogenesis. The ETS-related gene (ERG), an important regulator of normal haematopoiesis, is also highly expressed in T-ALL and acute myeloid leukaemia (AML). However, the transcriptional regulation of ERG in leukaemic cells remains poorly understood. In order to discover transcriptional regulators of ERG, we employed a quantitative mass spectrometry-based method to identify factors binding the 321 bp ERG +85 stem cell enhancer region in MOLT-4 T-ALL and KG-1 AML cells. Using this approach, we identified a number of known binders of the +85 enhancer in leukaemic cells along with previously unknown binders, including ETV6 and IKZF1. We confirmed that ETV6 and IKZF1 were also bound at the +85 enhancer in both leukaemic cells and in healthy human CD34+ haematopoietic stem and progenitor cells. Knockdown experiments confirmed that ETV6 and IKZF1 are transcriptional regulators not just of ERG, but also of a number of genes regulated by a densely interconnected network of seven transcription factors. At last, we show that ETV6 and IKZF1 expression levels are positively correlated with expression of a number of heptad genes in AML and high expression of all nine genes confers poorer overall prognosis. PMID:27604872

  17. A quantitative proteomics approach identifies ETV6 and IKZF1 as new regulators of an ERG-driven transcriptional network.

    PubMed

    Unnikrishnan, Ashwin; Guan, Yi F; Huang, Yizhou; Beck, Dominik; Thoms, Julie A I; Peirs, Sofie; Knezevic, Kathy; Ma, Shiyong; de Walle, Inge V; de Jong, Ineke; Ali, Zara; Zhong, Ling; Raftery, Mark J; Taghon, Tom; Larsson, Jonas; MacKenzie, Karen L; Van Vlierberghe, Pieter; Wong, Jason W H; Pimanda, John E

    2016-12-15

    Aberrant stem cell-like gene regulatory networks are a feature of leukaemogenesis. The ETS-related gene (ERG), an important regulator of normal haematopoiesis, is also highly expressed in T-ALL and acute myeloid leukaemia (AML). However, the transcriptional regulation of ERG in leukaemic cells remains poorly understood. In order to discover transcriptional regulators of ERG, we employed a quantitative mass spectrometry-based method to identify factors binding the 321 bp ERG +85 stem cell enhancer region in MOLT-4 T-ALL and KG-1 AML cells. Using this approach, we identified a number of known binders of the +85 enhancer in leukaemic cells along with previously unknown binders, including ETV6 and IKZF1. We confirmed that ETV6 and IKZF1 were also bound at the +85 enhancer in both leukaemic cells and in healthy human CD34(+) haematopoietic stem and progenitor cells. Knockdown experiments confirmed that ETV6 and IKZF1 are transcriptional regulators not just of ERG, but also of a number of genes regulated by a densely interconnected network of seven transcription factors. At last, we show that ETV6 and IKZF1 expression levels are positively correlated with expression of a number of heptad genes in AML and high expression of all nine genes confers poorer overall prognosis.

  18. hERG blocking potential of acids and zwitterions characterized by three thresholds for acidity, size and reactivity.

    PubMed

    Nikolov, Nikolai G; Dybdahl, Marianne; Jónsdóttir, Svava Ó; Wedebye, Eva B

    2014-11-01

    Ionization is a key factor in hERG K(+) channel blocking, and acids and zwitterions are known to be less probable hERG blockers than bases and neutral compounds. However, a considerable number of acidic compounds block hERG, and the physico-chemical attributes which discriminate acidic blockers from acidic non-blockers have not been fully elucidated. We propose a rule for prediction of hERG blocking by acids and zwitterionic ampholytes based on thresholds for only three descriptors related to acidity, size and reactivity. The training set of 153 acids and zwitterionic ampholytes was predicted with a concordance of 91% by a decision tree based on the rule. Two external validations were performed with sets of 35 and 48 observations, respectively, both showing concordances of 91%. In addition, a global QSAR model of hERG blocking was constructed based on a large diverse training set of 1374 chemicals covering all ionization classes, externally validated showing high predictivity and compared to the decision tree. The decision tree was found to be superior for the acids and zwitterionic ampholytes classes.

  19. Diverse Anhydrous Silicates in a Fine-Grained Rim in the Weakly Altered CM2 Chondrite Queen Elizabeth Range 97990: Evidence for the Localized Preservation of Pristine Nebular Dust in CM Chondrites.

    NASA Astrophysics Data System (ADS)

    Brearley, A. J.

    2016-08-01

    A fine-grained rim the QUE 97990 CM2 chondrite contains diverse submicron crystalline anhydrous silicates, including olivines, low-Ca and high Ca pyroxenes and represents a more pristine sample of nebular dust than is present in most CM chondrites.

  20. Kv11.1 (hERG)-induced cardiotoxicity: a molecular insight from a binding kinetics study of prototypical Kv11.1 (hERG) inhibitors

    PubMed Central

    Yu, Z; IJzerman, A P; Heitman, L H

    2015-01-01

    Background and Purpose Drug-induced arrhythmia due to blockade of the Kv11.1 channel (also known as the hERG K+ channel) is a frequent side effect. Previous studies have primarily focused on equilibrium parameters, i.e. affinity or potency, of drug candidates at the channel. The aim of this study was to determine the kinetics of the interaction with the channel for a number of known Kv11.1 blockers and to explore a possible correlation with the affinity or physicochemical properties of these compounds. Experimental Approach The affinity and kinetic parameters of 15 prototypical Kv11.1 inhibitors were evaluated in a number of [3H]-dofetilide binding assays. The lipophilicity (logKW-C8) and membrane partitioning (logKW-IAM) of these compounds were determined by means of HPLC analysis. Key Results A novel [3H]-dofetilide competition association assay was set up and validated, which allowed us to determine the binding kinetics of the Kv11.1 blockers used in this study. Interestingly, the compounds' affinities (Ki values) were correlated to their association rates rather than dissociation rates. Overall lipophilicity or membrane partitioning of the compounds were not correlated to their affinity or rate constants for the channel. Conclusions and Implications A compound's affinity for the Kv11.1 channel is determined by its rate of association with the channel, while overall lipophilicity and membrane affinity are not. In more general terms, our findings provide novel insights into the mechanism of action for a compound's activity at the Kv11.1 channel. This may help to elucidate how Kv11.1-induced cardiotoxicity is governed and how it can be circumvented in the future. PMID:25296617

  1. DFT studies on a high energy density cage compound 4-trinitroethyl-2,6,8,10,12-pentanitrohezaazaisowurtzitane.

    PubMed

    Zhang, Jian-ying; Du, Hong-chen; Wang, Fang; Gong, Xue-dong; Huang, Yin-sheng

    2011-06-23

    Polynitro cage compound 4-trinitroethyl-2,6,8,10,12-pentanitrohexaazaisowurtzitane has the same framework with but higher stability than CL-20 and is a potential new high energy density compound (HEDC). In this paper, the B3LYP/6-31G(d,p) method of density functional theory (DFT) has been used to study its heat of formation, IR spectrum, and thermodynamic properties. The stability of the compound was evaluated by the bond dissociation energies. The calculated results show that the first step of pyrolysis is the rupture of the N-NO(2) bond in the side chain and verify the experimental observation that the title compound has better stability than CL-20. The crystal structure obtained by molecular mechanics belongs to the P2(1)2(1)2(1) space group, with lattice parameters a = 12.59 Å, b = 10.52 Å, c = 12.89 Å, Z = 4, and ρ = 2.165 g·cm(-3). Both the detonation velocity of 9.767 km·s(-1) and the detonation pressure of 45.191 GPa estimated using the Kamlet-Jacobs equation are better than those of CL-20. Considering that this cage compound has a better detonation performance and stability than CL-20, it may be a superior HEDC.

  2. Moderate physical activity correlates with elevated leptin in physically active 10-12-year-old boys with normal BMI.

    PubMed

    Cicchella, Antonio; Stefanelli, Claudio; Jürimäe, Toivo; Saar, Meeli; Purge, Priit

    2013-10-01

    The aim of this study is to examine the relations between physical activity of differing intensity and duration with body energy-balance hormone leptin in 10-12-year-old boys (N = 94) who participated in moderate-to-vigorous physical activity at least four to five times per week. The boys reported their physical activity using a questionnaire. They had normal body mass index (BMI, kg/ m2), and were at Tanner Stage 2 of development. Boys were divided into three subgroups by leptin levels: normal serum leptin (M +/- .5 SD, n = 44, 1.2-3.9 ng/ml), low leptin (< or = M - .5 SD; n = 31, < 1.2 ng/ml), and high leptin (> or = M + .5 SD; n = 19, > 3.9 ng/ml). There were significant differences between subgroups in anthropometric parameters and serum leptin levels, but not in physical activity. A significant correlation was found between leptin and moderate physical activity of at least five times per week for at least 30 minutes each time in the high leptin group (r = .61). In conclusion, the correlations between physical activity and leptin are weak; only moderate physical activity was correlated with leptin levels in the high leptin

  3. TMPRSS2-ERG Fusion Gene Expression in Prostate Tumor Cells and Its Clinical and Biological Significance in Prostate Cancer Progression

    PubMed Central

    St. John, Jason; Powell, Katelyn; Conley-LaComb, M. Katie; Chinni, Sreenivasa R.

    2012-01-01

    TMPRSS2-Ets gene fusions were identified in prostate cancers where the promoter of transmembrane protease, serine 2 (TMPRSS2) fused with coding sequence of the erythroblastosis virus E26 (Ets) gene family members. TMPRSS2 is an androgen responsive transmembrane serine protease. Ets family members are oncogenic transcription factors that contain a highly conserved Ets DNA binding domain and an N-terminal regulatory domain. Fusion of these gene results in androgen dependent transcription of Ets factor in prostate tumor cells. The ERG is the most common fusion partner with TMPRSS2 promoter in prostate cancer patients. The high prevalence of these gene fusions, in particular TMPRSS2-ERG, makes them attractive as potential diagnostic and prognostic indicators, as well as making them a potential target for tailored therapies. This review focuses on the clinical and biological significance of TMPRSS2-ERG fusions and their role in PC development and progression. PMID:23264855

  4. The TMPRSS2-ERG gene fusion blocks XRCC4-mediated non-homologous end-joining repair and radiosensitizes prostate cancer cells to PARP inhibition

    PubMed Central

    Chatterjee, Payel; Choudhary, Gaurav S.; Alswillah, Turkeyah; Xiong, Xiahui; Heston, Warren D.; Magi-Galuzzi, Cristina; Zhang, Junran; Klein, Eric A.; Almasan, Alexandru

    2015-01-01

    Exposure to genotoxic agents, such as ionizing radiation (IR) produces DNA damage leading to DNA double-strand breaks (DSBs); IR toxicity is augmented when the DNA repair is impaired. We reported that radiosensitization by a PARP inhibitor (PARPi) was highly prominent in prostate cancer (PCa) cells expressing the TMPRSS2-ERG gene fusion protein. Here, we show that TMPRSS2-ERG blocks non-homologous end-joining (NHEJ) DNA repair by inhibiting DNA-PKcs. VCaP cells, which harbor TMPRSS2-ERG and PC3 cells that stably express it displayed γH2AX and 53BP1 foci constitutively, indicating persistent DNA damage that was absent if TMPRSS2-ERG was depleted by siRNA in VCaP cells. The extent of DNA damage was enhanced and associated with TMPRSS2-ERG’s ability to inhibit DNA-PKcs function, as indicated by its own phosphorylation (Thr2609, Ser2056) and that of its substrate, Ser1778-53BP1. DNA-PKcs deficiency caused by TMPRSS2-ERG destabilized critical NHEJ components on chromatin. Thus, XRCC4 was not recruited to chromatin, with retention of other NHEJ core factors being reduced. DNA-PKcs autophosphorylation was restored to the level of parental cells when TMPRSS2-ERG was depleted by siRNA. Following IR, TMPRSS2-ERG-expressing PC3 cells had elevated Rad51 foci and homologous recombination (HR) activity, indicating that HR compensated for defective NHEJ in these cells, hence addressing why TMPRSS2-ERG alone did not lead to radiosensitization. However, the presence of TMPRSS2-ERG, by inhibiting NHEJ DNA repair, enhanced PARPi-mediated radiosensitization. IR in combination with PARPi resulted in enhanced DNA damage in TMPRSS2-ERG-expressing cells. Thus, by inhibiting NHEJ, TMPRSS2-ERG provides a synthetic lethal interaction with PARPi in PCa patients expressing TMPRSS2-ERG. PMID:26026052

  5. Trafficking-deficient hERG K⁺ channels linked to long QT syndrome are regulated by a microtubule-dependent quality control compartment in the ER.

    PubMed

    Smith, Jennifer L; McBride, Christie M; Nataraj, Parvathi S; Bartos, Daniel C; January, Craig T; Delisle, Brian P

    2011-07-01

    The human ether-a-go-go related gene (hERG) encodes the voltage-gated K(+) channel that underlies the rapidly activating delayed-rectifier current in cardiac myocytes. hERG is synthesized in the endoplasmic reticulum (ER) as an "immature" N-linked glycoprotein and is terminally glycosylated in the Golgi apparatus. Most hERG missense mutations linked to long QT syndrome type 2 (LQT2) reduce the terminal glycosylation and functional expression. We tested the hypothesis that a distinct pre-Golgi compartment negatively regulates the trafficking of some LQT2 mutations to the Golgi apparatus. We found that treating cells in nocodazole, a microtubule depolymerizing agent, altered the subcellular localization, functional expression, and glycosylation of the LQT2 mutation G601S-hERG differently from wild-type hERG (WT-hERG). G601S-hERG quickly redistributed to peripheral compartments that partially colocalized with KDEL (Lys-Asp-Glu-Leu) chaperones but not calnexin, Sec31, or the ER golgi intermediate compartment (ERGIC). Treating cells in E-4031, a drug that increases the functional expression of G601S-hERG, prevented the accumulation of G601S-hERG to the peripheral compartments and increased G601S-hERG colocalization with the ERGIC. Coexpressing the temperature-sensitive mutant G protein from vesicular stomatitis virus, a mutant N-linked glycoprotein that is retained in the ER, showed it was not restricted to the same peripheral compartments as G601S-hERG at nonpermissive temperatures. We conclude that the trafficking of G601S-hERG is negatively regulated by a microtubule-dependent compartment within the ER. Identifying mechanisms that prevent the sorting or promote the release of LQT2 channels from this compartment may represent a novel therapeutic strategy for LQT2.

  6. In vitro and in vivo effects of 14alpha-demethylase (ERG11) depletion in Candida glabrata.

    PubMed

    Nakayama, H; Nakayama, N; Arisawa, M; Aoki, Y

    2001-11-01

    Sterol 14alpha-demethylase (ERG11) is the target enzyme of azole antifungals that are widely used for the treatment of fungal infections. Candida glabrata is known to be less susceptible to fluconazole than most Candida albicans strains, and the incidence of C. glabrata infection has been increasing mostly in conjunction with the use of azole antifungals. Recently, it has been reported that C. glabrata can rescue the defect of ergosterol biosynthesis by incorporating cholesterol from serum. To explore the effect of inactivating Erg11p in C. glabrata, we generated mutant strains in which the ERG11 gene was placed under the control of tetracycline-regulatable promoters. In these mutants, expression of the ERG11 gene can be repressed by doxycycline (DOX). All mutants showed a growth defect in the presence of DOX. The numbers of CFU of the mutants were lowered by only 1/10 with DOX treatment. In these mutants, accumulation of 4,14-dimethylzymosterol, which differs from an accumulated abnormal sterol detected in C. albicans and Saccharomyces cerevisiae treated with fluconazole, was observed by DOX treatment. Although such phenotypes were also observed in serum-containing media by DOX treatment, they were alleviated. Furthermore, the mutant could grow in DOX-treated mice without a severe reduction in the number of cells. Thus, depleting the expression of the ERG11 gene lowered the number of CFU by only 1/10 due to the accumulation of 4,14-demethylzymosterol in vitro, and it did not result in the defective growth of fungal cells in mice. These results suggested that Erg11p is not an ideal target molecule of antifungals for C. glabrata.

  7. Single doses of all-trans-N-retinylacetamide slow down the ERG amplitude recovery after bleaching in rats.

    PubMed

    Messias, Andre; Zrenner, Eberhart; Tzekov, Radouil; McGee, David; Peters, Tobias; Wilhelm, Barbara; Baryluk, Aneta; Kubota, Ryo; Gekeler, Florian

    2010-04-01

    To assess the safety and to quantify the effects of a single application of all-trans-N-retinylacetamide on the rat retina measured by electroretinography (ERG). Brown Norway rats were assigned to either a control group (n = 13) or to one of the three groups treated with a single intra-peritoneal dose of all-trans-N-retinylacetamide: 20 (n = 8), 5 (n = 7), or 1 mg/kg (n = 8). Full-field ERGs were performed 7 days before (baseline) and 12 h after treatment. Intensity-response relationship of b-wave amplitudes were evaluated in dark-adapted conditions using white stimuli (0.000003-0.3 cd.s/m(2)). Fast dynamics of rod sensitivity was assessed by a paired-flash paradigm; recovery dynamics of b-wave amplitudes after bleaching was followed for 70 min. Light-adapted ERGs were recorded for cone evaluation. No effects were found on either dark-adapted sensitivity or on fast rod recovery. However, drug treatment at 5 and 20 mg/kg significantly delayed ERG amplitude recovery after bleaching: 60 min after bleaching the b-wave amplitude was 21 + or - 9% (P < 0.05) and 66 + or - 10% (P < 0.05), respectively, compared to baseline. Recovery rates returned to normal 8 weeks after treatment. There were no changes in light-adapted ERG in any group. Systemic administration of a single dose of the visual cycle modulator all-trans-N-retinylacetamide reversibly delayed recovery of dark-adapted ERG amplitudes after bleaching, leaving other functions unchanged. This finding could make the compound potentially useful in experimental conditions or in specific diseases where the visual cycle is involved, such as retinitis pigmentosa or age-related macular degeneration.

  8. Monte Carlo method for predicting of cardiac toxicity: hERG blocker compounds.

    PubMed

    Gobbi, Marco; Beeg, Marten; Toropova, Mariya A; Toropov, Andrey A; Salmona, Mario

    2016-05-27

    The estimation of the cardiotoxicity of compounds is an important task for the drug discovery as well as for the risk assessment in ecological aspect. The experimental estimation of the above endpoint is complex and expensive. Hence, the theoretical computational methods are very attractive alternative of the direct experiment. A model for cardiac toxicity of 400 hERG blocker compounds (pIC50) is built up using the Monte Carlo method. Three different splits into the visible training set (in fact, the training set plus the calibration set) and invisible validation sets examined. The predictive potential is very good for all examined splits. The statistical characteristics for the external validation set are (i) the coefficient of determination r(2)=(0.90-0.93); and (ii) root-mean squared error s=(0.30-0.40).

  9. Functional interactions of voltage sensor charges with an S2 hydrophobic plug in hERG channels.

    PubMed

    Cheng, Yen May; Hull, Christina M; Niven, Christine M; Qi, Ji; Allard, Charlene R; Claydon, Tom W

    2013-09-01

    Human ether-à-go-go-related gene (hERG, Kv11.1) potassium channels have unusually slow activation and deactivation kinetics. It has been suggested that, in fast-activating Shaker channels, a highly conserved Phe residue (F290) in the S2 segment forms a putative gating charge transfer center that interacts with S4 gating charges, i.e., R362 (R1) and K374 (K5), and catalyzes their movement across the focused electric field. F290 is conserved in hERG (F463), but the relevant residues in the hERG S4 are reversed, i.e., K525 (K1) and R537 (R5), and there is an extra positive charge adjacent to R537 (i.e., K538). We have examined whether hERG channels possess a transfer center similar to that described in Shaker and if these S4 charge differences contribute to slow gating in hERG channels. Of five hERG F463 hydrophobic substitutions tested, F463W and F463Y shifted the conductance-voltage (G-V) relationship to more depolarized potentials and dramatically slowed channel activation. With the S4 residue reversals (i.e., K525, R537) taken into account, the closed state stabilization by F463W is consistent with a role for F463 that is similar to that described for F290 in Shaker. As predicted from results with Shaker, the hERG K525R mutation destabilized the closed state. However, hERG R537K did not stabilize the open state as predicted. Instead, we found the neighboring K538 residue to be critical for open state stabilization, as K538R dramatically slowed and right-shifted the voltage dependence of activation. Finally, double mutant cycle analysis on the G-V curves of F463W/K525R and F463W/K538R double mutations suggests that F463 forms functional interactions with K525 and K538 in the S4 segment. Collectively, these data suggest a role for F463 in mediating closed-open equilibria, similar to that proposed for F290 in Shaker channels.

  10. Thermal behavior and ice-table depth within the north polar erg of Mars

    USGS Publications Warehouse

    Putzig, Nathaniel E.; Mellon, Michael T.; Herkenhoff, Kenneth E.; Phillips, Roger J.; Davis, Brian J.; Ewer, Kenneth J.; Bowers, Lauren M.

    2014-01-01

    We fully resolve a long-standing thermal discrepancy concerning the north polar erg of Mars. Several recent studies have shown that the erg’s thermal properties are consistent with normal basaltic sand overlying shallow ground ice or ice-cemented sand. Our findings bolster that conclusion by thoroughly characterizing the thermal behavior of the erg, demonstrating that other likely forms of physical heterogeneity play only a minor role, and obviating the need to invoke exotic materials. Thermal inertia as calculated from orbital temperature observations of the dunes has previously been found to be more consistent with dust-sized materials than with sand. Since theory and laboratory data show that dunes will only form out of sand-sized particles, exotic sand-sized agglomerations of dust have been invoked to explain the low values of thermal inertia. However, the polar dunes exhibit the same darker appearance and color as that of dunes found elsewhere on the planet that have thermal inertia consistent with normal sand-sized basaltic grains, whereas Martian dust deposits are generally lighter and redder. The alternative explanation for the discrepancy as a thermal effect of a shallow ice table is supported by our analysis of observations from the Mars Global Surveyor Thermal Emission Spectrometer and the Mars Odyssey Thermal Emission Imaging System and by forward modeling of physical heterogeneity. In addition, our results exclude a uniform composition of dark dust-sized materials, and they show that the thermal effects of the dune slopes and bright interdune materials evident in high-resolution images cannot account for the erg’s thermal behavior.

  11. IRetinal Organization in the retinal degeneration 10 (rd10) Mutant Mouse: a Morphological and ERG Study

    PubMed Central

    Gargini, Cludia; Terzibasi, Eva; Mazzoni, Francesca; Strettoi, Enrica

    2008-01-01

    Retinal degeneration 10 (rd10) mice are a model of autosomal recessive Retinitis Pigmentosa (RP), identified by Chang et al. in 2002. These mice carry a spontaneous mutation of the rod-phosphodiesterase (PDE) gene, leading to a rod degeneration that starts around P18. Later, cones are also lost. Because of photoreceptor degeneration does not overlap with retinal development, and light responses can be recorded for about a month after birth, rd10 mice mimic typical human RP more closely than the well-known rd1 mutants. Aim of this study is to provide a comprehensive analysis of the morphology and function of the rd10 mouse retina during the period of maximum photoreceptor degeneration, thus contributing useful data for exploiting this novel model to study RP. We analyze the morphology and survival of retinal cells in rd10 mice of various ages with quantitative immunocytochemistry and confocal microscopy; we also study retinal function with the electroretinogram (ERG), recorded between P18 and P30. We find that photoreceptor death (peaking around P25) is accompanied and followed by dendritic retraction in bipolar and horizontal cells, which eventually undergo secondary degeneration. ERG reveals alterations in the physiology of the inner retina as early as P18 (before any obvious morphological change of inner neurons) and yet consistently with a reduced band amplification by bipolar cells. Thus, changes in the rd10 retina are very similar to what previously found in rd1 mutants. However, an overall slower decay of retinal structure and function predict that rd10 mice might become excellent models for rescue approaches. PMID:17111372

  12. PROCEEDINGS OF RIKEN BNL RESEARCH CENTER WORKSHOP, VOLUME 77, RBRC SCIENTIFIC REVIEW COMMITTEE MEETING, OCTOBER 10-12, 2005

    SciTech Connect

    SAMIOS, N.P.

    2005-10-10

    The eighth evaluation of the RIKEN BNL Research Center (RBRC) took place on October 10-12, 2005, at Brookhaven National Laboratory. The members of the Scientific Review Committee (SRC) were Dr. Jean-Paul Blaizot, Professor Makoto Kobayashi, Dr. Akira Masaike, Professor Charles Young Prescott (Chair), Professor Stephen Sharpe (absent), and Professor Jack Sandweiss. We are grateful to Professor Akira Ukawa who was appointed to the SRC to cover Professor Sharpe's area of expertise. In addition to reviewing this year's program, the committee, augmented by Professor Kozi Nakai, evaluated the RBRC proposal for a five-year extension of the RIKEN BNL Collaboration MOU beyond 2007. Dr. Koji Kaya, Director of the Discovery Research Institute, RIKEN, Japan, presided over the session on the extension proposal. In order to illustrate the breadth and scope of the RBRC program, each member of the Center made a presentation on higher research efforts. In addition, a special session was held in connection with the RBRC QCDSP and QCDOC supercomputers. Professor Norman H. Christ, a collaborator from Columbia University, gave a presentation on the progress and status of the project, and Professor Frithjof Karsch of BNL presented the first physics results from QCDOC. Although the main purpose of this review is a report to RIKEN Management (Dr. Ryoji Noyori, RIKEN President) on the health, scientific value, management and future prospects of the Center, the RBRC management felt that a compendium of the scientific presentations are of sufficient quality and interest that they warrant a wider distribution. Therefore we have made this compilation and present it to the community for its information and enlightenment.

  13. High Mobility Exceeding 80 cm2 V-1 s-1 in Polycrystalline Ta-Doped SnO2 Thin Films on Glass Using Anatase TiO2 Seed Layers

    NASA Astrophysics Data System (ADS)

    Nakao, Shoichiro; Yamada, Naoomi; Hitosugi, Taro; Hirose, Yasushi; Shimada, Toshihiro; Hasegawa, Tetsuya

    2010-03-01

    High-mobility Ta-doped SnO2 (TTO) thin films were grown on glass substrates by pulsed laser deposition using a seed-layer technique. The use of 10-nm-thick polycrystalline anatase TiO2 seed layers was found to lead to the preferred growth of (200)-oriented TTO films, resulting in a 30% increase in the carrier density and a more than two times increase in mobility, compared to films grown directly on the glass substrates. The highest mobility obtained was 83 cm2 V-1 s-1 with a resistivity of 2.8×10-4 Ω cm, whereas the film with the lowest resistivity of 1.8×10-4 Ω cm had a mobility of 60 cm2 V-1 s-1.

  14. Metal Acetylacetonate Series in Interface Engineering for Full Low-Temperature-Processed, High-Performance, and Stable Planar Perovskite Solar Cells with Conversion Efficiency over 16% on 1 cm(2) Scale.

    PubMed

    Chen, Wei; Xu, Leiming; Feng, Xiyuan; Jie, Jiansheng; He, Zhubing

    2017-02-14

    A series of metal acetylacetonates produced by a full low-temperature (below 100 °C) process are successfully employed to obtain both "multistable" and high-performance planar-inverted perovskite solar cells. All the three kinds of champion cells in small area exhibit over 18% in conversion-efficiency with negligible hysteresis, along with above 16% in conversion-efficiency for planar PSCs in an aperture area of over 1 cm(2) .

  15. Prostate cancer risk regions at 8q24 and 17q24 are differentially associated with somatic TMPRSS2:ERG fusion status.

    PubMed

    Luedeke, Manuel; Rinckleb, Antje E; FitzGerald, Liesel M; Geybels, Milan S; Schleutker, Johanna; Eeles, Rosalind A; Teixeira, Manuel R; Cannon-Albright, Lisa; Ostrander, Elaine A; Weikert, Steffen; Herkommer, Kathleen; Wahlfors, Tiina; Visakorpi, Tapio; Leinonen, Katri A; Tammela, Teuvo L J; Cooper, Colin S; Kote-Jarai, Zsofia; Edwards, Sandra; Goh, Chee L; McCarthy, Frank; Parker, Chris; Flohr, Penny; Paulo, Paula; Jerónimo, Carmen; Henrique, Rui; Krause, Hans; Wach, Sven; Lieb, Verena; Rau, Tilman T; Vogel, Walther; Kuefer, Rainer; Hofer, Matthias D; Perner, Sven; Rubin, Mark A; Agarwal, Archana M; Easton, Doug F; Al Olama, Ali Amin; Benlloch, Sara; Hoegel, Josef; Stanford, Janet L; Maier, Christiane

    2016-10-18

    Molecular and epidemiological differences have been described between TMPRSS2:ERG fusion-positive and fusion-negative prostate cancer (PrCa). Assuming two molecularly distinct subtypes, we have examined 27 common PrCa risk variants, previously identified in genome-wide association studies, for subtype specific associations in a total of 1221 TMPRSS2:ERG phenotyped PrCa cases. In meta-analyses of a discovery set of 552 cases with TMPRSS2:ERG data and 7650 unaffected men from five centers we have found support for the hypothesis that several common risk variants are associated with one particular subtype rather than with PrCa in general. Risk variants were analyzed in case-case comparisons (296 TMPRSS2:ERG fusion-positive versus 256 fusion-negative cases) and an independent set of 669 cases with TMPRSS2:ERG data was established to replicate the top five candidates. Significant differences (P < 0.00185) between the two subtypes were observed for rs16901979 (8q24) and rs1859962 (17q24), which were enriched in TMPRSS2:ERG fusion-negative (OR = 0.53, P = 0.0007) and TMPRSS2:ERG fusion-positive PrCa (OR = 1.30, P = 0.0016), respectively. Expression quantitative trait locus analysis was performed to investigate mechanistic links between risk variants, fusion status and target gene mRNA levels. For rs1859962 at 17q24, genotype dependent expression was observed for the candidate target gene SOX9 in TMPRSS2:ERG fusion-positive PrCa, which was not evident in TMPRSS2:ERG negative tumors. The present study established evidence for the first two common PrCa risk variants differentially associated with TMPRSS2:ERG fusion status. TMPRSS2:ERG phenotyping of larger studies is required to determine comprehensive sets of variants with subtype-specific roles in PrCa.

  16. Study of EWS/FLI-1 rearrangement in 18 cases of CK20+/CM2B4+ Merkel cell carcinoma using FISH and correlation to the differential diagnosis of Ewing sarcoma/peripheral neuroectodermal tumor.

    PubMed

    Fernandez-Flores, Angel; Suarez-Peñaranda, Jose M; Alonso, Soledad

    2013-10-01

    Merkel cell carcinoma (MCC) and primary cutaneous Ewing sarcoma/primitive neuroectodermal tumors (PCES/PNET) pose a challenging morphologic differential diagnosis. Approximately 90% of Ewing sarcoma/primitive neuroectodermal tumors (PNETs) have a specific translocation, t(11;22) (q24;q12). The EWS-friend leukemia integration-1 (FLI-1) fusion results in FLI-1 overexpression. EWS/FLI-1 rearrangement has been suggested as a useful tool in the diagnosis of PCES/PNET. In contrast, Merkel cell polyomavirus was found to be an infective agent related to the pathogenesis of MCC. Merkel cell polyomavirus can be immunohistochemically detected with the antibody CM2B4. To the best of our knowledge, there is no case of any cytokeratin (CK)20-/CM2B4+ PNET. The goal of our study was to investigate whether EWS/FLI-1 rearrangement was present in cases of MCC. We have studied 18 cases of MCC. To make sure that the cases investigated by fluorescent in situ hybridization were genuine MCC, we considered only CK20+/CM2B4+ cases. Six cases met this criterion. EWS/FLI-1 rearrangement was not evidenced in any of the 18 cases (including the 6 "genuine" cases of MCC). Although our findings were somewhat expected, we think that they fill a gap in the literature: the confirmation that MCC is devoid of the EWS/FLI-1 rearrangement.

  17. High-Level γ-Glutamyl-Hydrolase (GGH) Expression is Linked to Poor Prognosis in ERG Negative Prostate Cancer

    PubMed Central

    Melling, Nathaniel; Rashed, Masoud; Schroeder, Cornelia; Hube-Magg, Claudia; Kluth, Martina; Lang, Dagmar; Simon, Ronald; Möller-Koop, Christina; Steurer, Stefan; Sauter, Guido; Jacobsen, Frank; Büscheck, Franziska; Wittmer, Corinna; Clauditz, Till; Krech, Till; Tsourlakis, Maria Christina; Minner, Sarah; Huland, Hartwig; Graefen, Markus; Budäus, Lars; Thederan, Imke; Salomon, Georg; Schlomm, Thorsten; Wilczak, Waldemar

    2017-01-01

    γ-glutamyl-hydrolase (GGH) is a ubiquitously-expressed enzyme that regulates intracellular folate metabolism for cell proliferation, DNA synthesis, and repair. Employing GGH immunohistochemistry on a tissue microarray with 12,427 prostate cancers, we found that GGH expression was negative to low in normal prostate epithelium, whereas 88.3% of our 10,562 interpretable cancers showed GGH expression. GGH staining was considered as low intensity in 49.6% and as high intensity in 38.6% of cancers. High GGH expression was linked to the TMPRSS2:ERG-fusion positive subset of cancers (p < 0.0001), advanced pathological tumor stage, and high Gleason grade (p < 0.0001 each). Further analysis revealed that these associations were merely driven by the subset of ERG-negative cancers, High GGH expression was weakly linked to early biochemical recurrence in ERG negative cancers (p < 0.0001) and independent from established histo-pathological parameters. Moreover, GGH expression was linked to features of genetic instability, including presence of recurrent deletions at 3p, 5q, 6q, and 10q (PTEN, p ≤ 0.01 each), as well as to accelerated cell proliferation as measured by Ki67 immunohistochemistry (p < 0.0001). In conclusion, the results of our study identify GGH as an ERG subtype specific molecular marker with modest prognostic relevance, which may have clinical relevance if analyzed in combination with other molecular markers. PMID:28146062

  18. miRNA and TMPRSS2-ERG do not mind their own business in prostate cancer cells.

