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  1. The Committee met in the Columbia Room at the Holiday Inn

    U.S. Energy Information Administration (EIA) Indexed Site

    Friday, April 21, 1995 - - - The Committee met in the Columbia Room at the Holiday Inn Capitol, 550 C Street S.W., Washington, D.C., at 9:00 a.m., Timothy D. Mount, Chairman, presiding. PRESENT: TIMOTHY D. MOUNT, Chair DAVID R. BELLHOUSE CHARLES W. BISCHOFF BRENDA G. COX FAYE DUCHIN JOHN D. GRACE PHILIP HANSWER CALVIN KENT GRETA M. LJUNG JAMES L. O'BRIEN DANIEL A. RELLES BRADLEY O. SKARPNESS G. CAMPBELL WATKINS A-G-E-N-D-A Page No. Introductory Remarks, TIMOTHY MOUNT, Chairman 3 Announcement of

  2. Annual Review of BPA-Funded Projects in Natural and Artificial Propagation of Salmonids, March 27-29, 1985, Holiday Inn Airport, Portland, Oregon.

    SciTech Connect (OSTI)

    United States. Bonneville Power Administration.

    1985-04-01

    The Fish and Wildlife Division of Bonneville Power Administration (BPA) hosted a meeting for contractors to present the results of fiscal year 1984 research conducted to implement the Northwest Power Planning Council's Fish and Wildlife Program. The meeting focused on those projects specifically related to natural and artificial propagation of salmonids. The presentations were held at the Holiday Inn Airport in Portland, Oregon, on March 27-29, 1985. This document contains abstracts of the presentations from that meeting. Section 1 contains abstracts on artificial propagation, fish health, and downstream migration, and Section 2 contains abstracts on natural propagation and habitat improvement. The abstracts are indexed by BPA Project Number and by Fish and Wildlife Program Measure. The registered attendees at the meeting are listed alphabetically in Appendix A and by affiliation in Appendix B.

  3. Holiday Gift Drive

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Gift Drive Holiday Gift Drive Every year, Laboratory employees help fulfill the holiday wishes of children and seniors in our communities. In 2015, our employees donated more than 1,200 gifts to 23 nonprofit organizations to help Northern New Mexico children, senior citizens, and families have a brighter holiday season. May 7, 2015 Every holiday season, employees of Los Alamos National Laboratory donate and distribute gifts to families in need throughout Northern New Mexico. Contacts Annual Food

  4. Holiday Food Drive

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Food Drive Holiday Food Drive Laboratory employees helped donate 300 boxes of nonperishable food items and 360 frozen turkeys during the 2015 annual food drive. September 16, 2013 LANL employees organize food for the Holiday Food Drive. Contacts Annual Food & Holiday Gift Drives Mike Martinez (505) 699-3388 Community Relations & Partnerships (505) 665-4400 Email Helping feed Northern New Mexico families During the Laboratory's 2015 Annual Food Drive, employees and subcontract workers

  5. NERSC Holiday Schedule

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Holiday Schedule NERSC Holiday Schedule December 20, 2013 by Francesca Verdier All NERSC computing and storage systems will remain in full operation throughout the holiday season (Tuesday December 24 through Wednesday January 1). From Tuesday December 24 through Wednesday January 1, NERSC Consulting and Account Support services will be available *only* on Friday December 27 and Monday December 30, from 8:00 to 5:00 Pacific Time. Normal Consulting and Account Support schedules will resume on

  6. Berkeley Lab Holiday Schedule

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Thursday, January 1, 2015 New Year's Holiday Monday, January 19, 2015 Martin Luther King Day Monday, February 16, 2015 Presidents Day Friday, March 27, 2015 Cesar Chavez Day**...

  7. The Committee convened in the Clark Room, Holiday Inn Capitol...

    U.S. Energy Information Administration (EIA) Indexed Site

    ... On the one hand, you have people like Paul Joskow claiming in different ways that the benefits from restructuring and price competition may, in fact, be minimal. Joseph Stiglitz ...

  8. International Cooperation Holiday Cheer

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Cooperation Holiday Cheer RSL works with allies in major exercise. Beatty residents talk NNSS groundwater. NvE employees celebrated the holidays with host of activities. See page 6. See pages 4-5. NNSS Celebrates Successful 2013 With Major National Security Projects The Nevada National Security Site (NNSS) once again proved that it is at the forefront of National Security efforts with the completion of numerous high- profile projects in 2013 that have paved the way for an ambitious 2014 outlook.

  9. Post-Holiday Holiday Shopping | Department of Energy

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Post-Holiday Holiday Shopping Post-Holiday Holiday Shopping January 10, 2012 - 4:31pm Addthis Elizabeth Spencer Communicator, National Renewable Energy Laboratory Ah, January. It's still cold and it's still dark all the time, but now all the cheerful Christmas lights have come down so there's nothing to break up the monotony. What's there to do? Well, while it's a little late now, you might want to go do some post-holiday holiday shopping. Yes, I know. That sounds insane. Everyone is thoroughly

  10. Museum Closed for Thanksgiving Holiday

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Museum Closed for Thanksgiving Holiday Museum Closed for Thanksgiving Holiday WHEN: Nov 26, 2015 12:00 AM - 11:59 PM WHERE: Bradbury Science Museum 1350 Central Ave, Los Alamos, NM...

  11. Museum Closed for Christmas Holiday

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Museum Closed for Christmas Holiday Museum Closed for Christmas Holiday WHEN: Dec 25, 2015 12:00 AM - 11:59 PM WHERE: Bradbury Science Museum 1350 Central Ave, Los Alamos, NM 87544, USA CATEGORY: Bradbury INTERNAL: Calendar Login Museum Closed for Christmas Holiday Event Description The Bradbury Science Museum will be CLOSED for the Christmas holiday. The Bradbury Science Museum is open to the public every day except for Thanksgiving Day, Christmas Day and New Year's Day. Admission is always

  12. Museum Closed for Thanksgiving Holiday

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Museum Closed for Thanksgiving Holiday Museum Closed for Thanksgiving Holiday WHEN: Nov 26, 2015 12:00 AM - 11:59 PM WHERE: Bradbury Science Museum 1350 Central Ave, Los Alamos, NM 87544 USA CATEGORY: Bradbury INTERNAL: Calendar Login Museum Closed for Thanksgiving Holiday Event Description The museum will be CLOSED for the Thanksgiving holiday. The Bradbury Science Museum is open to the public every day except for Thanksgiving Day, Christmas Day and New Year's Day. Admission is always free.

  13. Leveraging Holidays and Other Events

    Broader source: Energy.gov [DOE]

    Better Buildings Residential Network Driving Demand Peer Exchange Call Series: Leveraging Holidays and Other Events, Call Slides and Discussion Summary, November 7, 2013.

  14. Riverside Inn Pool & Spa Low Temperature Geothermal Facility...

    Open Energy Info (EERE)

    Inn Pool & Spa Low Temperature Geothermal Facility Jump to: navigation, search Name Riverside Inn Pool & Spa Low Temperature Geothermal Facility Facility Riverside Inn Sector...

  15. Michelle L. Holiday | Department of Energy

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Michelle L. Holiday About Us Michelle L. Holiday - President, Michelle Holiday and Associates Michelle L. Holiday Michelle Holiday, an enrolled member of the Iowa Tribe of Oklahoma, has had extensive experience in tribal affairs, government relations, and the energy industry. From 1993 to 2001, She served in customer relations and public affairs roles for Southern California Edison (SCE). In 2002, she became the Senior Manager of Legislative and Native American Affairs for Edison International,

  16. Helping make the holidays happier

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Helping Make The Holidays Happier Community Connections: Your link to news and opportunities from Los Alamos National Laboratory Latest Issue:Mar. 2016 all issues All Issues » submit Helping make the holidays happier This year's LANL food drive collected enough donations to provide 11,600 meals for those in need. January 1, 2013 dummy image Read our archives Contacts Editor Linda Anderman Email Community Programs Office Kurt Steinhaus Email The contributions by employees included 164 turkeys

  17. GovCon Holiday Soiree

    Broader source: Energy.gov [DOE]

    The 6th Annual GovCon Holiday Soiree will be held at the Kennedy Center on Monday, December 8, from 5:30pm to 7:30pm. The event is expected to bring together more than 200 representatives of women-...

  18. holiday | National Nuclear Security Administration

    National Nuclear Security Administration (NNSA)

    holiday | National Nuclear Security Administration Facebook Twitter Youtube Flickr RSS People Mission Managing the Stockpile Preventing Proliferation Powering the Nuclear Navy Emergency Response Recapitalizing Our Infrastructure Countering Nuclear Terrorism About Our Programs Our History Who We Are Our Leadership Our Locations Budget Our Operations Library Bios Congressional Testimony Fact Sheets Newsletters Press Releases Photo Gallery Jobs Apply for Our Jobs Our Jobs Working at NNSA Blog Home

  19. The Saratoga Inn Pool & Spa Low Temperature Geothermal Facility...

    Open Energy Info (EERE)

    Saratoga Inn Pool & Spa Low Temperature Geothermal Facility Jump to: navigation, search Name The Saratoga Inn Pool & Spa Low Temperature Geothermal Facility Facility The Saratoga...

  20. River Inn Natural Hot Spring Pool & Spa Low Temperature Geothermal...

    Open Energy Info (EERE)

    Inn Natural Hot Spring Pool & Spa Low Temperature Geothermal Facility Jump to: navigation, search Name River Inn Natural Hot Spring Pool & Spa Low Temperature Geothermal Facility...

  1. Museum Closed for New Year's Holiday

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Museum Closed for New Year's Holiday Museum Closed for New Year's Holiday WHEN: Jan 01, 2016 12:00 AM - 11:59 PM WHERE: Bradbury Science Museum 1350 Central Ave, Los Alamos, NM 87544, USA CATEGORY: Bradbury INTERNAL: Calendar Login Museum Closed for New Year's Holiday Event Description The Bradbury Science Museum will be CLOSED for the New Year's holiday. The Bradbury Science Museum is open to the public every day except for Thanksgiving Day, Christmas Day and New Year's Day. Admission is always

  2. The Committee met in the Clark Room in the Holiday Inn Capitol...

    U.S. Energy Information Administration (EIA) Indexed Site

    ... That's government workers. We're now at about 375. You can ... We're trying to take the approach of informing our ... have a long knowledge of history of EIA and the survey. ...

  3. The Committee met in the Clark Room of the Capitol Holiday Inn...

    U.S. Energy Information Administration (EIA) Indexed Site

    ... on a transmission line, I can send that power across the country with very minimal losses. ... terms of the potential for collecting economic rents, it seems to me are pretty minimal. ...

  4. The Committee met in the Clark Room, Holiday Inn Capitol at ...

    U.S. Energy Information Administration (EIA) Indexed Site

    ... It becomes apples and oranges. How can they avoid getting into that quagmire unless they try and keep its impact as minimal as it is? CHAIRMAN WATKINS: Do you have a comment, ...

  5. The Committee met in the Columbia Room at the Holiday Inn...

    U.S. Energy Information Administration (EIA) Indexed Site

    ... That is a possibility also in this market. But again, that would require almost the most minimal amount of capacity to firm it up. You will not see reserved margins of 22 or 26 ...

  6. The Committee met in the Clark Room in the Holiday Inn Capitol...

    U.S. Energy Information Administration (EIA) Indexed Site

    ... funded by the American public and whose benefits ... in, say, The Journal of the American Medical Association ... has lots of health implications, and so to ...

  7. Tips to Save Energy During the Holidays | Department of Energy

    Broader source: Energy.gov (indexed) [DOE]

    Allison Casey Senior Communicator, NREL The winter holiday season has officially begun, and with it comes the frenzy of decorating, holiday gatherings, gift buying, and errand...

  8. EECBG Success Story: North Pole's Holiday Wish for an Energy...

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Pole's Holiday Wish for an Energy Efficient 2012 EECBG Success Story: North Pole's Holiday ... Wish for An Energy Efficient 2012 EECBG Success Story: Alaska Town Invests in Energy ...

  9. Holiday

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    in society. May 5 Tue 12:00 AM Cinco de Mayo United States and Mexico Cinco de Mayo (Spanish for "fifth of May") is a celebration held on May 5. Apr 13 Mon 12:00 AM Thomas...

  10. holiday

    National Nuclear Security Administration (NNSA)

    bloglabor-day-weekend-2015

    Labor Day is dedicated to the achievements of American workers and the contributions they made to the strength, prosperity, and well-being...

  11. A Factsheet on Holiday Fire Prevention

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    ach year fires occurring during the holiday season claim the lives of over 400 Americans, injure 1,650 more, and cause over $990 million in damage. According to the U. S. Fire Administration (USFA), there are simple life-saving steps you can take to ensure a safe and happy holiday. By following some of the outlined precautionary tips, individuals can greatly reduce their chances of becoming a holiday fire casualty. Preventing Christmas tree Fires Special fire safety precautions need to be taken

  12. Solar Field Powers Historic Garden Holiday Display

    Broader source: Energy.gov [DOE]

    Popular holiday attraction Longwood Gardens in Pennsylvania has commissioned an American-made 1.2 megawatt, 10.7-acre solar field as part of a goal to generate three megawatts of renewable energy by 2018.

  13. Code of Conduct regarding holiday gifts

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    questions about a specific event or item, please contact me at 665-3104, or Michelle Cantu, Ethics and Compliance Group Leader, at 667-7506. Have an enjoyable and safe Holiday...

  14. Fourth Friday Cancelled due to Thanksgiving Holidays

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Fourth Friday Cancelled due to Thanksgiving Holidays Fourth Friday Cancelled due to Thanksgiving Holidays WHEN: Nov 27, 2015 10:00 AM - 5:00 PM WHERE: Bradbury Science Museum 1350 Central Ave, Los Alamos, NM 87544, USA CONTACT: Linda Anderman (505) 665-9196 CATEGORY: Bradbury INTERNAL: Calendar Login Fourth Fridays Downtown Event Description Fourth Fridays Downtown will not take place during the weeks of Thanksgiving and Christmas. The Los Alamos Creative District sponsors Fourth Fridays, an

  15. I-N-N Electric Coop, Inc | Open Energy Information

    Open Energy Info (EERE)

    I-N-N Electric Coop, Inc Jump to: navigation, search Name: I-N-N Electric Coop, Inc Place: Alaska Phone Number: 800-571-1259 (In Alaska) or Outside Alaska: 907-571-1259 ...

  16. Buying an Appliance this Holiday Season? ENERGY STAR Products...

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Buying an Appliance this Holiday Season? ENERGY STAR Products will Save You Money and Energy All Year Buying an Appliance this Holiday Season? ENERGY STAR Products will Save You...

  17. Reduce Waste and Save Energy this Holiday Season

    Broader source: Energy.gov [DOE]

    Reduce waste and save energy this holiday season whether you're shopping, eating, partying, decorating, or wrapping.

  18. Fuel Cell Powers Up Festivities at Secretary Chu's Holiday Party |

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Department of Energy Powers Up Festivities at Secretary Chu's Holiday Party Fuel Cell Powers Up Festivities at Secretary Chu's Holiday Party December 16, 2011 - 11:25am Addthis A clean, efficient fuel cell powered the tree lights at the 2011 Energy Department holiday party. | Energy Department file photo. A clean, efficient fuel cell powered the tree lights at the 2011 Energy Department holiday party. | Energy Department file photo. Sunita Satyapal Sunita Satyapal Director, Fuel Cell

  19. How Do Holiday Lights Work? | Department of Energy

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    How Do Holiday Lights Work? How Do Holiday Lights Work? December 16, 2015 - 11:31am Addthis Daniel Wood Daniel Wood Data Visualization and Cartographic Specialist, Office of Public Affairs Sarah Gerrity Sarah Gerrity Former Multimedia Editor, Office of Public Affairs Want to learn more about Holiday Lights? Check out our recent post on the Top 5 Things You Didn't Know About Holiday Lights. Last year, we told you how incandescent holiday string lights work, but we left out an important topic: LED

  20. How Do Holiday Lights Work? | Department of Energy

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Do Holiday Lights Work? How Do Holiday Lights Work? December 16, 2015 - 11:31am Addthis Daniel Wood Daniel Wood Data Visualization and Cartographic Specialist, Office of Public Affairs Sarah Gerrity Sarah Gerrity Former Multimedia Editor, Office of Public Affairs Want to learn more about Holiday Lights? Check out our recent post on the Top 5 Things You Didn't Know About Holiday Lights. Last year, we told you how incandescent holiday string lights work, but we left out an important topic: LED

  1. Fountain Inn, South Carolina: Energy Resources | Open Energy...

    Open Energy Info (EERE)

    district.12 Registered Energy Companies in Fountain Inn, South Carolina Verde Biofuels Inc References US Census Bureau Incorporated place and minor civil...

  2. Holiday Gifts for Energy Efficiency | Department of Energy

    Energy Savers [EERE]

    Holiday Gifts for Energy Efficiency Holiday Gifts for Energy Efficiency December 18, 2013 - 3:58pm Addthis Energy-efficient light bulbs can make great energy-saving stocking stuffers. Energy-efficient light bulbs can make great energy-saving stocking stuffers. Erik Hyrkas Erik Hyrkas Media Relations Specialist, Office of Energy Efficiency & Renewable Energy How can I participate? Look for these energy-saving gifts while doing your holiday shopping this year. As the days get colder and

  3. Your Holidays...Brought to You by Fuel Cells

    Broader source: Energy.gov [DOE]

    A story about how fuel cells are helping bring the holidays to you is currently posted on the Energy Department's Blog.

  4. EECBG Success Story: South Carolina Community Lights Up the Season with Energy-Efficient Holiday Lights

    Broader source: Energy.gov [DOE]

    A South Carolina community is proving that energy efficiency can improve the holidays by reducing energy and maintenance costs, thanks to its new LED holiday lights. Learn more.

  5. Save Money with LED Holiday Light Strings | Department of Energy

    Energy Savers [EERE]

    Money with LED Holiday Light Strings Save Money with LED Holiday Light Strings December 1, 2009 - 11:19am Addthis Allison Casey Senior Communicator, NREL It's hard to believe, but the holidays are in full swing and many of you are starting to decorate your homes. Those of you who started around Halloween are probably feeling pretty smart because you got your decorating done while the weather was nice. I see you there, snacking on your Thanksgiving leftovers, laughing at me shivering in the cold

  6. SEP Success Story: Solar Field Powers Historic Garden Holiday...

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Photo courtesy of Longwood GardensW. Hill One of the most visited public gardens in the United States, and a popular attraction for elaborate holiday decorations, is now running...

  7. Meaningful Money Gifts at Holiday Time | The Ames Laboratory

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Meaningful Money Gifts at Holiday Time Give a small child a gift of money and they'll give you a sideways look and sheepish, "Thanks." They usually prefer boxes with big bows that...

  8. SRR Staff Send the Holidays to Soldiers Overseas

    Broader source: Energy.gov [DOE]

    A request for razors from a U.S. Army private serving in Afghanistan transformed into a full-scale holiday gift rescue operation by employees of the Savannah River Site’s liquid waste contractor,...

  9. SEP Success Story: Solar Field Powers Historic Garden Holiday Display

    Broader source: Energy.gov [DOE]

    One of the most visited public gardens in the United States, and a popular attraction for elaborate holiday decorations, is now running on solar power thanks to help from the Energy Department. Learn more.

  10. Tips to Save Energy During the Holidays | Department of Energy

    Broader source: Energy.gov (indexed) [DOE]

    Don't let your energy-saving efforts fall by the wayside amid all the festivities; the tips below will help you save energy and money even as you celebrate. Use LED Holiday Lights ...

  11. Energy-Efficient Holiday Decorating Tips | Department of Energy

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    With holidays almost upon us and home decorating to do, costs and energy savings should be on the brain. So as you put the finishing touches on that giant lighting display, be sure ...

  12. NREL Provides PV Holiday Lights for Christmas Tree

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Provides PV Holiday Lights for Christmas Tree For more information contact: George Douglas (303) 275-4096 Golden, Colo., December 2, 1997 -- National Renewable Energy Laboratory (NREL) engineers are showing off the power of photovoltaics in Washington, D.C. again this holiday season. They have installed an 8-kilowatt solar array on the Ellipse just south of the White House to help power lights on the National Christmas Tree. The tree lighting ceremony on Dec. 4 begins Washington's 1997 Pageant

  13. Energy Saving Holiday Kitchen Trivia | Department of Energy

    Office of Environmental Management (EM)

    Saving Holiday Kitchen Trivia Energy Saving Holiday Kitchen Trivia November 20, 2012 - 4:08pm Addthis Did you know that your dishwasher can be used as an oven in emergencies? | Photo courtesy of Chris Gunn, NREL. Did you know that your dishwasher can be used as an oven in emergencies? | Photo courtesy of Chris Gunn, NREL. Erik Hyrkas Erik Hyrkas Media Relations Specialist, Office of Energy Efficiency & Renewable Energy What does this mean for me? Knowing which kitchen appliances are most

  14. EM Waste Isolation Pilot Plant Team's Holiday Spirit Shines | Department

    Office of Environmental Management (EM)

    of Energy Waste Isolation Pilot Plant Team's Holiday Spirit Shines EM Waste Isolation Pilot Plant Team's Holiday Spirit Shines December 23, 2013 - 12:00pm Addthis Aspen Cass, a relative of an EM Carlsbad Field Office (CBFO) employee, holds donated coats with Farok Sharif (left), president and project manager of Nuclear Waste Partnership, the WIPP management and operating contractor, and Joe Franco, manager of CBFO. Aspen Cass, a relative of an EM Carlsbad Field Office (CBFO) employee, holds

  15. Two-Dimensional Electron Gas in Monolayer InN Quantum Wells....

    Office of Scientific and Technical Information (OSTI)

    Two-Dimensional Electron Gas in Monolayer InN Quantum Wells. Citation Details In-Document Search Title: Two-Dimensional Electron Gas in Monolayer InN Quantum Wells. Abstract not...

  16. It's Finally Time to Think about the Holidays! | Department of Energy

    Energy Savers [EERE]

    It's Finally Time to Think about the Holidays! It's Finally Time to Think about the Holidays! December 5, 2012 - 12:13pm Addthis Using LED holiday lights is just one of the ways that you can save energy and money this holiday season. | Photo courtesy of ©iStockphoto.com/AvailableLight Using LED holiday lights is just one of the ways that you can save energy and money this holiday season. | Photo courtesy of ©iStockphoto.com/AvailableLight Elizabeth Spencer Communicator, National Renewable

  17. Holiday Shopping and Electric Vehicles | GE Global Research

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Infrastructure Research: Holiday Shopping & Electric Vehicles Click to email this to a friend (Opens in new window) Share on Facebook (Opens in new window) Click to share (Opens in new window) Click to share on LinkedIn (Opens in new window) Click to share on Tumblr (Opens in new window) Infrastructure Research: Holiday Shopping & Electric Vehicles Jigar Shah 2014.12.03 With Thanksgiving behind us in the U.S., that can only mean one thing for all the bargain-hunters out there: deals

  18. LED Holiday Lights: Festive, Safe, and Efficient! | Department of Energy

    Office of Environmental Management (EM)

    LED Holiday Lights: Festive, Safe, and Efficient! LED Holiday Lights: Festive, Safe, and Efficient! December 2, 2010 - 4:34pm Addthis Andy Oare Andy Oare Former New Media Strategist, Office of Public Affairs Ed. Note cross posted from the Energy Savers Blog. Written by Allison Casey This week brings a day that many people look forward to all year -- and I'm not talking about Thanksgiving, or Black Friday. I'm talking about that magical day when it's finally okay to drag the dusty boxes from the

  19. LED Holiday Lights: Festive, Safe, and Efficient! | Department of Energy

    Office of Environmental Management (EM)

    LED Holiday Lights: Festive, Safe, and Efficient! LED Holiday Lights: Festive, Safe, and Efficient! November 23, 2010 - 6:30am Addthis Allison Casey Senior Communicator, NREL This week brings a day that many people look forward to all year-and I'm not talking about Thanksgiving, or Black Friday. I'm talking about that magical day when it's finally okay to drag the dusty boxes from the basement or attic, to lovingly sort and display your decorations, to untangle your light strings and set up the

  20. Save Energy on Appliances this Holiday Season | Department of Energy

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Save Energy on Appliances this Holiday Season Save Energy on Appliances this Holiday Season November 21, 2014 - 9:52am Addthis Buying ENERGY STAR appliances saves money and energy. | Jim Tetro, U.S. Department of Energy Buying ENERGY STAR appliances saves money and energy. | Jim Tetro, U.S. Department of Energy Erik Hyrkas Erik Hyrkas Media Relations Specialist, Office of Energy Efficiency & Renewable Energy How does it work? Use the ENERGY STAR Rebate Finder tool to find rebates and other

  1. Workers Across the Nuclear Security Enterprise Give Back This Holiday

    National Nuclear Security Administration (NNSA)

    Season | National Nuclear Security Administration Workers Across the Nuclear Security Enterprise Give Back This Holiday Season | National Nuclear Security Administration Facebook Twitter Youtube Flickr RSS People Mission Managing the Stockpile Preventing Proliferation Powering the Nuclear Navy Emergency Response Recapitalizing Our Infrastructure Countering Nuclear Terrorism About Our Programs Our History Who We Are Our Leadership Our Locations Budget Our Operations Library Bios Congressional

  2. Geothermal heating facilities for Frontier Inn, Susanville, California

    SciTech Connect (OSTI)

    Not Available

    1982-03-01

    The Frontier Inn, located in Susanville, California, is a 38 unit motel composed of six major sections (coffee shop, A frame units, apartments, back units, two story units and office). These sections were built over a number of years and exhibit widely varying types of construction. Space heating is provided by primarily electric resistance equipment with some propane use. Domestic hot water is provided primarily by propane with some electric resistance. The coffee shop uses fuel oil for both space and domestic hot water heating. The City of Susanville is currently in the process of installing a geothermal district heating system. Although the motel site is not located in the area of present construction activity, it is expected that the pipeline will be extended in the near future. This study examines the potential of retrofitting the existing heating facilities at the Frontier Inn to geothermal.

  3. #tipsEnergy: How to Save Energy During the Holidays | Department...

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    the Holidays December 21, 2012 - 2:20pm Addthis Rebecca Matulka Rebecca Matulka Former Digital Communications Specialist, Office of Public Affairs storify:http:storify.com...

  4. Y-12 employees help make holiday merrier for many | Y-12 National...

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    did the additional footwork needed to fulfill the holiday gift wishes of 497 children in Anderson, Roane, Morgan, Loudon, Scott and Monroe Counties. Coordinated this year...

  5. Top 5 Things You Didn't Know About Holiday Lights | Department of Energy

    Energy Savers [EERE]

    5 Things You Didn't Know About Holiday Lights Top 5 Things You Didn't Know About Holiday Lights December 5, 2014 - 4:49pm Addthis With a fiery past and a bright future, here are 5 things you probably didn’t know about holiday lights. | Graphic by <a href="/node/379579">Sarah Gerrity</a>, Energy Department. With a fiery past and a bright future, here are 5 things you probably didn't know about holiday lights. | Graphic by Sarah Gerrity, Energy Department. Pat Adams Pat

  6. Top 5 Things You Didn't Know About Holiday Lights | Department of Energy

    Broader source: Energy.gov (indexed) [DOE]

    With a fiery past and a bright future, here are 5 things you probably didn’t know about holiday lights. | Graphic by <a href="/node/379579">Sarah Gerrity</a>, Energy Department. With a fiery past and a bright future, here are 5 things you probably didn't know about holiday lights. | Graphic by Sarah Gerrity, Energy Department. Pat Adams Pat Adams Digital Content Specialist, Office of Public Affairs How does it work? Learn the science behind incandescent holiday lights

  7. They're Here! Winter, Holidays, and the New Year. How Will You Save Energy?

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    | Department of Energy They're Here! Winter, Holidays, and the New Year. How Will You Save Energy? They're Here! Winter, Holidays, and the New Year. How Will You Save Energy? December 22, 2009 - 10:11am Addthis Winter officially hit this week, and those of you on the east coast found out in a big way. Many of you are still shoveling out while trying to take care of those last-minute holiday preparations. (I'm actually kind of jealous. I love shoveling snow. I just hate driving in it.)

  8. How do You Save Energy During the Holidays? | Department of Energy

    Energy Savers [EERE]

    do You Save Energy During the Holidays? How do You Save Energy During the Holidays? December 20, 2011 - 9:03am Addthis Between family vacations, shopping, and cooking extravagant meals, energy conservation isn't usually at the top our list of things to do during the holiday season. But there are plenty of ways to save money & energy even now by doing little - and even fun - things to reduce our utility bills and use less energy overall. This week, Amanda showed us how she saves money and

  9. Buying an Appliance this Holiday Season? ENERGY STAR Products will Save You

    Energy Savers [EERE]

    Money and Energy All Year! | Department of Energy Buying an Appliance this Holiday Season? ENERGY STAR Products will Save You Money and Energy All Year! Buying an Appliance this Holiday Season? ENERGY STAR Products will Save You Money and Energy All Year! December 12, 2012 - 11:40am Addthis When shopping for appliances or electronics for the holidays, look for the ENERGY STAR® and EnergyGuide labels. | Photo by Dennis Schroeder, NREL 22090. When shopping for appliances or electronics for

  10. Video e-card offers holiday greetings from everyone at PPPL ...

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Video e-card offers holiday greetings from everyone at PPPL December 22, 2014 Tweet Widget Google Plus One Share on Facebook Staff of the U.S. Department of Energy's Princeton...

  11. Holiday Auction raises record amount -- $5,842 | The Ames Laboratory

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    loves his crisp Vince Dahl and Dave Boeke show off their holiday garb. Auctioneer Dale Meyer plies his trade. Julie Dredla looks to see who's bidding against her. What am I bid...

  12. Two-dimensional electron gas in monolayer InN quantum wells

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Pan, Wei; Dimakis, Emmanouil; Wang, George T.; Moustakas, Theodore D.; Tsui, Daniel C.

    2014-11-24

    We report in this letter experimental results that confirm the two-dimensional nature of the electron systems in monolayer InN quantum wells embedded in GaN barriers. The electron density and mobility of the two-dimensional electron system (2DES) in these InN quantum wells are 5×1015 cm-2 and 420 cm2 /Vs, respectively. Moreover, the diagonal resistance of the 2DES shows virtually no temperature dependence in a wide temperature range, indicating the topological nature of the 2DES.

  13. Lois Curfman McInnes, 2011 | U.S. DOE Office of Science (SC)

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Lois Curfman McInnes, 2011 The Ernest Orlando Lawrence Award Lawrence Award Home Nomination & Selection Guidelines Award Laureates 2010's 2000's 1990's 1980's 1970's 1960's Ceremony The Life of Ernest Orlando Lawrence Contact Information The Ernest Orlando Lawrence Award U.S. Department of Energy SC-2/Germantown Building 1000 Independence Ave., SW Washington, DC 20585 P: (301) 903-2411 E: Email Us 2010's Lois Curfman McInnes, 2011 Print Text Size: A A A FeedbackShare Page Computer,

  14. Growth mechanism and microstructure of low defect density InN (0001) In-face thin films on Si (111) substrates

    SciTech Connect (OSTI)

    Kehagias, Th.; Dimitrakopulos, G. P.; Koukoula, T.; Komninou, Ph.; Ajagunna, A. O.; Georgakilas, A.; Physics Department, University of Crete, P.O. Box 2208, 71003 Heraklion-Crete ; Tsagaraki, K.; Adikimenakis, A.