    PubMed

    Fayyaz, Sundas; Farooqi, Ammad Ahmad

    2013-05-01

    Oncogenic fusion proteins belong to an important class that disrupts gene expression networks in a cell. Astonishingly, fusion-positive prostate cancer cells enable the multi-gene regulatory capability of miRNAs to remodel the signal transduction landscape, enhancing or antagonizing the transmission of information to downstream effectors. Accumulating evidence substantiates the fact that miRNAs translate into dose-dependent responsiveness of cells to signaling regulators in transmembrane protease serine 2:ETS-related gene (TMPRSS2-ERG)-positive cells. Wide ranging signaling proteins are the targets for the degree of quantitative fluctuations imposed by miRNAs. miRNA signatures are aberrantly expressed in fusion-positive cancer cells, suggesting that they have a cumulative effect on tumor aggressiveness. It seems attractive to note that TMPRSS2:ERG fusion has a stronger effect as tumors positive for the oncogenic TMPRSS2:ERG have dysregulated oncomirs and tumor suppressor miRNA signature. It is undeniable that a comprehensive analysis of the prostate cancer microRNAome is necessary to uncover novel microRNAs and pathways associated with prostate cancer. Moreover, the identification and validation of miRNA signature in TMPRSS2-ERG-positive prostate cancer cells may help to identify novel molecular targets and pathways for personalized therapy.

  19. Norphlet Formation (Upper Jurassic) sand erg: depositional model for northeastern De Soto salt basin, eastern Gulf of Mexico

    SciTech Connect

    Kemmer, D.A.; Reagan, R.L.

    1987-05-01

    Available well control, seismic reflection geometries, and seismic modeling suggest the interpretation of a Norphlet Formation (Upper Jurassic) sand erg in the northeastern De Soto salt basin. Ranging in thickness from less than 100 ft to nearly 1000 ft, the Norphlet erg encompasses an area of approximately 700 mi/sup 2/. Separated from the major gas accumulation in the Norphlet in the Mobile Bay area by the offshore extension of the Pensacola arch, the Norphlet erg appears to be oriented transverse to the axis of the De Soto salt basin. Seismic signatures for the Smackover carbonate, Norphlet sand, and Louann Salt intervals are investigated using synthetic seismograms generated from six wells in the eastern Gulf of Mexico. General characteristics about the reflection coefficients from the major units in the interval are noted. The reflection coefficient information and synthetic seismograms are used to interpret seismic data on a regional basis. Two-dimensional, vertical-incidence, ray-trace modeling of the seismic data is done to aid the interpretation on a detailed basis. Interpreted Norphlet sandstone thicknesses and Louann Salt structures are combined to support the Norphlet Formation sand erg hypothesis.

  20. Proline Scan of the hERG Channel S6 Helix Reveals the Location of the Intracellular Pore Gate

    PubMed Central

    Thouta, Samrat; Sokolov, Stanislav; Abe, Yuki; Clark, Sheldon J.; Cheng, Yen M.; Claydon, Tom W.

    2014-01-01

    In Shaker-like channels, the activation gate is formed at the bundle crossing by the convergence of the inner S6 helices near a conserved proline-valine-proline motif, which introduces a kink that allows for electromechanical coupling with voltage sensor motions via the S4-S5 linker. Human ether-a-go-go-related gene (hERG) channels lack the proline-valine-proline motif and the location of the intracellular pore gate and how it is coupled to S4 movement is less clear. Here, we show that proline substitutions within the S6 of hERG perturbed pore gate closure, trapping channels in the open state. Performing a proline scan of the inner S6 helix, from Ile655 to Tyr667 revealed that gate perturbation occurred with proximal (I655P-Q664P), but not distal (R665P-Y667P) substitutions, suggesting that Gln664 marks the position of the intracellular gate in hERG channels. Using voltage-clamp fluorimetry and gating current analysis, we demonstrate that proline substitutions trap the activation gate open by disrupting the coupling between the voltage-sensing unit and the pore of the channel. We characterize voltage sensor movement in one such trapped-open mutant channel and demonstrate the kinetics of what we interpret to be intrinsic hERG voltage sensor movement. PMID:24606930

  1. Molecular Characterization and Clinical Impact of TMPRSS2-ERG Rearrangement on Prostate Cancer: Comparison between FISH and RT-PCR

    PubMed Central

    Fernández-Serra, A.; Rubio, L.; Calatrava, A.; Rubio-Briones, J.; Salgado, R.; Gil-Benso, R.; Espinet, B.; García-Casado, Z.; López-Guerrero, J. A.

    2013-01-01

    Prostate cancer (PCa) is a very heterogeneous disease, and there are constraints in its current diagnosis. Serum PSA levels, digital rectal examination (DRE), and histopathologic analysis often drive to overdiagnosis and overtreatment. Since 2005, the presence of the genetic rearrangement between transmembrane-serine protease gene (TMPRSS2) and the erythroblast transformation-specific (ETS) member ERG (v-ets erythroblastosis virus E26 oncogene homolog avian) has been demonstrated in almost half of PCa cases. Both FISH and RT-PCR are useful tools for detecting these rearrangements, but very few comparatives between both techniques have been published. In this study, we included FFPE tumors from 294 PCa patients treated with radical prostatectomy with more than 5 years of followup. We constructed a total of 20 tissue microarrays in order to perform break-apart and tricolor probe FISH approaches that were compared with RT-PCR, showing a concordance of 80.6% (P < 0.001). The presence of TMPRSS2-ERG rearrangement was observed in 56.6% of cases. No association between TMPRSS2-ERG status and clinicopathological parameters nor biochemical progression and clinical progression free survival was found. In conclusion, this study demonstrates that both FISH and RT-PCR are useful tools in the assessment of the TMPRSS2-ERG fusion gene status in PCa patients and that this genetic feature per se lacks prognostic value. PMID:23781502

  2. Molecular characterization and clinical impact of TMPRSS2-ERG rearrangement on prostate cancer: comparison between FISH and RT-PCR.

    PubMed

    Fernández-Serra, A; Rubio, L; Calatrava, A; Rubio-Briones, J; Salgado, R; Gil-Benso, R; Espinet, B; García-Casado, Z; López-Guerrero, J A

    2013-01-01

    Prostate cancer (PCa) is a very heterogeneous disease, and there are constraints in its current diagnosis. Serum PSA levels, digital rectal examination (DRE), and histopathologic analysis often drive to overdiagnosis and overtreatment. Since 2005, the presence of the genetic rearrangement between transmembrane-serine protease gene (TMPRSS2) and the erythroblast transformation-specific (ETS) member ERG (v-ets erythroblastosis virus E26 oncogene homolog avian) has been demonstrated in almost half of PCa cases. Both FISH and RT-PCR are useful tools for detecting these rearrangements, but very few comparatives between both techniques have been published. In this study, we included FFPE tumors from 294 PCa patients treated with radical prostatectomy with more than 5 years of followup. We constructed a total of 20 tissue microarrays in order to perform break-apart and tricolor probe FISH approaches that were compared with RT-PCR, showing a concordance of 80.6% (P < 0.001). The presence of TMPRSS2-ERG rearrangement was observed in 56.6% of cases. No association between TMPRSS2-ERG status and clinicopathological parameters nor biochemical progression and clinical progression free survival was found. In conclusion, this study demonstrates that both FISH and RT-PCR are useful tools in the assessment of the TMPRSS2-ERG fusion gene status in PCa patients and that this genetic feature per se lacks prognostic value.

  3. Enhancement of hERG channel activity by scFv antibody fragments targeted to the PAS domain

    PubMed Central

    Harley, Carol A.; Starek, Greg; Jones, David K.; Fernandes, Andreia S.; Robertson, Gail A.; Morais-Cabral, João H.

    2016-01-01

    The human human ether-à-go-go–related gene (hERG) potassium channel plays a critical role in the repolarization of the cardiac action potential. Changes in hERG channel function underlie long QT syndrome (LQTS) and are associated with cardiac arrhythmias and sudden death. A striking feature of this channel and KCNH channels in general is the presence of an N-terminal Per-Arnt-Sim (PAS) domain. In other proteins, PAS domains bind ligands and modulate effector domains. However, the PAS domains of KCNH channels are orphan receptors. We have uncovered a family of positive modulators of hERG that specifically bind to the PAS domain. We generated two single-chain variable fragments (scFvs) that recognize different epitopes on the PAS domain. Both antibodies increase the rate of deactivation but have different effects on channel activation and inactivation. Importantly, we show that both antibodies, on binding to the PAS domain, increase the total amount of current that permeates the channel during a ventricular action potential and significantly reduce the action potential duration recorded in human cardiomyocytes. Overall, these molecules constitute a previously unidentified class of positive modulators and establish that allosteric modulation of hERG channel function through ligand binding to the PAS domain can be attained. PMID:27516548

  4. Detection of early functional changes in diabetic retina using slow double-stimulation mfERG paradigm.

    PubMed

    Chan, Henry Ho-lung; Chu, Patrick Ho-wai; Lung, Jenny Chun-yee; Ho, Wing-cheung; Ting, Patrick Wai-ki; Sum, Rita Wing-man; Ng, Yiu-fai

    2011-11-01

    Aim Diabetes mellitus (DM) is a systemic disease with insufficient secretion of insulin or poor response to insulin. This typically causes poor control of blood glucose level leading to a range of complications. Early detection of the retinal function alteration in DM is needed. Methods A newly modified paradigm-slow double-stimulation multifocal electroretinogram (mfERG)-was introduced to measure early changes of retinal function in DM and to investigate changes in the adaptation mechanisms in the diabetic retina. The mfERG was measured by using a slow double-stimulation mfERG paradigm (M(1)M(2)OOO). Results The m1 amplitude of M(1) stimulation from diabetic subjects was significantly reduced in ring 1 in contrast to that of a control group. The m2 amplitude of M(2) stimulation from diabetic subjects was also significantly reduced in ring 1 and 2 as compared with those of the control group. The m1/m2 ratio which minimises intersubject variation shows a reasonable differentiation between the control and diabetic groups. There was a significant increase in the amplitude ratio from diabetic subjects in ring 2 and 3 as compared with those of the control group. Conclusions The present findings suggest that the new mfERG paradigm is a fast and sensitive test for the detection of early functional changes in the diabetic retina.

  5. Gain of chromosome 21 or amplification of chromosome arm 21q is one mechanism for increased ERG expression in acute myeloid leukemia.

    PubMed

    Weber, Simone; Haferlach, Claudia; Jeromin, Sabine; Nadarajah, Niroshan; Dicker, Frank; Noël, Louisa; Zenger, Melanie; Alpermann, Tamara; Kern, Wolfgang; Haferlach, Torsten; Schnittger, Susanne

    2016-02-01

    In acute myeloid leukemia (AML), acquired genomic gains and losses are common and lead to altered expression of genes located within or nearby the affected regions. Increased expression of the ETS-related transcription factor gene ERG has been described in myeloid malignancies with chromosomal rearrangements involving chromosome band 21q22, but also in cytogenetically normal AML, where it is associated with adverse prognosis. In this study, fluorescence in situ hybridization on interphase nuclei disclosed an amplification of the ERG gene (more than six copies) in 33 AML patients with structural rearrangements of 21q22. Array comparative genomic hybridization of these cases disclosed a minimal amplified region at the position 39.6-40.0 Mbp from pter that harbors ERG as the only gene. Analysis by quantitative real-time reverse transcription polymerase chain reaction revealed significantly higher ERG mRNA expression in these patients and in a group of 95 AML patients with complete or partial gain of chromosome 21 (three to six copies) compared with 351 AML patients without gain of chromosome 21. Quantification of ERG DNA copy numbers revealed a strong correlation with ERG mRNA expression. Furthermore, in patients with gain of chromosome 21, higher ERG expression was found to be associated with RUNX1 mutations. Our results suggest that acquired gain of chromosome 21 or amplification of chromosome arm 21q is one mechanism contributing to increased ERG expression in AML.

  6. Mutations of the XLRS1 gene cause abnormalities of photoreceptor as well as inner retinal responses of the ERG.

    PubMed

    Bradshaw, K; George, N; Moore, A; Trump, D

    1999-01-01

    Intensity-series rod and cone ERGs were recorded in 19 patients with XLRS and 26 control eyes. All patients were examined by one ophthalmologist and diagnosed on the basis of fundus appearance and evidence of the disease in other males in the same family. Mutations in the XLRS1 gene have been identified in 15 of the patients. Dark-adapted ERGs were significantly different from controls for all test conditions and for both a-wave and b-wave responses. Abnormalities were detectable in all patients but there was considerable variation in the severity of abnormality. One third of the patients had the dark-adapted 'negative-wave' response typically associated with inner retinal disorder, but about one third showed only mild depression of the b-wave while the remainder had abnormally low a-waves in addition to depressed b-waves. Light-adapted responses were also affected and both a-wave and b-wave responses differed significantly from controls, but the 'negative-wave' response was not seen in any patient. The severity of the ERG abnormality did not correlate with the classification of fundus appearance or patient age suggesting that retinal function is relatively stable throughout life. The severity of ERG abnormalities did not correlate with the type of mutation and responses could differ between affected males within the same family. These results indicate considerable heterogeneity of ERG response without clinical, age or genetic correlate. The abnormal a-wave responses indicate that photoreceptor as well as inner retinal layer function may be affected in XLRS, at least in some patients.

  7. Evidence for the hERG Liability of Antihistamines, Antipsychotics, and Anti-Infective Agents: A Systematic Literature Review From the ARITMO Project.

    PubMed

    Hazell, Lorna; Raschi, Emanuel; De Ponti, Fabrizio; Thomas, Simon H L; Salvo, Francesco; Ahlberg Helgee, Ernst; Boyer, Scott; Sturkenboom, Miriam; Shakir, Saad

    2016-12-26

    A systematic review was performed to categorize the hERG (human ether-a-go-go-related gene) liability of antihistamines, antipsychotics, and anti-infectives and to compare it with current clinical risk of torsade de pointes (TdP). Eligible studies were hERG assays reporting half-minimal inhibitory concentrations (IC50). A "hERG safety margin" was calculated from the IC50 divided by the peak human plasma concentration (free Cmax ). A margin below 30 defined hERG liability. Each drug was assigned an "uncertainty score" based on volume, consistency, precision, and internal and external validity of evidence. The hERG liability was compared to existing knowledge on TdP risk (www.credibledrugs.org). Of 1828 studies, 82 were eligible, allowing calculation of safety margins for 61 drugs. Thirty-one drugs (51%) had evidence of hERG liability including 6 with no previous mention of TdP risk (eg, desloratadine, lopinavir). Conversely, 16 drugs (26%) had no evidence of hERG liability including 6 with known, or at least conditional or possible, TdP risk (eg, chlorpromazine, sulpiride). The main sources of uncertainty were the validity of the experimental conditions used (antihistamines and antipsychotics) and nonuse of reference compounds (anti-infectives). In summary, hERG liability was categorized for 3 widely used drug classes, incorporating a qualitative assessment of the strength of available evidence. Some concordance with TdP risk was observed, although several drugs had hERG liability without evidence of clinical risk and vice versa. This may be due to gaps in clinical evidence, limitations of hERG/Cmax data, or other patient/drug-specific factors that contribute to real-life TdP risk.

  8. ERG A small-satellite mission to investigate the dynamics of the inner magnetosphere

    NASA Astrophysics Data System (ADS)

    Inner Magnetosphere Subgroup in Society Of Geomagnetism; Earth, Planetary; Space Sciences; Seki, K.; Miyoshi, Y.; Ieda, A.; Ono, T.; Iizima, M.; Nagatsuma, T.; Obara, T.; Takashima, T.; Asamura, K.; Kasaba, Y.; Matsuoka, A.; Saito, Y.; Saito, H.; Hirahara, M.; Tonegawa, Y.; Toyama, F.; Tanaka, M.; Nose, M.; Kasahara, Y.; Yumoto, K.; Kawano, H.; Yoshikawa, A.; Ebihara, Y.; Yukimatsu, A.; Sato, N.; Watanabe, S.

    2006-01-01

    The Earth’s inner magnetosphere (inside 10 Re) is a region where particle energy increases to the relativistic energy range. This region is very important as a laboratory where high-energy particle acceleration can be directly measured in a dipolar field configuration, as well as for human activities in space including space weather prediction. Despite abundant in situ satellite measurements, this region has been “missing” because of several difficulties arising from the measurements, such as high-energy particle contamination of low-energy particle measurement, protection against the possible incidence of radiation belt particles on the satellite, and the difficulties of measuring three-dimensional particles over a broad energy range, from a few electron volts to more than 10 MeV. In this paper, we address important scientific topics and propose a possible configuration of small satellites termed Energization and Radiation in Geospace (ERG), which would provide new insights into the dynamics of the inner magnetosphere and strongly contribute to the International Living With a Star project.

  9. Cone ERG Responses in Patients with Smith-Lemli-Opitz Syndrome (SLOS)

    PubMed Central

    Garry, Deirdre; Hansen, Ronald M.; Moskowitz, Anne; Elias, Ellen R.; Irons, Mira; Fulton, Anne B.

    2010-01-01

    Purpose To evaluate cone and cone-driven retinal function in patients with Smith-Lemli-Opitz syndrome (SLOS), a condition characterized by low cholesterol. Rod and rod-driven function in SLOS patients are known to be abnormal. Methods Electroretinographic (ERG) responses to full-field stimuli presented on a steady, rod suppressing background were recorded in 13 patients who had received long term cholesterol supplementation. Cone photoresponse sensitivity (SCONE) and saturated amplitude (RCONE) parameters were estimated using a model of the activation of phototransduction, and post-receptor b-wave and 30 Hz flicker responses were analyzed. The responses of the patients were compared to those of control subjects (N=13). Results Although average values of both SCONE and RCONE were lower than in controls, the differences were not statistically significant. Post-receptor b-wave amplitude and implicit time and flicker responses were normal. Conclusions The normal cone function contrasts with the significant abnormalities in rod function that were found previously in these same patients. Possibly cholesterol supplementation has a greater protective effect on cones than rods as has been demonstrated in the rat model of SLOS. PMID:20440536

  10. Sand rubification with time? The case of the Sinai - Negev erg

    NASA Astrophysics Data System (ADS)

    Roskin, J.; Rozenstein, O.; Tsoar, H.; Blumberg, D. G.; Porat, N.

    2012-04-01

    The redness index (RI) (RI = R2/(B*G3) of aeolian sand has been shown to be a promising qualitative spectroscopic method to define sand grain redness intensity, which reflects the extent of iron-oxide quartz grain coatings(1,2). Using the RI, this study investigates the relationship between redness intensity and optically stimulated luminescence (OSL) based depositional ages of sand samples taken from exposed and fully-drilled vegetated linear dunes in the northwestern Negev dunefield, Israel at the downwind end of the Sinai Negev erg. Sand redness intensity did not vary greatly along the Negev sand transport paths and dune sections dated to be active during the Late Pleistocene, late Holocene, and modern times. No correlation was found between RI intensity (i.e., redness) and the depositional age of the sand. The relatively uniform RI values and sedimentological properties along most of the dunes suggest that sand grain coating development, and consequent rubification, have probably been minimal since the Late Pleistocene. Although it is possible that RI developed rapidly following deposition in a wetter Late Pleistocene climate, the drier and less stormy Holocene does not seem conducive to sand-grain rubification. Based on analyses of northern Sinai sand samples, remote sensing, and previous studies, we suggest that the attributes of the sand grain RI have been inherited from upwind sources. We propose that the sand grain coatings are early diagenetic features that have been similarly red since their suggested aeolian departure from the middle and upper Nile Delta.

  11. Candida tropicalis Antifungal Cross-Resistance Is Related to Different Azole Target (Erg11p) Modifications

    PubMed Central

    Forastiero, A.; Mesa-Arango, A. C.; Alastruey-Izquierdo, A.; Alcazar-Fuoli, L.; Bernal-Martinez, L.; Pelaez, T.; Lopez, J. F.; Grimalt, J. O.; Gomez-Lopez, A.; Cuesta, I.; Zaragoza, O.

    2013-01-01

    Candida tropicalis ranks between third and fourth among Candida species most commonly isolated from clinical specimens. Invasive candidiasis and candidemia are treated with amphotericin B or echinocandins as first-line therapy, with extended-spectrum triazoles as acceptable alternatives. Candida tropicalis is usually susceptible to all antifungal agents, although several azole drug-resistant clinical isolates are being reported. However, C. tropicalis resistant to amphotericin B is uncommon, and only a few strains have reliably demonstrated a high level of resistance to this agent. The resistance mechanisms operating in C. tropicalis strains isolated from clinical samples showing resistance to azole drugs alone or with amphotericin B cross-resistance were elucidated. Antifungal drug resistance was related to mutations of the azole target (Erg11p) with or without alterations of the ergosterol biosynthesis pathway. The antifungal drug resistance shown in vitro correlated very well with the results obtained in vivo using the model host Galleria mellonella. Using this panel of strains, the G. mellonella model system was validated as a simple, nonmammalian minihost model that can be used to study in vitro-in vivo correlation of antifungals in C. tropicalis. The development in C. tropicalis of antifungal drug resistance with different mechanisms during antifungal treatment has potential clinical impact and deserves specific prospective studies. PMID:23877676

  12. Fully Enzymatic Membraneless Glucose|Oxygen Fuel Cell That Provides 0.275 mA cm(-2) in 5 mM Glucose, Operates in Human Physiological Solutions, and Powers Transmission of Sensing Data.

    PubMed

    Ó Conghaile, Peter; Falk, Magnus; MacAodha, Domhnall; Yakovleva, Maria E; Gonaus, Christoph; Peterbauer, Clemens K; Gorton, Lo; Shleev, Sergey; Leech, Dónal

    2016-02-16

    Coimmobilization of pyranose dehydrogenase as an enzyme catalyst, osmium redox polymers [Os(4,4'-dimethoxy-2,2'-bipyridine)2(poly(vinylimidazole))10Cl](+) or [Os(4,4'-dimethyl-2,2'-bipyridine)2(poly(vinylimidazole))10Cl](+) as mediators, and carbon nanotube conductive scaffolds in films on graphite electrodes provides enzyme electrodes for glucose oxidation. The recombinant enzyme and a deglycosylated form, both expressed in Pichia pastoris, are investigated and compared as biocatalysts for glucose oxidation using flow injection amperometry and voltammetry. In the presence of 5 mM glucose in phosphate-buffered saline (PBS) (50 mM phosphate buffer solution, pH 7.4, with 150 mM NaCl), higher glucose oxidation current densities, 0.41 mA cm(-2), are obtained from enzyme electrodes containing the deglycosylated form of the enzyme. The optimized glucose-oxidizing anode, prepared using deglycosylated enzyme coimmobilized with [Os(4,4'-dimethyl-2,2'-bipyridine)2(poly(vinylimidazole))10Cl](+) and carbon nanotubes, was coupled with an oxygen-reducing bilirubin oxidase on gold nanoparticle dispersed on gold electrode as a biocathode to provide a membraneless fully enzymatic fuel cell. A maximum power density of 275 μW cm(-2) is obtained in 5 mM glucose in PBS, the highest to date under these conditions, providing sufficient power to enable wireless transmission of a signal to a data logger. When tested in whole human blood and unstimulated human saliva maximum power densities of 73 and 6 μW cm(-2) are obtained for the same fuel cell configuration, respectively.

  13. Integrated Assessment of Hadley Centre (HadCM2) Climate-Change Impacts on Agricultural Productivity and Irrigation Water Supply in the Conterminous United States. Part II. Regional Agricultural Production in 2030 and 2095.

    SciTech Connect

    Izaurralde, R Cesar C.; Rosenberg, Norman J.; Brown, Robert A.; Thomson, Allison M.

    2003-06-30

    This study used scenarios of the HadCM2 GCM and the EPIC agroecosystem model to evaluate climate change impacts on crop yields and ecosystem processes. Baseline climate data were obtained from records for 1961-1990. The scenario runs for 2025-2034 and 2090-2099 were extracted from a HadCM2 run. EPIC was run on 204 representative farms under current climate and two 10-y periods centered on 2030 and 2095, each at CO2 concentrations of 365 and 560 ppm. Texas, New Mexico, Colorado, Utah, Arizona, and California are projected to experience significant temperature increases by 2030. Slight cooling is expected by 2030 in Alabama, Florida, Maine, Montana, Idaho, and Utah. Larger areas are projected to experience increased warming by 2095. Uniform precipitation increases are expected by 2030 in the NE. These increases are predicted to expand to the eastern half of the country by 2095. EPIC simulated yield increases for the Great Lakes, Corn Belt and Northeast regions. Simulated yields of irrigated corn yields were predicted to increase in almost all regions. Soybean yields could decrease in the Northern and Southern Plains, the Corn Belt, Delta, Appalachian, and Southeast regions and increase in the Lakes and Northeast regions. Simulated wheat yields exhibited upward yield trends under scenarios of climate change. National corn production in 2030 and 2095 could be affected by changes in three major producing regions. In 2030, corn production could increase in the Corn Belt and Lakes regions but decrease in the Northern Plains leading to an overall decrease in national production. National wheat production is expected to increase during both future periods. A proxy indicator was developed to provide a sense of where in the country, and when water would be available to satisfy change in irrigation demand for corn and alfalfa production as these are influenced by the HadCM2 scenarios and CO2-fertilization.

  14. Suggestions for Curriculum Development [And] Handbook High School Grades, Part D, 10-12. Environmental Education Interdependence: A Concept Approach. Revised.

    ERIC Educational Resources Information Center

    King, David C.; Wood, Jayne Millar

    Two booklets comprise the grades 10-12 component of a series of guides for incorporating environmental education into the existing curriculum. The guide and handbook emphasize a multidisciplinary approach, use the concept of interdependence as an organizing theme, and offer suggestions for using the local community as a resource. The guide…

  15. Reduced susceptibility to polyenes associated with a missense mutation in the ERG6 gene in a clinical isolate of Candida glabrata with pseudohyphal growth.

    PubMed

    Vandeputte, Patrick; Tronchin, Guy; Bergès, Thierry; Hennequin, Christophe; Chabasse, Dominique; Bouchara, Jean-Philippe

    2007-03-01

    Little information is available about the molecular mechanisms responsible for polyene resistance in pathogenic yeasts. A clinical isolate of Candida glabrata with a poor susceptibility to polyenes, as determined by disk diffusion method and confirmed by determination of MIC, was recovered from a patient treated with amphotericin B. Quantitative analysis of sterols revealed a lack of ergosterol and an accumulation of late sterol intermediates, suggesting a defect in the final steps of the ergosterol pathway. Sequencing of CgERG11, CgERG6, CgERG5, and CgERG4 genes revealed exclusively a unique missense mutation in CgERG6 leading to the substitution of a cysteine by a phenylalanine in the corresponding protein. In addition, real-time reverse transcription-PCR demonstrated an overexpression of genes encoding enzymes involved in late steps of the ergosterol pathway. Moreover, this isolate exhibited a pseudohyphal growth whatever the culture medium used, and ultrastructural changes of the cell wall of blastoconidia were seen consisting in a thinner inner layer. Cell wall alterations were also suggested by the higher susceptibility of growing cells to Calcofluor white. Additionally, complementation of this isolate with a wild-type copy of the CgERG6 gene restored susceptibility to polyenes and a classical morphology. Together, these results demonstrated that mutation in the CgERG6 gene may lead to a reduced susceptibility to polyenes and to a pseudohyphal growth due to the subsequent changes in sterol content of the plasma membrane.

  16. Novel intramolecular photoinduced electron transfer-based probe for the Human Ether-a-go-go-Related Gene (hERG) potassium channel.

    PubMed

    Liu, Zhenzhen; Zhou, Yubin; Du, Lupei; Li, Minyong

    2015-12-21

    Drug induced long QT syndrome is a high risk event in clinic, which mainly results from their high affinity to the Human Ether-a-go-go-Related Gene (hERG) potassium channel. Therefore, evaluation of the drug's inhibitory activity against the hERG potassium channel is a required step in drug discovery and development. In this study, we developed a series of novel conformation-mediated intramolecular photoinduced electron transfer fluorogenic probes for the hERG potassium channel. After careful evaluation, probes N4 and N6 showed good activity and may have a promising application in the cell-based hERG potassium channel inhibitory activity assay, as well as potential hERG-associated cardiotoxicity evaluation. Compared with other assay methods, such as patch clamp assay, radio-ligand competitive binding assay, fluorescence polarization and potential-sensitive fluorescent probes, this method is convenient and can also selectively measure the inhibitory activity in the native state of the hERG potassium channel. Meanwhile, these probes can also be used for hERG potassium channel imaging without complex washing steps.

  17. The yeast gene ERG6 is required for normal membrane function but is not essential for biosynthesis of the cell-cycle-sparking sterol.

    PubMed Central

    Gaber, R F; Copple, D M; Kennedy, B K; Vidal, M; Bard, M

    1989-01-01

    In Saccharomyces cerevisiae, methylation of the principal membrane sterol at C-24 produces the C-28 methyl group specific to ergosterol and represents one of the few structural differences between ergosterol and cholesterol. C-28 in S. cerevisiae has been suggested to be essential for the sparking function (W. J. Pinto and W. R. Nes, J. Biol. Chem. 258:4472-4476, 1983), a cell cycle event that may be required to enter G1 (C. Dahl, H.-P. Biemann, and J. Dahl, Proc. Natl. Acad. Sci. USA 84:4012-4016, 1987). The sterol biosynthetic pathway in S. cerevisiae was genetically altered to assess the functional role of the C-28 methyl group of ergosterol. ERG6, the putative structural gene for S-adenosylmethionine: delta 24-methyltransferase, which catalyzes C-24 methylation, was cloned, and haploid strains containing erg6 null alleles (erg6 delta 1 and erg6 delta ::LEU2) were generated. Although erg6 delta cells are unable to methylate ergosterol precursors at C-24, they exhibit normal vegatative growth, suggesting that C-28 sterols are not essential in S. cerevisiae. However, erg6 delta cells exhibit pleiotropic phenotypes that include defective conjugation, hypersensitivity to cycloheximide, resistance to nystatin, a severely diminished capacity for genetic transformation, and defective tryptophan uptake. These phenotypes reflect the role of ergosterol as a regulator of membrane permeability and fluidity. Genetic mapping experiments revealed that ERG6 is located on chromosome XIII, tightly linked to sec59. Images PMID:2677674

  18. Low level laser therapy (AlGaInP) applied at 5J/cm2 reduces the proliferation of Staphylococcus aureus MRSA in infected wounds and intact skin of rats*

    PubMed Central

    Silva, Daniela Conceição Gomes Gonçalves e; Plapler, Helio; da Costa, Mateus Matiuzzi; Silva, Silvio Romero Gonçalves e; de Sá, Maria da Conceição Aquino; Silva, Benedito Sávio Lima e

    2013-01-01

    BACKGROUND Laser therapy is a low cost, non-invasive procedure with good healing results. Doubts exist as to whether laser therapy action on microorganisms can justify research aimed at investigating its possible effects on bacteria-infected wounds. OBJECTIVE To assess the effect of low intensity laser on the rate of bacterial contamination in infected wounds in the skin of rats. METHODS An experimental study using 56 male Wistar rats. The animals were randomly divided into eight groups of seven each. Those in the "infected" groups were infected by Staphylococcus aureus MRSA in the dorsal region. Red laser diode (AlGaInP) 658nm, 5J/cm2 was used to treat the animals in the "treated" groups in scan for 3 consecutive days. Samples were drawn before inoculating bacteria and following laser treatment. For statistical analysis we used the nonparametric Wilcoxon (paired data) method with a significance level of p <0.05. RESULTS The statistical analysis of median values showed that the groups submitted to laser treatment had low bacterial proliferation. CONCLUSION The laser (AlGaInP), with a dose of 5J/cm2 in both intact skin and in wounds of rats infected with Staphylococcus aureus MRSA, is shown to reduce bacterial proliferation. PMID:23539003

  19. Regulation of the human ether-a-go-go-related gene (hERG) potassium channel by Nedd4 family interacting proteins (Ndfips).

    PubMed

    Kang, Yudi; Guo, Jun; Yang, Tonghua; Li, Wentao; Zhang, Shetuan

    2015-11-15

    The cardiac electrical disorder long QT syndrome (LQTS) pre-disposes affected individuals to ventricular arrhythmias and sudden death. Dysfunction of the human ether-a-go-go-related gene (hERG)-encoded rapidly activating delayed rectifier K(+) channel (IKr) is a major cause of LQTS. The expression of hERG channels is controlled by anterograde trafficking of newly synthesized channels to and retrograde degradation of existing channels from the plasma membrane. We have previously shown that the E3 ubiquitin (Ub) ligase Nedd4-2 (neural precursor cell expressed developmentally down-regulated protein 4-2) targets the PY motif of hERG channels to initiate channel degradation. Although both immature and mature hERG channels contain the PY motif, Nedd4-2 selectively mediates the degradation of mature hERG channels. In the present study, we demonstrate that Nedd4-2 is directed to specific cellular compartments by the Nedd4 family interacting proteins, Nedd4 family-interacting protein 1 (Ndfip1) and Ndfip2. Ndfip1 is primarily localized in the Golgi apparatus where it recruits Nedd4-2 to mediate the degradation of mature hERG proteins during channel trafficking to the plasma membrane. Although Ndfip2 directs Nedd4-2 to the Golgi apparatus, it also recruits Nedd4-2 to the multivesicular bodies (MVBs), which may impair MVB function and impede the degradation of mature hERG proteins mediated by Nedd4-2. These findings extend our understanding of hERG channel regulation and provide information which may be useful for the rescue of impaired hERG function in LQTS.