    2013-10-28

    Transmission electron microscopy has been employed to analyze the direct nucleation and growth, by plasma-assisted molecular beam epitaxy, of high quality InN (0001) In-face thin films on (111) Si substrates. Critical steps of the heteroepitaxial growth process are InN nucleation at low substrate temperature under excessively high N-flux conditions and subsequent growth of the main InN epilayer at the optimum conditions, namely, substrate temperature 400450 C and In/N flux ratio close to 1. InN nucleation occurs in the form of a very high density of three dimensional (3D) islands, which coalesce very fast into a low surface roughness InN film. The reduced reactivity of Si at low temperature and its fast coverage by InN limit the amount of unintentional Si nitridation by the excessively high nitrogen flux and good bonding/adhesion of the InN film directly on the Si substrate is achieved. The subsequent overgrowth of the main InN epilayer, in a layer-by-layer growth mode that enhances the lateral growth of InN, reduces significantly the crystal mosaicity and the density of threading dislocations is about an order of magnitude less compared to InN films grown using an AlN/GaN intermediate nucleation/buffer layer on Si. The InN films exhibit the In-face polarity and very smooth atomically stepped surfaces.

  15. Nanostructural and electronic properties of polytypes in InN nanocolumns

    SciTech Connect (OSTI)

    Kioseoglou, J.; Koukoula, T.; Komninou, Ph.; Kehagias, Th.; Georgakilas, A.; Androulidaki, M.

    2013-08-21

    Transmission electron microscopy techniques and density functional theory calculations were employed to investigate the nanostructural and electronic properties of InN polytypes observed in InN nanocolumns, grown on Si(111) by molecular beam epitaxy. Moir fringes and alternating hexagonal and cubic lattice stacking sequences along the c-axis, observed among the wurtzite layers, implied the presence of different structures embedded in the basic 2H structure of the nanocolumns. Quantitative electron diffraction analysis and high-resolution image simulations verified the coexistence of the wurtzite structure with the 4H, 6H, and the 3C zinc-blende structural polytypes. Total energies calculations established the 2H wurtzite structure as the most stable polytype. The band gap of all polytypes was found direct with the energies and the band gaps of the 4H (E{sub g} = 0.64 eV) and 6H (E{sub g} = 0.60 eV) structures calculated between the corresponding values of the 2H (E{sub g} = 0.75 eV) and 3C (E{sub g} = 0.49 eV) basic structures. Theoretical and experimental analysis showed that at the initial stages of growth InN nanocolumns were under tensile strain along both the basal plane and growth direction. Structural polytypes were then introduced in the form of embedded inclusions to accommodate the excess tensile strain along the growth direction, allowing the entire process of polymorphism to be the dominant strain relaxation mechanism of InN nanocolumns. Moreover, the lattice and energetic properties and band gap values of InN polytypes showed a linear dependence on hexagonality, while the presence of polytypes led to a characteristic broadening of the photoluminescence emission peak toward lower emission energies.

  16. Cut Gas Costs This Holiday Traveling Season with Three Easy Tips |

    Energy Savers [EERE]

    Department of Energy Cut Gas Costs This Holiday Traveling Season with Three Easy Tips Cut Gas Costs This Holiday Traveling Season with Three Easy Tips November 26, 2013 - 9:23am Addthis Turning off your engine while waiting in the parking lot is a great way to save money on gas. | Photo courtesy of Kristy Keel-Blackmon, NREL/21196. Turning off your engine while waiting in the parking lot is a great way to save money on gas. | Photo courtesy of Kristy Keel-Blackmon, NREL/21196. Jason

  17. Your Holidays ... Brought to You by Fuel Cells | Department of Energy

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Your Holidays ... Brought to You by Fuel Cells Your Holidays ... Brought to You by Fuel Cells December 19, 2013 - 11:44am Addthis Fuel cells, which work like batteries but don’t run down or need recharging, are ideal for powering material handling equipment, like forklifts and airport baggage carts, because they reduce recharging time and cut carbon pollution. This is helping them become more mainstream in the U.S., with more than 4,000 vehicles in operation in 2012, and this year, they

  18. Food and gift drives help make holidays brighter for regional families

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    STEM skills Community Connections: Your link to news and opportunities from Los Alamos National Laboratory Latest Issue:Mar. 2016 all issues All Issues » submit Food and gift drives help make holidays brighter for regional families Donations impact seven-county region February 1, 2016 Northern New Mexico children excitedly open their gifts, while volunteers get ready to deliver bicycles. Northern New Mexico children excitedly open their gifts, while volunteers get ready to deliver bicycles.

  19. Y-12 helps more than 600 kids for the holidays | National Nuclear Security

    National Nuclear Security Administration (NNSA)

    Administration helps more than 600 kids for the holidays | National Nuclear Security Administration Facebook Twitter Youtube Flickr RSS People Mission Managing the Stockpile Preventing Proliferation Powering the Nuclear Navy Emergency Response Recapitalizing Our Infrastructure Countering Nuclear Terrorism About Our Programs Our History Who We Are Our Leadership Our Locations Budget Our Operations Library Bios Congressional Testimony Fact Sheets Newsletters Press Releases Photo Gallery Jobs

  20. Whimsical SRS Relay Race Brings Joy to those in Need this Holiday Season |

    National Nuclear Security Administration (NNSA)

    National Nuclear Security Administration Whimsical SRS Relay Race Brings Joy to those in Need this Holiday Season | National Nuclear Security Administration Facebook Twitter Youtube Flickr RSS People Mission Managing the Stockpile Preventing Proliferation Powering the Nuclear Navy Emergency Response Recapitalizing Our Infrastructure Countering Nuclear Terrorism About Our Programs Our History Who We Are Our Leadership Our Locations Budget Our Operations Library Bios Congressional Testimony

  1. NNMCAB Board Minutes: November 2002 Santa Fe

    Broader source: Energy.gov [DOE]

    Minutes of the November 20, 2002 Board meeting at Holiday Inn Planning for SSAB Chairs Workshop on Transuranic Waste

  2. NNMCAB Board Minutes: June 2000 Taos

    Broader source: Energy.gov [DOE]

    Minutes of the June 28, 2000 Board meeting at Holiday Inn Presentation LANL, Procurement Process, Betty Romero

  3. Microsoft Word - 2015_LAHotel-Info-3.docx

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    postdoc:Postdoc Program:Career Fair:2015:2015_LAHotel-Info-3.docx Accommodations Info in Los Alamos There are many wonderful places to stay in Santa Fe as well as the surrounding areas but the following is a list of hotels in Los Alamos: Comfort Inn 2455 Trinity Dr. Los Alamos, NM 87544 505-661-1110 800-992-2694 Hampton Inn & Suites 124 State Rd. 4 Los Alamos, NM 87544 505-672-3838 Holiday Inn Express at Entrada Park 60 Entrada Dr. Los Alamos, NM 87544 505-661-2646 Motel 6 2175 Trinity Dr.

  4. 2012 BATTERIES GORDON RESEARCH CONFERENCE, MARCH 4-9, 2012

    SciTech Connect (OSTI)

    Stephen Harris

    2012-03-09

    The Gordon Research Conference on BATTERIES was held at Four Points Sheraton / Holiday Inn Express, Ventura, California, March 4-9, 2012. The Conference was well-attended with 176 participants. Gordon Research Conferences does not permit publication of meeting proceedings.

  5. Optical characterization of free electron concentration in heteroepitaxial InN layers using Fourier transform infrared spectroscopy and a 2 Multiplication-Sign 2 transfer-matrix algebra

    SciTech Connect (OSTI)

    Katsidis, C. C.; Ajagunna, A. O.; Georgakilas, A.

    2013-02-21

    Fourier Transform Infrared (FTIR) reflectance spectroscopy has been implemented as a non-destructive, non-invasive, tool for the optical characterization of a set of c-plane InN single heteroepitaxial layers spanning a wide range of thicknesses (30-2000 nm). The c-plane (0001) InN epilayers were grown by plasma-assisted molecular beam epitaxy (PAMBE) on GaN(0001) buffer layers which had been grown on Al{sub 2}O{sub 3}(0001) substrates. It is shown that for arbitrary multilayers with homogeneous anisotropic layers having their principal axes coincident with the laboratory coordinates, a 2 Multiplication-Sign 2 matrix algebra based on a general transfer-matrix method (GTMM) is adequate to interpret their optical response. Analysis of optical reflectance in the far and mid infrared spectral range has been found capable to discriminate between the bulk, the surface and interface contributions of free carriers in the InN epilayers revealing the existence of electron accumulation layers with carrier concentrations in mid 10{sup 19} cm{sup -3} at both the InN surface and the InN/GaN interface. The spectra could be fitted with a three-layer model, determining the different electron concentration and mobility values of the bulk and of the surface and the interface electron accumulation layers in the InN films. The variation of these values with increasing InN thickness could be also sensitively detected by the optical measurements. The comparison between the optically determined drift mobility and the Hall mobility of the thickest sample reveals a value of r{sub H} = 1.49 for the Hall factor of InN at a carrier concentration of 1.11 Multiplication-Sign 10{sup 19} cm{sup -3} at 300 Degree-Sign {Kappa}.

  6. Better Buildings Residential Network Driving Demand Peer Exchange Call Series: Leveraging Holidays and Other Events Call Slides and Discussion Summary, November 7, 2013

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Driving Demand Peer Exchange Call Series: Leveraging Holidays and Other Events Call Slides and Discussion Summary November 7, 2013 Agenda  Call Logistics and Introductions  Peer Exchange Call Overview and Announcements  Lessons Learned: Featured Speakers  California Center for Sustainable Energy  Minnesota Center for Energy and Environment  Discussion  How has your program leveraged (or considered leveraging) holidays or other seasonal events (e.g., Earth Day, winter

  7. Optical and structural characterization of nitrogen-rich InN: Transition from nearly intrinsic to strongly n-type degenerate with temperature

    SciTech Connect (OSTI)

    Hong Tran, Nhung; Huy Le, Binh; Fan, Shizhao; Zhao, Songrui; Mi, Zetian; Schmidt, Benjamin A.; Savard, Michel; Gervais, Guillaume; Butcher, Kenneth Scott A.

    2013-12-23

    We report on a detailed study of the structural and optical properties of nonstoichiometric nitrogen-rich InN grown on sapphire substrates, by migration enhanced afterglow deposition. The samples were polycrystalline, with the presence of InN dots. Unusually strong photoluminescence emission was measured at cryogenic temperatures, with the peak energy at ?0.68?eV. Detailed analysis further shows that the sample has very low residual electron density in the range of ?10{sup 16}?cm{sup ?3} at temperatures below 20?K.

  8. Electronic and thermoelectric properties of InN studied using ab initio density functional theory and Boltzmann transport calculations

    SciTech Connect (OSTI)

    Borges, P. D. E-mail: lscolfaro@txstate.edu; Scolfaro, L. E-mail: lscolfaro@txstate.edu

    2014-12-14

    The thermoelectric properties of indium nitride in the most stable wurtzite phase (w-InN) as a function of electron and hole concentrations and temperature were studied by solving the semiclassical Boltzmann transport equations in conjunction with ab initio electronic structure calculations, within Density Functional Theory. Based on maximally localized Wannier function basis set and the ab initio band energies, results for the Seebeck coefficient are presented and compared with available experimental data for n-type as well as p-type systems. Also, theoretical results for electric conductivity and power factor are presented. Most cases showed good agreement between the calculated properties and experimental data for w-InN unintentionally and p-type doped with magnesium. Our predictions for temperature and concentration dependences of electrical conductivity and power factor revealed a promising use of InN for intermediate and high temperature thermoelectric applications. The rigid band approach and constant scattering time approximation were utilized in the calculations.

  9. ERSUG: July 11 - 12, 1994 (Rockville, Maryland)

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    ERSUG: July 11 - 12, 1994 (Rockville, Maryland) Dates July 11 - 12, 1994 Location Holiday Inn, Crowne Plaza Rockville, Maryland Presentations Agenda ERSUG Meeting July 11-12, 1994 Holiday Inn, Crowne Plaza Rockville, MD The Energy Research Supercomputer Users' Group (ERSUG) will meet at the Holiday Inn, Crowne Plaza in Rockville, MD on July 11-12, 1994. In the past, this meeting has combined presentations describing work-in-progress at NERSC with lively user discussions in the areas of the

  10. NUG 1994

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    - 12, 1994 (Rockville, Maryland) Dates July 11 - 12, 1994 Location Holiday Inn, Crowne Plaza Rockville,... Read More ERSUGEXERSUG: Jan 12, 1994 (UCLA) Dates Jan 12, 1994...

  11. NNMCAB Board Minutes: April 2009 Santa Fe

    Broader source: Energy.gov [DOE]

    Minutes of the April 8, 2009 Board meeting at Holiday Inn Presentation Kerr Laboratory, Well Screen Analysis Report, Steven Acree, Richard Wilkin

  12. Holiday Gift Drive

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Disabilities Espanola Valley Toy Run Las Vegas Adult Protective Services La Tierra Montessori Los Alamos Family Council Ohkay Owingeh Boys and Girls Clubs Rio ArribaLos Alamos...

  13. Holiday Food Drive

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Community Programs Office (505) 665-4400 Email Get Expertise Helping feed Northern New Mexico families During the Laboratory's 2015 Annual Food Drive, employees and subcontract...

  14. NNMCAB Board Minutes: January 2009 Santa Fe

    Broader source: Energy.gov [DOE]

    Minutes of the January 28, 2009 Board meeting at Holiday Inn Presentation DOE, Implementation on NNMCAB Recommendations, Jeff Casalina Presentation LANL, Well Network Analysis and Characterization of Groundwater at LANL Site, Danny Katzman

  15. Northern New Mexico Citizens Advisory Board Meeting

    Office of Environmental Management (EM)

    bi-monthly meeting of the Northern New Mexico Citizens' Advisory Board (NNMCAB or CAB) meeting was held on September 30, 2009 at the Holiday Inn, 4048 Cerrillos Road Santa...

  16. Hydrogen and Fuel Cell Technical Advisory Committee Meeting

    Broader source: Energy.gov [DOE]

    The Hydrogen and Fuel Cell Technical Advisory Committee (HTAC) will hold its next meeting October 27–28, 2015, at the Holiday Inn Capitol in Washington, D.C.

  17. Agenda

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Agenda Agenda ERSUG Meeting July 11-12, 1994 Holiday Inn, Crowne Plaza Rockville, MD The Energy Research Supercomputer Users' Group (ERSUG) will meet at the Holiday Inn, Crowne Plaza in Rockville, MD on July 11-12, 1994. In the past, this meeting has combined presentations describing work-in-progress at NERSC with lively user discussions in the areas of the services and capabilities provided by NERSC. For this particular meeting, however, the focus will change somewhat. First, more emphasis will

  18. The Express

    Alternative Fuels and Advanced Vehicles Data Center [Office of Energy Efficiency and Renewable Energy (EERE)]

    site. We will let you know what is new through this newsletter. As you will find in this issue, briefs of various alternative fuels information are provided by category. We will also begin an addi- tional service, the AFDC Express. The Express will provide impor- tant information on late-breaking issues via e-mail and fax. If you would like to receive these, you should ensure that the Center has your correct information. For those of you who have finished this editorial of sorts, fear not, it is

  19. Relevant Links

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Relevant Links **Light Source Facilities Around the World** Advanced Materials Research Institiute(AMRI), UNO Area Hotels Chase Suite Hotel Baton Rouge Extended Stay America Holiday Inn South Baton Rouge Marriott Residence Inns Wyndham Garden Gulf Coast Protein Crystallography Consortium Health Physics Society Institute for Micromanufacturing, LA Tech University Interactions.org - Particle Physics News and Resources International Nuclear Information System (INIS) Light Sources.org The Louisiana

  20. Fourth Friday Cancelled due to Thanksgiving Holidays

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    open late until 6:00 PM offering access to exhibits and special activities for all ages, however it will not take place on Friday, November 27 due to the Thanksgiving...

  1. Today LED Holiday Lights, Tomorrow the World?

    SciTech Connect (OSTI)

    Gordon, Kelly L.

    2004-12-20

    This article for The APEM Advantage, the quarterly newsletter of the Association of Professional Energy Managers (APEM) describes the recent increase in the popularity of light emitting diode (LED) lighting and compares LED light output with that of incandescent and compact fluorescent lighting.

  2. NNMCAB Board Minutes: May 2003 Taos

    Broader source: Energy.gov [DOE]

    Minutes of the May 16, 2003 Board Retreat at Holiday Inn Don Fernando de Taos Presentation NNMCAB Fiscal Year 2003 Accomplishments Presentation DOE/LANL, Environmental Monitoring and Surveillance Groundwater and Surface Water, Ted Taylor Presentation DOE/LANL, Environmental Restoration, Ted Taylor

  3. High expression Zymomonas promoters

    DOE Patents [OSTI]

    Viitanen, Paul V.; Tao, Luan; Zhang, Yuying; Caimi, Perry G.; McCole, Laura : Zhang, Min; Chou, Yat-Chen; McCutchen, Carol M.; Franden, Mary Ann

    2011-08-02

    Identified are mutants of the promoter of the Z. mobilis glyceraldehyde-3-phosphate dehydrogenase gene, which direct improved expression levels of operably linked heterologous nucleic acids. These are high expression promoters useful for expression of chimeric genes in Zymomonas, Zymobacter, and other related bacteria.

  4. WebExpress

    Energy Science and Technology Software Center (OSTI)

    1998-10-25

    WEBEXPRESS is a web interface to the Express for Unix batch scheduling product by Tidal Software. It offers web access from any client on the network and increased functionality in some areas.

  5. Express Energy Efficiency Program

    Broader source: Energy.gov [DOE]

    The Express Energy Efficiency Program provides free installation of energy-saving products. This program travels around Wisconsin, community-by-community, to see if they are installing in your...

  6. TTW 12-1-05

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    , 2005 WIPP Quick Facts (As of 11-30-05) 4,110 Shipments received since opening 33,321 Cubic meters of waste disposed 72,865 Containers disposed in the underground Upcoming Holiday Parties WTS Kid's Party December 3 - 12:00 p.m. Mall Cinema Movies offered: Zathura (PG) and Harry Potter (PG-13) WTS Christmas Party December 3 - 5:30 p.m. Walter Gerrells Performing Arts & Exhibition Centre Annex CTAC Holiday Party December 12 - 6:00 p.m. Stevens Inn NMED issues draft permit The New Mexico

  7. Solar Express | Open Energy Information

    Open Energy Info (EERE)

    Express Place: Italy Product: A joint venture established to install some 11MW of photovoltaic generation capacity around the country. References: Solar Express1 This article...

  8. Express Power | Open Energy Information

    Open Energy Info (EERE)

    United Kingdom Zip: B1 2JB Sector: Renewable Energy Product: Express Power is a the sustainable energy company operating within the Express Park Group. It develops renewable...

  9. Sonoma Mission Inn Geothermal Area | Open Energy Information

    Open Energy Info (EERE)

    110C383.15 K 230 F 689.67 R 1 USGS Estimated Reservoir Volume: 1 km 1 USGS Mean Capacity: 6 MW 1 Click "Edit With Form" above to add content History and...

  10. Agenda12810 | U.S. DOE Office of Science (SC)

    Office of Science (SC) Website

    8, 2010 Nuclear Science Advisory Committee (NSAC) NSAC Home Meetings Members Charges/Reports Charter .pdf file (78KB) NP Committees of Visitors Federal Advisory Committees NP Home Meetings December 8, 2010 Print Text Size: A A A FeedbackShare Page DOE/NSF Nuclear Science Advisory Committee Meeting December 8, 2010 Where: Holiday Inn National Airport Hotel, Shenandoah Ballroom I & II, 2650 Jefferson Davis Highway, Arlington, VA, Phone Number: 703-684-7200. Purpose/Topics: Perspectives from

  11. Feasibility Study of Sustainable Distributed Generation Technologies for the Duck Valley Reservation

    Office of Environmental Management (EM)

    of Sustainable Distributed Generation Technologies for the Duck Valley Reservation Feasibility Study of Sustainable Distributed Generation Technologies for the Duck Valley Reservation Office of Energy Efficiency and Renewable Energy TRIBAL ENERGY PROGRAM FY2004 Program Review Meeting Denver West Holiday Inn Golden, Colorado Shoshone-Paiute Tribes of the Duck Valley Reservation CSHQA New West Technologies Idaho Department of Water Resources INEEL Feasibility Study of Sustainable Distributed

  12. Energy-Efficient Holiday Decorating Tips | Department of Energy

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    In addition to using 70% less energy than traditional bulbs, they're brighter, eco-friendly, and are safer, as they are much cooler than incandescent lights. In addition, they...

  13. Sales Tax Holiday for Energy-Efficient Appliances | Department...

    Broader source: Energy.gov (indexed) [DOE]

    each year for the following appliances*: Clothes washers Clothes dryers* Water heaters Trash compactors* Dishwashers Conventional ovens, ranges, and stoves* Air conditioners...

  14. Sales Tax Holiday for Energy-Efficient Appliances

    Broader source: Energy.gov [DOE]

    In order to qualify for the exemption, qualifying pieces of equipment must be designated as meeting or exceeding the efficiency requirements of the ENERGY STAR program. The law defines eligible...

  15. ENERGY STAR Sales Tax Holiday for Energy-Efficient Products

    Broader source: Energy.gov [DOE]

    Although the eligibility of some products is limited according to their sale price, there are no limitations on the total value or number of products exempt from sales tax.

  16. Holiday City-Berkeley, New Jersey: Energy Resources | Open Energy...

    Open Energy Info (EERE)

    City-Berkeley, New Jersey: Energy Resources Jump to: navigation, search Equivalent URI DBpedia Coordinates 39.9645797, -74.2707509 Show Map Loading map... "minzoom":false,"map...

  17. The Committee convened in the Clark Room of the Holiday ...

    U.S. Energy Information Administration (EIA) Indexed Site

    Measurement Model for Information Management Process (Agenda ... They've had several recently. The Wall Street Journal has ... in the industrial organization of its production by ...

  18. Leveraging Holidays and Other Events | Department of Energy

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    for Marketing Energy Efficiency The Solarize Guidebook: A community guide to collective purchasing of residential PV systems (Book), SunShot, U.S. Department of Energy (DOE

  19. Reduce Waste and Save Energy this Holiday Season | Department...

    Broader source: Energy.gov (indexed) [DOE]

    Wrap your gifts with recycled paper to reduce waste and save money. | Photo courtesy of istockphotodiane555 Wrap your gifts with recycled paper to reduce waste and save money. |...

  20. Express Farms Greenhouse Low Temperature Geothermal Facility...

    Open Energy Info (EERE)

    Express Farms Greenhouse Low Temperature Geothermal Facility Jump to: navigation, search Name Express Farms Greenhouse Low Temperature Geothermal Facility Facility Express Farms...

  1. Method of controlling gene expression

    DOE Patents [OSTI]

    Peters, Norman K. (Berkeley, CA); Frost, John W. (Menlo Park, CA); Long, Sharon R. (Palo Alto, CA)

    1991-12-03

    A method of controlling expression of a DNA segment under the control of a nod gene promoter which comprises administering to a host containing a nod gene promoter an amount sufficient to control expression of the DNA segment of a compound of the formula: ##STR1## in which each R is independently H or OH, is described.

  2. Baculovirus expression system and method for high throughput expression of genetic material

    DOE Patents [OSTI]

    Clark, Robin; Davies, Anthony

    2001-01-01

    The present invention provides novel recombinant baculovirus expression systems for expressing foreign genetic material in a host cell. Such expression systems are readily adapted to an automated method for expression foreign genetic material in a high throughput manner. In other aspects, the present invention features a novel automated method for determining the function of foreign genetic material by transfecting the same into a host by way of the recombinant baculovirus expression systems according to the present invention.

  3. Agenda112901 | U.S. DOE Office of Science (SC)

    Office of Science (SC) Website

    29-30, 2001 Nuclear Science Advisory Committee (NSAC) NSAC Home Meetings Members Charges/Reports Charter .pdf file (78KB) NP Committees of Visitors Federal Advisory Committees NP Home Meetings November 29-30, 2001 Print Text Size: A A A FeedbackShare Page DOE/NSF Nuclear Science Advisory Committee Meeting November 29-30, 2001 Holiday Inn 550 C Street, SW Washington, D.C. Preliminary Agenda November 29, 2001 Session 1 8:30 a.m. Welcome James Symons 8:40 a.m. Report from DOE 9:05 a.m. Report from

  4. Methods for monitoring multiple gene expression

    DOE Patents [OSTI]

    Berka, Randy; Bachkirova, Elena; Rey, Michael

    2013-10-01

    The present invention relates to methods for monitoring differential expression of a plurality of genes in a first filamentous fungal cell relative to expression of the same genes in one or more second filamentous fungal cells using microarrays containing Trichoderma reesei ESTs or SSH clones, or a combination thereof. The present invention also relates to computer readable media and substrates containing such array features for monitoring expression of a plurality of genes in filamentous fungal cells.

  5. Methods for monitoring multiple gene expression

    DOE Patents [OSTI]

    Berka, Randy (Davis, CA); Bachkirova, Elena (Davis, CA); Rey, Michael (Davis, CA)

    2008-06-01

    The present invention relates to methods for monitoring differential expression of a plurality of genes in a first filamentous fungal cell relative to expression of the same genes in one or more second filamentous fungal cells using microarrays containing Trichoderma reesei ESTs or SSH clones, or a combination thereof. The present invention also relates to computer readable media and substrates containing such array features for monitoring expression of a plurality of genes in filamentous fungal cells.

  6. Methods for monitoring multiple gene expression

    DOE Patents [OSTI]

    Berka, Randy (Davis, CA); Bachkirova, Elena (Davis, CA); Rey, Michael (Davis, CA)

    2012-05-01

    The present invention relates to methods for monitoring differential expression of a plurality of genes in a first filamentous fungal cell relative to expression of the same genes in one or more second filamentous fungal cells using microarrays containing Trichoderma reesei ESTs or SSH clones, or a combination thereof. The present invention also relates to computer readable media and substrates containing such array features for monitoring expression of a plurality of genes in filamentous fungal cells.

  7. PP-362 Champlain Hudson Power Express, Inc.

    Broader source: Energy.gov [DOE]

    Presidential Permit authorizing Champlain Hudson Power Express, Inc. (CHPEI) to construct, operate and maintain electric transmission facilities at the U.S. - Canada border.

  8. Express Package Shipping Services | The Ames Laboratory

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Express Package Shipping Services General Information: The materials handling office's express package shipping service offers overnight domestic letter and parcel service for official business only. Express international shipments are also offered. Many hazardous and non-hazardous materials may be shipped by express. This service is provided to Ames Laboratory personnel. Hours: Monday through Friday - 7:30 a.m. to 11:50 a.m. - 12:30 p.m. through 4:00 p.m. Cutoff: for 10:30 a.m. next day

  9. Xcel Energy - Express Energy Efficiency Program | Department...

    Broader source: Energy.gov (indexed) [DOE]

    t-products-appliancesexpr... State Wisconsin Program Type Rebate Program Rebate Amount Free. Summary The Express Energy Efficiency Program provides free installation of...

  10. Expression of eukaryotic polypeptides in chloroplasts

    DOE Patents [OSTI]

    Mayfield, Stephen P.

    2013-06-04

    The present invention relates to a gene expression system in eukaryotic and prokaryotic cells, preferably plant cells and intact plants. In particular, the invention relates to an expression system having a RB47 binding site upstream of a translation initiation site for regulation of translation mediated by binding of RB47 protein, a member of the poly(A) binding protein family. Regulation is further effected by RB60, a protein disulfide isomerase. The expression system is capable of functioning in the nuclear/cytoplasm of cells and in the chloroplast of plants. Translation regulation of a desired molecule is enhanced approximately 100 fold over that obtained without RB47 binding site activation.

  11. Café Systems Express Access Form

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Café Systems Express Access Form Job Aid 7/18/12 - rb Café Systems Express Access Form - Job Aid NEED HELP? Call 847-491-HELP consultant@northwestern.edu © 2012 Northwestern University Page 1 of 3 Follow these steps to complete the Café Systems Express Access Form. For access not indicated by the roles below, use the General Security Access Form instead. 1. Select Applicant Information checkboxes (as applicable).  Enter Date, NetID, Name, EMPLID (employee ID number), title, department

  12. Changes in gene expression following EMF exposure

    SciTech Connect (OSTI)

    Woloschak, G.E.; Paunesku, T.; Chang-Liu, C.M.; Loberg, L.; Gauger, J.; McCormick, D.

    1997-10-01

    Experiments were designed to examine the effects of electromagnetic field (EMF) exposure on specific gene expression, an effect that can be deleterious, beneficial, or neutral, depending on the long-term consequences; however, the proof of a reproducible, quantitative biological effect (such as change in gene expression) will lead to latter experiments aimed at determining the relative contribution of these changes to cellular consequences. Past work by ourselves and by others has shown that measures of gene expression are extremely sensitive indicators of the cellular and biological effects of ionizing radiation, with transcriptional changes being detected by exposure of cells to doses of {gamma}-rays as low as 0.01 cGy that have no pronounced cellular consequences. On the basis of this work, the authors hypothesized that measures of gene expression will be equally sensitive to EMF effects on cells.

  13. Materials Data on Er3InN (SG:221) by Materials Project

    DOE Data Explorer [Office of Scientific and Technical Information (OSTI)]

    Kristin Persson

    2014-11-02

    Computed materials data using density functional theory calculations. These calculations determine the electronic structure of bulk materials by solving approximations to the Schrodinger equation. For more information, see https://materialsproject.org/docs/calculations

  14. Materials Data on Ho3InN (SG:221) by Materials Project

    DOE Data Explorer [Office of Scientific and Technical Information (OSTI)]

    Kristin Persson

    2014-11-02

    Computed materials data using density functional theory calculations. These calculations determine the electronic structure of bulk materials by solving approximations to the Schrodinger equation. For more information, see https://materialsproject.org/docs/calculations

  15. Materials Data on Sc3InN (SG:221) by Materials Project

    DOE Data Explorer [Office of Scientific and Technical Information (OSTI)]

    Kristin Persson

    2014-11-02

    Computed materials data using density functional theory calculations. These calculations determine the electronic structure of bulk materials by solving approximations to the Schrodinger equation. For more information, see https://materialsproject.org/docs/calculations

  16. Materials Data on Dy3InN (SG:221) by Materials Project

    DOE Data Explorer [Office of Scientific and Technical Information (OSTI)]

    Kristin Persson

    2014-11-02

    Computed materials data using density functional theory calculations. These calculations determine the electronic structure of bulk materials by solving approximations to the Schrodinger equation. For more information, see https://materialsproject.org/docs/calculations

  17. Materials Data on InN (SG:186) by Materials Project

    DOE Data Explorer [Office of Scientific and Technical Information (OSTI)]

    Kristin Persson

    Computed materials data using density functional theory calculations. These calculations determine the electronic structure of bulk materials by solving approximations to the Schrodinger equation. For more information, see https://materialsproject.org/docs/calculations

  18. Materials Data on InN (SG:186) by Materials Project

    DOE Data Explorer [Office of Scientific and Technical Information (OSTI)]

    Kristin Persson

    2014-11-02

    Computed materials data using density functional theory calculations. These calculations determine the electronic structure of bulk materials by solving approximations to the Schrodinger equation. For more information, see https://materialsproject.org/docs/calculations

  19. Visualization and Analysis of 3D Gene Expression Data (Technical...

    Office of Scientific and Technical Information (OSTI)

    Technical Report: Visualization and Analysis of 3D Gene Expression Data Citation Details In-Document Search Title: Visualization and Analysis of 3D Gene Expression Data Recent...

  20. Rhodobacter System for the Expression of Membrane Proteins |...

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Rhodobacter System for the Expression of Membrane Proteins Technology available for licensing: A unique system for membrane protein expression makes it possible to obtain...

  1. V-188: Apache XML Security XPointer Expressions Processing Buffer...

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    8: Apache XML Security XPointer Expressions Processing Buffer Overflow Vulnerability V-188: Apache XML Security XPointer Expressions Processing Buffer Overflow Vulnerability June...

  2. Sales Tax Exemption for Energy-Efficient Products (Sales Tax Holiday)

    Broader source: Energy.gov [DOE]

    Virginia allows a four-day sales tax* exemption for dishwashers, clothes washers, air conditioners, ceiling fans, light bulb, dehumidifiers, programmable thermostat and refrigerators that meet fe...