  20. Prognostic value of ERG, PTEN, CRISP3 and SPINK1 in predicting biochemical recurrence in prostate cancer

    PubMed Central

    NOH, BYEONG-JOO; SUNG, JI-YOUN; KIM, YOUN WHA; CHANG, SUNG-GOO; PARK, YONG-KOO

    2016-01-01

    The established prognostic factors associated with prostatic adenocarcinoma are the Gleason score, pathological T staging and serum prostatic-specific antigen (PSA) level. However, these prognostic factors alone are not sufficient for predicting prognostic characteristics, including early stage or advanced prostate cancer, presence of metastasis or disease-related mortality. The purpose of the present study was to simultaneously evaluate the prognostic value and associations of four biomarkers, namely, transcriptional regulator ERG (ERG), phosphatase and tensin homolog (PTEN), cysteine-rich secretory protein 3 (CRISP3) and serine protease inhibitor Kazal type I (SPINK1), and to conduct risk stratification of prostate cancer for use in patient management. A total of 68 formalin-fixed, paraffin-embedded, prostate cancer samples from radical prostatectomies were obtained in the Kyung Hee University Hospital (Seoul, Korea) and were studied immunohistochemically for ERG, PTEN, CRISP3 and SPINK1 to determine the proportion and intensity of staining. SPINK1 expression was mutually exclusive of ERG expression (P=0.001). The loss of PTEN and high CRISP3 expression are unfavorable indicators for prostate cancer, as PTEN loss was associated with shorter biochemical recurrence (BCR) (P=0.039), and high CRISP3 expression was associated with increased BCR (P<0.001) and cancer-related mortalities (P=0.011). Using the combination of low PTEN and high CRISP3 expression enables attention to be focused on patients who exhibit a poor prognosis. Subgrouping of patients, into high-risk and low-risk categories, was correlated with BCR-free survival in prostate cancer upon multivariate analysis (P=0.030). Overall, low PTEN and high CRISP3 expression significantly characterize the subgroups of prostate cancer that have a poor prognosis for BCR. PMID:27284364

  1. ERG-28 controls BK channel trafficking in the ER to regulate synaptic function and alcohol response in C. elegans

    PubMed Central

    Oh, Kelly H; Haney, James J; Wang, Xiaohong; Chuang, Chiou-Fen; Richmond, Janet E; Kim, Hongkyun

    2017-01-01

    Voltage- and calcium-dependent BK channels regulate calcium-dependent cellular events such as neurotransmitter release by limiting calcium influx. Their plasma membrane abundance is an important factor in determining BK current and thus regulation of calcium-dependent events. In C. elegans, we show that ERG-28, an endoplasmic reticulum (ER) membrane protein, promotes the trafficking of SLO-1 BK channels from the ER to the plasma membrane by shielding them from premature degradation. In the absence of ERG-28, SLO-1 channels undergo aspartic protease DDI-1-dependent degradation, resulting in markedly reduced expression at presynaptic terminals. Loss of erg-28 suppressed phenotypic defects of slo-1 gain-of-function mutants in locomotion, neurotransmitter release, and calcium-mediated asymmetric differentiation of the AWC olfactory neuron pair, and conferred significant ethanol-resistant locomotory behavior, resembling slo-1 loss-of-function mutants, albeit to a lesser extent. Our study thus indicates that the control of BK channel trafficking is a critical regulatory mechanism for synaptic transmission and neural function. DOI: http://dx.doi.org/10.7554/eLife.24733.001 PMID:28168949

  2. Epidemiology, species distribution, antifungal susceptibility, and ERG11 mutations of Candida species isolated from pregnant Chinese Han women.

    PubMed

    Yang, L; Su, M Q; Ma, Y Y; Xin, Y J; Han, R B; Zhang, R; Wen, J; Hao, X K

    2016-04-28

    The widespread use of antifungal agents has led to increasing azole resistance in Candida species. A major azole-resistance mechanism involves point mutations in the ERG11 gene, which encodes cytochrome P450 lanosterol 14a-demethylase. In this study, vaginal swabs were obtained from 657 pregnant Chinese Han women and cultured appropriately. The open reading frame of the obtained fungal species were amplified by PCR and sequenced; additionally, the ERG11 gene of the isolated Candida species was amplified and sequenced, and the antifungal susceptibility of the isolated species was determined. The vaginal swabs of 124 women produced fungal cultures; five species of Candida were isolated from the patients, among which Candida albicans was predominant. Twelve C. albicans isolates (13.8%) were resistant to fluconazole and 2 (2.2%) were resistant to itraconazole. Seventeen mutations, including 9 silent and 8 missense mutations, were identified in the ERG11 gene of 31 C. albicans isolates. Our findings suggest that infection caused by C. albicans and non-C. albicansis common in Chinese Han women of reproductive age. Moreover, the relationship between Candida infection and certain epidemiological factors emphasizes the need to educate women about the precise diagnosis and punctual treatment of vaginitis.

  3. Presence of TMPRSS2-ERG is associated with alterations of the metabolic profile in human prostate cancer

    PubMed Central

    Hansen, Ailin Falkmo; Sandsmark, Elise; Rye, Morten Beck; Wright, Alan J.; Bertilsson, Helena; Richardsen, Elin; Viset, Trond; Bofin, Anna M.; Angelsen, Anders; Selnæs, Kirsten M.; Bathen, Tone Frost; Tessem, May-Britt

    2016-01-01

    TMPRSS2-ERG has been proposed to be a prognostic marker for prostate cancer. The aim of this study was to identify changes in metabolism, genes and biochemical recurrence related to TMPRSS2-ERG by using an integrated approach, combining metabolomics, transcriptomics, histopathology and clinical data in a cohort of 129 human prostate samples (41 patients). Metabolic analyses revealed lower concentrations of citrate and spermine comparing ERGhigh to ERGlow samples, suggesting an increased cancer aggressiveness of ERGhigh compared to ERGlow. These results could be validated in a separate cohort, consisting of 40 samples (40 patients), and magnetic resonance spectroscopy imaging (MRSI) indicated an in vivo translational potential. Alterations of gene expression levels associated with key enzymes in the metabolism of citrate and polyamines were in consistence with the metabolic results. Furthermore, the metabolic alterations between ERGhigh and ERGlow were more pronounced in low Gleason samples than in high Gleason samples, suggesting it as a potential tool for risk stratification. However, no significant difference in biochemical recurrence was detected, although a trend towards significance was detected for low Gleason samples. Using an integrated approach, this study suggests TMPRSS2-ERG as a potential risk stratification tool for inclusion of active surveillance patients. PMID:27276682

  4. Drug trapping in hERG K+ channels: (not) a matter of drug size?† †The authors declare no competing interests.

    PubMed Central

    Linder, Tobias; Bernsteiner, Harald; Saxena, Priyanka; Bauer, Florian; Erker, Thomas; Timin, Eugen; Hering, Steffen

    2016-01-01

    Inhibition of hERG K+ channels by structurally diverse drugs prolongs the ventricular action potential and increases the risk of torsade de pointes arrhythmias and sudden cardiac death. The capture of drugs behind closed channel gates, so-called drug trapping, is suggested to harbor an increased pro-arrhythmic risk. In this study, the trapping mechanisms of a trapped hERG blocker propafenone and a bulky derivative (MW: 647.24 g mol–1) were studied by making use of electrophysiological measurements in combination with molecular dynamics simulations. Our study suggests that the hERG cavity is able to accommodate very bulky compounds without disturbing gate closure.

  5. Optimization of piperidin-4-yl-urea-containing melanin-concentrating hormone receptor 1 (MCH-R1) antagonists: Reducing hERG-associated liabilities.

    PubMed

    Berglund, Susanne; Egner, Bryan J; Gradén, Henrik; Gradén, Joakim; Morgan, David G A; Inghardt, Tord; Giordanetto, Fabrizio

    2009-08-01

    The discovery and optimization of piperidin-4-yl-urea derivatives as MCH-R1 antagonists is herein described. Previous work around the piperidin-4-yl-amides led to the discovery of potent MCH-R1 antagonists. However, high affinity towards the hERG potassium channel proved to be an issue. Different strategies to increase hERG selectivity were implemented and resulted in the identification of piperidin-4-yl-urea compounds as potent MCH-R1 antagonists with minimized hERG inhibition.

  6. Phase-matched four-wave mixing of sub-100-TW/ cm2 femtosecond laser pulses in isolated air-guided modes of a hollow photonic-crystal fiber.

    PubMed

    Konorov, S O; Serebryannikov, E E; Akimov, D A; Ivanov, A A; Alfimov, M V; Zheltikov, A M

    2004-12-01

    Hollow-core photonic-crystal fibers are shown to allow propagation and nonlinear-optical frequency conversion of high-intensity ultrashort laser pulses in the regime of isolated guided modes confined in the hollow gas-filled fiber core. With a specially designed dispersion of such modes, the 3omega=2omega+2omega-omega four-wave mixing of fundamental (omega) and second-harmonic (2omega) sub-100- TW/ cm(2) femtosecond pulses of a Cr:forsterite laser can be phase matched in a hollow photonic-crystal fiber within a spectral band of more than 10 nm, resulting in the efficient generation of femtosecond pulses in a well-resolved higher-order air-guided mode of 417-nm radiation.

  7. A simple, rapid, low-cost technique for naked-eye detection of urine-isolated TMPRSS2:ERG gene fusion RNA

    PubMed Central

    Koo, Kevin M.; Wee, Eugene J. H.; Mainwaring, Paul N.; Trau, Matt

    2016-01-01

    The TMPRSS2:ERG gene fusion is one of a series of highly promising prostate cancer (PCa) biomarker alternatives to the controversial serum PSA. Current methods for detecting TMPRSS2:ERG are limited in terms of long processing time, high cost and the need for specialized equipment. Thus, there is an unmet need for less complex, faster, and cheaper methods to enable gene fusion detection in the clinic. We describe herein a simple, rapid and inexpensive assay which combines robust isothermal amplification technique with a novel visualization method for evaluating urinary TMPRSS2:ERG status at less than USD 5 and with minimal equipment. The assay is sensitive, and rapidly detects as low as 105 copies of TMPRSS2:ERG transcripts while maintaining high levels of specificity. PMID:27470540

  8. Azimuthal Structure of the Sand Erg that Encircles the North Polar Water-Ice Cap

    NASA Astrophysics Data System (ADS)

    Teodoro, L. A.; Elphic, R. C.; Eke, V. R.; Feldman, W. C.; Maurice, S.; Pathare, A.

    2011-12-01

    The sand erg that completely encircles the perennial water-ice cap that covers the Martian north geographic pole displays considerable azimuthal structure as seen in visible and near-IR images. Much of this structure is associated with the terminations of the many steep troughs that cut spiral the approximately 3 km thick polar ice cap. Other contributions come from the katabatic winds that spill over steep-sided edges of the cap, such as what bounds the largest set of dunes that comprise Olympia Undae. During the spring and summer months when these winds initiate from the higher altitudes that contain sublimating CO2 ice, which is very cold and dry, heat adiabatically when they compress as they lose altitude. These winds should then remove H2O moisture from the uppermost layer of the sand dunes that are directly in their path. Two likely locations where this desiccation may occur preferentially is at the termination of Chasma Boreale and the ice cap at Olympia Undae. We will search for this effect by sharpening the spatial structure of the epithermal neutron counting rates measured at northern high latitudes using the Mars Odyssey Neutron Spectrometer (MONS). The epithermal range of neutron energies is nearly uniquely sensitive to the hydrogen content of surface soils, which should likely be in the form of H2O/OH molecules/radicals. We therefore convert epithermal counting rates in terms of Water-Equivalent-Hydrogen, WEH. However, MONS counting-rate data have a FWHM of ~550 km., which is sufficiently broad to prevent a close association of WEH variability with images of geological features. In this study, we reduce spurious features in the instrument smeared neutron counting rates through deconvolution. We choose the PIXON numerical deconvolution technique for this purpose. This technique uses a statistical approach (Pina 2001, Eke 2001), which is capable of removing spurious features in the data in the presence of noise. We have previously carried out a detailed

  9. Long-Term Channel Block Is Required to Inhibit Cellular Transformation by Human Ether-à-Go-Go–Related Gene (hERG1) Potassium Channels

    PubMed Central

    Pier, David M.; Shehatou, George S. G.; Giblett, Susan; Pullar, Christine E.; Trezise, Derek J.; Pritchard, Catrin A.; Challiss, R. A. John

    2014-01-01

    Both human ether-à-go-go–related gene (hERG1) and the closely related human ether-à-go-go (hEAG1) channel are aberrantly expressed in a large proportion of human cancers. In the present study, we demonstrate that transfection of hERG1 into mouse fibroblasts is sufficient to induce many features characteristic of malignant transformation. An important finding of this work is that this transformation could be reversed by chronic incubation (for 2–3 weeks) with the hERG channel blocker dofetilide (100 nM), whereas more acute applications (for 1–2 days) were ineffective. The hERG1 expression resulted in a profound loss of cell contact inhibition, multiple layers of overgrowing cells, and high saturation densities. Cells also changed from fibroblast-like to a more spindle-shaped morphology, which was associated with a smaller cell size, a dramatic increase in cell polarization, a reduction in the number of actin stress fibers, and less punctate labeling of focal adhesions. Analysis of single-cell migration and scratch-wound closure clearly demonstrated that hERG1-expressing cells migrated more rapidly than vector-transfected control cells. In contrast to previous studies on hEAG1, there were no increases in rates of proliferation, or loss of growth factor dependency; however, hERG1-expressing cells were capable of substrate-independent growth. Allogeneic transplantation of hERG1-expressing cells into nude mice resulted in an increased incidence of tumors. In contrast to hEAG1, the mechanism of cellular transformation is dependent on ion conduction. Trafficking-deficient and conduction-deficient hERG1 mutants also prevented cellular transformation. These results provide evidence that hERG1 expression is sufficient to induce cellular transformation by a mechanism distinct from hEAG1. The most important conclusion of this study is that selective hERG1 channel blockers have therapeutic potential in the treatment of hERG1-expressing cancers. PMID:24830940

  10. Protein kinase C is necessary for recovery from the thyrotropin-releasing hormone-induced r-ERG current reduction in GH3 rat anterior pituitary cells

    PubMed Central

    Gómez-Varela, David; Giráldez, Teresa; la Peña, Pilar de; Dupuy, Silvia G; García-Manso, Diego; Barros, Francisco

    2003-01-01

    The biochemical cascade linking activation of phospholipase C-coupled thyrotropin-releasing hormone (TRH) receptors to rat ERG (r-ERG) channel modulation was studied in situ using perforated-patch clamped adenohypophysial GH3 cells and pharmacological inhibitors. To check the recent suggestion that Rho kinase is involved in the TRH-induced r-ERG current suppression, the hormonal effects were studied in cells pretreated with the Rho kinase inhibitors Y-27632 and HA-1077. The TRH-induced r-ERG inhibition was not significantly modified in the presence of the inhibitors. Surprisingly, the hormonal effects became irreversible in the presence of HA-1077 but not in the presence of the more potent Rho kinase inhibitor Y-27632. Further experiments indicated that the effect of HA-1077 correlated with its ability to inhibit protein kinase C (PKC). The hormonal effects also became irreversible in cells in which PKC activity was selectively impaired with GF109203X, Gö6976 or long-term incubation with phorbol esters. Furthermore, the reversal of the effects of TRH, but not its ability to suppress r-ERG currents, was blocked if diacylglycerol generation was prevented by blocking phospholipase C activity with U-73122. Our results suggest that a pathway involving an as yet unidentified protein kinase is the main cause of r-ERG inhibition in perforated-patch clamped GH3 cells. Furthermore, they demonstrate that although not necessary to trigger the ERG current reductions induced by TRH, an intracellular signal cascade involving phosphatidylinositol-4,5-bisphosphate hydrolysis by phospholipase C, activation of an α/βII conventional PKC and one or more dephosphorylation steps catalysed by protein phosphatase 2A, mediates recovery of ERG currents following TRH withdrawal. PMID:12562894

  11. Sterols of Saccharomyces cerevisiae erg6 Knockout Mutant Expressing the Pneumocystis carinii S-Adenosylmethionine:Sterol C-24 Methyltransferase (SAM:SMT)

    PubMed Central

    Kaneshiro, Edna S.; Johnston, Laura Q.; Nkinin, Stephenson W.; Romero, Becky I.; Giner, José-Luis

    2014-01-01

    The AIDS-associated lung pathogen Pneumocystis is classified as a fungus although Pneumocystis has several distinct features such as the absence of ergosterol, the major sterol of most fungi. The P. carinii S-adenosylmethionine:sterol C24-methyltransferase (SAM:SMT) enzyme, coded by the erg6 gene, transfers either one or two methyl groups to the C-24 position of the sterol side chain producing both C28 and C29 24-alkylsterols in approximately the same proportions whereas most fungal SAM:SMT transfer only one methyl group to the side chain. The sterol compositions of wild type Sacchromyces cerevisiae, the erg6 knockout mutant (Δerg6), and Δerg6 expressing the P. carinii or the S. cerevisiae erg6 gene were analyzed by a variety of chromatographic and spectroscopic procedures to examine functional complementation in the yeast expression system. Detailed sterol analyses were obtained using high performance liquid chromatography (HPLC) and proton nuclear magnetic resonance spectroscopy (1H-NMR). The P. carinii SAM:SMT in the Δerg6 restored its ability to produce the C28 sterol ergosterol as the major sterol, and also resulted in low levels of C29 sterols. This indicates that while the P. carinii SAM:SMT in the yeast Δerg6 cells was able to transfer a second methyl group to the side chain, the action of Δ24(28)-sterol reductase (coded by the erg4 gene) in the yeast cells prevented the formation and accumulation of as many C29 sterols as that found in P. carinii. PMID:25230683

  12. Evaluation of ERG and SPINK1 by Immunohistochemical Staining and Clinicopathological Outcomes in a Multi-Institutional Radical Prostatectomy Cohort of 1067 Patients

    PubMed Central

    Brooks, James D.; Wei, Wei; Hawley, Sarah; Auman, Heidi; Newcomb, Lisa; Boyer, Hilary; Fazli, Ladan; Simko, Jeff; Hurtado-Coll, Antonio; Troyer, Dean A.; Carroll, Peter R.; Gleave, Martin; Lance, Raymond; Lin, Daniel W.; Nelson, Peter S.; Thompson, Ian M.; True, Lawrence D.; Feng, Ziding; McKenney, Jesse K.

    2015-01-01

    Distinguishing between patients with early stage, screen detected prostate cancer who must be treated from those that can be safely watched has become a major issue in prostate cancer care. Identification of molecular subtypes of prostate cancer has opened the opportunity for testing whether biomarkers that characterize these subtypes can be used as biomarkers of prognosis. Two established molecular subtypes are identified by high expression of the ERG oncoprotein, due to structural DNA alterations that encode for fusion transcripts in approximately ½ of prostate cancers, and over-expression of SPINK1, which is purportedly found only in ERG-negative tumors. We used a multi-institutional prostate cancer tissue microarray constructed from radical prostatectomy samples with associated detailed clinical data and with rigorous selection of recurrent and non-recurrent cases to test the prognostic value of immunohistochemistry staining results for the ERG and SPINK1 proteins. In univariate analysis, ERG positive cases (419/1067; 39%) were associated with lower patient age, pre-operative serum PSA levels, lower Gleason scores (≤3+4=7) and improved recurrence free survival (RFS). On multivariate analysis, ERG status was not correlated with RFS, disease specific survival (DSS) or overall survival (OS). High-level SPINK1 protein expression (33/1067 cases; 3%) was associated with improved RFS on univariate and multivariate Cox regression analysis. Over-expression of either protein was not associated with clinical outcome. While expression of ERG and SPINK1 proteins was inversely correlated, it was not mutually exclusive since 3 (0.28%) cases showed high expression of both. While ERG and SPINK1 appear to identify discrete molecular subtypes of prostate cancer, only high expression of SPINK1 was associated with improved clinical outcome. However, by themselves, neither ERG nor SPINK1 appear to be useful biomarkers for prognostication of early stage prostate cancer. PMID

  13. Evaluation of ERG and SPINK1 by Immunohistochemical Staining and Clinicopathological Outcomes in a Multi-Institutional Radical Prostatectomy Cohort of 1067 Patients.

    PubMed

    Brooks, James D; Wei, Wei; Hawley, Sarah; Auman, Heidi; Newcomb, Lisa; Boyer, Hilary; Fazli, Ladan; Simko, Jeff; Hurtado-Coll, Antonio; Troyer, Dean A; Carroll, Peter R; Gleave, Martin; Lance, Raymond; Lin, Daniel W; Nelson, Peter S; Thompson, Ian M; True, Lawrence D; Feng, Ziding; McKenney, Jesse K

    2015-01-01

    Distinguishing between patients with early stage, screen detected prostate cancer who must be treated from those that can be safely watched has become a major issue in prostate cancer care. Identification of molecular subtypes of prostate cancer has opened the opportunity for testing whether biomarkers that characterize these subtypes can be used as biomarkers of prognosis. Two established molecular subtypes are identified by high expression of the ERG oncoprotein, due to structural DNA alterations that encode for fusion transcripts in approximately ½ of prostate cancers, and over-expression of SPINK1, which is purportedly found only in ERG-negative tumors. We used a multi-institutional prostate cancer tissue microarray constructed from radical prostatectomy samples with associated detailed clinical data and with rigorous selection of recurrent and non-recurrent cases to test the prognostic value of immunohistochemistry staining results for the ERG and SPINK1 proteins. In univariate analysis, ERG positive cases (419/1067; 39%) were associated with lower patient age, pre-operative serum PSA levels, lower Gleason scores (≤ 3+4=7) and improved recurrence free survival (RFS). On multivariate analysis, ERG status was not correlated with RFS, disease specific survival (DSS) or overall survival (OS). High-level SPINK1 protein expression (33/1067 cases; 3%) was associated with improved RFS on univariate and multivariate Cox regression analysis. Over-expression of either protein was not associated with clinical outcome. While expression of ERG and SPINK1 proteins was inversely correlated, it was not mutually exclusive since 3 (0.28%) cases showed high expression of both. While ERG and SPINK1 appear to identify discrete molecular subtypes of prostate cancer, only high expression of SPINK1 was associated with improved clinical outcome. However, by themselves, neither ERG nor SPINK1 appear to be useful biomarkers for prognostication of early stage prostate cancer.

  14. Urinary TMPRSS2:ERG and PCA3 in an active surveillance cohort: results from a baseline analysis in the Canary Prostate Active Surveillance Study

    PubMed Central

    Lin, Daniel W.; Newcomb, Lisa F.; Brown, Elissa C.; Brooks, James D.; Carroll, Peter R.; Feng, Ziding; Gleave, Martin E.; Lance, Raymond S.; Sanda, Martin G.; Thompson, Ian M.; Wei, John T.; Nelson, Peter S.

    2013-01-01

    Purpose Active surveillance is used to manage low risk prostate cancer. Both PCA3 and TMRPSS2-ERG are promising biomarkers that may be associated with aggressive disease. This study examines the correlation of these biomarkers with higher cancer volume and grade determined at the time of biopsy in an active surveillance cohort. Experimental Design Post-DRE urine was collected prospectively as part of the multi-institutional Canary Prostate Active Surveillance Study (PASS). PCA3 and TMPRSS2-ERG levels were analyzed in urine collected at study entry. Biomarker scores were correlated to clinical and pathologic variables. Results In 387 men, both PCA3 and TMPRSS2-ERG scores were significantly associated with higher volume disease. For a negative repeat biopsy, and 1–10%, 11–33%, ≥34% positive cores, median PCA3 and TMPRSS2-ERG scores increased incrementally (P < 0.005). Both PCA3 and TMPRSS2-ERG scores were also significantly associated with presence of high grade disease. For a negative repeat biopsy, Gleason 6 and Gleason ≥7 cancers, the median PCA3 and TMPRSS2-ERG scores also increased incrementally (P = 0.02 and P = 0.001, respectively). Using the marker scores as a continuous variables, the odds ratio for a biopsy in which cancer was detected versus a negative repeat biopsy (ref) on modeling was 1.41 (95% CI 1.07–1.85), P = 0.01 for PCA3 and 1.28 (95% CI 1.10–1.49), P = 0.001 for TMPRSS2-ERG. Conclusions For men on active surveillance both PCA3 and TMPRSS2-ERG appear to stratify risk of having aggressive cancer as defined by tumor volume or Gleason score. PMID:23515404

  15. hERG1 behaves as biomarker of progression to adenocarcinoma in Barrett's esophagus and can be exploited for a novel endoscopic surveillance

    PubMed Central

    Lastraioli, Elena; Lottini, Tiziano; Iorio, Jessica; Freschi, Giancarlo; Fazi, Marilena; Duranti, Claudia; Carraresi, Laura; Messerini, Luca; Taddei, Antonio; Ringressi, Maria Novella; Salemme, Marianna; Villanacci, Vincenzo; Vindigni, Carla; Tomezzoli, Anna; Mendola, Roberta La; Bencivenga, Maria; Compagnoni, Bruno; Chiudinelli, Mariella; Saragoni, Luca; Manzi, Ilaria; Manzoni, Giovanni De; Bechi, Paolo

    2016-01-01

    Barrett's esophagus (BE) is the only well-known precursor lesion of esophageal adenocarcinoma (EA). The exact estimates of the annual progression rate from BE to EA vary from 0.07% to 3.6%. The identification of BE patients at higher risk to progress to EA is hence mandatory, although difficult to accomplish. In search of novel BE biomarkers we analyzed the efficacy of hERG1 potassium channels in predicting BE progression to EA. Once tested by immunohistochemistry (IHC) on bioptic samples, hERG1 was expressed in BE, and its expression levels increased during progression from BE to esophageal dysplasia (ED) and EA. hERG1 was also over-expressed in the metaplastic cells arising in BE lesions obtained in different BE mouse models, induced either surgically or chemically. Furthermore, transgenic mice which over express hERG1 in the whole gastrointestinal tract, developed BE lesions after an esophago-jejunal anastomosis more frequently, compared to controls. A case-control study was performed on 104 bioptic samples from newly diagnosed BE patients further followed up for at least 10 years. It emerged a statistically significant association between hERG1 expression status and risk of progression to EA. Finally, a novel fluorophore- conjugated recombinant single chain variable fragment antibody (scFv-hERG1-Alexa488) was tested on freshly collected live BE biopsies: it could recognize hERG1 positive samples, perfectly matching IHC data. Overall, hERG1 can be considered a novel BE biomarker to be exploited for a novel endoscopic surveillance protocol, either in biopsies or through endoscopy, to identify those BE patients with higher risk to progress to EA. PMID:27517748

  16. Quantitative prediction of the arrhythmogenic effects of de novo hERG mutations in computational models of human ventricular tissues.

    PubMed

    Benson, Alan P; Al-Owais, Moza; Holden, Arun V

    2011-05-01

    Mutations to hERG which result in changes to the rapid delayed rectifier current I(Kr) can cause long and short QT syndromes and are associated with an increased risk of cardiac arrhythmias. Experimental recordings of I(Kr) reveal the effects of mutations at the channel level, but how these changes translate to the cell and tissue levels remains unclear. We used computational models of human ventricular myocytes and tissues to predict and quantify the effects that de novo hERG mutations would have on cell and tissue electrophysiology. Mutations that decreased I(Kr) maximum conductance resulted in an increased cell and tissue action potential duration (APD) and a long QT interval on the electrocardiogram (ECG), whereas those that caused a positive shift in the inactivation curve resulted in a decreased APD and a short QT. Tissue vulnerability to re-entrant arrhythmias was correlated with transmural dispersion of repolarisation, and any change to this vulnerability could be inferred from the ECG QT interval or T wave peak-to-end time. Faster I(Kr) activation kinetics caused cell APD alternans to appear over a wider range of pacing rates and with a larger magnitude, and spatial heterogeneity in these cellular alternans resulted in discordant alternans at the tissue level. Thus, from channel kinetic data, we can predict the tissue-level electrophysiological effects of any hERG mutations and identify how the mutation would manifest clinically, as either a long or short QT syndrome with or without an increased risk of alternans and re-entrant arrhythmias.

  17. The extremely high-energy plasma/particle sensor for electron (XEP-e) of the ERG satellite

    NASA Astrophysics Data System (ADS)

    Higashio, N.; Matsumoto, H.

    2015-12-01

    It is well known that satellites are always in danger in space and especially high-energy radiation damages them. One of the sources that cause them is the radiation belt (the Van Allen belt). It was thought to be static, but in the 1990s it rediscovered the radiation belt fluctuates greatly. There are some reasons to occur this phenomenon, but we have not understood a clear reason of this yet. On the other hand, it is well known that the energetic particle flux vary during geomagnetic disturbances and the relativistic electrons in the other radiation belt change with solar wind speed. Now we are trying to develop the satellite (ERG) to reveal this mechanism. ERG (Energization and Radiation in Geospace) satellite is the small space science platform for rapid investigation and test satellite of JAXA/ISAS. This satellite will be lanched in 2016. Our group is developing the instrument (the XEP-e) to measure high-energy electrons (400keV~20MeV), that is one of many ERG satellite instruments. The XEP-e (eXtremely high Energy Plasma/ particle sensor for electron) is consists of the 5 SSDs (Solid-State Silicon Detectors) and a GSO single crystal scintillator. It has one-way conic sight and an electric part is unified with a part of sensor that is covered with aluminum to protect from contaminationand and an anti-scintillator to detect it. The front part of the SSDs discriminate a radiation enters into the sensor and the back part of the plastic scintillator get the value of its energy. We can get the data of high-energy electron by using this sensor and it will be useful to reveal the detail of the radiation belt's fluctuation.

  18. Replacement of olivine by serpentine in the Queen Alexandra Range 93005 carbonaceous chondrite (CM2): Reactant-product compositional relations, and isovolumetric constraints on reaction stoichiometry and elemental mobility during aqueous alteration

    NASA Astrophysics Data System (ADS)

    Velbel, Michael A.; Tonui, Eric K.; Zolensky, Michael E.

    2015-01-01

    Isovolumetric replacement of euhedral and anhedral olivine by serpentine produced both centripetal and meshwork textures in the CM2 chondrites ALH 81002 and Nogoya. The compositions of these textural varieties of serpentine are uniform within narrow limits within each previously studied meteorite, independent of the composition of olivine being replaced, and different between the two meteorites. In QUE 93005 (CM2), coarse olivines of widely varying compositions (Fo<76-99) are replaced in a texturally similar manner by compositionally uniform serpentine (Mg0.73±0.05Fe0.27±0.05)3Si2O5(OH)4. The narrow compositional range of serpentine replacing coarse olivine indicates that the aqueous solution from which the serpentine formed was compositionally uniform on scales at least as large as the meteorite (∼2.5 cm in longest dimension). Isovolumetric textures and compositional observations constrain elemental redistribution from coarse olivine to serpentine and to surrounding phases during serpentinization. Regardless of olivine's composition, isovolumetric replacement of coarse olivines by serpentine of the observed composition released more Mg and Si from olivine than was required to form the serpentine. Excess Mg and Si released by olivine destruction and not retained in serpentine were exported from the replaced volume. Olivines with different Fa/Fo proportions contributed different amounts of Fe and Mg to the serpentine. Ferroan olivines released more Fe than required to form the serpentines replacing them, so some of the Fe released from ferroan olivine was exported from the replaced volumes. Forsteritic olivines released less Fe than required to form the serpentines replacing them, so some Fe was imported into the replaced volumes augmenting the small amount of Fe released from forsteritic olivine. In QUE 93005 Fo83.8 is the threshold composition between Fe-exporting and Fe-importing behavior in individual olivine-serpentine pairs, which released exactly the

  19. Components of gating charge movement and S4 voltage-sensor exposure during activation of hERG channels.

    PubMed

    Wang, Zhuren; Dou, Ying; Goodchild, Samuel J; Es-Salah-Lamoureux, Zeineb; Fedida, David

    2013-04-01

    The human ether-á-go-go-related gene (hERG) K(+) channel encodes the pore-forming α subunit of the rapid delayed rectifier current, IKr, and has unique activation gating kinetics, in that the α subunit of the channel activates and deactivates very slowly, which focuses the role of IKr current to a critical period during action potential repolarization in the heart. Despite its physiological importance, fundamental mechanistic properties of hERG channel activation gating remain unclear, including how voltage-sensor movement rate limits pore opening. Here, we study this directly by recording voltage-sensor domain currents in mammalian cells for the first time and measuring the rates of voltage-sensor modification by [2-(trimethylammonium)ethyl] methanethiosulfonate chloride (MTSET). Gating currents recorded from hERG channels expressed in mammalian tsA201 cells using low resistance pipettes show two charge systems, defined as Q(1) and Q(2), with V(1/2)'s of -55.7 (equivalent charge, z = 1.60) and -54.2 mV (z = 1.30), respectively, with the Q(2) charge system carrying approximately two thirds of the overall gating charge. The time constants for charge movement at 0 mV were 2.5 and 36.2 ms for Q(1) and Q(2), decreasing to 4.3 ms for Q(2) at +60 mV, an order of magnitude faster than the time constants of ionic current appearance at these potentials. The voltage and time dependence of Q2 movement closely correlated with the rate of MTSET modification of I521C in the outermost region of the S4 segment, which had a V(1/2) of -64 mV and time constants of 36 ± 8.5 ms and 11.6 ± 6.3 ms at 0 and +60 mV, respectively. Modeling of Q(1) and Q(2) charge systems showed that a minimal scheme of three transitions is sufficient to account for the experimental findings. These data point to activation steps further downstream of voltage-sensor movement that provide the major delays to pore opening in hERG channels.