  3. They're Here! Winter, Holidays, and the New Year. How Will You...

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    This entry from last year will help you make SMART energy-saving resolutions that last longer than the average new-year gym membership, so check it out and tell us in the comments ...

  4. Sales Tax Exemption for Energy-Efficient Products (Sales Tax Holiday)

    Broader source: Energy.gov [DOE]

    In the past few years, the Georgia legislature has traditionally allowed an annual state and local sales tax exemption on Energy Star products of $1,500 or less per product, purchased for non...

  5. How Do You Save Money on Summer Holidays? | Department of Energy

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    the 4th of July. She mentioned several ways you can save energy by doing the things you probably did anyway -- like leaving the house or barbequing. But now that the 4th...

  6. North Pole’s Holiday Wish for An Energy Efficient 2012

    Broader source: Energy.gov [DOE]

    The city of North Pole, Alaska, is hoping to use $100,000 in Energy Efficiency and Conservation Block Grant funds to improve the energy efficiency of several key city facilities.

  7. EECBG Success Story: North Pole's Holiday Wish for an Energy Efficient 2012

    Broader source: Energy.gov [DOE]

    Up in the iconic community of North Pole, Alaska, the city leaders have made their energy efficiency upgrade wish list, and now state auditors are “checking it twice” to see which projects the city will be able to complete in 2012 using funds from the state’s Energy Efficiency and Conservation Block Grant (EECBG) as part of the American Recovery and Reinvestment Act. This can be critical in a city that sees winter temperatures as cold as -67F. Learn more.

  8. What Do You Think of Your LED Holiday Lights? | Department of...

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    savings, but some folks have also discovered that the lights have a slightly different color or reflect off of ornaments in a different way. Change can be difficult, especially...

  9. Gene expression profiles in irradiated cancer cells

    SciTech Connect (OSTI)

    Minafra, L.; Bravat, V.; Russo, G.; Ripamonti, M.; Gilardi, M. C.

    2013-07-26

    Knowledge of the molecular and genetic mechanisms underlying cellular response to radiation may provide new avenues to develop innovative predictive tests of radiosensitivity of tumours and normal tissues and to improve individual therapy. Nowadays very few studies describe molecular changes induced by hadrontherapy treatments, therefore this field has to be explored and clarified. High-throughput methodologies, such as DNA microarray, allow us to analyse mRNA expression of thousands of genes simultaneously in order to discover new genes and pathways as targets of response to hadrontherapy. Our aim is to elucidate the molecular networks involved in the sensitivity/resistance of cancer cell lines subjected to hadrontherapy treatments with a genomewide approach by using cDNA microarray technology to identify gene expression profiles and candidate genes responsible of differential cellular responses.

  10. Application of multidisciplinary analysis to gene expression.

    SciTech Connect (OSTI)

    Wang, Xuefel; Kang, Huining; Fields, Chris; Cowie, Jim R.; Davidson, George S.; Haaland, David Michael; Sibirtsev, Valeriy; Mosquera-Caro, Monica P.; Xu, Yuexian; Martin, Shawn Bryan; Helman, Paul; Andries, Erik; Ar, Kerem; Potter, Jeffrey; Willman, Cheryl L.; Murphy, Maurice H.

    2004-01-01

    Molecular analysis of cancer, at the genomic level, could lead to individualized patient diagnostics and treatments. The developments to follow will signal a significant paradigm shift in the clinical management of human cancer. Despite our initial hopes, however, it seems that simple analysis of microarray data cannot elucidate clinically significant gene functions and mechanisms. Extracting biological information from microarray data requires a complicated path involving multidisciplinary teams of biomedical researchers, computer scientists, mathematicians, statisticians, and computational linguists. The integration of the diverse outputs of each team is the limiting factor in the progress to discover candidate genes and pathways associated with the molecular biology of cancer. Specifically, one must deal with sets of significant genes identified by each method and extract whatever useful information may be found by comparing these different gene lists. Here we present our experience with such comparisons, and share methods developed in the analysis of an infant leukemia cohort studied on Affymetrix HG-U95A arrays. In particular, spatial gene clustering, hyper-dimensional projections, and computational linguistics were used to compare different gene lists. In spatial gene clustering, different gene lists are grouped together and visualized on a three-dimensional expression map, where genes with similar expressions are co-located. In another approach, projections from gene expression space onto a sphere clarify how groups of genes can jointly have more predictive power than groups of individually selected genes. Finally, online literature is automatically rearranged to present information about genes common to multiple groups, or to contrast the differences between the lists. The combination of these methods has improved our understanding of infant leukemia. While the complicated reality of the biology dashed our initial, optimistic hopes for simple answers from microarrays, we have made progress by combining very different analytic approaches.

  11. Expression of multiple proteins in transgenic plants

    DOE Patents [OSTI]

    Vierstra, Richard D.; Walker, Joseph M.

    2002-01-01

    A method is disclosed for the production of multiple proteins in transgenic plants. A DNA construct for introduction into plants includes a provision to express a fusion protein of two proteins of interest joined by a linking domain including plant ubiquitin. When the fusion protein is produced in the cells of a transgenic plant transformed with the DNA construction, native enzymes present in plant cells cleave the fusion protein to release both proteins of interest into the cells of the transgenic plant. Since the proteins are produced from the same fusion protein, the initial quantities of the proteins in the cells of the plant are approximately equal.

  12. Visualization and Analysis of 3D Gene Expression Data (Technical...

    Office of Scientific and Technical Information (OSTI)

    Technical Report: Visualization and Analysis of 3D Gene Expression Data Citation Details In-Document Search Title: Visualization and Analysis of 3D Gene Expression Data You are...

  13. Induction of gene expression using a high concentration sugar mixture

    DOE Patents [OSTI]

    England, George R.; Kelley, Aaron; Mitchinson, Colin

    2015-05-19

    Described herein is a composition useful for inducing expression of genes whose expression is under control of an inducible promoter sequence and methods for the compositions preparation and use.

  14. Ice Particle Projected Area- and Mass-dimension Expressions

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    m-D and A-D expressions in BMPs is described in this paper. Figure 1. The m-D expression (black curve) for synoptic ice clouds between -20C and -40C based on SCPP m-D...

  15. Induction of gene expression using a high concentration sugar mixture

    DOE Patents [OSTI]

    England, George R.; Kelley, Aaron; Mitchinson, Colin

    2010-05-11

    Described herein is a composition useful for inducing expression of genes whose expression is under control of an inducible promoter sequence and methods for the compositions preparation and use.

  16. Reduced expression of Autographa californica nucleopolyhedrovirus ORF34, an essential gene, enhances heterologous gene expression

    SciTech Connect (OSTI)

    Salem, Tamer Z.; Department of Microbial Molecular Biology, AGERI, Agricultural Research Center, Giza 12619; Division of Biomedical Sciences, Zewail University, Zewail City of Science and Technology, Giza 12588 ; Zhang, Fengrui; Thiem, Suzanne M.

    2013-01-20

    Autographa californica multiple nucleopolyhedrovirus ORF34 is part of a transcriptional unit that includes ORF32, encoding a viral fibroblast growth factor (FGF) and ORF33. We identified ORF34 as a candidate for deletion to improve protein expression in the baculovirus expression system based on enhanced reporter gene expression in an RNAi screen of virus genes. However, ORF34 was shown to be an essential gene. To explore ORF34 function, deletion (KO34) and rescue bacmids were constructed and characterized. Infection did not spread from primary KO34 transfected cells and supernatants from KO34 transfected cells could not infect fresh Sf21 cells whereas the supernatant from the rescue bacmids transfection could recover the infection. In addition, budded viruses were not observed in KO34 transfected cells by electron microscopy, nor were viral proteins detected from the transfection supernatants by western blots. These demonstrate that ORF34 is an essential gene with a possible role in infectious virus production.

  17. New expression for collisionless magnetic reconnection rate (Journal

    Office of Scientific and Technical Information (OSTI)

    Article) | SciTech Connect New expression for collisionless magnetic reconnection rate Citation Details In-Document Search Title: New expression for collisionless magnetic reconnection rate For 2D, symmetric, anti-parallel, collisionless magnetic reconnection, new expressions for the reconnection rate in the electron diffusion region are introduced. It is shown that these expressions can be derived in just a few simple steps from a physically intuitive starting point; the derivations are

  18. Express Primer Tool for high-throughput gene cloning and expression

    Energy Science and Technology Software Center (OSTI)

    2002-12-01

    A tool to assist in the design of primers for DNA amplification. The Express Primer web-based tool generates primer sequences specifically for the generation of expression clones for both lab scale and high-throughput projects. The application is designed not only to allow the user complete flexibility to specify primer design parameters but also to minimize the amount of manual intervention needed to generate a large number of primers for simultaneous amplification of multiple target genes.more » The Express Primer Tool enables the user to specify various experimental parameters (e.g. optimal Tm, Tm range, maximum Tm difference) for single or multiple candidate sequence(s) in FASTA format input as a flat text (ASCII) file. The application generates condidate primers, selects optimal primer pairs, and writes the forward and reverse primers pairs to an Excel file that is suitable for electronic submission to a synthesis facility. The program parameters emphasize high-throughput but allow for target atrition at various stages of the project.« less

  19. Repressor-mediated tissue-specific gene expression in plants

    DOE Patents [OSTI]

    Meagher, Richard B. (Athens, GA); Balish, Rebecca S. (Oxford, OH); Tehryung, Kim (Athens, GA); McKinney, Elizabeth C. (Athens, GA)

    2009-02-17

    Plant tissue specific gene expression by way of repressor-operator complexes, has enabled outcomes including, without limitation, male sterility and engineered plants having root-specific gene expression of relevant proteins to clean environmental pollutants from soil and water. A mercury hyperaccumulation strategy requires that mercuric ion reductase coding sequence is strongly expressed. The actin promoter vector, A2pot, engineered to contain bacterial lac operator sequences, directed strong expression in all plant vegetative organs and tissues. In contrast, the expression from the A2pot construct was restricted primarily to root tissues when a modified bacterial repressor (LacIn) was coexpressed from the light-regulated rubisco small subunit promoter in above-ground tissues. Also provided are analogous repressor operator complexes for selective expression in other plant tissues, for example, to produce male sterile plants.

  20. Estrogen induced {beta}-1,4-galactosyltransferase 1 expression regulates

    Office of Scientific and Technical Information (OSTI)

    proliferation of human breast cancer MCF-7 cells (Journal Article) | SciTech Connect Estrogen induced {beta}-1,4-galactosyltransferase 1 expression regulates proliferation of human breast cancer MCF-7 cells Citation Details In-Document Search Title: Estrogen induced {beta}-1,4-galactosyltransferase 1 expression regulates proliferation of human breast cancer MCF-7 cells Highlights: Black-Right-Pointing-Pointer We examined the regulation and biological functions of B4GALT1 expression induced

  1. DOE Seeks Expression of Interest for Carlsbad Technical Support Services |

    Energy Savers [EERE]

    Department of Energy Expression of Interest for Carlsbad Technical Support Services DOE Seeks Expression of Interest for Carlsbad Technical Support Services July 10, 2014 - 12:00pm Addthis Media Contact Lynette Chafin, 513-246-0461, Lynette.Chafin@emcbc.doe.gov Cincinnati - The U.S. Department of Energy (DOE) Environmental Management Consolidated Business Center (EMCBC) today issued a Sources Sought Synopsis/Expression of Interest seeking small business concerns with the capabilities to

  2. Small, Fast S-Expression Library, Version 1.0

    Energy Science and Technology Software Center (OSTI)

    2004-12-02

    This software is a library for C and C++ programmers to use for fast and efficient parsing and processing of symbolic expressions, or s-expressions. An s-expression is a text-representation of a tree data structure, and is the basis for the syntax of the LISP and Scheme programming languages. Multiple similar libraries exist, but this one was designed from the ground up for speed, efficiency, and simplicity.

  3. Alternative Fuels Data Center: Massachusetts Fleet Braun's Express

    Alternative Fuels and Advanced Vehicles Data Center [Office of Energy Efficiency and Renewable Energy (EERE)]

    Celebrates 10 Years of Petroleum Reduction Success Massachusetts Fleet Braun's Express Celebrates 10 Years of Petroleum Reduction Success to someone by E-mail Share Alternative Fuels Data Center: Massachusetts Fleet Braun's Express Celebrates 10 Years of Petroleum Reduction Success on Facebook Tweet about Alternative Fuels Data Center: Massachusetts Fleet Braun's Express Celebrates 10 Years of Petroleum Reduction Success on Twitter Bookmark Alternative Fuels Data Center: Massachusetts Fleet

  4. New methods for tightly regulated gene expression and highly...

    Office of Scientific and Technical Information (OSTI)

    utilizing the site-specific recombination system from the Streptomyces phage C31. Host strains expressing the C31 integrase gene and carrying an appropriate recombination ...

  5. Using Rhodobacter Bacteria to Express Membrane Proteins (ANL...

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    For example, eukaryotic expression systems are costly, and a system that uses the E. coli bacterium can cause host toxicity or produce insoluble aggregates. However, a unique...

  6. U-058: Apache Struts Conversion Error OGNL Expression Injection...

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    in Apache Struts. A remote user can execute arbitrary commands on the target system. PLATFORM: Apache Struts 2.x ABSTRACT: Apache Struts Conversion Error OGNL Expression...

  7. Estrogen induced {beta}-1,4-galactosyltransferase 1 expression...

    Office of Scientific and Technical Information (OSTI)

    cancer MCF-7 cells Citation Details In-Document Search Title: Estrogen induced beta-1,4-galactosyltransferase 1 expression regulates proliferation of human breast cancer ...

  8. ANALYTIC EXPRESSIONS FOR THE LIGHT-SCATTERING CROSS SECTION

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    of r and m, with many "wiggles" on many scales, and its computation, though straightforward, is time intensive. A simple expression for Q sp would simplify calculation of...

  9. Massively parallel processor networks with optical express channels

    DOE Patents [OSTI]

    Deri, R.J.; Brooks, E.D. III; Haigh, R.E.; DeGroot, A.J.

    1999-08-24

    An optical method for separating and routing local and express channel data comprises interconnecting the nodes in a network with fiber optic cables. A single fiber optic cable carries both express channel traffic and local channel traffic, e.g., in a massively parallel processor (MPP) network. Express channel traffic is placed on, or filtered from, the fiber optic cable at a light frequency or a color different from that of the local channel traffic. The express channel traffic is thus placed on a light carrier that skips over the local intermediate nodes one-by-one by reflecting off of selective mirrors placed at each local node. The local-channel-traffic light carriers pass through the selective mirrors and are not reflected. A single fiber optic cable can thus be threaded throughout a three-dimensional matrix of nodes with the x,y,z directions of propagation encoded by the color of the respective light carriers for both local and express channel traffic. Thus frequency division multiple access is used to hierarchically separate the local and express channels to eliminate the bucket brigade latencies that would otherwise result if the express traffic had to hop between every local node to reach its ultimate destination. 3 figs.

  10. Massively parallel processor networks with optical express channels

    DOE Patents [OSTI]

    Deri, Robert J. (Pleasanton, CA); Brooks, III, Eugene D. (Livermore, CA); Haigh, Ronald E. (Tracy, CA); DeGroot, Anthony J. (Castro Valley, CA)

    1999-01-01

    An optical method for separating and routing local and express channel data comprises interconnecting the nodes in a network with fiber optic cables. A single fiber optic cable carries both express channel traffic and local channel traffic, e.g., in a massively parallel processor (MPP) network. Express channel traffic is placed on, or filtered from, the fiber optic cable at a light frequency or a color different from that of the local channel traffic. The express channel traffic is thus placed on a light carrier that skips over the local intermediate nodes one-by-one by reflecting off of selective mirrors placed at each local node. The local-channel-traffic light carriers pass through the selective mirrors and are not reflected. A single fiber optic cable can thus be threaded throughout a three-dimensional matrix of nodes with the x,y,z directions of propagation encoded by the color of the respective light carriers for both local and express channel traffic. Thus frequency division multiple access is used to hierarchically separate the local and express channels to eliminate the bucket brigade latencies that would otherwise result if the express traffic had to hop between every local node to reach its ultimate destination.

  11. Workshop on Program for Elimination of Requirements Marginal to Safety: Proceedings

    SciTech Connect (OSTI)

    Dey, M. [Nuclear Regulatory Commission, Washington, DC (United States). Div. of Safety Issue Resolution; Arsenault, F.; Patterson, M.; Gaal, M. [SCIENTECH, Inc., Rockville, MD (United States)

    1993-09-01

    These are the proceedings of the Public Workshop on the US Nuclear Regulatory Commission`s Program for Elimination of Requirements Marginal to Safety. The workshop was held at the Holiday Inn, Bethesda, on April 27 and 28, 1993. The purpose of the workshop was to provide an opportunity for public and industry input to the program. The workshop addressed the institutionalization of the program to review regulations with the purpose of eliminating those that are marginal. The objective is to avoid the dilution of safety efforts. One session was devoted to discussion of the framework for a performance-based regulatory approach. In addition, panelists and attendees discussed scope, schedules and status of specific regulatory items: containment leakage testing requirements, fire protection requirements, requirements for environmental qualification of electrical equipment, requests for information under 10CFR50.54(f), requirements for combustible gas control systems, and quality assurance requirements.

  12. Northern New Mexico Citizens Advisory Board Meeting

    Office of Environmental Management (EM)

    CERTIFIED-- NNMCAB Meeting Minutes 1-28-09 1 2 3 4 5 6 7 8 9 10 Northern New Mexico Citizens' Advisory Board Meeting January 28, 2009 2:00 p.m. to 8:00 p.m. Holiday Inn Santa Fe 4048 Cerrillos Road Santa Fe, New Mexico MINUTES In Attendance: 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 NNMCAB Members- 1. J.D Campbell, NNMCAB Chair 2. Ralph Phelps, NNMCAB Vice Chair 3. Gerry Maestas, WM Committee Chair 4. Peter Baston 5. Donald Dayton 6.

  13. Northern New Mexico Citizens Advisory Board Meeting

    Office of Environmental Management (EM)

    CAB Meeting Minutes 5-22-08 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 Northern New Mexico Citizens' Advisory Board Meeting May 22, 2008 9:00 a.m. to 4:00 p.m. Holiday Inn 4048 Cerrillos Road, Santa Fe Santa Fe, New Mexico In Attendance: NNMCAB Members- 1. J.D Campbell, NNMCAB Chair 2. Pam Henline, EMSR Committee Chair 3. Ralph Phelps, WM Committee Chair 4. Larry Rapagnani, WM Committee Vice Chair 5. Donald

  14. Northern New Mexico Citizens Advisory Board Meeting

    Office of Environmental Management (EM)

    9-30-09 Chair Certified 1 2 3 4 5 6 7 8 Northern New Mexico Citizens' Advisory Board Meeting September 30, 2009 2:00 p.m. to 8:00 p.m. Holiday Inn, 4048 Cerrillos Road Santa Fe, New Mexico MINUTES Attending: 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 NNMCAB Members- 1. J.D Campbell, NNMCAB Chair 2. Ralph Phelps, NNMCAB Vice Chair 3. Mike Loya, EMSR Committee Chair 4. Pam Henline, EMSR Committee Vice Chair 5. Gerry Maestas, WM Committee

  15. Combined hairpin-antisense compositions and methods for modulating expression

    DOE Patents [OSTI]

    Shanklin, John; Nguyen, Tam Huu

    2015-11-24

    A nucleotide construct comprising a nucleotide sequence that forms a stem and a loop, wherein the loop comprises a nucleotide sequence that modulates expression of a target, wherein the stem comprises a nucleotide sequence that modulates expression of a target, and wherein the target modulated by the nucleotide sequence in the loop and the target modulated by the nucleotide sequence in the stem may be the same or different. Vectors, methods of regulating target expression, methods of providing a cell, and methods of treating conditions comprising the nucleotide sequence are also disclosed.

  16. Combined hairpin-antisense compositions and methods for modulating expression

    DOE Patents [OSTI]

    Shanklin, John; Nguyen, Tam

    2014-08-05

    A nucleotide construct comprising a nucleotide sequence that forms a stem and a loop, wherein the loop comprises a nucleotide sequence that modulates expression of a target, wherein the stem comprises a nucleotide sequence that modulates expression of a target, and wherein the target modulated by the nucleotide sequence in the loop and the target modulated by the nucleotide sequence in the stem may be the same or different. Vectors, methods of regulating target expression, methods of providing a cell, and methods of treating conditions comprising the nucleotide sequence are also disclosed.

  17. Chicago Solar Express Reduces Costs, Wait Times | Department of Energy

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Chicago Solar Express Reduces Costs, Wait Times Chicago Solar Express Reduces Costs, Wait Times October 28, 2014 - 10:48am Addthis The Solar Express program in Chicago, Illinois-funded through a SunShot Initiative Rooftop Solar Challenge (RSC) I award of $750,000-is making it faster, easier, and cheaper for residents to go solar by cutting long wait times and fees for solar permits. Residents of Chicago can now acquire permits for their residential solar installations in one day, compared to 30

  18. DOE - Office of Legacy Management -- American Railway Express Office - NY

    Office of Legacy Management (LM)

    0-03 Railway Express Office - NY 0-03 FUSRAP Considered Sites Site: American Railway Express Office (NY.0-03 ) Eliminated from consideration under FUSRAP Designated Name: Not Designated Alternate Name: None Location: American Railway Express (Downtown) , New York , New York NY.0-03-1 Evaluation Year: 1987 NY.0-03-1 Site Operations: None - Involved with a fire during transport of uranium scrap. NY.0-03-2 Site Disposition: Eliminated - Potential for contamination remote NY.0-03-1 NY.0-03-2

  19. Array design and expression evaluation in POOMA II

    SciTech Connect (OSTI)

    Karmesin, S.; Crotinger, J.; Cummings, J.; Haney, S.; Humphrey, W.; Reynders, J.; Smith, S.; Williams, T.J.

    1998-12-31

    POOMA is a templated C++ class library for use in the development of large-scale scientific simulations on serial and parallel computers. POOMA II is a new design and implementation of POOMA intended to add richer capabilities and greater flexibility to the framework. The new design employs a generic Array class that acts as an interface to, or view on, a wide variety of data representation objects referred to as engines. This design separates the interface and the representation of multidimensional arrays. The separation is achieved using compile-time techniques rather than virtual functions, and thus code efficiency is maintained. POOMA II uses PETE, the Portable Expression Template Engine, to efficiently represent complex mathematical expressions involving arrays and other objects. The representation of expressions is kept separate from expression evaluation, allowing the use of multiple evaluator mechanisms that can support nested where-block constructs, hardware-specific optimizations and different run-time environments.

  20. Global Profiling of Shewanella oneidensis MR-1: Expression of Hypothetical

    Office of Scientific and Technical Information (OSTI)

    Genes and Improved functional annotations (Journal Article) | SciTech Connect Global Profiling of Shewanella oneidensis MR-1: Expression of Hypothetical Genes and Improved functional annotations Citation Details In-Document Search Title: Global Profiling of Shewanella oneidensis MR-1: Expression of Hypothetical Genes and Improved functional annotations The y-proteobacterium Shewanella oneidensis strain MR-1 is a respiratory versatile organism that can reduce a wide range of organics, metals,

  1. Fluorescence-based optimization of human bitter taste receptor expression

    Office of Scientific and Technical Information (OSTI)

    in Saccharomyces cerevisiae (Journal Article) | SciTech Connect Fluorescence-based optimization of human bitter taste receptor expression in Saccharomyces cerevisiae Citation Details In-Document Search Title: Fluorescence-based optimization of human bitter taste receptor expression in Saccharomyces cerevisiae Human TAS2 receptors (hTAS2Rs) perceive bitter tastants, but few studies have explored the structure-function relationships of these receptors. In this paper, we report our trials on

  2. New methods for tightly regulated gene expression and highly efficient

    Office of Scientific and Technical Information (OSTI)

    chromosomal integration of cloned genes for Methanosarcina species (Journal Article) | SciTech Connect New methods for tightly regulated gene expression and highly efficient chromosomal integration of cloned genes for Methanosarcina species Citation Details In-Document Search Title: New methods for tightly regulated gene expression and highly efficient chromosomal integration of cloned genes for Methanosarcina species A highly efficient method for chromosomal integration of cloned DNA into

  3. Advanced method for high-throughput expression of mutated eukaryotic

    Office of Scientific and Technical Information (OSTI)

    membrane proteins in Saccharomyces cerevisiae (Journal Article) | SciTech Connect SciTech Connect Search Results Journal Article: Advanced method for high-throughput expression of mutated eukaryotic membrane proteins in Saccharomyces cerevisiae Citation Details In-Document Search Title: Advanced method for high-throughput expression of mutated eukaryotic membrane proteins in Saccharomyces cerevisiae Crystallization of eukaryotic membrane proteins is a challenging, iterative process. The

  4. Final EIS for Champlain Hudson Power Express Transmission Project Now

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Available | Department of Energy Final EIS for Champlain Hudson Power Express Transmission Project Now Available Final EIS for Champlain Hudson Power Express Transmission Project Now Available August 8, 2014 - 4:44pm Addthis The U.S. Department of Energy (DOE) has prepared a Final Environmental Impact Statement (EIS) pursuant to the National Environmental Policy Act (NEPA) of 1969, as amended (42 U.S.C. 4321 et seq.), the Council on Environmental Quality (CEQ) NEPA regulations (40 CFR

  5. Gene co-expression network analysis in Rhodobacter capsulatus and application to comparative expression analysis of Rhodobacter sphaeroides

    SciTech Connect (OSTI)

    Pena-Castillo, Lourdes; Mercer, Ryan; Gurinovich, Anastasia; Callister, Stephen J.; Wright, Aaron T.; Westbye, Alexander; Beatty, J. T.; Lang, Andrew S.

    2014-08-28

    The genus Rhodobacter contains purple nonsulfur bacteria found mostly in freshwater environments. Representative strains of two Rhodobacter species, R. capsulatus and R. sphaeroides, have had their genomes fully sequenced and both have been the subject of transcriptional profiling studies. Gene co-expression networks can be used to identify modules of genes with similar expression profiles. Functional analysis of gene modules can then associate co-expressed genes with biological pathways, and network statistics can determine the degree of module preservation in related networks. In this paper, we constructed an R. capsulatus gene co-expression network, performed functional analysis of identified gene modules, and investigated preservation of these modules in R. capsulatus proteomics data and in R. sphaeroides transcriptomics data. Results: The analysis identified 40 gene co-expression modules in R. capsulatus. Investigation of the module gene contents and expression profiles revealed patterns that were validated based on previous studies supporting the biological relevance of these modules. We identified two R. capsulatus gene modules preserved in the protein abundance data. We also identified several gene modules preserved between both Rhodobacter species, which indicate that these cellular processes are conserved between the species and are candidates for functional information transfer between species. Many gene modules were non-preserved, providing insight into processes that differentiate the two species. In addition, using Local Network Similarity (LNS), a recently proposed metric for expression divergence, we assessed the expression conservation of between-species pairs of orthologs, and within-species gene-protein expression profiles. Conclusions: Our analyses provide new sources of information for functional annotation in R. capsulatus because uncharacterized genes in modules are now connected with groups of genes that constitute a joint functional annotation. We identified R. capsulatus modules enriched with genes for ribosomal proteins, porphyrin and bacteriochlorophyll anabolism, and biosynthesis of secondary metabolites to be preserved in R. sphaeroides whereas modules related to RcGTA production and signalling showed lack of preservation in R. sphaeroides. In addition, we demonstrated that network statistics may also be applied within-species to identify congruence between mRNA expression and protein abundance data for which simple correlation measurements have previously had mixed results.

  6. Assessment of Normal Variability in Peripheral Blood Gene Expression

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Campbell, Catherine; Vernon, Suzanne D.; Karem, Kevin L.; Nisenbaum, Rosane; Unger, Elizabeth R.

    2002-01-01

    Peripheral blood is representative of many systemic processes and is an ideal sample for expression profiling of diseases that have no known or accessible lesion. Peripheral blood is a complex mixture of cell types and some differences in peripheral blood gene expression may reflect the timing of sample collection rather than an underlying disease process. For this reason, it is important to assess study design factors that may cause variability in gene expression not related to what is being analyzed. Variation in the gene expression of circulating peripheral blood mononuclear cells (PBMCs) from three healthy volunteers sampled three times onemore » day each week for one month was examined for 1,176 genes printed on filter arrays. Less than 1% of the genes showed any variation in expression that was related to the time of collection, and none of the changes were noted in more than one individual. These results suggest that observed variation was due to experimental variability.« less

  7. Recombinant cells that highly express chromosomally-integrated heterologous gene

    DOE Patents [OSTI]

    Ingram, Lonnie O.; Ohta, Kazuyoshi; Wood, Brent E.

    2007-03-20

    Recombinant host cells are obtained that comprise (A) a heterologous, polypeptide-encoding polynucleotide segment, stably integrated into a chromosome, which is under transcriptional control of an endogenous promoter and (B) a mutation that effects increased expression of the heterologous segment, resulting in enhanced production by the host cells of each polypeptide encoded by that segment, relative to production of each polypeptide by the host cells in the absence of the mutation. The increased expression thus achieved is retained in the absence of conditions that select for cells displaying such increased expression. When the integrated segment comprises, for example, ethanol-production genes from an efficient ethanol producer like Zymomonas mobilis, recombinant Escherichia coli and other enteric bacterial cells within the present invention are capable of converting a wide range of biomass-derived sugars efficiently to ethanol.

  8. Recombinant cells that highly express chromosomally-integrated heterologous genes

    DOE Patents [OSTI]

    Ingram, Lonnie O.; Ohta, Kazuyoshi; Wood, Brent E.

    2000-08-22

    Recombinant host cells are obtained that comprise (A) a heterologous, polypeptide-encoding polynucleotide segment, stably integrated into a chromosome, which is under transcriptional control of an endogenous promoter and (B) a mutation that effects increased expression of the heterologous segment, resulting in enhanced production by the host cells of each polypeptide encoded by that segment, relative to production of each polypeptide by the host cells in the absence of the mutation. The increased expression thus achieved is retained in the absence of conditions that select for cells displaying such increased expression. When the integrated segment comprises, for example, ethanol-production genes from an efficient ethanol producer like Zymomonas mobilis, recombinant Escherichia coli and other enteric bacterial cells within the present invention are capable of converting a wide range of biomass-derived sugars efficiently to ethanol.

  9. Recombinant cells that highly express chromosomally-integrated heterologous genes

    DOE Patents [OSTI]

    Ingram, L.O.; Ohta, Kazuyoshi; Wood, B.E.

    1998-10-13

    Recombinant host cells are obtained that comprise (A) a heterologous, polypeptide-encoding polynucleotide segment, stably integrated into a chromosome, which is under transcriptional control of an endogenous promoter and (B) a mutation that effects increased expression of the heterologous segment, resulting in enhanced production by the host cells of each polypeptide encoded by that segment, relative to production of each polypeptide by the host cells in the absence of the mutation. The increased expression thus achieved is retained in the absence of conditions that select for cells displaying such increased expression. When the integrated segment comprises, for example, ethanol-production genes from an efficient ethanol producer like Zymomonas mobilis, recombinant Escherichia coli and other enteric bacterial cells within the present invention are capable of converting a wide range of biomass-derived sugars efficiently to ethanol. 13 figs.

  10. Recombinant cells that highly express chromosomally-integrated heterologous genes

    DOE Patents [OSTI]

    Ingram, Lonnie O.; Ohta, Kazuyoshi; Wood, Brent E.