  20. Pharmacological and electrophysiological characterization of nine, single nucleotide polymorphisms of the hERG-encoded potassium channel

    PubMed Central

    Männikkö, R; Overend, G; Perrey, C; Gavaghan, CL; Valentin, J-P; Morten, J; Armstrong, M; Pollard, CE

    2010-01-01

    Background and purpose: Potencies of compounds blocking KV11.1 [human ether-ago-go-related gene (hERG)] are commonly assessed using cell lines expressing the Caucasian wild-type (WT) variant. Here we tested whether such potencies would be different for hERG single nucleotide polymorphisms (SNPs). Experimental approach: SNPs (R176W, R181Q, Del187-189, P347S, K897T, A915V, P917L, R1047L, A1116V) and a binding-site mutant (Y652A) were expressed in Tet-On CHO-K1 cells. Potencies [mean IC50; lower/upper 95% confidence limit (CL)] of 48 hERG blockers was estimated by automated electrophysiology [IonWorks™ HT (IW)]. In phase one, rapid potency comparison of each WT-SNP combination was made for each compound. In phase two, any compound-SNP combinations from phase one where the WT upper/lower CL did not overlap with those of the SNPs were re-examined. Electrophysiological WT and SNP parameters were determined using conventional electrophysiology. Key results: IW detected the expected sixfold potency decrease for propafenone in Y652A. In phase one, the WT lower/upper CL did not overlap with those of the SNPs for 77 compound-SNP combinations. In phase two, 62/77 cases no longer yielded IC50 values with non-overlapping CLs. For seven of the remaining 15 cases, there were non-overlapping CLs but in the opposite direction. For the eight compound-SNP combinations with non-overlapping CLs in the same direction as for phase 1, potencies were never more than twofold apart. The only statistically significant electrophysiological difference was the voltage dependence of activation of R1047L. Conclusion and implications: Potencies of hERG channel blockers defined using the Caucasian WT sequence, in this in vitro assay, were representative of potencies for common SNPs. This article is part of a themed section on QT safety. To view this issue visit http://www3.interscience.wiley.com/journal/121548564/issueyear?year=2010 PMID:19673885

  1. Physics: Grades 10-12.

    ERIC Educational Resources Information Center

    Instructional Objectives Exchange, Los Angeles, CA.

    The physics objectives are geared to use in college preparatory, high school physics courses and are based on the three most common physics curricula: (1) Physical Science Study Committee (PSSC); (2) The Project Physics Course; and (3) Modern Physics by Dull, Metcalf, and Williams. Since many of the sample items can be answered in various ways,…

  2. Study of the 20,22Ne+20,22Ne and 10,12,13,14,15C+12C Fusion Reactions with MUSIC

    NASA Astrophysics Data System (ADS)

    Avila, M. L.; Rehm, K. E.; Almaraz-Calderon, S.; Carnelli, P. F. F.; DiGiovine, B.; Esbensen, H.; Hoffman, C. R.; Jiang, C. L.; Kay, B. P.; Lai, J.; Nusair, O.; Pardo, R. C.; Santiago-Gonzalez, D.; Talwar, R.; Ugalde, C.

    2016-05-01

    A highly efficient MUlti-Sampling Ionization Chamber (MUSIC) detector has been developed for measurements of fusion reactions. A study of fusion cross sections in the 10,12,13,14,15C+12C and 20,22Ne+20,22Ne systems has been performed at ATLAS. Experimental results and comparison with theoretical predictions are presented. Furthermore, results of direct measurements of the 17O(α, n)20Ne, 23Ne(α, p)26Mg and 23Ne(α, n)26Al reactions will be discussed.

  3. Key comparison BIPM.RI(I)-K6 of the standards for absorbed dose to water at 10 g cm-2 of the NPL, United Kingdom and the BIPM in accelerator photon beams

    NASA Astrophysics Data System (ADS)

    Picard, S.; Burns, D. T.; Roger, P.; Duane, S.; Bass, G. A.; Manning, J. W.; Shipley, D. R.

    2015-01-01

    A comparison of the dosimetry for accelerator photon beams was carried out between the National Physical Laboratory (NPL) and the Bureau International des Poids et Mesures (BIPM) from 23 September to 7 October 2014. The comparison was based on the determination of absorbed dose to water at 10 g cm-2 for three radiation qualities at the NPL. The results, reported as ratios of the NPL and the BIPM evaluations (and with the combined standard uncertainties given in parentheses), are 1.0000(62) at 6 MV, 0.9999(70) at 10 MV and 0.9993(80) at 25 MV. This result is the seventh in the on-going BIPM.RI(I)-K6 series of comparisons. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCRI, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

  4. Key comparison BIPM.RI(I)-K6 of the standards for absorbed dose to water at 10 g cm-2 of the NMIJ, Japan and the BIPM in accelerator photon beams

    NASA Astrophysics Data System (ADS)

    Picard, S.; Burns, D. T.; Roger, P.; Shimizu, M.; Morishita, Y.; Kato, M.; Tanaka, T.; Kurosawa, T.; Saito, N.

    2016-01-01

    A comparison of the dosimetry for accelerator photon beams was carried out between the National Metrology Institute of Japan (NMIJ) and the Bureau International des Poids et Mesures (BIPM) from 9 to 23 April 2015. The comparison was based on the determination of absorbed dose to water at 10 g cm-2 for three radiation qualities at the NMIJ. The results, reported as ratios of the NMIJ and the BIPM evaluations (and with the combined standard uncertainties given in parentheses), are 0.9966 (47) at 6 MV, 0.9965 (60) at 10 MV and 0.9953 (50) at 15 MV. This result is the eighth in the on-going BIPM.RI(I)-K6 series of comparisons. Main text To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCRI, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

  5. Enhanced therapeutic anti-inflammatory effect of betamethasone upon topical administration with low frequency, low intensity (20 kHz, 100 mW/cm2) ultrasound exposure on carrageenan-induced arthritis in mice model

    PubMed Central

    Cohen, Gadi; Natsheh, Hiba; Sunny, Youhan; Bawiec, Christopher R.; Touitou, Elka; Lerman, Melissa A.; Lazarovici, Philip; Lewin, Peter A.

    2015-01-01

    The purpose of this work was to investigate whether low frequency, low intensity (LFLI, 20 kHz, <100 mW/cm2, spatial-peak, temporal-peak) ultrasound (US), delivered by a light-weight (<100g), tether-free, fully wearable, battery powered applicator is capable of reducing inflammation in a mouse model of Rheumatoid Arthritis (RA). The therapeutic, acute, anti-inflammatory effect was estimated by the relative swelling induced in mice hind limb paws. In an independent, indirect approach, the inflammation was bio-imaged by measuring glycolytic activity with near infrared labeled 2-deoxy-glucose (2DG). The outcome of the experiments indicated that the combination of US exposure with topical application of 0.1% w/w betamethasone gel, exhibited statistically significant (p<0.05) enhanced anti-inflammatory properties in comparison with the drug or US treatment alone. The present study underscores the potential benefits of LFLI US assisted drug delivery. However, the proof of concept presented indicates the need for additional experiments to systematically evaluate and optimize the potential of, and the conditions for, safe, LFLI ultrasound promoted non-invasive drug delivery. PMID:26003010

  6. Vortex pinning properties in Co-doped BaFe2As2 thin films with a high critical current density over 2 MA cm-2 at 9 T

    NASA Astrophysics Data System (ADS)

    Yuan, Pusheng; Xu, Zhongtang; Wang, Dongliang; Zhang, Ming; Li, Jianqi; Ma, Yanwei

    2017-02-01

    Similar to other high-temperature superconductors, the pinning centers largely determine the critical current density (J c) in Co-doped BaFe2As2 (Ba122:Co). Therefore, understanding the vortex pinning mechanism of high-J c Ba122:Co thin films is important for the applications of Ba122:Co. Herein we report the pinning of the naturally grown defects in Ba122:Co thin films with a high critical current density. The transport J c of thin films is up to 2.6 MA cm-2 in 9 T at 4.2 K, which is the highest value in iron-based superconductors. Microstructure investigations reveal a high density of ab-planar defects (stacking faults) and localized vertical defects present in the sample. The Dew-Hughes mode analyses prove that pinning centers by surface defects and by point defects are responsible for H//ab and H//c, respectively. Therefore, the high J c in strong magnetic field for both H//ab and H//c are related to surface and point defects which act as the pinning centers in Ba122:Co films.

  7. cis-Acting Elements within the Candida albicans ERG11 Promoter Mediate the Azole Response through Transcription Factor Upc2p▿

    PubMed Central

    Oliver, Brian G.; Song, Jia L.; Choiniere, Jake H.; White, Theodore C.

    2007-01-01

    The azole antifungal drugs are used to treat infections caused by Candida albicans and other fungi. These drugs interfere with the biosynthesis of ergosterol, the major sterol in fungal cells, by inhibiting an ergosterol biosynthetic enzyme, lanosterol 14 α-demethylase, encoded by the ERG11 gene. In vitro, these drugs as well as other ergosterol biosynthesis inhibitors increase ERG11 mRNA expression by activation of the ERG11 promoter. The signal for this activation most likely is the depletion of ergosterol, the end product of the pathway. To identify cis-acting regulatory elements that mediate this activation, ERG11 promoter fragments have been fused to the luciferase reporter gene from Renilla reniformis. Promoter deletions and linker scan mutations localized the region important for azole induction to a segment from bp −224 to −251 upstream of the start codon, specifically two 7-bp sequences separated by 13 bp. These sequences form an imperfect inverted repeat. The region is recognized by the transcription factor Upc2p and functions as an enhancer of transcription, as it can be placed upstream of a heterologous promoter in either direction, resulting in the azole induction of that promoter. The promoter constructs are not azole inducible in the upc2/upc2 homozygous deletion, demonstrating that Upc2p controls the azole induction of ERG11. These results identify an azole-responsive enhancer element (ARE) in the ERG11 promoter that is controlled by the Upc2p transcription factor. No other ARE is present in the promoter. Thus, this ARE and Upc2p are necessary and sufficient for azole induction of ERG11. PMID:17951521

  8. A novel four-color fluorescence in situ hybridization assay for the detection of TMPRSS2 and ERG rearrangements in prostate cancer.

    PubMed

    Qu, Xiaoyu; Randhawa, Grace; Friedman, Cynthia; O'Hara-Larrivee, Siobhan; Kroeger, Kathleen; Dumpit, Ruth; True, Larry; Vakar-Lopez, Funda; Porter, Christopher; Vessella, Robert; Nelson, Peter; Fang, Min

    2013-01-01

    Since the identification of the TMPRSS2-ERG rearrangement as the most common fusion event in prostate cancer, various methods have been developed to detect this rearrangement and to study its prognostic significance. We report a novel four-color fluorescence in situ hybridization (FISH) assay that detects not only the typical TMPRSS2-ERG fusion but also alternative rearrangements of the TMPRSS2 or ERG gene. We validated this assay on fresh, frozen, or formalin-fixed paraffin-embedded prostate cancer specimens, including cell lines, primary prostate cancer tissues, xenograft tissues derived from metastatic prostate cancer, and metastatic tissues from castration-resistant prostate cancer (CRPC) patients. When compared with either reverse transcription-polymerase chain reaction or the Gen-Probe method as the technical reference, analysis using the four-color FISH assay demonstrated an analytical sensitivity of 94.5% (95% confidence interval [CI] 0.80-0.99) and specificity of 100% (95% CI 0.89-1.00) for detecting the TMPRSS2-ERG fusion. The TMPRSS2-ERG fusion was detected in 41% and 43% of primary prostate cancer (n = 59) and CRPC tumors (n = 82), respectively. Rearrangements other than the typical TMPRSS2-ERG fusion were confirmed by karyotype analysis and found in 7% of primary cancer and 13% of CRPC tumors. Successful karyotype analyses are reported for the first time on four of the xenograft samples, complementing the FISH results. Analysis using the four-color FISH assay provides sensitive detection of TMPRSS2 and ERG gene rearrangements in prostate cancer.

  9. Improved method of detecting the ERG gene rearrangement in prostate cancer using combined dual-color chromogenic and silver in situ hybridization.

    PubMed

    Braun, Martin; Stomper, Julia; Boehm, Diana; Vogel, Wenzel; Scheble, Veit J; Wernert, Nicolas; Shaikhibrahim, Zaki; Fend, Falko; Kristiansen, Glen; Perner, Sven

    2012-07-01

    The recently detected TMPRSS2-ERG fusion gene was revealed as a recurrent and prevalent prostate cancer (PCa)-specific event, potentially qualifying it for clinical use. To detect this alteration, fluorescence in situ hybridization (FISH) is the method of choice. However, FISH has some disadvantages for widespread adoption in clinical practice. Subsequently, chromogenic in situ hybridization, which uses organic chromogens, and enzymatic metallography silver in situ hybridization have emerged as promising bright-field alternatives. Compared with chromogenic in situ hybridization, silver in situ hybridization signals are very distinct and superior with regard to signal clarity and resolution, but the method excludes multicolor protocols. Based on the ERG break-apart FISH assay, we established a dual-color ERG break-apart assay using combined chromogenic in situ hybridization and silver in situ hybridization (CS-ISH) and compared these results with those obtained by FISH. We assessed 178 PCa and 10 benign specimens for their ERG rearrangement status by applying dual-color FISH and CS-ISH ERG break-apart assays to consecutive sections. We observed a highly significant concordance (97.7%) between FISH- and CS-ISH-based results (Pearson's correlation coefficient = 0.955, P < 0.001). Our findings demonstrate that the ERG rearrangement status can reliably be assessed by CS-ISH. Further, the CS-ISH technique combines the accuracy and precision of FISH with the ease of bright-field microscopy. This tool allows a much broader spectrum of applications in which to study the biological role and clinical use of ERG rearrangements in PCa.

  10. Bag1 Co-chaperone Promotes TRC8 E3 Ligase-dependent Degradation of Misfolded Human Ether a Go-Go-related Gene (hERG) Potassium Channels.

    PubMed

    Hantouche, Christine; Williamson, Brittany; Valinsky, William C; Solomon, Joshua; Shrier, Alvin; Young, Jason C

    2017-02-10

    Cardiac long QT syndrome type 2 is caused by mutations in the human ether a go-go-related gene (hERG) potassium channel, many of which cause misfolding and degradation at the endoplasmic reticulum instead of normal trafficking to the cell surface. The Hsc70/Hsp70 chaperones assist the folding of the hERG cytosolic domains. Here, we demonstrate that the Hsp70 nucleotide exchange factor Bag1 promotes hERG degradation by the ubiquitin-proteasome system at the endoplasmic reticulum to regulate hERG levels and channel activity. Dissociation of hERG complexes containing Hsp70 and the E3 ubiquitin ligase CHIP requires the interaction of Bag1 with Hsp70, but this does not involve the Bag1 ubiquitin-like domain. The interaction with Bag1 then shifts hERG degradation to the membrane-anchored E3 ligase TRC8 and its E2-conjugating enzyme Ube2g2, as determined by siRNA screening. TRC8 interacts through the transmembrane region with hERG and decreases hERG functional expression. TRC8 also mediates degradation of the misfolded hERG-G601S disease mutant, but pharmacological stabilization of the mutant structure prevents degradation. Our results identify TRC8 as a previously unknown Hsp70-independent quality control E3 ligase for hERG.

  11. Histopathological changes in retinas and F-ERG features of streptozotocin-induced diabetic rats treated with ozone

    PubMed Central

    Xie, Ting-Yu; Li, Qin; Chen, Xue-Yi

    2016-01-01

    AIM To study the histopathological changes in the retina and flash electroretinogram (F-ERG) features of ozone-treated streptozotocin (STZ)-induced diabetic rats. METHODS Seventy male Sprague Dawley rats were grouped as follows: blank group (GB, n=10), model control group (GM, n=18), ozone group (GO3, n=19), and oxygen group (GO2, n=18). The model was induced by single intraperitoneal injection of STZ. Ozone or oxygen enteroclysm was given twice per week for 4wk. F-ERG and histopathological examinations were performed one month after treatment. RESULTS Under dark adaption, as compared to GB, the other groups each had differential decreases in the a-wave amplitudes (P<0.05); the latencies were delayed in GM, GO2, and GO3 rats (P<0.05). Similar results were observed under light adaption, with the exception that the a-wave of the amplitudes (F=0.28, P>0.05). There were significant differences in the apoptosis index among the groups (P<0.05). Under ozone treatment, apoptosis was decreased in GO3 as compared to GM and GO2. CONCLUSION Ozone administration alleviates nerve damage and reduces pathology and apoptosis in the retinas of diabetic rats. PMID:27366680

  12. Integration of tissue metabolomics, transcriptomics and immunohistochemistry reveals ERG- and gleason score-specific metabolomic alterations in prostate cancer.

    PubMed

    Meller, Sebastian; Meyer, Hellmuth-A; Bethan, Bianca; Dietrich, Dimo; Maldonado, Sandra González; Lein, Michael; Montani, Matteo; Reszka, Regina; Schatz, Philipp; Peter, Erik; Stephan, Carsten; Jung, Klaus; Kamlage, Beate; Kristiansen, Glen

    2016-01-12

    Integrated analysis of metabolomics, transcriptomics and immunohistochemistry can contribute to a deeper understanding of biological processes altered in cancer and possibly enable improved diagnostic or prognostic tests. In this study, a set of 254 metabolites was determined by gas-chromatography/liquid chromatography-mass spectrometry in matched malignant and non-malignant prostatectomy samples of 106 prostate cancer (PCa) patients. Transcription analysis of matched samples was performed on a set of 15 PCa patients using Affymetrix U133 Plus 2.0 arrays. Expression of several proteins was immunohistochemically determined in 41 matched patient samples and the association with clinico-pathological parameters was analyzed by an integrated data analysis. These results further outline the highly deregulated metabolism of fatty acids, sphingolipids and polyamines in PCa. For the first time, the impact of the ERG translocation on the metabolome was demonstrated, highlighting an altered fatty acid oxidation in TMPRSS2-ERG translocation positive PCa specimens. Furthermore, alterations in cholesterol metabolism were found preferentially in high grade tumors, enabling the cells to create energy storage. With this integrated analysis we could not only confirm several findings from previous metabolomic studies, but also contradict others and finally expand our concepts of deregulated biological pathways in PCa.

  13. Sedimentary cycles in a Mesoproterozoic aeolian erg-margin succession: Mangabeira Formation, Espinhaço Supergroup, Brazil

    NASA Astrophysics Data System (ADS)

    Bállico, M. B.; Scherer, C. M. S.; Mountney, N. P.; Souza, E. G.; Reis, A. D.; Raja Gabaglia, G. P.; Magalhães, A. J. C.

    2017-03-01

    Aeolian systems were abundant and widespread in the early Proterozoic, post-2.2 Ga. However, the majority of aeolian successions of such great age are intensely deformed and are preserved only in a fragmentary state meaning that, hitherto, few attempts have been made to apply a sequence stratigraphic approach to determine mechanisms of aeolian construction, accumulation and preservation in such systems. The Mangabeira Formation is a well preserved Mesoproterozoic erg successions covering part of the São Francisco Craton, northeastern Brazil. The lower unit of the Mangabeira Formation ( 500 m thick) comprises aeolian deposits of dune, interdune, and sand-sheet origin, as well as some of waterlain origin. These deposits are organized into vertically stacked depositional cycles, each 6 to 20 m thick, and characterized by aeolian sandsheet and waterlain deposits succeeded by aeolian dune and interdune deposits indicative of a drying-upward trend. Aeolian cross-strata exhibit a mean dip direction to the north. Each of these cycles likely arose in response to climatic oscillation from relatively humid to arid conditions, possibly related to orbital forcing. The lower unit of the Mangabeira Formation comprises up to 14 erg sequences. The accumulation and preservation of each was determined by the relative rate of water-table rise and the availability of sand for aeolian transport, both of which changed through time, resulting in the preservation of a succession of repeated drying-upward cycles.

  14. Brain Gym To Increase Academic Performance Of Children Aged 10-12 Years Old ( Experimental Study in Tembalang Elementary School and Pedalangan Elementary School Semarang)

    NASA Astrophysics Data System (ADS)

    Marpaung, M. G.; Sareharto, T. P.; Purwanti, A.; Hermawati, D.

    2017-02-01

    Academic performance becomes an important determinant of individual quality. it is determined by the function of affective, cognitive, psychomotor, and intelligence. Brain gym can improve learning processes and integrate all areas that related to the learning process. To prove the effect of brain gym towards academic performance of children aged 10-12 years. This study was a quasy experiment study with one group pre and post test design. Samples (n=18 male=7 and female=11) were taken from five and six grader and conducted in Tembalang and Pedalangan Elementary School, Semarang. Pretest were administered, followed by brain gym, and post test administered in the end of study. The measurement of Intelligence Quotient pre and post test using Culture Fair Intelligence Test Scale 2. Among the 18 subjects (male=7 and female=11) the average of academic performance and IQ score after brain gym showed improvement. The Improvement of IQ score with Culture Fair Test Scale 2 was analyzed by Dependent T test showed significant results (p=0,000). The improvement of Bahasa score was analyzed by Wilcoxon test showed significant results (p=0,001), an unsignificant result were shown in Mathematics p=0,079 and natural sciences p=0,306. Brain gym can increase academic performance of children aged 10-12 years old.

  15. Autoantibodies against oncogenic ERG protein in prostate cancer: potential use in diagnosis and prognosis in a panel with C-MYC, AMACR and HERV-K Gag

    PubMed Central

    Rastogi, Anshu; Ali, Amina; Tan, Shyh-Han; Banerjee, Sreedatta; Chen, Yongmei; Cullen, Jennifer; Xavier, Charles P.; Mohamed, Ahmed A.; Ravindranath, Lakshmi; Srivastav, Jigisha; Young, Denise; Sesterhenn, Isabell A.; Kagan, Jacob; Srivastava, Sudhir; McLeod, David G.; Rosner, Inger L.; Petrovics, Gyorgy; Dobi, Albert; Srivastava, Shiv; Srinivasan, Alagarsamy

    2016-01-01

    Overdiagnosis and overtreatment of prostate cancer (CaP) is attributable to widespread reliance on PSA screening in the US. This has prompted us and others to search for improved biomarkers for CaP, to facilitate early detection and disease stratification. In this regard, autoantibodies (AAbs) against tumor antigens could serve as potential candidates for diagnosis and prognosis of CaP. Towards this, our goals were: i) To investigate whether AAbs against ERG oncoprotein (overexpressed in 25-50% of Caucasian American and African American CaP) are present in the sera of CaP patients; ii) To evaluate an AAb panel to enhance CaP detection. The results using an enzyme-linked immunosorbent assay (ELISA) showed that anti-ERG AAbs are present in a significantly higher proportion in the sera of CaP patients compared to healthy controls (p = 0.0001). Furthermore, a panel of AAbs against ERG, AMACR and human endogenous retrovirus-K Gag successfully differentiated CaP patient sera from healthy controls (AUC = 0.791). These results demonstrate for the first time that anti-ERG AAbs are present in the sera of CaP patients. In addition, the data also suggest that AAbs against ERG together with AMACR and HERV-K Gag may be a useful panel of biomarkers for diagnosis and prognosis of CaP. PMID:28191285

  16. Structure of the Cyclic Nucleotide-Binding Homology Domain of the hERG Channel and Its Insight into Type 2 Long QT Syndrome

    PubMed Central

    Li, Yan; Ng, Hui Qi; Li, Qingxin; Kang, CongBao

    2016-01-01

    The human ether-à-go-go related gene (hERG) channel is crucial for the cardiac action potential by contributing to the fast delayed-rectifier potassium current. Mutations in the hERG channel result in type 2 long QT syndrome (LQT2). The hERG channel contains a cyclic nucleotide-binding homology domain (CNBHD) and this domain is required for the channel gating though molecular interactions with the eag domain. Here we present solution structure of the CNBHD of the hERG channel. The structural study reveals that the CNBHD adopts a similar fold to other KCNH channels. It is self-liganded and it contains a short β-strand that blocks the nucleotide-binding pocket in the β-roll. Folding of LQT2-related mutations in this domain was shown to be affected by point mutation. Mutations in this domain can cause protein aggregation in E. coli cells or induce conformational changes. One mutant-R752W showed obvious chemical shift perturbation compared with the wild-type, but it still binds to the eag domain. The helix region from the N-terminal cap domain of the hERG channel showed unspecific interactions with the CNBHD. PMID:27025590

  17. Comparative study of the structure and interaction of the pore helices of the hERG and Kv1.5 potassium channels in model membranes.

    PubMed

    Beaugrand, Maïwenn; Arnold, Alexandre A; Bourgault, Steve; Williamson, Philip T F; Marcotte, Isabelle

    2017-03-17

    The hERG channel is a voltage-gated potassium channel found in cardiomyocytes that contributes to the repolarization of the cell membrane following the cardiac action potential, an important step in the regulation of the cardiac cycle. The lipids surrounding K(+) channels have been shown to play a key role in their regulation, with anionic lipids shown to alter gating properties. In this study, we investigate how anionic lipids interact with the pore helix of hERG and compare the results with those from Kv1.5, which possesses a pore helix more typical of K(+) channels. Circular dichroism studies of the pore helix secondary structure reveal that the presence of the anionic lipid DMPS within the bilayer results in a slight unfolding of the pore helices from both hERG and Kv1.5, albeit to a lesser extent for Kv1.5. In the presence of anionic lipids, the two pore helices exhibit significantly different interactions with the lipid bilayer. We demonstrate that the pore helix from hERG causes significant perturbation to the order in lipid bicelles, which contrasts with only small changes observed for Kv1.5. These observations suggest that the atypical sequence of the pore helix of hERG may play a key role in determining how anionic lipids influence its gating.

  18. U.S. Regional Agricultural Production in 2030 and 2095: Response to CO2 Fertilization and Hadley Climate Model (HadCM2) Projections of Greenhouse-Forced Climatic Change

    SciTech Connect

    NJ Rosenberg; RC Izaurralde: RA Brown

    1999-11-19

    Research activities underway to evaluate potential consequences of climate change and variability on the agriculture, water resources, and other U.S. sectors were mandated by the Global Change Research Act of 1990. These activities are being carried out in a public-private partnership under the guidance of the U.S. Global Change Research Program. Researchers at Pacific Northwest National Laboratory (PNNL) have been using integrated assessment methodologies to appraise the possible impacts of global warming and climatic variability on the behavior of managed and natural systems. This interim PNNL report contributes to the U.S. National Assessment process with an analysis of the modeled impacts of climatic changes projected by the Hadley/UKMO (HadCM2) general circulation model on agricultural productivity and selected environmental variables. The construction of climatic data for the simulation runs followed general guidelines established by the U.S. National Assessment Synthesis Team. The baseline climate data were obtained from national records for the period 1961 - 1990. The scenario runs for two future periods (2025 - 2030 and 2090 - 2099) were extracted from results of a HadCM2 run distributed at a half-degree spatial resolution. The Erosion Productivity Impact Calculator (EPIC) was used to simulate the behavior of 204 "representative farms" (i.e., soil-climate-management combinations) under baseline climate, the two future periods and their combinations with two levels of atmospheric C02 concentrations (365 and 560 ppm). Analysis of simulation results identified areas in Texas, New Mexico, Colorado, Utah, Arizona, and California that would experience large temperature increases by 2030. Slight cooling is expected by 2030 in parts of Alabama, Florida, Maine, Montana, Idaho, and Utah. Larger areas will experience increased warming by 2095. Uniform precipitation increases are expected by 2030 in the north eastern quarter of the country. These uniform precipitation

  19. Low-voltage back-gated atmospheric pressure chemical vapor deposition based graphene-striped channel transistor with high-κ dielectric showing room-temperature mobility > 11,000 cm(2)/V·s.

    PubMed

    Smith, Casey; Qaisi, Ramy; Liu, Zhihong; Yu, Qingkai; Hussain, Muhammad Mustafa

    2013-07-23

    Utilization of graphene may help realize innovative low-power replacements for III-V materials based high electron mobility transistors while extending operational frequencies closer to the THz regime for superior wireless communications, imaging, and other novel applications. Device architectures explored to date suffer a fundamental performance roadblock due to lack of compatible deposition techniques for nanometer-scale dielectrics required to efficiently modulate graphene transconductance (gm) while maintaining low gate capacitance-voltage product (CgsVgs). Here we show integration of a scaled (10 nm) high-κ gate dielectric aluminum oxide (Al2O3) with an atmospheric pressure chemical vapor deposition (APCVD)-derived graphene channel composed of multiple 0.25 μm stripes to repeatedly realize room-temperature mobility of 11,000 cm(2)/V·s or higher. This high performance is attributed to the APCVD graphene growth quality, excellent interfacial properties of the gate dielectric, conductivity enhancement in the graphene stripes due to low tox/Wgraphene ratio, and scaled high-κ dielectric gate modulation of carrier density allowing full actuation of the device with only ±1 V applied bias. The superior drive current and conductance at Vdd = 1 V compared to other top-gated devices requiring undesirable seed (such as aluminum and poly vinyl alcohol)-assisted dielectric deposition, bottom gate devices requiring excessive gate voltage for actuation, or monolithic (nonstriped) channels suggest that this facile transistor structure provides critical insight toward future device design and process integration to maximize CVD-based graphene transistor performance.

  20. Particle-size fractionation of aeolian sand along a climatic and geomorphic gradient of the Sinai-Negev erg

    NASA Astrophysics Data System (ADS)

    Roskin, Joel; Katra, Itzhak; Blumberg, Dan G.

    2015-04-01

    This study examines changes in the aeolian sand fractions along the west-east aeolian transport path of the northern Sinai Peninsula - northwestern (NW) Negev erg of Egypt and Israel. This erg originates from the Nile Delta and is composed of currently active linear (seif) dunes in northern Sinai (its western part), and currently stabilized vegetated linear dunes (VLDs) in the NW Negev dunefield (its eastern part). Sand samples from the Nile Delta, northern Sinai and NW Negev were analyzed for particle-size distribution and sand grain morphology in accordance to their Eastern Mediterranean INQUA Dunes Atlas luminescence and radiocarbon chronologies. Linear seif dunes differ from VLDs in their vegetation cover, linearity, and dynamics. Although both are continuous landforms with similar orientations and sand-grain roundness values, the linear dunes of Sinai are coarser-grained than the Negev VLDs. The VLDs have a significantly higher proportion of very fine sand (125-50 μm) content and a varying but lower sand fining ratio defined as the ratio of fine sand percentage to very fine sand percentage. Very fine sands are suggested to have been winnowed by saltation and low suspension from source deposits and sand sheets. Detailed semi-quantitative examinations of sand grains by a SEM of a Negev VLD shows that most grains do not exhibit features that can be attributed to aeolian abrasion by sand grain-grain collisions. From these observations we infer that fractionation of sand was a major process leading to downwind fining along the studied aeolian transport path. We suggest that the very fine sand fraction of Nile Delta and Sinai sands has been transported downwind since the late middle Pleistocene. In the late Pleistocene, sand reached the NW Negev in the form of VLDs due to last-glacial period windiness of intensities unprecedented today and probably larger sediment supply. Generally current and inferred past decreasing wind velocities and increasing precipitation

  1. Stabilization of the Activated hERG Channel Voltage Sensor by Depolarization Involves the S4-S5 Linker.

    PubMed

    Thouta, Samrat; Hull, Christina M; Shi, Yu Patrick; Sergeev, Valentine; Young, James; Cheng, Yen M; Claydon, Thomas W

    2017-01-24

    Slow deactivation of hERG channels is critical for preventing cardiac arrhythmia yet the mechanistic basis for the slow gating transition is unclear. Here, we characterized the temporal sequence of events leading to voltage sensor stabilization upon membrane depolarization. Progressive increase in step depolarization duration slowed voltage-sensor return in a biphasic manner (τfast = 34 ms, τslow = 2.5 s). The faster phase of voltage-sensor return slowing correlated with the kinetics of pore opening. The slower component occurred over durations that exceeded channel activation and was consistent with voltage sensor relaxation. The S4-S5 linker mutation, G546L, impeded the faster phase of voltage sensor stabilization without attenuating the slower phase, suggesting that the S4-S5 linker is important for communications between the pore gate and the voltage sensor during deactivation. These data also demonstrate that the mechanisms of pore gate-opening-induced and relaxation-induced voltage-sensor stabilization are separable. Deletion of the distal N-terminus (Δ2-135) accelerated off-gating current, but did not influence the relative contribution of either mechanism of stabilization of the voltage sensor. Lastly, we characterized mode-shift behavior in hERG channels, which results from stabilization of activated channel states. The apparent mode-shift depended greatly on recording conditions. By measuring slow activation and deactivation at steady state we found the "true" mode-shift to be ∼15 mV. Interestingly, the "true" mode-shift of gating currents was ∼40 mV, much greater than that of the pore gate. This demonstrates that voltage sensor return is less energetically favorable upon repolarization than pore gate closure. We interpret this to indicate that stabilization of the activated voltage sensor limits the return of hERG channels to rest. The data suggest that this stabilization occurs as a result of reconfiguration of the pore gate upon opening by

  2. A large number of novel Ergtoxin-like genes and ERG K+-channels blocking peptides from scorpions of the genus Centruroides.

    PubMed

    Corona, Miguel; Gurrola, Georgina B; Merino, Enrique; Cassulini, Rita Restano; Valdez-Cruz, Norma A; García, Blanca; Ramírez-Domínguez, Martha E; Coronas, Fredy I V; Zamudio, Fernando Z; Wanke, Enzo; Possani, Lourival D

    2002-12-04

    Twenty-three novel sequences similar to Ergtoxin (ErgTx) were obtained by direct sequencing of peptides or deduced from gene cloned using cDNAs of venomous glands of Centruroides (C.) elegans, C. exilicauda, C. gracilis, C. limpidus limpidus, C. noxius and C. sculpturatus. These peptides have from 42 to 47 amino acid residues cross-linked by four disulfide bridges. They share sequence similarities (60-98% compared with ErgTx1) and were shown to block ERG K(+)-channels of F-11 clone (N18TG-2xrat DRG) cultured cells. An unrooted phylogenetic tree analysis of these peptides showed that they conform at least five different subfamilies, of which three are novel subfamilies.

  3. Utility of PTEN and ERG immunostaining for distinguishing high-grade PIN from intraductal carcinoma of the prostate on needle biopsy.