    1998-01-01

    Recombinant host cells are obtained that comprise (A) a heterologous, polypeptide-encoding polynucleotide segment, stably integrated into a chromosome, which is under transcriptional control of an endogenous promoter and (B) a mutation that effects increased expression of the heterologous segment, resulting in enhanced production by the host cells of each polypeptide encoded by that segment, relative to production of each polypeptide by the host cells in the absence of the mutation. The increased expression thus achieved is retained in the absence of conditions that select for cells displaying such increased expression. When the integrated segment comprises, for example, ethanol-production genes from an efficient ethanol producer like Zymomonas mobilis, recombinant Escherichia coli and other enteric bacterial cells within the present invention are capable of converting a wide range of biomass-derived sugars efficiently to ethanol.

  11. Explicit Expressions for 3D Boundary Integrals in Potential Theory

    SciTech Connect (OSTI)

    Nintcheu Fata, Sylvain

    2009-01-01

    On employing isoparametric, piecewise linear shape functions over a flat triangular domain, exact expressions are derived for all surface potentials involved in the numerical solution of three-dimensional singular and hyper-singular boundary integral equations of potential theory. These formulae, which are valid for an arbitrary source point in space, are represented as analytic expressions over the edges of the integration triangle. They can be used to solve integral equations defined on polygonal boundaries via the collocation method or may be utilized as analytic expressions for the inner integrals in the Galerkin technique. Also, the constant element approximation can be directly obtained with no extra effort. Sample problems solved by the collocation boundary element method for the Laplace equation are included to validate the proposed formulae.

  12. Metaproteomics reveals abundant transposase expression in mutualistic endosymbionts

    SciTech Connect (OSTI)

    Kleiner, Manuel; Young, Jacque C; Shah, Manesh B; Verberkmoes, Nathan C; Dubilier, Nicole

    2013-01-01

    Transposases, enzymes that catalyze the movement of mobile genetic elements, are the most abundant genes in nature. While many bacteria encode an abundance of transposases in their genomes, the current paradigm is that transposase gene expression is tightly regulated and generally low due to its severe mutagenic effects. In the current study, we detected the highest number of transposase proteins ever reported in bacteria, in symbionts of the gutless marine worm Olavius algarvensis using metaproteomics. At least 26 different transposases from 12 different families were detected and genomic and proteomic analyses suggest many of these are active. This high expression of transposases indicates that the mechanisms for their tight regulation have been disabled or destroyed. Based on recent studies on other symbionts and pathogens that showed high transposase transcription, we speculate that abundant transposase expression might be common in symbionts and pathogens.

  13. Robust expression of a bioactive mammalian protein in Chlamydomonas chloroplast

    DOE Patents [OSTI]

    Mayfield, Stephen P

    2015-01-13

    Methods and compositions are disclosed to engineer chloroplast comprising heterologous mammalian genes via a direct replacement of chloroplast Photosystem II (PSII) reaction center protein coding regions to achieve expression of recombinant protein above 5% of total protein. When algae is used, algal expressed protein is produced predominantly as a soluble protein where the functional activity of the peptide is intact. As the host algae is edible, production of biologics in this organism for oral delivery of proteins/peptides, especially gut active proteins, without purification is disclosed.

  14. Robust expression of a bioactive mammalian protein in chlamydomonas chloroplast

    DOE Patents [OSTI]

    Mayfield, Stephen P.

    2010-03-16

    Methods and compositions are disclosed to engineer chloroplast comprising heterologous mammalian genes via a direct replacement of chloroplast Photosystem II (PSII) reaction center protein coding regions to achieve expression of recombinant protein above 5% of total protein. When algae is used, algal expressed protein is produced predominantly as a soluble protein where the functional activity of the peptide is intact. As the host algae is edible, production of biologics in this organism for oral delivery or proteins/peptides, especially gut active proteins, without purification is disclosed.

  15. A novel cantharidin analog N-Benzylcantharidinamide reduces the expression

    Office of Scientific and Technical Information (OSTI)

    of MMP-9 and invasive potentials of Hep3B via inhibiting cytosolic translocation of HuR (Journal Article) | SciTech Connect A novel cantharidin analog N-Benzylcantharidinamide reduces the expression of MMP-9 and invasive potentials of Hep3B via inhibiting cytosolic translocation of HuR Citation Details In-Document Search Title: A novel cantharidin analog N-Benzylcantharidinamide reduces the expression of MMP-9 and invasive potentials of Hep3B via inhibiting cytosolic translocation of HuR

  16. Multiclass cancer diagnosis using tumor gene expression signatures

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Ramaswamy, S.; Tamayo, P.; Rifkin, R.; Mukherjee, S.; Yeang, C. -H.; Angelo, M.; Ladd, C.; Reich, M.; Latulippe, E.; Mesirov, J. P.; et al

    2001-12-11

    The optimal treatment of patients with cancer depends on establishing accurate diagnoses by using a complex combination of clinical and histopathological data. In some instances, this task is difficult or impossible because of atypical clinical presentation or histopathology. To determine whether the diagnosis of multiple common adult malignancies could be achieved purely by molecular classification, we subjected 218 tumor samples, spanning 14 common tumor types, and 90 normal tissue samples to oligonucleotide microarray gene expression analysis. The expression levels of 16,063 genes and expressed sequence tags were used to evaluate the accuracy of a multiclass classifier based on a supportmore » vector machine algorithm. Overall classification accuracy was 78%, far exceeding the accuracy of random classification (9%). Poorly differentiated cancers resulted in low-confidence predictions and could not be accurately classified according to their tissue of origin, indicating that they are molecularly distinct entities with dramatically different gene expression patterns compared with their well differentiated counterparts. Taken together, these results demonstrate the feasibility of accurate, multiclass molecular cancer classification and suggest a strategy for future clinical implementation of molecular cancer diagnostics.« less

  17. Gene expression analysis of precision-cut human liver slices indicates stable expression of ADME-Tox related genes

    SciTech Connect (OSTI)

    Elferink, M.G.L., E-mail: m.g.l.elferink@rug.nl [Department of Pharmacokinetics, Toxicology and Targeting, Groningen Research Institute for Pharmacy, University of Groningen (Netherlands); Olinga, P. [Department of Pharmacokinetics, Toxicology and Targeting, Groningen Research Institute for Pharmacy, University of Groningen (Netherlands); van Leeuwen, E.M.; Bauerschmidt, S.; Polman, J. [Molecular Design and Informatics, MSD, Oss (Netherlands); Schoonen, W.G. [Toxicology and Drug Disposition, MSD, Oss (Netherlands); Heisterkamp, S.H. [Biostatistics and Research Decision Sciences MSD, Oss (Netherlands); Bioinformatics Centre, University of Groningen (Netherlands); Groothuis, G.M.M. [Department of Pharmacokinetics, Toxicology and Targeting, Groningen Research Institute for Pharmacy, University of Groningen (Netherlands)

    2011-05-15

    In the process of drug development it is of high importance to test the safety of new drugs with predictive value for human toxicity. A promising approach of toxicity testing is based on shifts in gene expression profiling of the liver. Toxicity screening based on animal liver cells cannot be directly extrapolated to humans due to species differences. The aim of this study was to evaluate precision-cut human liver slices as in vitro method for the prediction of human specific toxicity by toxicogenomics. The liver slices contain all cell types of the liver in their natural architecture. This is important since drug-induced toxicity often is a multi-cellular process. Previously we showed that toxicogenomic analysis of rat liver slices is highly predictive for rat in vivo toxicity. In this study we investigated the levels of gene expression during incubation up to 24 h with Affymetrix microarray technology. The analysis was focused on a broad spectrum of genes related to stress and toxicity, and on genes encoding for phase-I, -II and -III metabolizing enzymes and transporters. Observed changes in gene expression were associated with cytoskeleton remodeling, extracellular matrix and cell adhesion, but for the ADME-Tox related genes only minor changes were observed. PCA analysis showed that changes in gene expression were not associated with age, sex or source of the human livers. Slices treated with acetaminophen showed patterns of gene expression related to its toxicity. These results indicate that precision-cut human liver slices are relatively stable during 24 h of incubation and represent a valuable model for human in vitro hepatotoxicity testing despite the human inter-individual variability.

  18. Emerging Use of Gene Expression Microarrays in Plant Physiology

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Wullschleger, Stan D.; Difazio, Stephen P.

    2003-01-01

    Microarrays have become an important technology for the global analysis of gene expression in humans, animals, plants, and microbes. Implemented in the context of a well-designed experiment, cDNA and oligonucleotide arrays can provide highthroughput, simultaneous analysis of transcript abundance for hundreds, if not thousands, of genes. However, despite widespread acceptance, the use of microarrays as a tool to better understand processes of interest to the plant physiologist is still being explored. To help illustrate current uses of microarrays in the plant sciences, several case studies that we believe demonstrate the emerging application of gene expression arrays in plant physiology weremore » selected from among the many posters and presentations at the 2003 Plant and Animal Genome XI Conference. Based on this survey, microarrays are being used to assess gene expression in plants exposed to the experimental manipulation of air temperature, soil water content and aluminium concentration in the root zone. Analysis often includes characterizing transcript profiles for multiple post-treatment sampling periods and categorizing genes with common patterns of response using hierarchical clustering techniques. In addition, microarrays are also providing insights into developmental changes in gene expression associated with fibre and root elongation in cotton and maize, respectively. Technical and analytical limitations of microarrays are discussed and projects attempting to advance areas of microarray design and data analysis are highlighted. Finally, although much work remains, we conclude that microarrays are a valuable tool for the plant physiologist interested in the characterization and identification of individual genes and gene families with potential application in the fields of agriculture, horticulture and forestry.« less

  19. Transgenic rabbit that expresses a functional human lipoprotein (a)

    DOE Patents [OSTI]

    Rouy, Didier (Thiais, FR); Duverger, Nicolas (Paris, FR); Emmanuel, Florence (Aubervilliers, FR); Denefle, Patrice (Saint Maur, FR); Houdebine, Louis-Marie (Buc, FR); Viglietta, Celine (Versailles, FR); Rubin, Edward M. (Berkeley, CA); Hughes, Steven D. (Oakland, CA)

    2003-01-01

    A transgenic rabbit which has in its genomic DNA sequences that encode apolipoprotein (a) and apolipoprotein B polypeptides which are capable of combining to produce lipoprotein (a), a process for creating such a rabbit, and the use of the rabbit to identify compounds which are effective in the treatment of human diseases which are associated with, induced and/or exacerbated by Lp(a) expression.

  20. Imaging gene expression in real-time using aptamers

    SciTech Connect (OSTI)

    Shin, Il Chung

    2011-12-13

    Signal transduction pathways are usually activated by external stimuli and are transient. The downstream changes such as transcription of the activated genes are also transient. Real-time detection of promoter activity is useful for understanding changes in gene expression, especially during cell differentiation and in development. A simple and reliable method for viewing gene expression in real time is not yet available. Reporter proteins such as fluorescent proteins and luciferase allow for non-invasive detection of the products of gene expression in living cells. However, current reporter systems do not provide for real-time imaging of promoter activity in living cells. This is because of the long time period after transcription required for fluorescent protein synthesis and maturation. We have developed an RNA reporter system for imaging in real-time to detect changes in promoter activity as they occur. The RNA reporter uses strings of RNA aptamers that constitute IMAGEtags (Intracellular MultiAptamer GEnetic tags), which can be expressed from a promoter of choice. The tobramycin, neomycin and PDC RNA aptamers have been utilized for this system and expressed in yeast from the GAL1 promoter. The IMAGEtag RNA kinetics were quantified by RT-qPCR. In yeast precultured in raffinose containing media the GAL1 promoter responded faster than in yeast precultured in glucose containing media. IMAGEtag RNA has relatively short half-life (5.5 min) in yeast. For imaging, the yeast cells are incubated with their ligands that are labeled with fluorescent dyes. To increase signal to noise, ligands have been separately conjugated with the FRET (Frster resonance energy transfer) pairs, Cy3 and Cy5. With these constructs, the transcribed aptamers can be imaged after activation of the promoter by galactose. FRET was confirmed with three different approaches, which were sensitized emission, acceptor photobleaching and donor lifetime by FLIM (fluorescence lifetime imaging microscopy). Real-time transcription was measured by FLIM-FRET, which was detected by the decrease in donor lifetime resulting from ligand binding to IMAGEtags that were newly synthesized from the activated GAL1 promoter. The FRET signal was specific for transcribed IMAGEtags.

  1. Population genetic variation in gene expression is associated withphenotypic variation in Saccharomyces cerevisiae

    SciTech Connect (OSTI)

    Fay, Justin C.; McCullough, Heather L.; Sniegowski, Paul D.; Eisen, Michael B.

    2004-02-25

    The relationship between genetic variation in gene expression and phenotypic variation observable in nature is not well understood. Identifying how many phenotypes are associated with differences in gene expression and how many gene-expression differences are associated with a phenotype is important to understanding the molecular basis and evolution of complex traits. Results: We compared levels of gene expression among nine natural isolates of Saccharomyces cerevisiae grown either in the presence or absence of copper sulfate. Of the nine strains, two show a reduced growth rate and two others are rust colored in the presence of copper sulfate. We identified 633 genes that show significant differences in expression among strains. Of these genes,20 were correlated with resistance to copper sulfate and 24 were correlated with rust coloration. The function of these genes in combination with their expression pattern suggests the presence of both correlative and causative expression differences. But the majority of differentially expressed genes were not correlated with either phenotype and showed the same expression pattern both in the presence and absence of copper sulfate. To determine whether these expression differences may contribute to phenotypic variation under other environmental conditions, we examined one phenotype, freeze tolerance, predicted by the differential expression of the aquaporin gene AQY2. We found freeze tolerance is associated with the expression of AQY2. Conclusions: Gene expression differences provide substantial insight into the molecular basis of naturally occurring traits and can be used to predict environment dependent phenotypic variation.

  2. Methods and constructs for expression of foreign proteins in photosynthetic organisms

    DOE Patents [OSTI]

    Laible, Philip D. (Villa Park, IL); Hanson, Deborah K. (Downers Grove, IL)

    2002-01-01

    A method for expressing and purifying foreign proteins in photosynthetic organisms comprising the simultaneous expression of both the heterologous protein and a means for compartmentalizing or sequestering of the protein.

  3. Computational expressions for signals in frequency-modulation spectroscopy

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Di Rosa, Michael D.; Reiten, M. T.

    2015-05-25

    In this study, general expressions for the signals in frequency-modulation spectroscopy (FMS) appear in the literature but are often reduced to simple analytical equations following the assumption of a weak modulation index. This is little help to the experimentalist who wants to predict signals for modulation depths of the order of unity or greater, where strong FMS signals reside. Here, we develop general formulas for FMS signals in the case of an absorber with a Voigt line shape and then link these expressions to an example and existing numerical code for the line shape. The resulting computational recipe is easymore » to implement and exercised here to show where the larger FMS signals are found over the coordinates of modulation index and modulation frequency. One can also estimate from provided curves the in-phase FMS signal over a wide range of modulation parameters at either the Lorentzian-broadening or Doppler-broadening limit, or anywhere in between by interpolation.« less

  4. Advanced radioisotope power source options for Pluto Express

    SciTech Connect (OSTI)

    Underwood, M.L.

    1995-12-31

    In the drive to reduce mass and cost, Pluto Express is investigating using an advanced power conversion technology in a small Radioisotope Power Source (RPS) to deliver the required mission power of 74 W(electric) at end of mission. Until this year the baseline power source under consideration has been a Radioisotope Thermoelectric Generator (RTG). This RTG would be a scaled down GPHS RTG with an inventory of 6 General Purpose Heat Sources (GPHS) and a mass of 17.8 kg. High efficiency, advanced technology conversion options are being examined to lower the power source mass and to reduce the amount of radioisotope needed. Three technologies are being considered as the advanced converter technology: the Alkali Metal Thermal-to-Electric Converter (AMTEC), Thermophotovoltaic (TPV) converters, and Stirling Engines. Conceptual designs for each of these options have been prepared. Each converter would require only 2 GPHSs to provide the mission power and would have a mass of 6.1, 7.2, and 12.4 kg for AMTEC, TPV, and Stirling Engines respectively. This paper reviews the status of each technology and the projected performance of an advanced RPS based on each technology. Based on the projected performance and spacecraft integration issues, Pluto Express would prefer to use the AMTEC based RPS. However, in addition to technical performance, selection of a power technology will be based on many other factors.