    PubMed

    Morais, Carlos L; Han, Jeong S; Gordetsky, Jennifer; Nagar, Michael S; Anderson, Ann E; Lee, Stephen; Hicks, Jessica L; Zhou, Ming; Magi-Galluzzi, Cristina; Shah, Rajal B; Epstein, Jonathan I; De Marzo, Angelo M; Lotan, Tamara L

    2015-02-01

    Intraductal carcinoma of the prostate and high-grade prostatic intraepithelial neoplasia (PIN) have markedly different implications for patient care but can be difficult to distinguish in needle biopsies. In radical prostatectomies, we demonstrated that PTEN and ERG immunostaining may be helpful to resolve this differential diagnosis. Here, we tested whether these markers are diagnostically useful in the needle biopsy setting. Separate or combined immunostains were applied to biopsies containing morphologically identified intraductal carcinoma, PIN, or borderline intraductal proliferations more concerning than PIN but falling short of morphologic criteria for intraductal carcinoma. Intraductal carcinoma occurring with concurrent invasive tumor showed the highest rate of PTEN loss, with 76% (38/50) lacking PTEN and 58% (29/50) expressing ERG. Of biopsies containing isolated intraductal carcinoma, 61% (20/33) showed PTEN loss and 30% (10/33) expressed ERG. Of the borderline intraductal proliferations, 52% (11/21) showed PTEN loss and 27% (4/15) expressed ERG. Of the borderline cases with PTEN loss, 64% (7/11) had carcinoma in a subsequent needle biopsy specimen, compared with 50% (5/10) of PTEN-intact cases. In contrast, none of the PIN cases showed PTEN loss or ERG expression (0/19). On needle biopsy, PTEN loss is common in morphologically identified intraductal carcinoma yet is very rare in high-grade PIN. Borderline intraductal proliferations, especially those with PTEN loss, have a high rate of carcinoma on resampling. If confirmed in larger prospective studies, these results suggest that PTEN and ERG immunostaining may provide a useful ancillary assay to distinguish intraductal carcinoma from high-grade PIN in this setting.

  4. hERG1 positivity and Glut-1 negativity identifies high-risk TNM stage I and II colorectal cancer patients, regardless of adjuvant chemotherapy

    PubMed Central

    Muratori, Leonardo; Petroni, Giulia; Antonuzzo, Lorenzo; Boni, Luca; Iorio, Jessica; Lastraioli, Elena; Bartoli, Gianluca; Messerini, Luca; Di Costanzo, Francesco; Arcangeli, Annarosa

    2016-01-01

    Background The identification of early-stage colorectal cancer (CRC) with high risk of progression is one major clinical challenge, mainly due to lack of validated biomarkers. The aims of the present study were to analyze the prognostic impact of three molecular markers belonging to the ion channels and transporters family: the ether-à-go-go-related gene 1 (hERG1) and the calcium-activated KCa3.1 potassium channels, as well as the glucose transporter 1 (Glut-1); and to define the impact of adjuvant chemotherapy in conjunction with the abovementioned biomarkers, in a cohort of radically resected stage I–III CRC patients. Patients and methods The expressions of hERG1, KCa3.1, and Glut-1 were tested by immunohistochemistry on 162 surgical samples of nonmetastatic, stage I–III CRC patients. The median follow-up was 32 months. The association between biological markers, clinicopathological features, and survival outcomes was investigated by evaluating both disease-free survival and overall survival. Results Although no prognostic valence emerged for KCa3.1, evidence of a negative impact of hERG1 expression on survival outcomes was provided. On the contrary, Glut-1 expression had a positive impact. According to the results of the multivariate analysis, patients were stratified in four risk groups, based on TNM stage and hERG1/Glut-1 expression. After adjusting for adjuvant therapy, stage I and II, Glut-1-negative, and hERG1-positive patients showed the worst survival experience. Conclusion This study strongly indicates that the combination of hERG1 positivity and Glut-1 negativity behaves as a prognostic biomarker in radically resected CRC patients. This combination identifies a group of stage I and II CRC patients with a bad prognosis, even worse than that of stage III patients, regardless of adjuvant therapy accomplishment. PMID:27789963

  5. Evidence for a High-Pressure Phase Transition of ε-2,4,6,8,10,12-hexanitrohexaazaisowurtzitane (CL-20) Using Vibrational Spectroscopy

    SciTech Connect

    Ciezak, J.; Jenkins, T; Liu, Z

    2007-01-01

    The high-pressure response of {epsilon}-2,4,6,8,10,12-hexanitrohexaazaisowurtizane (CL-20) has been examined to 27 GPa in diamond anvil cells using vibrational spectroscopy. The results reveal evidence of an {epsilon}{yields}{Upsilon} pressure-induced phase transition between 4.1 and 6.4 GPa and suggest the existence of a {Upsilon}{yields}{zeta} transition near 18.7 GPa. Several Raman and infrared frequencies were found to decrease in intensity as the phase boundaries are approached. An anomalous intensity increase was noted in the C-N-C infrared mode that is believed to result from an increase in the Raman cross-section due to a stronger interlayer coupling under pressure.

  6. Thermal Nondestructive Evaluation Report: Inspection of the Refurbished Manipulator Arm System in the Manipulator Development Facility at Johnson Space Center 10-12 January 2001

    NASA Technical Reports Server (NTRS)

    Cramer, K. Elliott

    2002-01-01

    On 4 December 2002, a failure of the Refurbished Manipulator Arm System (RMAS) occurred in the Manipulator Development Facility (MDF) at Johnson Space Center. When the Test Director commanded a should pitch maneuver to lift the arm from its payload bay pedestal, the yaw controls failed. This, coupled with a gravitational forces (due to the angle of the shoulder joint with respect to vertical), resulted in uncontrolled arm motion. The shoulder yaw joint moved approximately 20 degrees, causing the extended arm to strike and severely damage the port side MDF catwalk handrails. The arm motion stopped after impact with the handrails. On 10-12 January 2001, inspections were performed on the port face of the lower and upper arms of the RMAS using a infrared thermography developed at Langley Research Center. This paper presents the results of those nondestructive inspections and provides a complete description of the anomalies found and their locations.

  7. Reading comprehension among typically developing Swedish-speaking 10-12-year-olds: examining subgroups differentiated in terms of language and decoding skills.

    PubMed

    Asberg, Jakob; Carlsson, Marika; Oderstam, Ann-Marie; Miniscalco, Carmela

    2010-12-01

    Based on data from 156 typically developing 10-12-year-olds from Sweden, reading comprehension skills were studied in three subgroups: those classified with specific poor word decoding skills (n = 10), those with specific poor language comprehension (n = 12), and those with mixed difficulties in word decoding and language comprehension (n = 11). The mixed poor group achieved significantly lower scores than both specific groups in reading comprehension, and was the only group displaying poor reading comprehension test results relative to the performance of the full sample. Results are indicative of the necessity of a combined effect of poor word decoding and language in reading comprehension difficulties for this group. Implications and limitations are discussed.

  8. Stereospecificity of an enzymatic monoene 1,4-dehydrogenation reaction: conversion of (Z)-11-tetradecenoic acid into (E,E)-10,12-tetradecadienoic acid.

    PubMed

    Rodríguez, Sergio; Clapés, Pere; Camps, Francisco; Fabriàs, Gemma

    2002-04-05

    In this article, we report the first stereochemical study of an enzymatic 1,4-dehydrogenation reaction, namely, the transformation of (Z)-11-tetradecenoic acid into (E,E)-10,12-tetradecadienoic acid, involved in the sex pheromone biosynthesis of the moth Spodoptera littoralis. The investigation was carried out using the labeled substrates (R)-[10-(2)H]- and (S)-[10-(2)H]-tridecanoic acids ((R)-2 and (S)-2, respectively) and (R)-[2,2,3,3,13-(2)H(5)]- and (S)-[2,2,3,3,13-(2)H(5)]-tetradecanoic acids ((R)-1 and (S)-1, respectively). Probes (R)-2 and (S)-2 were prepared as described in a previous article.(1) The synthesis of the pentadeuterated chiral substrates (R)-1 and (S)-1 was accomplished by kinetic resolution of the racemic 12-tridecyn-2-ol (6) with immobilized porcine pancreatic lipase. The enantiomerically pure alcohols (R)-6 and (S)-6 were transformed into the final acids (S)-1 and (R)-1, respectively, by a sequence of well-established reactions. The analyses of methanolyzed lipidic extracts from glands incubated separatedly with each individual probe showed that in the transformation of (Z)-11-tetradecenoic acid into (E,E)-10,12-tetradecadienoic acid, both pro-(R) hydrogen atoms at C-10 and C-13 are removed from the substrate. This is the first example reported of a desaturase with pro-(R)/pro-(R) stereospecificities that gives rise to (E)-double bonds. A mechanistic explanation for the stereochemical outcome of this reaction is advanced.

  9. [PhSiO1.5]8,10,12 as nanoreactors for non-enzymatic introduction of ortho, meta or para-hydroxyl groups to aromatic molecules.

    PubMed

    Bahrami, Mozhgan; Zhang, Xingwen; Ehsani, Morteza; Jahani, Yousef; Laine, Richard M

    2017-03-20

    Traditional electrophilic bromination follows long established "rules": electron-withdrawing substituents cause bromination selective for meta positions, whereas electron-donating substituents favor ortho and para bromination. In contrast, in the [PhSiO1.5]8,10,12 silsesquioxanes, the cages act as bulky, electron withdrawing groups equivalent to CF3; yet bromination under mild conditions, without a catalyst, greatly favors ortho substitution. Surprisingly, ICl iodination without a catalyst favors (>90%) para substitution [p-IC6H4SiO1.5]8,10,12. Finally, nitration and Friedel-Crafts acylation and sulfonylation are highly meta selective, >80%. In principle, the two halogenation formats coupled with the traditional electrophilic reactions provide selective functionalization at each position on the aromatic ring. Furthermore, halogenation serves as a starting point for the synthesis of two structural isomers of practical utility, i.e. in drug prospecting. The o-bromo and p-iodo compounds are easily modified by catalytic cross-coupling to append diverse functional groups. Thereafter, F(-)/H2O2 treatment cleaves the Si-C bonds replacing Si with OH. This represents a rare opportunity to introduce hydroxyl groups to aromatic rings, a process not easily accomplished using traditional organic synthesis methods. The as-produced phenol provides additional opportunities for modification. Each cage can be considered a nanoreactor generating 8-12 product molecules. Examples given include syntheses of 4,2'-R,OH-stilbenes and 4,4'-R,OH-stilbenes (R = Me, CN). Unoptimized cleavage of the Br/I derivatives yields 55-85% phenol. Unoptimized cleavage of the stilbene derivatives yields 35-40% (3-5 equivalents of phenol) in the preliminary studies presented here. In contrast, meta R-phenol yields are 80% (7-10 mol per cage).

  10. EPS Biomarkers Improve Stratification of NCCN Active Surveillance Candidates: Performance of Secretion Capacity and TMPRSS2:ERG Models

    PubMed Central

    Whelan, Christopher; Kawachi, Mark; Smith, David D.; Linehan, Jennifer; Babilonia, Gail; Mejia, Rosa; Wilson, Timothy; Smith, Steven

    2014-01-01

    Purpose Active surveillance (AS) is a viable patient option for prostate cancer where a clinical determination of low-risk and presumably organ-confined disease can be made. In an effort to standardize risk stratification schemes, the National Comprehensive Cancer Network (NCCN) has provided guidelines for the AS option. Our purpose was to determine the effectiveness of expressed prostatic secretion (EPS) biomarkers in detecting occult risk factors in NCCN AS candidates. Materials and Methods EPS specimens were obtained prior to Robot-Assisted Radical Prostatectomy (RARP). Secretion capacity biomarkers: total RNA and EPS specimen volume were measured by standard techniques. RNA expression biomarkers: TXNRD1-mRNA, PSA-mRNA, TMPRSS2:ERG fusion mRNA and PCA3-mRNAs were measured by quantitative reverse-transcription PCR. Results Of the 528 patients from whom EPS was collected, 216 were eligible for AS under NCCN guidelines. Variable Selection in logistic regression identified two models, one featuring Type III and Type VI TMPRSS2:ERG variants, and one featuring two secretion capacity biomarkers. Of the two high performing models, the secretion capacity model was the most effective in detecting patients within this group that were upstaged or both upstaged and upgraded. It reduced the risk of upstaging in patients with a negative test by nearly 8 fold, and reduced the risk of being both upstaged and upgraded by about 5 fold, while doubling the prevalence upstaging in the positive test group. Conclusions Non-invasive EPS testing may improve patient acceptance of AS by dramatically reducing the presence of occult risk factors among patients eligible for AS under NCCN guidelines. PMID:23669563

  11. Arabidopsis ERG28 Tethers the Sterol C4-Demethylation Complex to Prevent Accumulation of a Biosynthetic Intermediate That Interferes with Polar Auxin Transport[C][W

    PubMed Central

    Mialoundama, Alexis Samba; Jadid, Nurul; Brunel, Julien; Di Pascoli, Thomas; Heintz, Dimitri; Erhardt, Mathieu; Mutterer, Jérôme; Bergdoll, Marc; Ayoub, Daniel; Van Dorsselaer, Alain; Rahier, Alain; Nkeng, Paul; Geoffroy, Philippe; Miesch, Michel; Camara, Bilal; Bouvier, Florence

    2013-01-01

    Sterols are vital for cellular functions and eukaryotic development because of their essential role as membrane constituents. Sterol biosynthetic intermediates (SBIs) represent a potential reservoir of signaling molecules in mammals and fungi, but little is known about their functions in plants. SBIs are derived from the sterol C4-demethylation enzyme complex that is tethered to the membrane by Ergosterol biosynthetic protein28 (ERG28). Here, using nonlethal loss-of-function strategies focused on Arabidopsis thaliana ERG28, we found that the previously undetected SBI 4-carboxy-4-methyl-24-methylenecycloartanol (CMMC) inhibits polar auxin transport (PAT), a key mechanism by which the phytohormone auxin regulates several aspects of plant growth, including development and responses to environmental factors. The induced accumulation of CMMC in Arabidopsis erg28 plants was associated with diagnostic hallmarks of altered PAT, including the differentiation of pin-like inflorescence, loss of apical dominance, leaf fusion, and reduced root growth. PAT inhibition by CMMC occurs in a brassinosteroid-independent manner. The data presented show that ERG28 is required for PAT in plants. Furthermore, it is accumulation of an atypical SBI that may act to negatively regulate PAT in plants. Hence, the sterol pathway offers further prospects for mining new target molecules that could regulate plant development. PMID:24326590

  12. Functional Characterization of the Protein Product of the Prostate Carcinoma Gene Fusion TMPRSS2:ERG Using the Proteomic and Microarray Analyses

    DTIC Science & Technology

    2009-07-01

    Guava ™ FACS analyzer with the ViaCount™ kit (Millipore). As shown in Fig. 7, VCaP cells infected with ERG or ERGa once again failed to proliferate...and the cell numbers in each series were determined using the Guava ViaCount™ kit. Average values of the triplicate samples are shown. Milestone

  13. A convenient approach to 10-12 g/ g ICP-MS limits for Th and U in aurubis electrolytic NA-ESN brand copper

    NASA Astrophysics Data System (ADS)

    Leonard, Douglas S.

    2014-06-01

    Inductively-coupled-plasma mass spectroscopy is a powerful technique for measuring trace levels of radioactive contaminants, specifically Th and U, in materials for use in construction of low-background rare-event detectors such as double beta decay and dark matter detectors. I describe here a technique for measuring Th and U contamination in copper by using direct acid digestion and dilution without further chemical processing, achieving results comparable to those achieved in previous work [1, 2] which utilized more complex chemical pre-concentration techniques. A convenient research-oriented analysis environment is described as well. Results are presented for measurements of three samples from the production line of electrolytically-purified, LME (London Metal Exchange) grade A, NA-ESN Aurubis copper. Purified samples showed levels consistent with zero contamination for both elements, while weak, but inconclusive, indications of contamination were present for the unpurified anode copper. The best limits achieved are near 1•10-12 g/ g (95% CL) for both Th and U measured for copper from the cathode of the purification process.

  14. Characterization of 10,12-pentacosadiynoic acid Langmuir–Blodgett monolayers and their use in metal–insulator–metal tunnel devices

    PubMed Central

    Sharma, Saumya; Khawaja, Mohamad; Ram, Manoj K; Goswami, D Yogi

    2014-01-01

    Summary The characterization of Langmuir–Blodgett thin films of 10,12-pentacosadiynoic acid (PDA) and their use in metal–insulator–metal (MIM) devices were studied. The Langmuir monolayer behavior of the PDA film was studied at the air/water interface using surface tension–area isotherms of polymeric and monomeric PDA. Langmuir–Blodgett (LB, vertical deposition) and Langmuir–Schaefer (LS, horizontal deposition) techniques were used to deposit the PDA film on various substrates (glass, quartz, silicon, and nickel-coated film on glass). The electrochemical, electrical and optical properties of the LB and LS PDA films were studied using cyclic voltammetry, current–voltage characteristics (I–V), and UV–vis and FTIR spectroscopies. Atomic force microscopy measurements were performed in order to analyze the surface morphology and roughness of the films. A MIM tunnel diode was fabricated using a PDA monolayer assembly as the insulating barrier, which was sandwiched between two nickel layers. The precise control of the thickness of the insulating monolayers proved critical for electron tunneling to take place in the MIM structure. The current–voltage characteristics of the MIM diode revealed tunneling behavior in the fabricated Ni–PDA LB film–Ni structures. PMID:25551052

  15. Frequency-agile, rapid scanning spectroscopy: absorption sensitivity of 2 × 10-12 cm-1 Hz-1/2 with a tunable diode laser

    NASA Astrophysics Data System (ADS)

    Long, D. A.; Truong, G.-W.; van Zee, R. D.; Plusquellic, D. F.; Hodges, J. T.

    2014-03-01

    We present ultrasensitive measurements of molecular absorption using frequency-agile rapid scanning, cavity ring-down spectroscopy with an external-cavity diode laser. A microwave source that drives an electro-optic phase modulator with a bandwidth of 20 GHz generates pairs of sidebands on the probe laser. The optical cavity provides for high sensitivity and filters the carrier and all but a single, selected sideband. Absorption spectra were acquired by stepping the tunable sideband from mode-to-mode of the ring-down cavity at a rate that was limited only by the cavity decay time. This approach allows for scanning rates of 8 kHz per cavity resonance, a minimum detectable absorption coefficient of 1.7 × 10-11 cm-1 after only 20 ms of averaging, and a noise-equivalent absorption coefficient of 1.7 × 10-12 cm-1 Hz-1/2. By comparison with cavity-enhanced laser absorption spectrometers reported in the literature, the present system is, to the best of our knowledge, among the most sensitive and has by far the highest spectrum scanning rate.

  16. Geochemical Characterization of Bitumen Carbonate from Grosmont Formation, Alberta: Well 10-12-93-24W4 and 11-33-94-22W4

    NASA Astrophysics Data System (ADS)

    Seol, J.; Kil, Y. W.; Kim, J. H.; Choi, J.

    2014-12-01

    Carbonate bitumen, one of the unconventional oils, has been attracting attention as an energy resource, which can substitute petroleum resources. Various researchers have been investigating carbonate rocks to exploit bitumen reservoirs. Grosmont Formation, located in northern Alberta, is one of the largest carbonate reservoirs in the world. We conducted inorganic geochemical analysis about two-carbonate rock cores of Grosmont Formation (well 10-12-93-24W4, 11-33-94-22W4), obtained from Core Research Center (CRC) in Canada to investigate the characteristics and origin of dolomite in the Grosmont Formation. Grosmont Formation consists of four carbonate units, UG3, UG2, UG1 and LG, with three shale beds, SB3, SB2 and SB1. Major and trace elements, and isotope compositions of dolomites from UG and LG units indicate that UG dolomites were formed within a near surface environment under evaporitic condition, whereas LG dolomites were formed in a diagenetic environment. UG and LG dolomites might be formed at temperature ranging from 43.9 °C to 56.7 °C with Devonian seawater.

  17. Complexes of DNA Bases and Watson-Crick Base Pairs Interaction with Neutral Silver Agn (n = 8, 10, 12) Clusters: A DFT and TDDFT Study.

    PubMed

    Srivastava, Ruby

    2017-03-21

    We study the binding of the neutral Agn (n = 8, 10, 12) to the DNA base- adenine (A), guanine (G) and Watson-Crick -adenine-thymine (AT), guanine-cytosine (GC) pairs. Geometries of complexes were optimized at the DFT level using the hybrid B3LYP functional. LANL2DZ effective core potential (ECP) was used for silver and 6-31+G(**) was used for all other atoms. NBO charges were analyzed using the Natural population analysis. The absorption properties of Agn-A,G/WC complexes were also studied using time-dependent density functional theory (TDDFT). The absorption spectra for these complexes show wavelength in the visible region. It was revealed that silver clusters interact more strongly with WC pairs than with isolated DNA complexes. Furthermore, it was found that the electronic charge transferred from silver to isolated DNA clusters are less than the electronic charge transferred from silver to the Agn-WC complexes. The vertical ionization potential, vertical electron affinity, hardness and electrophilicity index of Agn-DNA/WC complexes have also been discussed.

  18. The Prevalence of Spine Deformities and Flat Feet among 10-12 Year Old Children Who Train Basketball--Cross-Sectional Study.

    PubMed

    Puzovic, Vladimir; Rotim, Kresimir; Jurisic, Vladimir; Samardzic, Miroslav; Zivkovic, Bojana; Savic, Andrija; Rasulic, Lukas

    2015-09-01

    The aim of this study is to estimate the prevalence of spine and feet deformities among children who are regularly involved in basketball trainings, as well as finding differences in the prevalence of those deformities between children of different gender and age. The study included a total of 64 children, of which 43 were boys and 21 were girls, ages 10-12. All subjects have been regularly participating in basketball trainings for at least one year. Postural disorder is defined as an irregularity in posture of the spine and feet, and it is assessed by visual methods from the front, side and rear side of the body. The prevalence of spinal deformities in our group was 53.13%. The boys had a significantly higher prevalence than girls, 65.1% compared to 28.57% (p=0.006). There was no significant difference in prevalence of spine deformities between children of different ages. The prevalence of feet deformities was 64.06%. There was a statistically significant difference between the sexes, where boys had a significantly greater prevalence of the feet deformities than girls, 83.7% compared to 23.81% (p=0.001). Flat feet were the most common in 10 year old children (85.71%). In conclusion, it can be said that despite regular participation in basketball training, subjects in this study have high prevalence of deformities; especially boys who stand out with the high prevalence of flat feet.

  19. Expression of DNA ligase IV is linked to poor prognosis and characterizes a subset of prostate cancers harboring TMPRSS2:ERG fusion and PTEN deletion.

    PubMed

    Grupp, Katharina; Roettger, Laura; Kluth, Martina; Hube-Magg, Claudia; Simon, Ronald; Lebok, Patrick; Minner, Sarah; Tsourlakis, Maria Christina; Koop, Christina; Graefen, Markus; Adam, Meike; Haese, Alexander; Wittmer, Corinna; Sauter, Guido; Wilczak, Waldemar; Huland, Hartwig; Schlomm, Thorsten; Steurer, Stefan; Krech, Till

    2015-09-01

    DNA ligases are essential for the maintenance of genome integrity as they are indispensable for DNA replication, recombination and repair. The present study was undertaken to gain insights into the prevalence and clinical significance of ligase IV (LIG4) expression in prostate cancer. A total of 11,152 prostate cancer specimens were analyzed by immunohistochemistry for LIG4 expression. Results were compared to follow-up data, ERG status and deletions at PTEN, 3p13, 5q21 and 6q15. LIG4 expression was predominantly localized in the nucleus of the cells with increased intensities in malignant as compared to benign prostate epithelium. In prostate cancer, LIG4 expression was found in 91% of interpretable tumors, including 12% cancers with weak, 23% with moderate and 56% with strong LIG4 positivity. Strong LIG4 expression was tightly linked to advanced Gleason score (P<0.0001) and positive nodal involvement (P=0.03). There was a remarkable accumulation of strong LIG4 expression in tumors harboring TMPRSS2:ERG fusion and PTEN deletions (P<0.0001 each). High LIG4 expression was also tightly related to early biochemical recurrence when all tumors (P<0.0001) or the subsets of ERG-negative (P=0.0004) or ERG-positive prostate cancers (P=0.006) were analyzed. Multivariate analysis including parameters that are available before surgery demonstrated independent association with biochemical recurrence for advanced Gleason grade on biopsy, high preoperative PSA level, high clinical stage (P<0.0001 each) and for LIG4 immunostaining (P=0.03). Our study identifies LIG4 as a predictor of an increased risk for early PSA recurrence in prostate cancer. Moreover, the strong association with TMPRSS2:ERG fusion and PTEN deletions suggest important interactions between these pathways in prostate cancers.

  20. Reduced CD147 expression is linked to ERG fusion-positive prostate cancers but lacks substantial impact on PSA recurrence in patients treated by radical prostatectomy.

    PubMed

    Grupp, Katharina; Höhne, Thorsten Simon; Prien, Kristina; Hube-Magg, Claudia; Tsourlakis, Maria Christina; Sirma, Hüseyin; Pham, Taher; Heinzer, Hans; Graefen, Markus; Michl, Uwe; Simon, Ronald; Wilczak, Waldemar; Izbicki, Jakob; Sauter, Guido; Minner, Sarah; Schlomm, Thorsten; Steurer, Stefan

    2013-10-01

    The extracellular matrix metalloproteinase inducer CD147 has been suggested as a prognostic marker in prostate cancer. CD147 expression was analyzed by immunohistochemistry on a tissue microarray containing 11,152 prostate cancer specimens. Results were compared to tumor phenotype, biochemical recurrence, ERG status and deletions on PTEN, 3p13, 6q15 and 5q21. CD147 expression was strong in benign prostatic glands and often reduced in prostate cancers. CD147 immunostaining was found in 71.7% of 7628 interpretable cases. CD147 staining was considered strong in 34.6%, moderate in 24.3% and weak in 12.8% of cancers while 28.3% did not show any CD147 reactivity. Reduced CD147 staining was strongly associated with both TMPRSS2-ERG-rearrangement and ERG expression (p<0.0001 each). Within the subgroups of ERG positive and negative cancers, deletions of PTEN, 3p13, 6q15 and 5q21 were unrelated to the CD147 expression status. Decreased CD147 expression was significantly linked to high preoperative PSA values, high Gleason grade, advanced tumor stage (p<0.0001 each), and positive lymph node involvement (p=0.0026) in all cancers. There was a marginal, but statistically significant, association of reduced CD147 expression with early biochemical recurrence (p=0.0296). The significant reduction of CD147 expression in ERG positive prostate cancer provides further evidence for marked biological differences between "fusion type" and "non-fusion type" prostate cancer. Despite a weak association with PSA recurrence, CD147 cannot be considered a relevant prognostic biomarker.

  1. ERG deregulation induces IGF-1R expression in prostate cancer cells and affects sensitivity to anti-IGF-1R agents.

    PubMed

    Mancarella, Caterina; Casanova-Salas, Irene; Calatrava, Ana; Ventura, Selena; Garofalo, Cecilia; Rubio-Briones, José; Magistroni, Vera; Manara, Maria Cristina; López-Guerrero, José Antonio; Scotlandi, Katia

    2015-06-30

    Identifying patients who may benefit from targeted therapy is an urgent clinical issue in prostate cancer (PCa). We investigated the molecular relationship between TMPRSS2-ERG (T2E) fusion gene and insulin-like growth factor receptor (IGF-1R) to optimize the use of IGF-1R inhibitors.IGF-1R was analyzed in cell lines and in radical prostatectomy specimens in relation to T2E status. ERG binding to IGF-1R promoter was evaluated by chromatin immunoprecipitation (ChIP). Sensitivity to anti-IGF-1R agents was evaluated alone or in combination with anti-androgen abiraterone acetate in vitro at basal levels or upon ERG modulation.IGF-1R analysis performed in PCa cells or clinical samples showed that T2E expression correlated with higher IGF-1R expression at mRNA and protein levels. Genetic modulation of ERG directly affected IGF-1R protein levels in vitro. ChIP analysis showed that ERG binds IGF-1R promoter and that promoter occupancy is higher in T2E-positive cells. IGF-1R inhibition was more effective in cell lines expressing the fusion gene and combination of IGF-1R inhibitors with abiraterone acetate produced synergistic effects in T2E-expressing cells.Here, we provide the rationale for use of T2E fusion gene to select PCa patients for anti-IGF-1R treatments. The combination of anti-IGF-1R-HAbs with an anti-androgen therapy is strongly advocated for patients expressing T2E.

  2. ERG deregulation induces IGF-1R expression in prostate cancer cells and affects sensitivity to anti-IGF-1R agents

    PubMed Central

    Mancarella, Caterina; Casanova-Salas, Irene; Calatrava, Ana; Ventura, Selena; Garofalo, Cecilia; Rubio-Briones, José; Magistroni, Vera; Manara, Maria Cristina; López-Guerrero, José Antonio; Scotlandi, Katia

    2015-01-01

    Identifying patients who may benefit from targeted therapy is an urgent clinical issue in prostate cancer (PCa). We investigated the molecular relationship between TMPRSS2-ERG (T2E) fusion gene and insulin-like growth factor receptor (IGF-1R) to optimize the use of IGF-1R inhibitors. IGF-1R was analyzed in cell lines and in radical prostatectomy specimens in relation to T2E status. ERG binding to IGF-1R promoter was evaluated by chromatin immunoprecipitation (ChIP). Sensitivity to anti-IGF-1R agents was evaluated alone or in combination with anti-androgen abiraterone acetate in vitro at basal levels or upon ERG modulation. IGF-1R analysis performed in PCa cells or clinical samples showed that T2E expression correlated with higher IGF-1R expression at mRNA and protein levels. Genetic modulation of ERG directly affected IGF-1R protein levels in vitro. ChIP analysis showed that ERG binds IGF-1R promoter and that promoter occupancy is higher in T2E-positive cells. IGF-1R inhibition was more effective in cell lines expressing the fusion gene and combination of IGF-1R inhibitors with abiraterone acetate produced synergistic effects in T2E-expressing cells. Here, we provide the rationale for use of T2E fusion gene to select PCa patients for anti-IGF-1R treatments. The combination of anti-IGF-1R-HAbs with an anti-androgen therapy is strongly advocated for patients expressing T2E. PMID:25906745

  3. A complicated story of frost and wind: Present-day gully activity within the north polar erg, Mar

    NASA Astrophysics Data System (ADS)

    Diniega, Serina; Hansen, Candice; Allen, Amanda; Grisby, Nathan; Li, Zheyu Joey

    2016-10-01

    Analyses of high-resolution observations have shown that the dunes within the martian north polar erg (AKA Olympia Undae) are currently very active on seasonal and yearly timescales, with 20-60% of the dunes within five polar dune fields undergoing the formation of alcove-apron features each Mars year. Previous studies have hypothesized formation mechanisms, based on observations of when new alcove and alcove-apron features form within an individual field through one Mars year. However, results are ill-constrained (and thus different hypotheses have been proposed) as the polar hood and winter night mean very few images are taken during the actual period of activity. In this study, we mitigate this limitation by examining several fields over several Mars years -- thus bringing aggregated results as well as detailed correlation checks against environmental conditions and seasonal processes to bear on the problem. From this, we propose a new process that appears consistent with all observations: (1) small alcoves form along the dune brink in the autumn (under the polar hood) due to instabilities induced by the night-formation and morning-sublimation of frost. As autumn progresses, the seasonal frost layer builds over the altered dune slope. (2) In the early spring, sublimation activity is concentrated and/or enhanced over these alcoves, causing further erosion and the formation of larger alcove-apron features. From the planform dimensions of the newly formed alcoves, we can estimate the volume of sand moved down the dune slope during one period of activity and estimate the aeolian sediment flux by looking at how quickly the alcoves are erased. We find that, over a Mars year, the amount of material moved via alcove-apron activity and via the wind are both comparable to aeolian sand flux estimates over dunes within lower-latitude regions of Mars. Thus it appears that the formation of alcoves-aprons is a significant mechanism for dune advancement and evolution within the

  4. NS1643 Interacts around L529 of hERG to Alter Voltage Sensor Movement on the Path to Activation

    PubMed Central

    Guo, Jiqing; Cheng, Yen May; Lees-Miller, James P.; Perissinotti, Laura L.; Claydon, Tom W.; Hull, Christina M.; Thouta, Samrat; Roach, Daniel E.; Durdagi, Serdar; Noskov, Sergei Y.; Duff, Henry J.

    2015-01-01

    Activators of hERG1 such as NS1643 are being developed for congenital/acquired long QT syndrome. Previous studies identify the neighborhood of L529 around the voltage-sensor as a putative interacting site for NS1643. With NS1643, the V1/2 of activation of L529I (−34 ± 4 mV) is similar to wild-type (WT) (−37 ± 3 mV; P > 0.05). WT and L529I showed no difference in the slope factor in the absence of NS1643 (8 ± 0 vs. 9 ± 0) but showed a difference in the presence of NS1643 (9 ± 0.3 vs. 22 ± 1; P < 0.01). Voltage-clamp-fluorimetry studies also indicated that in L529I, NS1643 reduces the voltage-sensitivity of S4 movement. To further assess mechanism of NS1643 action, mutations were made in this neighborhood. NS1643 shifts the V1/2 of activation of both K525C and K525C/L529I to hyperpolarized potentials (−131 ± 4 mV for K525C and −120 ± 21 mV for K525C/L529I). Both K525C and K525C/K529I had similar slope factors in the absence of NS1643 (18 ± 2 vs. 34 ± 5, respectively) but with NS1643, the slope factor of K525C/L529I increased from 34 ± 5 to 71 ± 10 (P < 0.01) whereas for K525C the slope factor did not change (18 ± 2 at baseline and 16 ± 2 for NS1643). At baseline, K525R had a slope factor similar to WT (9 vs. 8) but in the presence of NS1643, the slope factor of K525R was increased to 24 ± 4 vs. 9 ± 0 mV for WT (P < 0.01). Molecular modeling indicates that L529I induces a kink in the S4 voltage-sensor helix, altering a salt-bridge involving K525. Moreover, docking studies indicate that NS1643 binds to the kinked structure induced by the mutation with a higher affinity. Combining biophysical, computational, and electrophysiological evidence, a mechanistic principle governing the action of some activators of hERG1 channels is proposed. PMID:25809253

  5. The human ether-a'-go-go related gene (hERG) K+ channel blockade by the investigative selective-serotonin reuptake inhibitor CONA-437: limited dependence on S6 aromatic residues.

    PubMed

    Alexandrou, A J; Milnes, J T; Sun, S Z; Fermini, B; Kim, S C; Jenkinson, S; Leishman, D J; Witchel, H J; Hancox, J C; Leaney, J L

    2014-08-01

    Diverse non-cardiac drugs adversely influence cardiac electrophysiology by inhibiting repolarising K(+) currents mediated by channels encoded by the human ether-a-go-go-related gene (hERG). In this study, pharmacological blockade of hERG K(+) channel current (I(hERG)) by a novel investigative serotonin-selective reuptake inhibitor (SSRI), CONA-437, was investigated. Whole-cell patch-clamp measurements of I(hERG) were made from human embryonic kidney (HEK 293) cells expressing wild-type (WT) or mutant forms of the hERG channel. With a step-ramp voltage-command, peak I(hERG) was inhibited with an IC(50) of 1.34 μM at 35 ±1°C; the IC(50) with the same protocol was not significantly different at room temperature. Voltage-command waveform selection had only a modest effect on the potency of I(hERG) block: the IC50 with a ventricular action potential command was 0.72 μM. I(hERG) blockade developed rapidly with time following membrane depolarisation and showed a weak dependence on voltage, accompanied by a shift of ≈ -5 mV in voltage-dependence of activation. There was no significant effect of CONA-437 on voltage-dependence of I(hERG) inactivation, though at some voltages an apparent acceleration of the time-course of inactivation was observed. Significantly, mutation of the S6 aromatic amino acid residues Y652 and F656 had only a modest effect on I(hERG) blockade by CONA-437 (a 3-4 fold shift in affinity). CONA-437 at up to 30 μM had no significant effect on either Nav1.5 sodium channels or L-type calcium channels. In conclusion, the novel SSRI CONA-437 is particularly notable as a gating-dependent hERG channel inhibitor for which neither S6 aromatic amino-acid constituent of the canonical drug binding site on the hERG channel appears obligatory for I(hERG) inhibition to occur.

  6. Engineering Review Group (ERG) and Geologic Review Group (GRG) report on brine migration at the Deaf Smith County site salt repository horizon

    SciTech Connect

    Not Available

    1986-01-01

    In April 1986, ONWI requested the ERG and GRG to jointly address the status of current knowledge of, and ONWI approach to further characterization of, the geohydrology of the candidate repository horizon of the potential site in Deaf Smith County, Texas. Specifically, the ERG-GRG was asked to evaluate the status of understanding of the hydrogeology of the Lower San Andres Unit 4 (LSA-4) evaporite section and identify any major gaps in the data; evaluate the current understanding of the chemistry and movement of brines in the LSA-4 salt and associated interbeds; develop recommendations for estimating the upper limit quantity of brines, and modeling the brine movement, with respect to the emplaced HLW packages; and identify questions concerning the chemistry of the brines and recommend a technical approach to addressing these questions. 19 refs.

  7. Regulation of the human ether-a-go-go-related gene (hERG) channel by Rab4 protein through neural precursor cell-expressed developmentally down-regulated protein 4-2 (Nedd4-2).

    PubMed

    Cui, Zhi; Zhang, Shetuan

    2013-07-26

    The human ether-a-go-go-related gene (hERG) encodes the pore-forming α-subunit of the rapidly activating delayed rectifier K(+) channel in the heart, which plays a critical role in cardiac action potential repolarization. Dysfunction of IKr causes long QT syndrome, a cardiac electrical disorder that predisposes affected individuals to fatal arrhythmias and sudden death. The homeostasis of hERG channels in the plasma membrane depends on a balance between protein synthesis and degradation. Our recent data indicate that hERG channels undergo enhanced endocytic degradation under low potassium (hypokalemia) conditions. The GTPase Rab4 is known to mediate rapid recycling of various internalized proteins to the plasma membrane. In the present study, we investigated the effect of Rab4 on the expression level of hERG channels. Our data revealed that overexpression of Rab4 decreases the expression level of hERG in the plasma membrane. Rab4 does not affect the expression level of the Kv1.5 or EAG K(+) channels. Mechanistically, our data demonstrate that overexpression of Rab4 increases the expression level of endogenous Nedd4-2, a ubiquitin ligase that targets hERG but not Kv1.5 or EAG channels for ubiquitination and degradation. Nedd4-2 undergoes self- ubiquitination and degradation. Rab4 interferes with Nedd4-2 degradation, resulting in an increased expression level of Nedd4-2, which targets hERG. In summary, the present study demonstrates a novel pathway for hERG regulation; Rab4 decreases the hERG density at the plasma membrane by increasing the endogenous Nedd4-2 expression.

  8. Complete genome sequence of Arthrobacter sp. ERGS1:01, a putative novel bacterium with prospective cold active industrial enzymes, isolated from East Rathong glacier in India.

    PubMed

    Kumar, Rakshak; Singh, Dharam; Swarnkar, Mohit Kumar; Singh, Anil Kumar; Kumar, Sanjay

    2015-11-20

    We report the complete genome sequence of Arthrobacter sp. ERGS1:01, a novel bacterium which produces industrial enzymes at low temperature. East Rathong glacier in Sikkim Himalayas is untouched and unexplored for microbial diversity though it has a rich source of glaciers, alpine and meadows. Genome sequence has provided the basis for understanding its adaptation under harsh condition of Himalayan glacier, its ability to produce cold active industrial enzymes and has unlocked opportunities for microbial bioprospection from East Rathong glacier.

  9. Comparison of the effects of DC031050, a class III antiarrhythmic agent, on hERG channel and three neuronal potassium channels

    PubMed Central

    Li, Ping; Sun, Hai-feng; Zhou, Ping-zheng; Ma, Chao-ying; Hu, Guo-yuan; Jiang, Hua-liang; Li, Min; Liu, Hong; Gao, Zhao-bing

    2012-01-01

    Aim: This study was conducted to test the selectivity of DC031050 on cardiac and neuronal potassium channels. Methods: Human ether-à-go-go related gene (hERG), KCNQ and Kv1.2 channels were expressed in CHO cells. The delayed rectifier potassium current (IK) was recorded from dissociated hippocampal pyramidal neurons of neonatal rats. Whole-cell voltage patch clamp was used to record the voltage-activated potassium currents. Drug-containing solution was delivered using a RSC-100 Rapid Solution Changer. Results: Both DC031050 and dofetilide potently inhibited hERG currents with IC50 values of 2.3±1.0 and 17.9±1.2 nmol/L, respectively. DC031050 inhibited the IK current with an IC50 value of 2.7±1.5 μmol/L, which was >1000 times the concentration required to inhibit hERG current. DC031050 at 3 μmol/L did not significantly affect the voltage-dependence of the steady activation, steady inactivation of IK, or the rate of IK from inactivation. Intracellular application of DC031050 (5 μmol/L) was insufficient to inhibit IK. DC031050 up to 10 μmol/L had no effects on KCNQ2 and Kv1.2 channel currents. Conclusion: DC031050 is a highly selective hERG potassium channel blocker with a substantial safety margin of activity over neuronal potassium channels, thus holds significant potential for therapeutic application as a class III antiarrhythmic agent. PMID:22609836

  10. Development of recombinant cell line co-expressing mutated Nav1.5, Kir2.1, and hERG for the safety assay of drug candidates.

    PubMed

    Fujii, Masato; Ohya, Susumu; Yamamura, Hisao; Imaizumi, Yuji

    2012-07-01

    To provide a high-throughput screening method for human ether-a-go-go-gene-related gene (hERG) K(+) channel inhibition, a new recombinant cell line, in which single action potential (AP)-induced cell death was produced by gene transfection. Mutated human cardiac Na(+) channel Nav1.5 (IFM/Q3), which shows extremely slow inactivation, and wild-type inward rectifier K(+) channel, Kir2.1, were stably co-expressed in HEK293 cells (IFM/Q3+Kir2.1). In IFM/Q3+Kir2.1, application of single electrical stimulation (ES) elicited a long AP lasting more than 30 s and led cells to die by more than 70%, whereas HEK293 co-transfected with wild-type Nav1.5 and Kir2.1 fully survived. The additional expression of hERG K(+) channels in IFM/Q3+Kir2.1 shortened the duration of evoked AP and thereby markedly reduced the cell death. The treatment of the cells with hERG channel inhibitors such as nifekalant, E-4031, cisapride, terfenadine, and verapamil, recovered the prolonged AP and dose-dependently facilitated cell death upon ES. The EC(50) values to induce the cell death were 3 µM, 19 nM, 17 nM, 74 nM, and 3 µM, respectively, whereas 10 µM nifedipine did not induce cell death. Results indicate the high utility of this cell system for hERG K(+) channel safety assay.

  11. Analysis of the ergosterol biosynthesis pathway cloning, molecular characterization and phylogeny of lanosterol 14 α-demethylase (ERG11) gene of Moniliophthora perniciosa

    PubMed Central

    de Oliveira Ceita, Geruza; Vilas-Boas, Laurival Antônio; Castilho, Marcelo Santos; Carazzolle, Marcelo Falsarella; Pirovani, Carlos Priminho; Selbach-Schnadelbach, Alessandra; Gramacho, Karina Peres; Ramos, Pablo Ivan Pereira; Barbosa, Luciana Veiga; Pereira, Gonçalo Amarante Guimarães; Góes-Neto, Aristóteles

    2014-01-01

    The phytopathogenic fungus Moniliophthora perniciosa (Stahel) Aime & Philips-Mora, causal agent of witches’ broom disease of cocoa, causes countless damage to cocoa production in Brazil. Molecular studies have attempted to identify genes that play important roles in fungal survival and virulence. In this study, sequences deposited in the M. perniciosa Genome Sequencing Project database were analyzed to identify potential biological targets. For the first time, the ergosterol biosynthetic pathway in M. perniciosa was studied and the lanosterol 14α-demethylase gene (ERG11) that encodes the main enzyme of this pathway and is a target for fungicides was cloned, characterized molecularly and its phylogeny analyzed. ERG11 genomic DNA and cDNA were characterized and sequence analysis of the ERG11 protein identified highly conserved domains typical of this enzyme, such as SRS1, SRS4, EXXR and the heme-binding region (HBR). Comparison of the protein sequences and phylogenetic analysis revealed that the M. perniciosa enzyme was most closely related to that of Coprinopsis cinerea. PMID:25505843

  12. Discovery of high-affinity ligands of sigma1 receptor, ERG2, and emopamil binding protein by pharmacophore modeling and virtual screening.

    PubMed

    Laggner, Christian; Schieferer, Claudia; Fiechtner, Birgit; Poles, Gloria; Hoffmann, Rémy D; Glossmann, Hartmut; Langer, Thierry; Moebius, Fabian F

    2005-07-28

    ERG2, emopamil binding protein (EBP), and sigma-1 receptor (sigma(1)) are enzymes of sterol metabolism and an enzyme-related protein, respectively, that share high affinity for various structurally diverse compounds. To discover novel high-affinity ligands, pharmacophore models were built with Catalyst based upon a series of 23 structurally diverse chemicals exhibiting K(i) values from 10 pM to 100 microM for all three proteins. In virtual screening experiments, we retrieved drugs that were previously reported to bind to one or several of these proteins and also tested 11 new hits experimentally, of which three, among them raloxifene, had affinities for sigma(1) or EBP of <60 nM. When used to search a database of 3525 biochemicals of intermediary metabolism, a slightly modified ERG2 pharmacophore model successfully retrieved 10 substrate candidates among the top 28 hits. Our results indicate that inhibitor-based pharmacophore models for sigma(1), ERG2, and EBP can be used to screen drug and metabolite databases for chemically diverse compounds and putative endogenous ligands.

  13. Origin of the Sinai-Negev erg, Egypt and Israel: mineralogical and geochemical evidence for the importance of the Nile and sea level history

    USGS Publications Warehouse

    Muhs, Daniel R.; Roskin, Joel; Tsoar, Haim; Skipp, Gary; Budahn, James R.; Sneh, Amihai; Porat, Naomi; Stanley, Jean-Daniel; Katra, Itzhak; Blumberg, Dan G.

    2013-01-01

    The Sinai–Negev erg occupies an area of 13,000 km2 in the deserts of Egypt and Israel. Aeolian sand of this erg has been proposed to be derived from the Nile Delta, but empirical data supporting this view are lacking. An alternative source sediment is sand from the large Wadi El Arish drainage system in central and northern Sinai. Mineralogy of the Negev and Sinai dunes shows that they are high in quartz, with much smaller amounts of K-feldspar and plagioclase. Both Nile Delta sands and Sinai wadi sands, upstream of the dunes, also have high amounts of quartz relative to K-feldspar and plagioclase. However, Sinai wadi sands have abundant calcite, whereas Nile Delta sands have little or no calcite. Overall, the mineralogical data suggest that the dunes are derived dominantly from the Nile Delta, with Sinai wadi sands being a minor contributor. Geochemical data that proxy for both the light mineral fraction (SiO2/10–Al2O3 + Na2O + K2O–CaO) and heavy mineral fraction (Fe2O3–MgO–TiO2) also indicate a dominant Nile Delta source for the dunes. Thus, we report here the first empirical evidence that the Sinai–Negev dunes are derived dominantly from the Nile Delta. Linkage of the Sinai–Negev erg to the Nile Delta as a source is consistent with the distribution of OSL ages of Negev dunes in recent studies. Stratigraphic studies show that during the Last Glacial period, when dune incursions in the Sinai–Negev erg began, what is now the Nile Delta area was characterized by a broad, sandy, minimally vegetated plain, with seasonally dry anastomosing channels. Such conditions were ideal for providing a ready source of sand for aeolian transport under what were probably much stronger glacial-age winds. With the post-glacial rise in sea level, the Nile River began to aggrade. Post-glacial sedimentation has been dominated by fine-grained silts and clays. Thus, sea level, along with favorable climatic conditions, emerges as a major influence on the timing of dune

  14. Education For All: A Committment and an Opportunity. National EFA Coordinators Meeting under the Sub-Regional EFA Forum for East and Southeast Asia Final Report (2nd, Bangkok, Thailand, December 10-12, 2001).

    ERIC Educational Resources Information Center

    United Nations Educational, Scientific, and Cultural Organization, Bangkok (Thailand). Regional Office for Education in Asia and the Pacific.

    The working group of Sub-Regional Forum (SRF) and the Thematic Working Group (TWG) on Education for All (EFA) organized the second meeting of the SRF for East and Southeast Asia and the National EFA Coordinators in Bangkok, Thailand December 10-12, 2001. The meeting offered an opportunity for EFA coordinators to reflect on the outcomes of the EFA…

  15. Tank testing of a 2500-cm2 solar panel

    NASA Technical Reports Server (NTRS)

    Bever, R. S.; Staskus, J.

    1981-01-01

    A 50 cm by 50 cm solar array panel test patch was investigated for spacecraft charging and arcing effects. Bombardment with monochromatic electron was carried out. Some objectives of the test were: (1) to estimate at what voltage of electron bombardment arcing would be probable; (2) to find whether the arc's energy would be tolerable or damagingly large; (3) to try and separate thermal and photoeffects; and, (4) to see whether materials used were such as to minimize arcing. Some conclusions were: In sunlight the tracking data relay satellite's solar panel which has ceria glass on the front and conductive paint on the backside is probably a good design for reducing charge-up. In a geomagnetic substorm simulated in testing there will be arcing at the interconnects during eclipse and transitions into and out of eclipse in testing especially in view of the very cold temperatures that will be reached by this lightweight array. Ceria-doped glass is preferred to fused silica glass for reducing charge build up. The Kapton bare patch should still be conductively painted. The differential voltages on the panel determine when arcing first begins, and the electron beam voltages vary depending upon whether the metallic structure is directly grounded or semifloating.

  16. Tank testing of a 2500-cm2 solar panel

    SciTech Connect

    Bever, R.S.; Staskus, J.

    1981-10-01

    A 50 cm by 50 cm solar array panel test patch was investigated for spacecraft charging and arcing effects. Bombardment with monochromatic electron was carried out. Some objectives of the test were: (1) to estimate at what voltage of electron bombardment arcing would be probable (2) to find whether the arc's energy would be tolerable or damagingly large (3) to try and separate thermal and photoeffects and, (4) to see whether materials used were such as to minimize arcing. Some conclusions were: In sunlight the tracking data relay satellite's solar panel which has ceria glass on the front and conductive paint on the backside is probably a good design for reducing charge-up. In a geomagnetic substorm simulated in testing there will be arcing at the interconnects during eclipse and transitions into and out of eclipse in testing especially in view of the very cold temperatures that will be reached by this lightweight array. Ceria-doped glass is preferred to fused silica glass for reducing charge build up. The Kapton bare patch should still be conductively painted. The differential voltages on the panel determine when arcing first begins, and the electron beam voltages vary depending upon whether the metallic structure is directly grounded or semifloating.

  17. Amorphous nanostructuralization in HOPG by 1014 W cm-2 laser

    NASA Astrophysics Data System (ADS)

    NISHIMURA, Yasuhiko; KITAGAWA, Yoneyoshi; MORI, Yoshitaka; ISHII, Katsuhiro; HANAYAMA, Ryohei; AZUMA, Hirozumi; HIOKI, Tatsumi; NISHI, Teppei; MOTOHIRO, Tomoyoshi; KOMEDA, Osamu; SEKINE, Takashi; SATO, Nakahiro; KURITA, Takashi; KAWASHIMA, Toshiyuki; KAN, Hirofumi; SUNAHARA, Atsushi; SENTOKU, Yasuhiko; MIURA, Eisuke

    2016-05-01

    This reports provide an amorphous nanostructuralization technique on the surface modification in Highly Oriented Pyrolytic Graphite (HOPG) by using a femtosecond laser. We showed, for the first time, that the surface of HOPG is changed to the amorphous nanostructuralization graphite by using a femtosecond laser-driven compression technique. Our results also suggest that the HOPG surface is changed until the deeper area from the surface by the laser-driven shock wave. A single shot of a femtosecond laser beam (1.27 ∼ 1.33×1014 Wcm∼2 in intensity, with 2 mm-diameter, and 110 fs in pulse width) is irradiated under the vacuum ambience onto a 2 mm-thick of HOPG. The calculated impact pressures on a sample was 8.3 ∼ 8.7 GPa. Crystal structure in the HOPG were analyzed using a Raman spectroscopy and an X-ray diffraction, those analyzing depth from the surface were 50 nm and 350 μm, respectively.

  18. A nonsense mutation in the ERG6 gene leads to reduced susceptibility to polyenes in a clinical isolate of Candida glabrata.

    PubMed

    Vandeputte, Patrick; Tronchin, Guy; Larcher, Gérald; Ernoult, Emilie; Bergès, Thierry; Chabasse, Dominique; Bouchara, Jean-Philippe

    2008-10-01

    Unlike the molecular mechanisms that lead to azole drug resistance, the molecular mechanisms that lead to polyene resistance are poorly documented, especially in pathogenic yeasts. We investigated the molecular mechanisms responsible for the reduced susceptibility to polyenes of a clinical isolate of Candida glabrata. Sterol content was analyzed by gas-phase chromatography, and we determined the sequences and levels of expression of several genes involved in ergosterol biosynthesis. We also investigated the effects of the mutation harbored by this isolate on the morphology and ultrastructure of the cell, cell viability, and vitality and susceptibility to cell wall-perturbing agents. The isolate had a lower ergosterol content in its membranes than the wild type, and the lower ergosterol content was found to be associated with a nonsense mutation in the ERG6 gene and induction of the ergosterol biosynthesis pathway. Modifications of the cell wall were also seen, accompanied by increased susceptibility to cell wall-perturbing agents. Finally, this mutation, which resulted in a marked fitness cost, was associated with a higher rate of cell mortality. Wild-type properties were restored by complementation of the isolate with a centromeric plasmid containing a wild-type copy of the ERG6 gene. In conclusion, we have identified the molecular event responsible for decreased susceptibility to polyenes in a clinical isolate of C. glabrata. The nonsense mutation detected in the ERG6 gene of this isolate led to a decrease in ergosterol content. This isolate may constitute a useful tool for analysis of the relevance of protein trafficking in the phenomena of azole resistance and pseudohyphal growth.

  19. 3D-SDAR modeling of hERG potassium channel affinity: A case study in model design and toxicophore identification.

    PubMed

    Stoyanova-Slavova, Iva B; Slavov, Svetoslav H; Buzatu, Dan A; Beger, Richard D; Wilkes, Jon G

    2017-03-01

    A dataset of 237 human Ether-à-go-go Related Gene (hERG) potassium channel inhibitors (180 of which were used for model building and validation, whereas 57 constituted the "true" external prediction set) collected from 22 literature sources was modeled by 3D-SDAR. To produce reliable and reproducible classification models for hERG blocking, the initial set of 180 chemicals was split into two subsets: a balanced modeling set consisting of 118 compounds and an unbalanced validation set comprised of 62 compounds. A PLS bagging-like algorithm written in Matlab was used to process the data and assign each compound to one of the two (hERG+ or hERG-) activity classes. The best predictive model evaluated on the basis of a fully randomized hold-out test set (comprising 20% of the modeling set) used 4 latent variables and a grid of 6ppm×6ppm×1Å in the C-C region, 6ppm×30ppm×1Å in the C-N region, and 30ppm×30ppm×1Å in the N-N region. An overall accuracy of 0.84 was obtained for both the hold-out test set and the validation set. Further, an external prediction set consisting of 57 drugs and drug derivatives was used to estimate the true predictive power of the reported 3D-SDAR model - a slight reduction of the overall accuracy down to 0.77 was observed. 3D-SDAR map of the most frequently occurring bins and their projection on the standard coordinate space of the chemical structures allowed identification of a three-center toxicophore composed of two aromatic rings and an amino group. A U test along the distance axis of the most frequently occurring 3D-SDAR bins was used to set the distance limits of the toxicophore. This toxicophore was found to be similar to an earlier reported phospholipidosis (PLD) toxicophore.

  20. Quantitative structure-activity relationship of phenoxyphenyl-methanamine compounds with 5HT2A, SERT, and hERG activities.

    PubMed

    Mente, Scot; Gallaschun, Randall; Schmidt, Anne; Lebel, Lorrie; Vanase-Frawley, Michelle; Fliri, Anton

    2008-12-01

    QSAR models have been used to evaluate activities for compounds in the phenoxyphenyl-methanamine (PPMA) class of compounds. These models utilize Hammett-type donating-withdrawing substituent values as well as simple parameters to describe substituent size and elucidate the SAR of the 'A' and 'B' rings. Using this methodology, intuitive QSAR relationships were found for the three biological activities with R(2) values of 0.73, 0.45, and 0.58 for 5HT(2A), SerT, and hERG activities.

  1. Proceedings of REMR (Repair, Evaluation, Maintenance, and Rehabilitation Research Program) Workshop on Assessment of the Stability of Concrete Structures on Rock Held in Vicksburg, Mississippi on 10-12 September 1985.

    DTIC Science & Technology

    1987-01-01

    art methods of stobilitv aralysis of concrete hydraul ic ;tructures or, rock fouinctions. .2 Scope. The scope ,i this report includes a review of...FOLLOWING COVPILATION REPORT : TITLE: Proceedings of REMR (Repair, Evaluation, Maintenance, and Rehabilitation Research Program) Workshop Assessment of...the Stability of Concrete Structures on Roc)( Held in Vicksburg, Mississippi on 10-12 September 1985. To ORDER THE COMPLETE COMPILATION REPORT , USE AD

  2. A novel homologous dominant selection marker for genetic transformation of Penicillium chrysogenum: overexpression of squalene epoxidase-encoding ergA.

    PubMed

    Sigl, Claudia; Handler, Monika; Sprenger, Georg; Kürnsteiner, Hubert; Zadra, Ivo

    2010-11-01

    Genetic engineering requires genetic selection markers. For generation of biosafe strains in industrial applications, homologous dominant selection markers allowing "self-cloning" are best suited but scarce. Here we describe a novel homologous dominant genetic selection system for the filamentous fungus Penicillium chrysogenum based on overexpression of the P. chrysogenum squalene epoxidase-encoding ergA gene, which confers resistance against terbinafine. Terbinafine (TRB) is a potent antifungal drug used in therapy of fungal infections. Overexpression of ergA was driven by the P. chrysogenum endoxylanase xylP promoter that is highly inducible by xylose. The suitability of the novel selection marker cassette for genetic manipulation was proven by its use for targeted deletion of the transcription factor nosA in P. chrysogenum. NosA-deficiency did not affect growth rates on solid or in liquid media, conidiation in light or darkness, and resistance to hydrogen peroxide. However, NosA-deficiency significantly decreased penicillin productivity. As TRB inhibits the growth of a variety of fungal species, this novel selection marker is expected to be suitable for genetic engineering of diverse fungal species.

  3. Overexpression of erg20 gene encoding farnesyl pyrophosphate synthase has contrasting effects on activity of enzymes of the dolichyl and sterol branches of mevalonate pathway in Trichoderma reesei.

    PubMed

    Piłsyk, Sebastian; Perlińska-Lenart, Urszula; Górka-Nieć, Wioletta; Graczyk, Sebastian; Antosiewicz, Beata; Zembek, Patrycja; Palamarczyk, Grażyna; Kruszewska, Joanna S

    2014-07-10

    The mevalonate pathway is the most diverse metabolic route resulting in the biosynthesis of at least 30,000 isoprenoid compounds, many of which, such as sterols or dolichols, are indispensable for living cells. In the filamentous fungus Trichoderma of major biotechnological interest isoprenoid metabolites are also involved in the biocontrol processes giving the mevalonate pathway an additional significance. On the other hand, little is known about genes coding for enzymes of the mevalonate pathway in Trichoderma. Here, we present cloning and functional analysis of the erg20 gene from Trichoderma reesei coding for farnesyl pyrophosphate (FPP) synthase (EC 2.5.1.10), an enzyme located at the branching point of the mevalonate pathway. Expression of the gene in a thermosensitive erg20-2 mutant of Saccharomyces cerevisiae impaired in the FPP synthase activity suppressed the thermosensitive phenotype. The same gene overexpressed in T. reesei significantly enhanced the FPP synthase activity and also stimulated the activity of cis-prenyltransferase, an enzyme of the dolichyl branch of the mevalonate pathway. Unexpectedly, the activity of squalene synthase from the other, sterol branch, was significantly decreased without, however, affecting ergosterol level.

  4. Principles of a New Protocol for Prediction of Azole Resistance in Candida albicans Infections on the Basis of ERG11 Polymorphisms.

    PubMed

    Caban, Monika; Strapagiel, Dominik; Dziadek, Jarosław; Korycka-Machała, Małgorzata; Grzelak, Agnieszka

    2016-08-01

    In recent years, Candida albicans infections treatment has become a growing problem because, among others, pathogenic strains are capable to develop resistance to the administered drugs. The elaboration of rapid and accurate method of resistance assessment is an important goal of many studies. They aim to avoid inappropriate dosage or drug choice, which may be life threatening in case of severe candidiasis. Here we propose a new protocol to predict C. albicans infections. The resistance prediction is based on high-resolution melt (HRM) analysis of ERG11 gene, especially, at the particularly unstable regions. Two statistically significant nucleotide polymorphisms were detected among twenty-seven strains isolated from saliva, one of which was silent mutation (Glu266Asp, Leu480Leu). We propose also HRM analysis as a convenient, simple and inexpensive method of preliminary selection of C. albicans DNA samples that vary in ERG11 nucleotide sequence within presumed region. Taken together, our study provides firm basis for the development of fast, simple and reliable methodology for diagnosis of C. albicans infections.

  5. NMR solution structure of the N-terminal domain of hERG and its interaction with the S4-S5 linker

    SciTech Connect

    Li, Qingxin; Gayen, Shovanlal; Chen, Angela Shuyi; Huang, Qiwei; Raida, Manfred; Kang, CongBao

    2010-12-03

    Research highlights: {yields} The N-terminal domain (NTD, eag domain) containing 135 residues of hERG was expressed and purified from E. coli cells. {yields} Solution structure of NTD was determined with NMR spectroscopy. {yields} The alpha-helical region (residues 13-23) was demonstrated to possess the characteristics of an amphipathic helix. {yields} NMR titration confirmed the interaction between NTD and the peptide from the S4-S5 linker. -- Abstract: The human Ether-a-go-go Related Gene (hERG) potassium channel mediates the rapid delayed rectifier current (IKr) in the cardiac action potential. Mutations in the 135 amino acid residue N-terminal domain (NTD) cause channel dysfunction or mis-translocation. To study the structure of NTD, it was overexpressed and purified from Escherichia coli cells using affinity purification and gel filtration chromatography. The purified protein behaved as a monomer under purification conditions. Far- and near-UV, circular dichroism (CD) and solution nuclear magnetic resonance (NMR) studies showed that the purified protein was well-folded. The solution structure of NTD was obtained and the N-terminal residues 13-23 forming an amphipathic helix which may be important for the protein-protein or protein-membrane interactions. NMR titration experiment also demonstrated that residues from 88 to 94 in NTD are important for the molecular interaction with the peptide derived from the S4-S5 linker.

  6. Growth, Characterization and Device Development in Monocrystalline Diamond Films

    DTIC Science & Technology

    1990-09-30

    silicon substrate surfaces. The aspect ratio of diamond grown on single crystals of different substrate materials correlates with their surface free ...to check the feasibility of such considerations for the present research. The surface free energy of diamond is estimated to be 3387 ergs/cm 2, [Il...while the surface free energies of Si, Ni, Mo, Ta, and W are 1457 ergs/cm2 , 2072 ergs/cm 2, 2463 ergs/cm 2 , 2628 ergs/cm 2 and 3111 ergs/cm 2

  7. 2-Amino-N-pyrimidin-4-ylacetamides as A2A receptor antagonists: 2. Reduction of hERG activity, observed species selectivity, and structure-activity relationships.

    PubMed

    Slee, Deborah H; Moorjani, Manisha; Zhang, Xiaohu; Lin, Emily; Lanier, Marion C; Chen, Yongsheng; Rueter, Jaimie K; Lechner, Sandra M; Markison, Stacy; Malany, Siobhan; Joswig, Tanya; Santos, Mark; Gross, Raymond S; Williams, John P; Castro-Palomino, Julio C; Crespo, María I; Prat, Maria; Gual, Silvia; Díaz, José-Luis; Jalali, Kayvon; Sai, Yang; Zuo, Zhiyang; Yang, Chun; Wen, Jenny; O'Brien, Zhihong; Petroski, Robert; Saunders, John

    2008-03-27

    Previously we have described a series of novel A 2A receptor antagonists with excellent water solubility. As described in the accompanying paper, the antagonists were first optimized to remove an unsubstituted furyl moiety, with the aim of avoiding the potential metabolic liabilities that can arise from the presence of an unsubstituted furan. This effort identified a series of potent and selective methylfuryl derivatives. Herein, we describe the further optimization of this series to increase potency, maintain selectivity for the human A 2A vs the human A 1 receptor, and minimize activity against the hERG channel. In addition, the observed structure-activity relationships against both the human and the rat A 2A receptor are reported.

  8. Optical Memory Stores 10 12sup. Bits

    NASA Technical Reports Server (NTRS)

    1982-01-01

    Optical Mass Memory has separate recorder (write) and reproducer (read) modules. Data are recorded on fiches and stored in a carrousel. Fische is retrieved from carrousel by transporter in under 10 seconds. Input to optical memory is standard TV camera. TV monitor at memory output displays stored video images when they are retrieved from fisches. Input to the optical memory can also be taken from pseudorandom sequence generator.

  9. General Business: Grades 10-12.

    ERIC Educational Resources Information Center

    Instructional Objectives Exchange, Los Angeles, CA.

    Thirty-five objectives and related test items assessing general business skills taught in grades 10 through 12 are included in this collection. Each objective is stated in operational terms and identified by a subject area within the hood category of general business. Objectives include the desired behavior and subject content so that students are…

  10. Field Ecology, Grades 10-12.

    ERIC Educational Resources Information Center

    Long, David C.; Powell, Nancy A.

    Presented are activities which provide a program of first-hand experiences in field ecology for groups of three to five students under the leadership of a teacher or adult volunteer. Concentration is centered upon study of the effects of environment on living plants and animals and the interdependence of members of ecosystems. Each investigation…

  11. A new C-terminal hERG mutation A915fs+47X associated with symptomatic LQT2 and auditory-trigger syncope

    PubMed Central

    Christé, Georges; Thériault, Olivier; Chahine, Mohamed; Millat, Gilles; Rodriguez-Lafrasse, Claire; Rousson, Robert; Deschênes, Isabelle; Ficker, Eckhard; Chevalier, Philippe

    2008-01-01

    Background A novel mutation of hERG (A915fs+47X) was discovered in a 32 year-old woman with torsades de pointes, long QTc interval (515 ms) and syncope upon auditory trigger. Objective We explored whether the properties of this mutation could explain the pathology. Methods Western blots and sedimentation analysis of del/WT hERG were used to analyze protein expression, assembly and trafficking. Whole-cell A915fs+47X (del) and wild-type (WT) currents were recorded in transiently transfected COS7 cells or Xenopus oocytes. Results The tail current density at −40 mV after a 2 s depolarization to +40 mV in COS7 cells expressing del was 36% of that for WT. Inactivation was 1.9-fold to 2.8-fold faster in del versus WT between −60 and +60 mV. In the range −60 to −10 mV, we found that a non-deactivating fraction of current was increased in del at the expense of a rapidly deactivating fraction, with a slowly deactivating fraction being unchanged. In Xenopus oocytes, expression of del alone produced 38% of WT currents whereas co-expression of 1/2 WT + 1/2 del produced 49.8%. Furthermore, the expression of del protein at the cell surface was reduced by about 50%. This suggests that a partial trafficking defect of del contributes to the reduction in del current densities and to the dominant negative effect when co-expressed with WT. In model simulations, the mutation causes a 10% prolongation of action potential duration. Conclusions Decreased current levels due to a trafficking defect may explain the long QT syndrome observed in our patient. PMID:18984536

  12. Preface: Proceedings of the ESF Exploratory Workshop on Glassy Liquids under Pressure: Fundamentals and Applications (Ustroń, Poland, 10-12 October 2007) Proceedings of the ESF Exploratory Workshop on Glassy Liquids under Pressure: Fundamentals and Applications (Ustroń, Poland, 10-12 October 2007)

    NASA Astrophysics Data System (ADS)

    Drozd-Rzoska, Aleksandra; Rzoska, Sylwester J.; Tamarit, Josep Ll

    2008-06-01

    This preface focuses on the importance of pressure studies for explaining the glass transitions puzzle. Subsequently, some issues related to the European Science Foundation Exploratory Workshop (ESF EW) Glassy Liquids under Pressure: Fundamentals and Applications (Ustroń, Poland, 10-12 October 2007) are recalled. Most liquids crystallize on 'normal' cooling at the melting temperature Tm. However, some liquids can skip crystallization and undergo supercooling down to glass temperature Tg. Turnbull 1 proposed an empirical link between these temperatures indicating good glass forming ability (GFA) for Tg/Tm « 2/3. Values of the GFA factor Tg/Tm → 1/2 were suggested for 'poor' glass formers, where crystallization is difficult to avoid. Recently, the significance of the pressure dependence of the GFA factor was also noted 2. Reaching the glass transition is associated with a series of phenomena, namely 3: (i) the thermal expansion coefficient at constant pressure changes smoothly from values common for a liquid to those of a crystal, showing anomalous behaviour near Tg, (ii) viscosity reaches a value of η = 1013 P and the structural relaxation time τ ≈ 100 s, (iii) the specific heat drop occurs, giving rise to the famous Kauzmann paradox. On cooling towards glass transition, the 'pretransitional' behaviour can be observed for dynamic properties even well above Tg + 100 K 3. This includes the non-Arrhenius evolution of such magnitudes as viscosity, primary (structural-, α-) relaxation time, electric conductivity or diffusion coefficient associated with increasingly non-Debye distribution of relaxation times 3. Such behaviour is associated with short-time scale relaxation processes. The most characteristic is the secondary (β-) relaxation 4, 5 which merges with the 'structural' dynamics near τ(TB) = 10-7+/-1s, the hypothetically universal (magic) time-scale 6. Below TB the split in the evolution of the translation and orientation related properties occurs 4, 5

  13. Strain distribution in Zr64.13Cu15.75Ni10.12Al10 bulk metallic glass investigated by in situ tensile tests under synchrotron radiation

    NASA Astrophysics Data System (ADS)

    Stoica, M.; Das, J.; Bednarcik, J.; Franz, H.; Mattern, N.; Wang, W. H.; Eckert, J.

    2008-07-01

    We report on the evolution of the atomic-scale strain tensor of ductile Zr64.13Cu15.75Ni10.12Al10 bulk metallic glass under tensile loading by using x-ray synchrotron radiation. The same kind of samples was previously investigated under compressive loading and revealed yielding at 1690 MPa together with large deformability of up to 160% strain. In tension the samples fracture at a lower stress, 1500 MPa, with no sign of yielding or plastic deformation. With no macroplasticity observed under tension, large differences in the elastic constants obtained from the strain tensor and from ultrasonic sound velocity measurements are revealed. This paper presents in detail the measuring procedure as well as the calculation of the tensile tensor and pair distribution functions of Zr64.13Cu15.75Ni10.12Al10 at different stages of deformation. The results are discussed in comparison with other reported data obtained from x-ray diffraction measurements using synchrotron radiation.

  14. The Novel PIM1 Inhibitor NMS-P645 Reverses PIM1-Dependent Effects on TMPRSS2/ERG Positive Prostate Cancer Cells And Shows Anti-Proliferative Activity in Combination with PI3K Inhibition

    PubMed Central

    Mologni, Luca; Magistroni, Vera; Casuscelli, Francesco; Montemartini, Marisa; Gambacorti-Passerini, Carlo

    2017-01-01

    PIM1 is over-expressed in multiple tumors, including prostate cancer (PCa). PIM1 upregulation is mediated by direct binding of the ERG transcription factor to its promoter. About 50% of PCa cases are characterized by the presence of the TMPRSS2/ERG fusion, leading to ERG over-expression and thus to PIM1 transcriptional activation. PIM kinases are considered as weak oncogenes, but when combined with additional genetic alterations can induce strong transforming effects. Here we show anti-proliferative activity of the newly described PIM1 inhibitor NMS-P645 in combination with the PI3K inhibitor GDC-0941 in TMPRSS2/ERG positive and negative PCa cells. Treatment with NMS-P645 alone can reverse PIM1-mediated pro-survival signals in prostate cells, such as activation of STAT3 through Tyr705 phosphorylation and resistance to taxane-based treatments, but does not exert a strong anti-tumoral effect. However, the simultaneous treatment with NMS-P645 and GDC-0941 induces a significant anti-proliferative response in PCa cells. These results support the use of combination strategies with PIM and PI3K inhibitors as effective treatment for PCa cases. PMID:28123608

  15. Full-field ERGs obtained using a contact lens electrode with built-in high intensity white light-emitting diodes in beagle dogs can be applied to toxicological assessments.

    PubMed

    Sasaki, Shoji; Yamashita, Haruhiro; Yagi, Kumiko; Iwaki, Yoshinobu; Kimura, Masaaki

    2006-10-10

    We investigated full-field ERGs in beagle dogs using a contact lens electrode with built-in LED. Experiment 1 was performed to determine the appropriate conditions for stimulus intensity and background illumination. We found that full-field ERGs could be recorded under the following conditions: stimulus intensity: -2.5logcd*s/m(2) in rod responses (RRs), 1.2logcd*s/m(2) in maximal responses (MRs), oscillatory potentials (OPs), cone responses (CRs), 30-Hz flicker responses (FRs), and background illumination: more than 25cd/m(2) in CRs and FRs. Experiment 2 was performed to apply full-field ERGs in beagle dogs to the detection of retinal toxicities. A dog was given one 30mg/kg dose of sodium iodate (NaIO(3)) intravenously. ERGs were recorded before administration and 1, 3, 5, 8, 24h, 7 and 14 days after administration of NaIO(3). The RRs disappeared completely at 1h when MRs and OPs decreased. On the other hand, CRs and FRs were recorded even at 8h. All responses disappeared at 24h. These findings indicate that retinal toxicity by NaIO(3) is first expressed in rods, followed by cones. These results suggest that full-field ERGs in beagle dogs using an LED contact lens can be used to evaluate toxic effects on rods and cones separately, with the potential to prove more useful than conventional methods for toxicological assessments of developing pharmaceuticals, and can be applied to it.

  16. The physiological roles of membrane ergosterol as revealed by the phenotypes of syr1/erg3 null mutant of Saccharomyces cerevisiae.

    PubMed

    Hemmi, K; Julmanop, C; Hirata, D; Tsuchiya, E; Takemoto, J Y; Miyakawa, T

    1995-03-01

    Ergosterol is a major sterol component of fungal plasma membranes. The effects of disrupting the Saccharomyces cerevisiae SYR1/ERG3 gene, which encodes sterol C-5 desaturase, an enzyme of ergosterol biosynthesis pathway, were markedly different for different S. cerevisiae strains and growth temperatures. The null mutation of SYR1 (delta syr1) in strain RAY-3A had only a slight effect on the growth rate at 28 degrees C. However, at this temperature, the same mutation caused poor growth in strain KA-311A and no growth in strain W303-1A. The delta syr1 disruptant of these strains were able to grow at 37 degrees C, as well as their parental strains. Moreover, the growth of the delta syr1 disruptant of W303-1A and KA-311A strains were severely inhibited at 16 degrees C. These results indicated that ergosterol is essential for growth at low temperatures, and the effects of the gene disruption are variable by the genetic background. The growth defect at low temperatures appeared to be due to the defect of tryptophan uptake in the delta syr1 mutants. The delta syr1 mutants were sensitive to a wide variety of drugs, chemicals, and ions, suggesting that yeast ergosterol is important as permeability barrier against various chemical stresses.

  17. hERG1/Kv11.1 activation stimulates transcription of p21waf/cip in breast cancer cells via a calcineurin-dependent mechanism

    PubMed Central

    Perez-Neut, Mathew; Rao, Vidhya R.; Gentile, Saverio

    2016-01-01

    The function of Kv11.1 is emerging in breast cancer biology, as a growing body of evidence indicates that the hERG1/Kv11.1 potassium channel is aberrantly expressed in several cancer types including breast cancers. The biological effects of Kv11.1 channel blockers and their associated side effects are very well known but the potential use of Kv11.1 activators as an anticancer strategy are still unexplored. In our previous work, we have established that stimulation of the Kv11.1 potassium channel activates a senescent-like program that is characterized by a significant increase in tumor suppressor protein levels, such as p21waf/cip and p16INK4A. In this study we investigated the mechanism linking Kv11.1 stimulation to augmentation of p21waf/cip protein level. We have demonstrated that the Kv11.1 channel activator NS1643 activates a calcineurin-dependent transcription of p21waf/cip and that this event is fundamental for the inhibitory effect of NS1643 on cell proliferation. Our results reveal a novel mechanism by which stimulation of Kv11.1 channel leads to transcription of a potent tumor suppressor and suggest a potential therapeutic use for Kv11.1 channel activators. PMID:25945833

  18. NMR solution structure of the N-terminal domain of hERG and its interaction with the S4-S5 linker.

    PubMed

    Li, Qingxin; Gayen, Shovanlal; Chen, Angela Shuyi; Huang, Qiwei; Raida, Manfred; Kang, Congbao

    2010-12-03

    The human Ether-à-go-go Related Gene (hERG) potassium channel mediates the rapid delayed rectifier current (IKr) in the cardiac action potential. Mutations in the 135 amino acid residue N-terminal domain (NTD) cause channel dysfunction or mis-translocation. To study the structure of NTD, it was overexpressed and purified from Escherichia coli cells using affinity purification and gel filtration chromatography. The purified protein behaved as a monomer under purification conditions. Far- and near-UV, circular dichroism (CD) and solution nuclear magnetic resonance (NMR) studies showed that the purified protein was well-folded. The solution structure of NTD was obtained and the N-terminal residues 13-23 forming an amphipathic helix which may be important for the protein-protein or protein-membrane interactions. NMR titration experiment also demonstrated that residues from 88 to 94 in NTD are important for the molecular interaction with the peptide derived from the S4-S5 linker.

  19. Trichodiene Production in a Trichoderma harzianum erg1-Silenced Strain Provides Evidence of the Importance of the Sterol Biosynthetic Pathway in Inducing Plant Defense-Related Gene Expression.

    PubMed

    Malmierca, M G; McCormick, S P; Cardoza, R E; Monte, E; Alexander, N J; Gutiérrez, S

    2015-11-01

    Trichoderma species are often used as biocontrol agents against plant-pathogenic fungi. A complex molecular interaction occurs among the biocontrol agent, the antagonistic fungus, and the plant. Terpenes and sterols produced by the biocontrol fungus have been found to affect gene expression in both the antagonistic fungus and the plant. The terpene trichodiene (TD) elicits the expression of genes related to tomato defense and to Botrytis virulence. We show here that TD itself is able to induce the expression of Botrytis genes involved in the synthesis of botrydial (BOT) and also induces terpene gene expression in Trichoderma spp. The terpene ergosterol, in addition to its role as a structural component of the fungal cell membranes, acts as an elicitor of defense response in plants. In the present work, using a transformant of T. harzianum, which is silenced in the erg1 gene and accumulates high levels of squalene, we show that this ergosterol precursor also acts as an important elicitor molecule of tomato defense-related genes and induces Botrytis genes involved in BOT biosynthesis, in both cases, in a concentration-dependent manner. Our data emphasize the importance of a balance of squalene and ergosterol in fungal interactions as well as in the biocontrol activity of Trichoderma spp.

  20. Endoglin expression in the endothelium is regulated by Fli-1, Erg, and Elf-1 acting on the promoter and a -8-kb enhancer.

    PubMed

    Pimanda, John E; Chan, W Y Iris; Donaldson, Ian J; Bowen, Mark; Green, Anthony R; Göttgens, Berthold

    2006-06-15

    Angiogenesis is critical to the growth and regeneration of tissue but is also a key component of tumor growth and chronic inflammatory disorders. Endoglin plays a key role in angiogenesis by modulating cellular responses to transforming growth factor-beta (TGF-beta) signaling and is upregulated in proliferating endothelial cells. To gain insights into the transcriptional hierarchies that govern endoglin expression, we used a combination of comparative genomic, biochemical, and transgenic approaches. Both the promoter and a region 8 kb upstream of exon 1 were active in transfection assays in endothelial cells. In transgenic mice, the promoter directed low-level expression to a subset of endothelial cells. By contrast, inclusion of the -8 enhancer resulted in robust endothelial activity with additional staining in developing ear mesenchyme. Subsequent molecular analysis demonstrated that both the -8 enhancer and the promoter depend on conserved Ets sites, which were bound in endothelial cells in vivo by Fli-1, Erg, and Elf-1. This study therefore establishes the transcriptional framework within which endoglin functions during angiogenesis.

  1. Acute and Chronic Toxicity, Cytochrome P450 Enzyme Inhibition, and hERG Channel Blockade Studies with a Polyherbal, Ayurvedic Formulation for Inflammation

    PubMed Central

    Dey, Debendranath; Chaskar, Sunetra; Athavale, Nitin; Chitre, Deepa

    2015-01-01

    Ayurvedic plants are known for thousands of years to have anti-inflammatory and antiarthritic effect. We have recently shown that BV-9238, a proprietary formulation of Withania somnifera, Boswellia serrata, Zingiber officinale, and Curcuma longa, inhibits LPS-induced TNF-alpha and nitric oxide production from mouse macrophage and reduces inflammation in different animal models. To evaluate the safety parameters of BV-9238, we conducted a cytotoxicity study in RAW 264.7 cells (0.005–1 mg/mL) by MTT/formazan method, an acute single dose (2–10 g/kg bodyweight) toxicity study and a 180-day chronic study with 1 g and 2 g/kg bodyweight in Sprague Dawley rats. Some sedation, ptosis, and ataxia were observed for first 15–20 min in very high acute doses and hence not used for further chronic studies. At the end of 180 days, gross and histopathology, blood cell counts, liver and renal functions were all at normal levels. Further, a modest attempt was made to assess the effects of BV-9238 (0.5 µg/mL) on six major human cytochrome P450 enzymes and 3H radioligand binding assay with human hERG receptors. BV-9238 did not show any significant inhibition of these enzymes at the tested dose. All these suggest that BV-9238 has potential as a safe and well tolerated anti-inflammatory formulation for future use. PMID:25893199

  2. [The Influence of the Functioning of Brain Regulatory Systems onto the Voluntary Regulation of Cognitive Performance in Children. Report 2. Neuropsychological and Electrophysiological Assessment of Brain Regulatory Functions in Children Aged 10-12 with Learning Difficulties].

    PubMed

    Semenova, O A; Machinskaya, R I

    2015-01-01

    A total number of 172 children aged 10-12 were electrophysiologically and neuropsychologically assessed in order to analyze the influence of the functioning of brain regulatory systems onto the voluntary regulation of cognitive performance during the preteen years. EEG patterns associated with the nonoptimal functioning of brain regulatory systems, particularly fronto-thalamic, limbic and fronto-striatal structures were significantly more often observed in children with learning and behavioral difficulties, as compared to the control group. Neuropsychological assessment showed that the nonoptimal functioning of different brain regulatory systems specifically affect the voluntary regulation of cognitive performance. Children with EEG patterns of fronto-thalamic nonoptimal functioning demonstrated poor voluntary regulation such as impulsiveness and difficulties in continuing the same algorithms. Children with EEG patterns of limbic nonoptimal functioning showed a less pronounced executive dysfunction manifested only in poor switching between program units within a task. Children with EEG patterns of fronto-striatal nonoptimal functioning struggled with such executive dysfunctions as motor and tactile perseverations and emotional-motivational deviations such as poor motivation and communicative skills.

  3. DFT studies on a high-energy density cage compound 1, 3, 5, 7, 9, 11-hexo(N(CH3)NO2)-2, 4, 6, 8, 10, 12-hexaazatetracyclo[5, 5, 0, 0, 0] dodecane

    NASA Astrophysics Data System (ADS)

    Li, Xiao-Hong; Yong, Yong-Liang; Zhang, Xian-Zhou

    2014-04-01

    The infrared and Raman spectra, heat of formation (HOF) and thermodynamic properties were investigated by B3LYP/6-31G** method for a new designed polynitro cage compound 1,3,5,7,9,11-hexo(N(CH3)NO2)-2,4,6,8,10,12-hexaazatetracyclo[5,5,0,0,0]dodecane. The detonation velocity (D) and pressure (P) were predicted by the Kamlet-Jacobs equations based on the theoretical density and condensed HOF. The bond dissociation energies and bond orders for the weakest bonds were analysed to investigate the thermal stability of the title compound. The computational result shows that the detonation velocity and pressure of the title compound are superior to those of hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX), but inferior to those of 1,3,5,7-tetranitro-1,3,5,7-tetraazacyclooctane (HMX) and hexanitrohexaazaisowurtzitane (HNIW). And the analysis of thermal stability shows that the first step of pyrolysis is the rupture of the N7-NO2 bond. The crystal structure obtained by molecular mechanics belongs to the P21 space group, with the lattice parameters Z = 2, a = 11.8246 Å, b = 10.4632 Å, c = 15.9713 Å, ρ = 1.98 g cm-3.

  4. Lacustrine sediments and Holocene climates in the southern Sahara: the example of paleolakes in the Grand Erg of Bilma (Zoo Baba and Dibella, eastern Niger)

    NASA Astrophysics Data System (ADS)

    Grunert, J.; Baumhauer, R.; Völkel, J.

    Two endoreic depressions of the central Erg of Bilma/eastern Republic of Niger are described here. Special interest is given to widespread lacustrine sediments (diatomites) in the depressions, indicating paleolakes with an extension of some 10 km 2. The minimum depth of the lakes has been found to be 25 m (Zoo Baba) and 36 m (Dibella). At Dibella there is evidence of two lacustrine phases of Early Holocene (9785-7890 years B.P.) and of Mid-Holocene age (about 6500 years B.P.). At Zoo Baba only one lacustrine phase of Mid-Holocene age (about 6000-5370 years B.P.) can be proven for Zoo Baba. The difference between the Early- and Mid-Holocene diatomites Di I and Di II is based not only on radiocarbon data but also on geomorphological characteristics. Despite the small distance of only 70 km between the two depressions, the reconstruction of pluvial water chemistry particularly by analyzing the diatoms shows great differences. The Mid-Holocene Zoo Baba lake was a freshwater lake; the Early- and Mid-Holocene Dibella lakes obviously contained brackish water with great fluctuations of salinity. An explanation may be given by the varying groundwater influence during the pluvial periods. It is supposed that Zoo Baba has been touched by the large Kaouar/Bilma aquifer while Dibella has been influenced by a smaller local aquifer. Any groundwater supply from Paleo-Chad can be excluded. Both the rising of groundwater table and of lake levels must have been induced by a largely increased annual precipitation rate (today 20-40 mm only), combined with a very effective reduction in evaporation rate. Based on lacrustine sediments, paleosoils and geomorphological features (fossil landslides of Zoo Baba), the Early Holocene annual precipitation rate is estimated at least at 300-400 mm, the Mid-Holocene rate at 200-300 mm. The special character of precipitations cannot be reconstructed.

  5. Total petroleum systems of the Grand Erg/Ahnet Province, Algeria and Morocco; the Tanezzuft-Timimoun, Tanezzuft-Ahnet, Tanezzuft-Sbaa, Tanezzuft Mouydir, Tanezzuft-Benoud, and Tanezzuft-Bechar/Abadla

    USGS Publications Warehouse

    Klett, T.R.

    2000-01-01

    Undiscovered, conventional oil and gas resources were assessed within total petroleum systems of the Grand Erg/Ahnet Province (2058) as part of the U.S. Geological Survey World Petroleum Assessment 2000. The majority of the Grand Erg/ Ahnet Province is in western Algeria; a very small portion extends into Morocco. The province includes the Timimoun Basin, Ahnet Basin, Sbaa Basin, Mouydir Basin, Benoud Trough, Bechar/Abadla Basin(s), and part of the Oued Mya Basin. Although several petroleum systems may exist within each of these basins, only seven ?composite? total petroleum systems were identified. Each total petroleum system occurs in a separate basin, and each comprises a single assessment unit. The main source rocks are the Silurian Tanezzuft Formation (or lateral equivalents) and Middle to Upper Devonian mudstone. Maturation history and the major migration pathways from source to reservoir are unique to each basin. The total petroleum systems were named after the oldest major source rock and the basin in which it resides. The estimated means of the undiscovered conventional petroleum volumes in total petroleum systems of the Grand Erg/ Ahnet Province are as follows: [MMBO, million barrels of oil; BCFG, billion cubic feet of gas; MMBNGL, million barrels of natural gas liquids] Total Petroleum System MMBO BCFG MMBNGL Tanezzuft-Timimoun 31 1,128 56 Tanezzuft-Ahnet 34 2,973 149 Tanezzuft-Sbaa 162 645 11 Tanezzuft-Mouydir 12 292 14 Tanezzuft-Benoud 72 2,541 125 Tanezzuft-Bechar/Abadla 16 441 22

  6. Low-visibility light-intensity laser-triggered release of entrapped calcein from 1,2-bis (tricosa-10,12-diynoyl)-sn-glycero-3-phosphocholine liposomes is mediated through a type I photoactivation pathway.

    PubMed

    Yavlovich, Amichai; Viard, Mathias; Gupta, Kshitij; Sine, Jessica; Vu, Mylinh; Blumenthal, Robert; Tata, Darrell B; Puri, Anu

    2013-01-01

    We recently reported on the physical characteristics of photo-triggerable liposomes containing dipalmitoylphosphatidylcholine (DPPC), and 1,2-bis (tricosa-10,12-diynoyl)-sn-glycero-3-phosphocholine (DC(8,9)PC) carrying a photo agent as their payload. When exposed to a low-intensity 514 nm wavelength (continuous-wave) laser light, these liposomes were observed to release entrapped calcein green (Cal-G; Ex/Em 490/517 nm) but not calcein blue (Cal-B; Ex/Em 360/460 nm). In this study, we have investigated the mechanism for the 514 nm laser-triggered release of the Cal-G payload using several scavengers that are known specifically to inhibit either type I or type II photoreaction pathways. Liposomes containing DPPC:DC(8,9)PC: distearoylphosphatidylethanolamine (DSPE)-polyethylene glycol (PEG)-2000 (86:10:04 mole ratio) were loaded either with fluorescent (calcein) or nonfluorescent ((3)H-inulin) aqueous markers. In addition, a non-photo-triggerable formulation (1-palmitoyl-2-oleoyl phosphatidylcholine [POPC]:DC(8,9)PC:DSPE-PEG2000) was also studied with the same payloads. The 514 nm wavelength laser exposure on photo-triggerable liposomes resulted in the release of Cal-G but not that of Cal-B or (3)H-inulin, suggesting an involvement of a photoactivated state of Cal-G due to the 514 nm laser exposure. Upon 514 nm laser exposures, substantial hydrogen peroxide (H2O2, ≈100 μM) levels were detected from only the Cal-G loaded photo-triggerable liposomes but not from Cal-B-loaded liposomes (≤10 μM H2O2). The Cal-G release from photo-triggerable liposomes was found to be significantly inhibited by ascorbic acid (AA), resulting in a 70%-80% reduction in Cal-G release. The extent of AA-mediated inhibition of Cal-G release from the liposomes also correlated with the consumption of AA. No AA consumption was detected in the 514 nm laser-exposed Cal B-loaded liposomes, thus confirming a role of photoactivation of Cal-G in liposome destabilization. Inclusion of 100 mM K3Fe

  7. Acute alteration of cardiac ECG, action potential, I{sub Kr} and the human ether-a-go-go-related gene (hERG) K{sup +} channel by PCB 126 and PCB 77

    SciTech Connect

    Park, Mi-Hyeong; Park, Won Sun; Jo, Su-Hyun

    2012-07-01

    Polychlorinated biphenyls (PCBs) have been known as serious persistent organic pollutants (POPs), causing developmental delays and motor dysfunction. We have investigated the effects of two PCB congeners, 3,3′,4,4′-tetrachlorobiphenyl (PCB 77) and 3,3′,4,4′,5-pentachlorobiphenyl (PCB 126) on ECG, action potential, and the rapidly activating delayed rectifier K{sup +} current (I{sub Kr}) of guinea pigs' hearts, and hERG K{sup +} current expressed in Xenopus oocytes. PCB 126 shortened the corrected QT interval (QTc) of ECG and decreased the action potential duration at 90% (APD{sub 90}), and 50% of repolarization (APD{sub 50}) (P < 0.05) without changing the action potential duration at 20% (APD{sub 20}). PCB 77 decreased APD{sub 20} (P < 0.05) without affecting QTc, APD{sub 90}, and APD{sub 50}. The PCB 126 increased the I{sub Kr} in guinea-pig ventricular myocytes held at 36 °C and hERG K{sup +} current amplitude at the end of the voltage steps in voltage-dependent mode (P < 0.05); however, PCB 77 did not change the hERG K{sup +} current amplitude. The PCB 77 increased the diastolic Ca{sup 2+} and decreased Ca{sup 2+} transient amplitude (P < 0.05), however PCB 126 did not change. The results suggest that PCB 126 shortened the QTc and decreased the APD{sub 90} possibly by increasing I{sub Kr}, while PCB 77 decreased the APD{sub 20} possibly by other modulation related with intracellular Ca{sup 2+}. The present data indicate that the environmental toxicants, PCBs, can acutely affect cardiac electrophysiology including ECG, action potential, intracellular Ca{sup 2+}, and channel activity, resulting in toxic effects on the cardiac function in view of the possible accumulation of the PCBs in human body. -- Highlights: ► PCBs are known as serious environmental pollutants and developmental disruptors. ► PCB 126 shortened QT interval of ECG and action potential duration. ► PCB 126 increased human ether-a-go-go-related K{sup +} current and I{sub Kr}. ► PCB

  8. The NuSTAR Extragalactic Surveys: The Number Counts of Active Galactic Nuclei and the Resolved Fraction of the Cosmic X-Ray Background

    NASA Astrophysics Data System (ADS)

    Harrison, F. A.; Aird, J.; Civano, F.; Lansbury, G.; Mullaney, J. R.; Ballantyne, D. R.; Alexander, D. M.; Stern, D.; Ajello, M.; Barret, D.; Bauer, F. E.; Baloković, M.; Brandt, W. N.; Brightman, M.; Boggs, S. E.; Christensen, F. E.; Comastri, A.; Craig, W. W.; Del Moro, A.; Forster, K.; Gandhi, P.; Giommi, P.; Grefenstette, B. W.; Hailey, C. J.; Hickox, R. C.; Hornstrup, A.; Kitaguchi, T.; Koglin, J.; Luo, B.; Madsen, K. K.; Mao, P. H.; Miyasaka, H.; Mori, K.; Perri, M.; Pivovaroff, M.; Puccetti, S.; Rana, V.; Treister, E.; Walton, D.; Westergaard, N. J.; Wik, D.; Zappacosta, L.; Zhang, W. W.; Zoglauer, A.

    2016-11-01

    We present the 3-8 keV and 8-24 keV number counts of active galactic nuclei (AGNs) identified in the Nuclear Spectroscopic Telescope Array (NuSTAR) extragalactic surveys. NuSTAR has now resolved 33%-39% of the X-ray background in the 8-24 keV band, directly identifying AGNs with obscuring columns up to ˜ {10}25 {{cm}}-2. In the softer 3-8 keV band the number counts are in general agreement with those measured by XMM-Newton and Chandra over the flux range 5× {10}-15 ≲ S(3-8 keV)/{erg} {{{s}}}-1 {{cm}}-2 ≲ {10}-12 probed by NuSTAR. In the hard 8-24 keV band NuSTAR probes fluxes over the range 2× {10}-14 ≲ S(8-24 keV)/{erg} {{{s}}}-1 {{cm}}-2 ≲ {10}-12, a factor ˜100 fainter than previous measurements. The 8-24 keV number counts match predictions from AGN population synthesis models, directly confirming the existence of a population of obscured and/or hard X-ray sources inferred from the shape of the integrated cosmic X-ray background. The measured NuSTAR counts lie significantly above simple extrapolation with a Euclidian slope to low flux of the Swift/BAT 15-55 keV number counts measured at higher fluxes (S(15-55 keV) ≳ 10-11 {erg} {{{s}}}-1 {{cm}}-2), reflecting the evolution of the AGN population between the Swift/BAT local (z\\lt 0.1) sample and NuSTAR’s z˜ 1 sample. CXB synthesis models, which account for AGN evolution, lie above the Swift/BAT measurements, suggesting that they do not fully capture the evolution of obscured AGNs at low redshifts.

  9. X-Ray Monitoring of GRBs with Lobster Eye Telescopes

    SciTech Connect

    Sveda, L.; Pina, L.; Hudec, R.; Inneman, A.; Pizzichini, G.

    2004-09-28

    We present here the soft X-ray All-Sky Monitor (ASM). It is based on the current technological capabilities, sensitive in the {approx} 0.1 - 10.0 keV range with angular resolution of {approx} 3 - 4 arcmin, and has a limiting detectable flux {approx} 10-12 erg/s/cm2 for daily scans in the mentioned energy range. The ASM will play a key role in studying transient X-ray sources like XRBs, GRBs, XRFs, X-ray novae, as well as in the study of the long term variability of X-ray sources like XRBs, AGN, or stellar X-ray flares.

  10. The BMW-Chandra Serendipitous Source Catalog

    NASA Astrophysics Data System (ADS)

    Romano, P.; Campana, S.; Mignani, R. P.; Moretti, A.; Mottini, M.; Panzera, M. R.; Tagliaferri, G.

    2004-08-01

    We present the BMW-Chandra source catalog drawn from all Chandra ACIS-I pointed observations with an exposure time in excess of 10 ks public as of March 2003 (136 observations). Using the wavelet detection algorithm developed by Lazzati et al. (1999) and Campana et al. (1999), which can characterize point-like as well as extended sources, we identified 21325 sources. Among them, 16758 are serendipitous, i.e. not associated with the targets of the pointings, and do not require a non-automated analysis. This makes our catalog the largest compilation of Chandra sources to date. The 0.5--10 keV absorption corrected fluxes of these sources range from ˜ 3× 10-16 to 9×10-12 erg cm-2 s-1 with a median of 7× 10-15 erg cm-2 s-1. The catalog consists of count rates and relative errors in three energy bands (total, 0.5--7 keV; soft, 0.5--2 keV; and hard band, 2--7 keV), and source positions relative to the highest signal-to-noise detection among the three bands. The wavelet algorithm also provides an estimate of the extension of the source which we refined with a σ -clipping method. We report on the main properties of the sources in our catalog, such as sky coverage ( ˜ 8 deg2 at a limiting flux of ˜ 10-13 erg cm-2 s-1) and cosmological log N--log S for a subset at high Galactic latitude (∣ b ∣ > 20o) for a flux as low as ˜ 1.5 × 10-15 erg cm-2 s-1. Support for this work was provided by the Italian MIUR.

  11. The Brera Multi-scale Wavelet HRI Cluster Survey. I. Selection of the sample and number counts

    NASA Astrophysics Data System (ADS)

    Moretti, A.; Guzzo, L.; Campana, S.; Lazzati, D.; Panzera, M. R.; Tagliaferri, G.; Arena, S.; Braglia, F.; Dell'Antonio, I.; Longhetti, M.

    2004-12-01

    We describe the construction of the Brera Multi-scale Wavelet (BMW) HRI Cluster Survey, a deep sample of serendipitous X-ray selected clusters of galaxies based on the ROSAT HRI archive. This is the first cluster catalog exploiting the high angular resolution of this instrument. Cluster candidates are selected on the basis of their X-ray extension only, a parameter which is well measured by the BMW wavelet detection algorithm. The survey includes 154 candidates over a total solid angle of ˜160 deg2 at 10-12 erg s-1 cm-2 and ˜80 deg2 at 1.8×10-13 erg s-1 cm-2. At the same time, a fairly good sky coverage in the faintest flux bins (3-5 × 10-14 erg s-1 cm-2) gives this survey the capability of detecting a few clusters with z˜ 1-1.2, depending on evolution. We present the results of extensive Monte Carlo simulations, providing a complete statistical characterization of the survey selection function and contamination level. We also present a new estimate of the surface density of clusters of galaxies down to a flux of 3× 10-14 erg s-1 cm-2, which is consistent with previous measurements from PSPC-based samples. Several clusters with redshifts up to z=0.92 have already been confirmed, either by cross-correlation with existing PSPC surveys or from early results of an ongoing follow-up campaign. Overall, these results indicate that the excellent HRI PSF (5 arcsec FWHM on axis) more than compensates for the negative effect of the higher instrumental background on the detection of high-redshift clusters. In addition, it allows us to detect compact clusters that could be lost at lower resolution, thus potentially providing an important new insight into cluster evolution. Partially based on observations taken at ESO and TNG telescopes.

  12. Benchmarking the ERG valve tip and MRI Interventions Smart Flow neurocatheter convection-enhanced delivery system's performance in a gel model of the brain: employing infusion protocols proposed for gene therapy for Parkinson's disease.

    PubMed

    Sillay, Karl; Schomberg, Dominic; Hinchman, Angelica; Kumbier, Lauren; Ross, Chris; Kubota, Ken; Brodsky, Ethan; Miranpuri, Gurwattan

    2012-04-01

    Convection-enhanced delivery (CED) is an advanced infusion technique used to deliver therapeutic agents into the brain. CED has shown promise in recent clinical trials. Independent verification of published parameters is warranted with benchmark testing of published parameters in applicable models such as gel phantoms, ex vivo tissue and in vivo non-human animal models to effectively inform planned and future clinical therapies. In the current study, specific performance characteristics of two CED infusion catheter systems, such as backflow, infusion cloud morphology, volume of distribution (mm(3)) versus the infused volume (mm(3)) (Vd/Vi) ratios, rate of infusion (µl min(-1)) and pressure (mmHg), were examined to ensure published performance standards for the ERG valve-tip (VT) catheter. We tested the hypothesis that the ERG VT catheter with an infusion protocol of a steady 1 µl min(-1) functionality is comparable to the newly FDA approved MRI Interventions Smart Flow (SF) catheter with the UCSF infusion protocol in an agarose gel model. In the gel phantom models, no significant difference was found in performance parameters between the VT and SF catheter. We report, for the first time, such benchmark characteristics in CED between these two otherwise similar single-end port VT with stylet and end-port non-stylet infusion systems. Results of the current study in agarose gel models suggest that the performance of the VT catheter is comparable to the SF catheter and warrants further investigation as a tool in the armamentarium of CED techniques for eventual clinical use and application.

  13. Benchmarking the ERG valve tip and MRI Interventions Smart Flow neurocatheter convection-enhanced delivery system's performance in a gel model of the brain: employing infusion protocols proposed for gene therapy for Parkinson's disease

    NASA Astrophysics Data System (ADS)

    Sillay, Karl; Schomberg, Dominic; Hinchman, Angelica; Kumbier, Lauren; Ross, Chris; Kubota, Ken; Brodsky, Ethan; Miranpuri, Gurwattan

    2012-04-01

    Convection-enhanced delivery (CED) is an advanced infusion technique used to deliver therapeutic agents into the brain. CED has shown promise in recent clinical trials. Independent verification of published parameters is warranted with benchmark testing of published parameters in applicable models such as gel phantoms, ex vivo tissue and in vivo non-human animal models to effectively inform planned and future clinical therapies. In the current study, specific performance characteristics of two CED infusion catheter systems, such as backflow, infusion cloud morphology, volume of distribution (mm3) versus the infused volume (mm3) (Vd/Vi) ratios, rate of infusion (µl min-1) and pressure (mmHg), were examined to ensure published performance standards for the ERG valve-tip (VT) catheter. We tested the hypothesis that the ERG VT catheter with an infusion protocol of a steady 1 µl min-1 functionality is comparable to the newly FDA approved MRI Interventions Smart Flow (SF) catheter with the UCSF infusion protocol in an agarose gel model. In the gel phantom models, no significant difference was found in performance parameters between the VT and SF catheter. We report, for the first time, such benchmark characteristics in CED between these two otherwise similar single-end port VT with stylet and end-port non-stylet infusion systems. Results of the current study in agarose gel models suggest that the performance of the VT catheter is comparable to the SF catheter and warrants further investigation as a tool in the armamentarium of CED techniques for eventual clinical use and application.

  14. Ion Irradiation Experiments on the Murchison CM2 Carbonaceous Chondrite: Simulating Space Weathering of Primitive Asteroids

    NASA Technical Reports Server (NTRS)

    Keller, L. P.; Christoffersen, R.; Dukes, C. A.; Baragiola, R. A.; Rahman, Z.

    2015-01-01

    Remote sensing observations show that space weathering processes affect all airless bodies in the Solar System to some degree. Sample analyses and lab experiments provide insights into the chemical, spectroscopic and mineralogic effects of space weathering and aid in the interpretation of remote- sensing data. For example, analyses of particles returned from the S-type asteroid Itokawa by the Hayabusa mission revealed that space-weathering on that body was dominated by interactions with the solar wind acting on LL ordinary chondrite-like materials [1, 2]. Understanding and predicting how the surface regoliths of primitive carbonaceous asteroids respond to space weathering processes is important for future sample return missions (Hayabusa 2 and OSIRIS-REx) that are targeting objects of this type. Here, we report the results of our preliminary ion irradiation experiments on a hydrated carbonaceous chondrite with emphasis on microstructural and infrared spectral changes.

  15. Coordinated Chemical and Isotopic Imaging of Bells (CM2) Meteorite Matrix

    NASA Technical Reports Server (NTRS)

    Clemett, S. J.; Messenger, S.; Naklamura-Messenger, K.; Thomas-Keprta, K. L.

    2014-01-01

    Meteoritic organic matter is a complex conglomeration of species formed in distinct environments and processes in circumstellar space, the interstellar medium, the Solar Nebula and asteroids. Consequently meteorites constitute a unique record of primordial organic chemical evolution. While bulk chemical analysis has provided a detailed description of the range and diversity of organic species present in carbonaceous chondrites, there is little information as to how these species are spatially distributed and their relationship to the host mineral matrix. The distribution of organic phases is nevertheless critical to understanding parent body processes. The CM and CI chondrites all display evidence of low temperature (< 350K) aqueous alteration that may have led to aqueous geochromatographic separation of organics and synthesis of new organics coupled to aqueous mineral alteration. Here we present the results of the first coordinated in situ isotopic and chemical mapping study of the Bells meteorite using a newly developed two-step laser mass spectrometer (mu-L(sup 2)MS) capable of measuring a broad range of organic compounds.

  16. W/Z production cross sections and asymmetries at E(CM) = 2-TeV

    SciTech Connect

    Bellavance, Angela M.; /Nebraska U.

    2005-06-01

    The most recent results for W and Z boson production cross sections and asymmetries are presented from the CDF and D0 collaborations using Run II data taken at the Fermi National Accelerator Laboratory (FNAL) Tevatron. Data set sizes range from 72 pb{sup -1} to 226 pb{sup -1}, and results range from published to preliminary. Results presented agree with the Standard Model and world averages within errors.

  17. Design of a 10**36 CM-2 S-1 Super-B Factory

    SciTech Connect

    Biagini, M.E.; Boni, R.; Boscolo, M.; Demma, T.; Drago, A.; Guiducci, S.; Raimondi, P.; Tomassini, S.; Zobov, M.; Bertsche, Kirk J.; Novokhatski, A.; Seeman, J.; Sullivan, M.; Wienands, U.; Wittmer, W.; Bettoni, S.; Paoloni, E.; Marchiori, G.; Bogomyagkov, A.; Koop, I.; Levichev, E.; /Novosibirsk, IYF

    2011-10-24

    Parameters have been studied for a high luminosity e{sup +}e{sup -} collider operating at the Upsilon 4S that would deliver a luminosity of 1 to 4 x 10{sup 36}/cm{sup 2}/s. This collider, called a Super-B Factory, would use a combination of linear collider and storage ring techniques. In this scheme an electron beam and a positron beam are stored in low-emittance damping rings similar to those designed for a Linear Collider (LC) or the next generation light source. A LC style interaction region is included in the ring to produce sub-millimeter vertical beta functions at the collision point. A large crossing angle (+/- 24 mrad) is used at the collision point to allow beam separation. A crab-waist scheme is used to reduce the hourglass effect and restore peak luminosity. Beam currents of 1.8 A at 4 x 7 GeV in 1251 bunches can produce a luminosity of 10{sup 36}/cm{sup 2}/s with upgrade possibilities. Such a collider would produce an integrated luminosity of about 10,000 fb{sup -1} (10 ab{sup -1}) in a running year (10{sup 7} sec) at the {gamma}(4S) resonance. Further possibilities include having longitudinally polarized e- at the IR and operating at the J/Psi and Psi beam energies.

  18. 44 CFR 10.12 - Pre-implementation actions.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... shall provide further guidance, commensurate with their programs and organization, for integration of... information, upon request, as specified in 40 CFR 1505.3. This does not, however, include standing or...

  19. 44 CFR 10.12 - Pre-implementation actions.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... Pre-implementation actions. (a) Decision-making. The Regional Administrator shall ensure that... integrated into the decision-making process. Because of the diversity of FEMA, it is not feasible to describe in this part the decision-making process for each of the various FEMA programs. Proposals and...

  20. 44 CFR 10.12 - Pre-implementation actions.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... Pre-implementation actions. (a) Decision-making. The Regional Administrator shall ensure that... integrated into the decision-making process. Because of the diversity of FEMA, it is not feasible to describe in this part the decision-making process for each of the various FEMA programs. Proposals and...

  1. 44 CFR 10.12 - Pre-implementation actions.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... Pre-implementation actions. (a) Decision-making. The Regional Administrator shall ensure that... integrated into the decision-making process. Because of the diversity of FEMA, it is not feasible to describe in this part the decision-making process for each of the various FEMA programs. Proposals and...

  2. Modern Europe: An Instructional Guide, Grades 10-12.

    ERIC Educational Resources Information Center

    Los Angeles City Schools, CA. Div. of Instructional Planning and Services.

    Emphasizing Europe's role in the growth of Western institutions and ideals, this guide stresses the development of skills necessary for active participation in democratic processes. An introductory unit on the Renaissance and the Reformation is followed by units on (1) the political events fostering the growth of the modern national state and the…

  3. Business Education--Secretarial Skills. Grades 10-12.

    ERIC Educational Resources Information Center

    Instructional Objectives Exchange, Los Angeles, CA.

    The secretarial skills collection is composed of 47 objectives and related test items designed for business education courses in grades 10 through 12. Objectives are organized into the following categories: (1) typing; (2) shorthand; (3) office machines; (4) office practice; and (5) business English and communication. Each objective contains three…

  4. Revised Guidelines for Comprehensive Health Education, Grades 10-12.

    ERIC Educational Resources Information Center

    Nebraska State Dept. of Education, Lincoln. Div. of Instructional Services.

    These health curriculum guidelines were designed for teachers of secondary students. Four major topic areas are covered: 1) anatomy, physiology, tissues, systems, heredity, physical activity, and nutriition; 2) family structures, functions, and responsibilities; 3) values, stress, and drug abuse; and 4) the environment, disease control, cancer,…

  5. The Project Physics Course (Modularized) for Grades 10-12.

    ERIC Educational Resources Information Center

    Flint, William

    This report was produced by the Sedro-Woolley Project which has the goal of infusing environmental education into the whole curriculum of a school district. Included are assumptions which the author believes are appropriate to environmental education; a relating of these assumptions to some topics of chemistry and physics; an outline of specific…

  6. General Math 10-12 [Instructional Objectives Exchange].

    ERIC Educational Resources Information Center

    California Univ., Los Angeles. Center for the Study of Evaluation.

    This collection of 123 objectives and related evaluation items is for general mathematics in grades 10 through 12. The content has been organized into nine major categories: sets; numbers, numerals, and numeration systems; operations and their properties; measurements; per cents; geometry; probability and statistics; logic; and applications and…

  7. Business Education--Bookkeeping. Grades 10-12.

    ERIC Educational Resources Information Center

    Instructional Objectives Exchange, Los Angeles, CA.

    This collection contains 17 objectives and related test items for bookkeeping, grades ten through twelve. The first ten objectives represent the bookkeeping cycle: journalizing, posting, preparing a trial balance, preparing a financial statement from the worksheet, making closing entries, balancing and ruling accounts, and preparing and…

  8. 44 CFR 10.12 - Pre-implementation actions.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... decision. In those cases requiring environmental impact statements, the Regional Administrator at the time... decision and describes the alternatives and relevant factors considered as specified in 40 CFR 1505.2. The... will be identified in the environmental impact statement and made available to...

  9. Microwave differential dilatometer measures 10 - 12 m, at 1 Hz

    NASA Astrophysics Data System (ADS)

    Aschero, G.; Mango, F.; Gizdulich, P.

    1996-12-01

    To check and measure the converse piezoelectric effect in bone samples, we had to detect displacements in the range of 1-100 pm with three kinds of restrictions: (1) the biological nature of the samples imposes severe limits in selecting a suitable device and method; (2) such a method has to take into account some clinical applications to which the experiment is devoted; (3) the piezoelectric behavior of bone samples is particularly interesting at low frequencies, around 1 Hz. For such reasons we modified an existing dilatometer based on a microwave differential spectrometer. A 14 GHz klystron, linearly modulated in frequency by a triangular 50 Hz voltage applied to the repeller, is connected, via magic T, to two identical cavities tunable around 14 GHz and whose resonance curves are recorded by crystal detectors. When one of the two cavities changes its height according to the length variations of the sample, its resonance frequency varies resulting in a shift of the resonant curve with respect to the resonance curve of the other cavity acting as reference. The comparison between the cavities' responses is performed by a pulse technique transforming the frequency shifts into time intervals, that are then converted into dc voltages. The differential character of this measurement avoids the need for the microwave source stabilization. The relative shift in frequency is measured with an accuracy better than 500 Hz. This accuracy allows us to measure displacements smaller than 7 nm in the cavity's height. After 2 h of warmup, thanks to the differential arrangement of the system, thermal or other drifts are not detectable within a lapse of time of 12 h. This feature allows coherent signal averaging over long periods. With a piezoelectric ceramic stack moving 100 pm in square wave fashion at 50 mHz we found that the signal to noise ratio was 20 dB after 1000 cycles of signal averaging, when our bandpass filter was tuned at 1 Hz. In conclusion, this system can detect periodic displacements as small as 1 pm in a short time and reliably. Due to the operational simplicity and stability, at room temperature and humidity, the device is suitable for dilatometric measurements on biological samples.

  10. INTEGRAL/IBIS 7-year All-Sky Hard X-ray Survey. I. Image reconstruction

    NASA Astrophysics Data System (ADS)

    Krivonos, R.; Revnivtsev, M.; Tsygankov, S.; Sazonov, S.; Vikhlinin, A.; Pavlinsky, M.; Churazov, E.; Sunyaev, R.

    2010-09-01

    This paper is the first in a series devoted to the hard X-ray whole sky survey performed by the INTEGRAL observatory over seven years. Here we present an improved method for image reconstruction with the IBIS coded mask telescope. The main improvements are related to the suppression of systematic effects that strongly limit sensitivity in the region of the Galactic plane (GP), especially in the crowded field of the Galactic center (GC). We extended the IBIS/ISGRI background model to take into account the Galactic ridge X-ray emission (GRXE). To suppress residual systematic artifacts on a reconstructed sky image, we applied nonparametric sky image filtering based on wavelet decomposition. The implemented modifications of the sky reconstruction method decrease the systematic noise in the ~20 Ms deep field of GC by ~44%, and practically remove it from the high-latitude sky images. New observational data sets, along with an improved reconstruction algorithm, allow us to conduct the hard X-ray survey with the best currently available minimal sensitivity 3.7 × 10-12 erg s-1 cm-2 ~ 0.26 mCrab in the 17-60 keV band at a 5σ detection level. The survey covers 90% of the sky down to the flux limit of 6.2 × 10-11 erg s-1 cm-2 (~4.32 mCrab) and 10% of the sky area down to the flux limit of 8.6 × 10-12 erg s-1 cm-2 (~0.60 mCrab). Based on observations with INTEGRAL, an ESA project with the instruments and science data center funded by ESA member states (especially the PI countries: Denmark, France, Germany, Italy, Switzerland, Spain), Czech Republic, and Poland, and with the participation of Russia and the USA.

  11. Retinal architecture and mfERG

    PubMed Central

    Schnurman, Zane S.; Frohman, Teresa C.; Beh, Shin C.; Conger, Darrel; Conger, Amy; Saidha, Shiv; Galetta, Steven; Calabresi, Peter A.; Green, Ari J.

    2014-01-01

    Objective: To describe a novel neurophysiologic signature of the retinal ganglion cell and to elucidate its relationship to abnormalities in validated structural and functional measures of the visual system. Methods: We used multifocal electroretinogram–generated optic nerve head component (ONHC) responses from normal subjects (n = 18), patients with multiple sclerosis (MS) (n = 18), and those with glaucoma (n = 3). We then characterized the relationship between ONHC response abnormalities and performance on low-contrast visual acuity, multifocal visual-evoked potential–induced cortical responses, and average and quadrant retinal nerve fiber layer (RNFL) thicknesses, as measured by spectral-domain optical coherence tomography. Results: Compared with the eyes of normal subjects, the eyes of patients with MS exhibited an increased number of abnormal or absent ONHC responses (p < 0.0001). For every 7-letter reduction in low-contrast letter acuity, there were corresponding 4.6 abnormal ONHC responses at 2.5% contrast (p < 0.0001) and 6.6 abnormalities at the 1.25% contrast level (p < 0.0001). Regarding average RNFL thickness, for each 10-μm thickness reduction, we correspondingly observed 6.8 abnormal ONHC responses (p = 0.0002). The most robust association was between RNFL thinning in the temporal quadrant and ONHC response abnormalities (p < 0.0001). Conclusion: Further characterization of ONHC abnormalities (those that are reversible and irreversible) may contribute to the development of novel neurotherapeutic strategies aimed at achieving neuroprotective, and perhaps even neurorestorative, effects in disorders that target the CNS in general, and MS in particular. PMID:24789865

  12. Discovery of 5-Chloro-1-(5-chloro-2-(methylsulfonyl)benzyl)-2-imino-1,2-dihydropyridine-3-carboxamide (TAK-259) as a Novel, Selective, and Orally Active α1D Adrenoceptor Antagonist with Antiurinary Frequency Effects: Reducing Human Ether-a-go-go-Related Gene (hERG) Liabilities.

    PubMed

    Sakauchi, Nobuki; Kohara, Yasuhisa; Sato, Ayumu; Suzaki, Tomohiko; Imai, Yumi; Okabe, Yuichi; Imai, Shigemitsu; Saikawa, Reiko; Nagabukuro, Hiroshi; Kuno, Haruhiko; Fujita, Hisashi; Kamo, Izumi; Yoshida, Masato

    2016-04-14

    A novel structural class of iminopyridine derivative 1 was identified as a potent and selective human α1D adrenoceptor (α1D adrenergic receptor; α1D-AR) antagonist against α1A- and α1B-AR through screening of an in-house compound library. From initial structure-activity relationship studies, we found lead compound 9m with hERG K(+) channel liability. To develop analogues with reduced hERG K(+) channel inhibition, a combination of site-directed mutagenesis and docking studies was employed. Further optimization led to the discovery of (R)-9s and 9u, which showed antagonistic activity by a bladder strip test in rats with bladder outlet obstruction, as well as ameliorated cystitis-induced urinary frequency in rats. Ultimately, 9u was selected as a clinical candidate. This is the first study to show the utility of iminopyridine derivatives as selective α1D-AR antagonists and evaluate their effects in vivo.

  13. The Fermi-LAT view of the colliding wind binaries

    NASA Astrophysics Data System (ADS)

    Pshirkov, M. S.

    2016-03-01

    Colliding wind binaries (CWBs) have been considered as a possible high-energy γ-ray sources for some time, however no system other than η Car has been detected. In the Letter, a sample of seven CWBs (WR 11, WR 70, WR 137, WR 140, WR 146, WR 147) which, by means of theoretic modelling, were deemed most promising candidates, was analysed using almost 7 yr of the Fermi-Large Area Telescope (LAT) data. WR 11 (γ2 Vel) was detected at 6.1σ confidence level with a photon flux in 0.1-100 GeV range (1.8 ± 0.6) × 10-9 ph cm-2 s-1 and an energy flux (2.7 ± 0.5) × 10-12 erg cm-2 s-1. At the adopted distance d = 340 pc this corresponds to a luminosity L = (3.7 ± 0.7) × 1031 erg s-1. This luminosity amounts to ˜6 × 10-6 fraction of the total wind kinetic power and ˜1.6 × 10-4 fraction of the power injected into the wind-wind interaction region of this system. Upper limits were set on the high energy flux from the WR 70 and WR 140 systems.

  14. The Brera multi-scale wavelet Chandra survey. I. Serendipitous source catalogue

    NASA Astrophysics Data System (ADS)

    Romano, P.; Campana, S.; Mignani, R. P.; Moretti, A.; Mottini, M.; Panzera, M. R.; Tagliaferri, G.

    2008-09-01

    We present the BMW-Chandra source catalogue drawn from essentially all Chandra ACIS-I pointed observations with an exposure time in excess of 10 ks public as of March 2003 (136 observations). Using the wavelet detection algorithm developed by Lazzati et al. (1999) and Campana et al. (1999), which can characterise both point-like and extended sources, we identified 21 325 sources. Among them, 16 758 are serendipitous, i.e. not associated with the targets of the pointings, and do not require a non-automated analysis. This makes our catalogue the largest compilation of Chandra sources to date. The 0.5-10 keV absorption corrected fluxes of these sources range from ˜3× 10-16 to 9×10-12 erg cm-2 s-1 with a median of 7× 10-15 erg cm-2 s-1. The catalogue consists of count rates and relative errors in three energy bands (total, 0.5-7 keV; soft, 0.5-2 keV; and hard, 2-7 keV), and source positions relative to the highest signal-to-noise detection among the three bands. The wavelet algorithm also provides an estimate of the extension of the source. We include information drawn from the headers of the original files, as well, and extracted source counts in four additional energy bands, SB1 (0.5-1 keV), SB2 (1-2 keV), HB1 (2-4 keV), and HB2 (4-7 keV). We computed the sky coverage for the full catalogue and for a subset at high Galactic latitude (mid b mid > 20°). The complete catalogue provides a sky coverage in the soft band (0.5-2 keV, S/N =3) of 8 deg2 at a limiting flux of ˜10-13 erg cm-2 s-1, and 2 deg2 at a limiting flux of ˜10-15 erg cm-2 s-1. Furthermore, we present the results of the cross-match with existing catalogues at different wavelengths (FIRST, IRAS, 2MASS, GSC2, and ChaMP). The total numbers of matches with the FIRST, IRASPSC, 2MASS, and GSC2 catalogues obtained after a closest-distance selection are 13, 87, 6700, and 4485, respectively. The catalogue is only available in electronic form at the CDS via anonymous ftp to cdsarc.u-strasbg.fr (130

  15. Key comparison BIPM.RI(I)-K6 of the standards for absorbed dose to water at 5 g cm-2 and 7 g cm-2 of the NPL, United Kingdom and the BIPM in accelerator photon beams

    NASA Astrophysics Data System (ADS)

    Picard, S.; Burns, D. T.; Roger, P.; Duane, S.; Bass, G. A.; Manning, J. W.; Shipley, D. R.

    2015-01-01

    A comparison of the dosimetry for accelerator photon beams was carried out between the National Physical Laboratory (NPL) and the Bureau International des Poids et Mesures (BIPM) in two periods from September to November 2013. The comparison was based on the determination of absorbed dose to water for three radiation qualities at the NPL. The results, reported as ratios of the NPL and the BIPM evaluations (and with the combined standard uncertainties given in parentheses), are 0.9973(62) at 6 MV, 0.9995(66) at 10 MV and 0.9957(81) at 25 MV. This result is the sixth in the on-going BIPM.RI(I)-K6 series of comparisons. Main text. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCRI, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

  16. Fine-Grained Rims in the Allan Hills 81002 and Lewis Cliff 90500 CM2 Meteorites: Their Origin and Modification

    NASA Technical Reports Server (NTRS)

    Hua, X.; Wang, J.; Buseck, P. R.

    2002-01-01

    Antarctic CM meteorites Allan Hills (ALH) 8 1002 and Lewis Cliff (LEW) 90500 contain abundant fine-grained rims (FGRs) that surround a variety of coarse-grained objects. FGRs from both meteorites have similar compositions and petrographic features, independent of their enclosed objects. The FGRs are chemically homogeneous at the 10 m scale for major and minor elements and at the 25 m scale for trace elements. They display accretionary features and contain large amounts of volatiles, presumably water. They are depleted in Ca, Mn, and S but enriched in P. All FGRs show a slightly fractionated rare earth element (REE) pattern, with enrichments of Gd and Yb and depletion of Er. Gd is twice as abundant as Er. Our results indicate that those FGRs are not genetically related to their enclosed cores. They were sampled from a reservoir of homogeneously mixed dust, prior to accretion to their parent body. The rim materials subsequently experienced aqueous alteration under identical conditions. Based on their mineral, textural, and especially chemical similarities, we conclude that ALH 8 1002 and LEW 90500 likely have a similar or identical source.

  17. Massively parallel hypercube FFTs: CM-2 implementation and error analysis of a parallel trigonometric factor generation method

    SciTech Connect

    Tong, C.H. ); Swarztrauber, P.N. )

    1991-08-01

    On parallel computers, the way the data elements are mapped to the processors may have a large effect on the timing performance of a given algorithm. In our previous paper, we have examined a few mapping strategies for the ordered radix-2 DIF (decimation-in-frequency) Fast Fourier Transform. In particular, we have shown how reduction of communication can be achieved by combining the order and computational phases through the use of i-cycles. A parallel methods was also presented for computing the trigonometric factors which requires neither trigonometric function evaluation nor interprocessor communication. This paper first reviews some of the experimental results on the Connection Machine to demonstrate the importance of reducing communication in a parallel algorithm. The emphasis of this paper, however, is on analyzing the numerical stability of the proposed method for generating the trigonometric factors and showing how the error can be improved. 16 refs., 12 tabs.

  18. Fluorescent imaging over an ultra-large field-of-view of 532 cm2 using a flatbed scanner

    NASA Astrophysics Data System (ADS)

    Göröcs, Zoltán. S.; Ling, Yuye; Yu, Meng D.; Karahalios, Dimitri; Mogharabi, Kian; Lu, Kenny; Wei, Qingshan; Ozcan, Aydogan

    2014-03-01

    We introduce a fluorescent imaging method that is capable of detecting fluorescent micro-particles over an ultra-wide field of view of 19 cm × 28 cm using a modified flatbed scanner. We added a custom-designed absorbing emission filter, a computer controlled two dimensional LED array, and modified the driver of the scanner to maximize the sensitivity, exposure time, and gain for fluorescent detection of micro-objects. This high-throughput fluorescent imaging device used in conjunction with a microfluidic sample holder enables rapid screening of fluorescent micro-objects inside more than 2.2mL of optically dense media (i.e., whole blood) within 5 minutes. The device is sensitive enough to detect fluorescently labeled cells, and generates images that have an effective pixel count of 2.2 Giga-pixels.

  19. 53Mn-53Cr dating of aqueously formed carbonates in the CM2 lithology of the Sutter's Mill carbonaceous chondrite

    NASA Astrophysics Data System (ADS)

    Jilly, Christine E.; Huss, Gary R.; Krot, Alexander N.; Nagashima, Kazuhide; Yin, Qing-Zhu; Sugiura, Naoji

    2014-11-01

    Radiometric dating of secondary minerals can be used to constrain the timing of aqueous alteration on meteoritic parent bodies. Dolomite is a well-documented secondary mineral in CM chondrites, and is thought to have formed by precipitation from an aqueous fluid on the CM parent body within several million years of accretion. The petrographic context of crosscutting dolomite veins indicates that aqueous alteration occurred in situ, rather than in the nebular setting. Here, we present 53Mn-53Cr systematics for dolomite grains in Sutter's Mill section SM51-1. The Mn-Cr isotope data show well-resolved excesses of 53Cr correlated with 55Mn/52Cr ratio, which we interpret as evidence for the in situ decay of radioactive 53Mn. After correcting for the relative sensitivities of Mn and Cr using a synthetic Mn- and Cr-bearing calcite standard, the data yield an isochron with slope corresponding to an initial 53Mn/55Mn ratio of 3.42 ± 0.86 × 10-6. The reported error includes systematic uncertainty from the relative sensitivity factor. When calculated relative to the U-corrected Pb-Pb absolute age of the D'Orbigny angrite, Sutter's Mill dolomites give a formation age between 4564.8 and 4562.2 Ma (2.4-5.0 Myr after the birth of the solar system). This age is contemporaneous with previously reported ages for secondary carbonates in CM and CI chondrites. Consistent carbonate precipitation ages between the carbonaceous chondrite groups suggest that aqueous alteration was a common process during the early stages of parent body formation, probably occurring via heating from internal 26Al decay. The high-precision isochron for Sutter's Mill dolomite indicates that late-stage processing did not reach temperatures that were high enough to further disturb the Mn-Cr isochron.

  20. Compositional Zoning and Mn-Cr Systematics in Carbonates from the Y791198 CM2 Carbonaceous Chondrite

    NASA Technical Reports Server (NTRS)

    Brearley, Adrian J.; Hutcheon, Ian D.; Browning, Lauren

    2001-01-01

    Cathodoluminescence and microprobe analyses show that carbonates in Y791198 exhibit complex zoning. Cr-Mn dating suggests formation of carbonates 10 Ma after CAI formation Additional information is contained in the original extended abstract..

  1. Spectral characteristics of iron-bearing phyllosilicates: Comparison to Orgueil (CI1), Murchison and Murray (CM2)

    USGS Publications Warehouse

    Calvin, W.M.; King, T.V.V.

    1997-01-01

    Phyllosilicate alteration minerals are commonly found in low petrologic types of carbonaceous chondrites. Previous spectral studies have examined Mg-bearing phyllosilicates with limited success in matching the spectral properties of CM and CI chondrites. Transmission electron microscope and other analytical techniques suggest that Fe-bearing clays are more abundant in CI and CM chondrites than magnesian varieties. Here, we present the results of an examination of the reflectance spectra of Fe-phyllosilicates, including serpentines and berthierines, of which the latter were formerly known as septechlorites. We have measured the diffuse reflectance spectra of powdered samples from 0.3 to 25 ??m. We find that these minerals provide a better spectral match to many of the features seen in CI and CM chondrites, and simple linear combinations of the spectra of both Fe- and Mg-phyllosilicates closely approximate the spectra of CM and CI chondrites.

  2. TeV gamma rays from the blazar H 1426+428 and the diffuse extragalactic background radiation

    NASA Astrophysics Data System (ADS)

    Aharonian, F.; Akhperjanian, A.; Barrio, J.; Beilicke, M.; Bernlöhr, K.; Börst, H.; Bojahr, H.; Bolz, O.; Contreras, J.; Cornils, R.; Cortina, J.; Denninghoff, S.; Fonseca, V.; Girma, M.; Gonzalez, J.; Götting, N.; Heinzelmann, G.; Hermann, G.; Heusler, A.; Hofmann, W.; Horns, D.; Jung, I.; Kankanyan, R.; Kestel, M.; Kettler, J.; Kohnle, A.; Konopelko, A.; Kornmeyer, H.; Kranich, D.; Krawczynski, H.; Lampeitl, H.; Lopez, M.; Lorenz, E.; Lucarelli, F.; Magnussen, N.; Mang, O.; Meyer, H.; Mirzoyan, R.; Moralejo, A.; Ona, E.; Padilla, L.; Panter, M.; Plaga, R.; Plyasheshnikov, A.; Pühlhofer, G.; Rauterberg, G.; Röhring, A.; Rhode, W.; Robrade, J.; Rowell, G.; Sahakian, V.; Samorski, M.; Schilling, M.; Schröder, F.; Sevilla, I.; Siems, M.; Stamm, W.; Tluczykont, M.; Völk, H. J.; Wiedner, C. A.; Wittek, W.

    2002-03-01

    The detection of TeV gamma -rays from the blazar H 1426+428 at an integral flux level of (4 +/- 2stat +/- 1syst) x 10-12 erg cm-2 s-1 above 1 TeV with the HEGRA imaging atmospheric Cherenkov telescope system is reported. H 1426+428 is located at a redshift of z = 0.129, which makes it the most distant source detected in TeV gamma -rays so far. The TeV radiation is expected to be strongly absorbed by the diffuse extragalactic background radiation (DEBRA). The observed energy spectrum of TeV photons is in good agreement with an intrinsic power law spectrum of the source ~ E-1.9 corrected for DEBRA absorption. Statistical errors as well as uncertainties about the intrinsic source spectrum, however, do not permit strong statements about the density of the DEBRA infrared photon field.

  3. A Search for Extreme-Ultraviolet Emission from Comets with the Cosmic Hot Interstellar Plasma Spectrometer (CHIPS)

    NASA Astrophysics Data System (ADS)

    Sasseen, T. P.; Hurwitz, M.; Lisse, C. M.; Kharchenko, V.; Christian, D.; Wolk, S. J.; Sirk, M. M.; Dalgarno, A.

    2006-10-01

    We have obtained EUV spectra between 90 and 255 Å of the comets C/2002 T7 (LINEAR), C/2001 Q4 (NEAT), and C/2004 Q2 (Machholz) near their perihelion passages in 2004 with the Cosmic Hot Interstellar Plasma Spectrometer (CHIPS). We obtained contemporaneous data on NEAT with the Chandra ACIS instrument, marking the first simultaneous EUV and X-ray spectral observations of a comet. The total CHIPS/EUV observing times were 337 ks for NEAT, 234 ks for LINEAR, and 483 ks for Machholz, and for both CHIPS and Chandra we calculate we have captured all the comet flux in the instrument field of view. We set upper limits on solar wind charge-exchange emission lines of O, C, N, Ne, and Fe occurring in the spectral bandpass of CHIPS. The spectrum of NEAT obtained with Chandra can be reproduced by modeling emission lines of C, N, O, Mg, Fe, Si, S, and Ne solar wind ions. The measured X-ray emission-line intensities are consistent with our predictions from a solar wind charge-exchange model. The model predictions for the EUV emission-line intensities are determined from the intensity ratios of the cascading X-ray and EUV photons arising in the charge-exchange processes. They are compatible with the measured limits on the intensities of the EUV lines. For NEAT, we measured a total X-ray flux of 3.7×10-12 ergs cm-2 s-1 and derive from model predictions a total EUV flux of 1.5×10-12 ergs cm-2 s-1. The CHIPS observations occurred predominantly while the satellite was on the dayside of Earth. For much of the observing time, CHIPS performed observations at smaller solar angles than it was designed for, and EUV emission from the Sun scattered into the instrument limited the sensitivity of the EUV measurements.

  4. Francais: Programme d'etudes du secondaire deuxieme cycle (10-12). Francais langue premiere (French: High School Curriculum [10-12]. French as a Native Language).