  5. Simple and compact expressions for neutrino oscillation probabilities in matter

    SciTech Connect (OSTI)

    Minakata, Hisakazu; Parke, Stephen J.

    2015-05-07

    We reformulate perturbation theory for neutrino oscillations in matter with an expansion parameter related to the ratio of the solar to the atmospheric ?m2 scales. Unlike previous works, use a renormalized basis in which certain first-order effects are taken into account in the zeroth-order Hamiltonian. Using this perturbation theory we derive extremely compact expressions for the neutrino oscillations probabilities in matter. We find, for example, that the ?e disappearance probability at this order is of a simple two flavor form with an appropriately identified mixing angle and ?m2. Furthermore, despite exceptional simplicity in their forms they accommodate all order effects ?13 and the matter potential.

  6. Bioluminescent reporters for catabolic gene expression and pollutant bioavailability

    SciTech Connect (OSTI)

    Heitzer, A.; DiGrazia, P.M.; Sayler, G.S. . Center for Environmental Biotechnology); Burlage, R.S. )

    1991-01-01

    The application of visualized catabolic nah-gene expression using a luxCDABE gene fusion provides a valuable method to measure quantitatively and specifically naphthalene and salicylate bioavailability. It has been demonstrated that the physiological state of the test culture together with the intrinsic regulation mechanisms of the naphthalene degradation pathway as well as the physiological aspects of the lux gene fusion have to be taken into account. The method presented provides a high potential for in situ bioprocess monitoring. In addition, the results obtained with immobilized cells provide a basis for the development of biosensors for environmental applications in specific pollutant monitoring in waste streams and soil slurry systems but, as a general method, also for more conventional biotechnological process control. 8 refs., 2 figs., 1 tab.

  7. Human AZU-1 gene, variants thereof and expressed gene products

    DOE Patents [OSTI]

    Chen, Huei-Mei; Bissell, Mina

    2004-06-22

    A human AZU-1 gene, mutants, variants and fragments thereof. Protein products encoded by the AZU-1 gene and homologs encoded by the variants of AZU-1 gene acting as tumor suppressors or markers of malignancy progression and tumorigenicity reversion. Identification, isolation and characterization of AZU-1 and AZU-2 genes localized to a tumor suppressive locus at chromosome 10q26, highly expressed in nonmalignant and premalignant cells derived from a human breast tumor progression model. A recombinant full length protein sequences encoded by the AZU-1 gene and nucleotide sequences of AZU-1 and AZU-2 genes and variant and fragments thereof. Monoclonal or polyclonal antibodies specific to AZU-1, AZU-2 encoded protein and to AZU-1, or AZU-2 encoded protein homologs.

  8. Analytical expression for the electric potential in the plasma sheath near an arc-cathode

    SciTech Connect (OSTI)

    Askari, S.; Minoo, H.

    2008-04-15

    An expression for the spatial dependence of the electric potential in a collisionless plasma sheath near an electron-emitting cathode is presented. The applicability of this expression for an arc cathode is demonstrated. Comparison with the numerical solutions of the model equations indicates that the sheath thickness and potential variation predicted by this expression are accurate in a wide range of the electron-emission yield.

  9. EIS-0450: TransWest Express Transmission Project; Wyoming, Colorado, Utah,

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    and Nevada | Department of Energy 0: TransWest Express Transmission Project; Wyoming, Colorado, Utah, and Nevada EIS-0450: TransWest Express Transmission Project; Wyoming, Colorado, Utah, and Nevada Summary This EIS, prepared jointly by DOE's Western Area Power Administration and the Department of the Interior's Bureau of Land Management (Wyoming State Office), evaluates the potential environmental impacts of granting a right-of-way for the TransWest Express Transmission Project and amending

  10. Calibration Model Assignments expressed as U3O8, Summary Table ES-1 |

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Department of Energy Model Assignments expressed as U3O8, Summary Table ES-1 Calibration Model Assignments expressed as U3O8, Summary Table ES-1 Calibration Model Assignments expressed as U3O8, Summary Table ES-1 PDF icon Calibration Model Assignments expressed as U3O8, Summary Table ES-1 More Documents & Publications Field Calibration Facilities for Environmental Measurement of Radium, Thorium, and Potassium (October 2013) Offshore Wind Market and Economic Analysis Report 2013 Natural

  11. High density growth of T7 expression strains with auto-induction option

    DOE Patents [OSTI]

    Studier, F. William

    2010-04-27

    Methods for promoting and suppressing auto-induction of transcription of cloned DNA in cultures of T7 expression strains are disclosed.

  12. Seismic expressions of Monterey Formation diagenesis: examples from offshore California

    SciTech Connect (OSTI)

    Roy, M.B.

    1988-03-01

    Diagenesis of the diatomaceous rocks in the Monterey Formation in California coastal and offshore basins involves changes from amorphous biogenic silica to a stable crystalline quartz facies. In the intermediate stage, the transformation undergoes passage from the Opal-A to the Opal-CT phase. Associated with this diagenetic process is a marked increase in bulk densities between the different silica phases, owing to loss of porosity from compaction and solution recrystallization caused by increase in burial load and other physical factors. The sharp density contrast between the silica phases is manifested by an acoustic impedance boundary that may be expressed on seismic records. This seismic event can be distinct and independent of structural configuration, and in many places cuts through stratigraphic boundaries. Several examples of seismic records from offshore California demonstrate the diagenetically caused reflection cutting through Monterey and post-Monterey formations. Current and future exploration efforts in offshore California will continue to center on the widespread Monterey Formation. In addition to being the main source rock, the Monterey is also the reservoir rock. Recent discoveries indicate that oil production is mainly from the highly permeable, fractured, silica-rich sections. It is therefore important to recognize the diagenetic boundaries on seismic records and to delineate the more brittle quartz-rich facies where the reservoir quality is expected to be better than the intermediate Opal-A or Opal-CT facies. Furthermore, these boundaries could also provide good diagenetic traps off the flanks of structures where updip unaltered impermeable rocks hinder fluid migration.

  13. Sandia National Laboratories: MTEM 2014: Malware Technical Exchange...

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Pyramid North Andaluz Hotel (downtown ABQ) Candlewood Suites ABQ Courtyard Albuquerque Airport Embassy Suites ABQ Hilton Garden Inn Uptown Hyatt Place Uptown Residence Inn Airport...

  14. Orthogonal control of expression mean and variance by epigenetic features at different genomic loci

    SciTech Connect (OSTI)

    Dey, Siddharth S.; Foley, Jonathan E.; Limsirichai, Prajit; Schaffer, David V.; Arkin, Adam P.

    2015-05-05

    While gene expression noise has been shown to drive dramatic phenotypic variations, the molecular basis for this variability in mammalian systems is not well understood. Gene expression has been shown to be regulated by promoter architecture and the associated chromatin environment. However, the exact contribution of these two factors in regulating expression noise has not been explored. Using a dual-reporter lentiviral model system, we deconvolved the influence of the promoter sequence to systematically study the contribution of the chromatin environment at different genomic locations in regulating expression noise. By integrating a large-scale analysis to quantify mRNA levels by smFISH and protein levels by flow cytometry in single cells, we found that mean expression and noise are uncorrelated across genomic locations. Furthermore, we showed that this independence could be explained by the orthogonal control of mean expression by the transcript burst size and noise by the burst frequency. Finally, we showed that genomic locations displaying higher expression noise are associated with more repressed chromatin, thereby indicating the contribution of the chromatin environment in regulating expression noise.

  15. Orthogonal control of expression mean and variance by epigenetic features at different genomic loci

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Dey, Siddharth S.; Foley, Jonathan E.; Limsirichai, Prajit; Schaffer, David V.; Arkin, Adam P.

    2015-05-05

    While gene expression noise has been shown to drive dramatic phenotypic variations, the molecular basis for this variability in mammalian systems is not well understood. Gene expression has been shown to be regulated by promoter architecture and the associated chromatin environment. However, the exact contribution of these two factors in regulating expression noise has not been explored. Using a dual-reporter lentiviral model system, we deconvolved the influence of the promoter sequence to systematically study the contribution of the chromatin environment at different genomic locations in regulating expression noise. By integrating a large-scale analysis to quantify mRNA levels by smFISH andmore » protein levels by flow cytometry in single cells, we found that mean expression and noise are uncorrelated across genomic locations. Furthermore, we showed that this independence could be explained by the orthogonal control of mean expression by the transcript burst size and noise by the burst frequency. Finally, we showed that genomic locations displaying higher expression noise are associated with more repressed chromatin, thereby indicating the contribution of the chromatin environment in regulating expression noise.« less

  16. Cul4A overexpression associated with Gli1 expression in malignant pleural mesothelioma

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Yang, Yi -Lin; Ni, Jian; Hsu, Ping -Chih; Mao, Jian -Hua; Hsieh, David; Xu, Angela; Chan, Geraldine; Au, Alfred; Xu, Zhidong; Jablons, David M.; et al

    2015-07-27

    Malignant pleural mesothelioma (mesothelioma) is a highly aggressive cancer without an effective treatment. Cul4A, a scaffold protein that recruits substrates for degradation, is amplified in several human cancers, including mesothelioma. We have recently shown that Cul4A plays an oncogenic role in vitro and in a mouse model. In this study, we analysed clinical mesothelioma tumours and found moderate to strong expression of Cul4A in 70.9% (51/72) of these tumours, as shown by immunohistochemistry. In 72.2% mesothelioma tumours with increased Cul4A copy number identified by fluorescence in situ hybridization analysis, Cul4A protein expression was moderate to strong. Similarly, Cul4A was overexpressedmore » and Cul4A copy number was increased in human mesothelioma cell lines. Because Gli1 is highly expressed in human mesothelioma cells, we compared Cul4A and Gli1 expression in mesothelioma tumours and found their expression associated (P < 0.05, chi-square). In mesothelioma cell lines, inhibiting Cul4A by siRNA decreased Gli1 expression, suggesting that Gli1 expression is, at least in part, regulated by Cul4A in mesothelioma cells. Our results suggest a linkage between Cul4A and Gli1 expression in human mesothelioma.« less

  17. Transcriptomic analysis in the developing zebrafish embryo after compound exposure: Individual gene expression and pathway regulation

    SciTech Connect (OSTI)

    Hermsen, Sanne A.B.; Pronk, Tessa E.; Brandhof, Evert-Jan van den; Ven, Leo T.M. van der; Piersma, Aldert H.

    2013-10-01

    The zebrafish embryotoxicity test is a promising alternative assay for developmental toxicity. Classically, morphological assessment of the embryos is applied to evaluate the effects of compound exposure. However, by applying differential gene expression analysis the sensitivity and predictability of the test may be increased. For defining gene expression signatures of developmental toxicity, we explored the possibility of using gene expression signatures of compound exposures based on commonly expressed individual genes as well as based on regulated gene pathways. Four developmental toxic compounds were tested in concentration-response design, caffeine, carbamazepine, retinoic acid and valproic acid, and two non-embryotoxic compounds, D-mannitol and saccharin, were included. With transcriptomic analyses we were able to identify commonly expressed genes, which were mostly development related, after exposure to the embryotoxicants. We also identified gene pathways regulated by the embryotoxicants, suggestive of their modes of action. Furthermore, whereas pathways may be regulated by all compounds, individual gene expression within these pathways can differ for each compound. Overall, the present study suggests that the use of individual gene expression signatures as well as pathway regulation may be useful starting points for defining gene biomarkers for predicting embryotoxicity. - Highlights: The zebrafish embryotoxicity test in combination with transcriptomics was used. We explored two approaches of defining gene biomarkers for developmental toxicity. Four compounds in concentration-response design were tested. We identified commonly expressed individual genes as well as regulated gene pathways. Both approaches seem suitable starting points for defining gene biomarkers.

  18. Jefferson Lab Activities Group Invites Young and Not-So-Young...

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Holiday Events Holiday Party on Dec. 12 for Jefferson Lab Children and the Young at Heart The JAG Holiday Party is scheduled for Saturday, Dec. 12, 10 a.m.-1 p.m. in the CEBAF...

  19. Interleukin-2 (IL-2) dependent expression of biologically relevant IL-2 receptors: uncoupling of anti-T3 induced receptor expression with cyclosporin

    SciTech Connect (OSTI)

    Gitter, A.B.D.; Labus, J.M.; Butler, L.D.

    1986-03-01

    Human peripheral blood T cell expression of IL-2 receptors (IL-2R), detected by both immunocytofluorometry and /sup 125/I-IL-2 binding, was studied using lymphocytes stimulated with monoclonal anti-T3 antibodies (Leu-4, OKT3). Lymphocytes, isolated from healthy individuals, were prescreened and classified as Leu-4 responders or non-responders according to 72 h /sup 3/H-thymidine incorporation experiments. Leu-4 non-responder lymphocytes, though capable of normal IL-2R expression and IL-2 secretion when cultured with OKT3 (IgG2a), expressed little to no IL-2R nor secreted IL-2 when stimulated with Leu-4 (IgG1). In addition, the amount of IL-2 secreted by Leu-4 stimulated, Leu-4 responder cells, was one-third- to one-fifth of that detected when OKT3 was used as the stimulant. The addition of recombinant IL-2 (rIL-2) to a Leu-4 stimulated, Leu-4 non-responder lymphocyte culture, resulted in the expression of IL-2R and cellular proliferation, indicating that IL-2 upregulated its biologically relevant receptor. As expected, cyclosporin-A (CSA) inhibited the secretion of IL-2 and subsequent proliferation of Leu-4 stimulated, Leu-4 responder cells. Unexpectedly, however, the expression of IL-2R was also blocked. Exogenous rIL-2 partially reversed the effect of CSA on IL-2R expression and proliferation. The results indicate that IL-2 may provide an additional, required signal for optimal IL-2R expression.

  20. Sodium arsenite represses the expression of myogenin in C2C12 mouse myoblast cells through histone modifications and altered expression of Ezh2, Glp, and Igf-1

    SciTech Connect (OSTI)

    Hong, Gia-Ming; Present address: The University of Chicago, Section of Hematology Bain, Lisa J.

    2012-05-01

    Arsenic is a toxicant commonly found in water systems and chronic exposure can result in adverse developmental effects including increased neonatal death, stillbirths, and miscarriages, low birth weight, and altered locomotor activity. Previous studies indicate that 20 nM sodium arsenite exposure to C2C12 mouse myocyte cells delayed myoblast differentiation due to reduced myogenin expression, the transcription factor that differentiates myoblasts into myotubes. In this study, several mechanisms by which arsenic could alter myogenin expression were examined. Exposing differentiating C2C12 cells to 20 nM arsenic increased H3K9 dimethylation (H3K9me2) and H3K9 trimethylation (H3K9me3) by 3-fold near the transcription start site of myogenin, which is indicative of increased repressive marks, and reduced H3K9 acetylation (H3K9Ac) by 0.5-fold, indicative of reduced permissive marks. Protein expression of Glp or Ehmt1, a H3-K9 methyltransferase, was also increased by 1.6-fold in arsenic-exposed cells. In addition to the altered histone remodeling status on the myogenin promoter, protein and mRNA levels of Igf-1, a myogenic growth factor, were significantly repressed by arsenic exposure. Moreover, a 2-fold induction of Ezh2 expression, and an increased recruitment of Ezh2 (3.3-fold) and Dnmt3a (? 2-fold) to the myogenin promoter at the transcription start site (? 40 to + 42), were detected in the arsenic-treated cells. Together, we conclude that the repressed myogenin expression in arsenic-exposed C2C12 cells was likely due to a combination of reduced expression of Igf-1, enhanced nuclear expression and promoter recruitment of Ezh2, and altered histone remodeling status on myogenin promoter (? 40 to + 42). -- Highlights: ? Igf-1 expression is decreased in C2C12 cells after 20 nM arsenite exposure. ? Arsenic exposure alters histone remodeling on the myogenin promoter. ? Glp expression, a H3K9 methyltransferase, was increased in arsenic-exposed cells. ? Ezh2 and Dnmt3a localization to the myogenin promoter is induced by arsenic.

  1. Enhanced memory effect via quantum confinement in 16?nm InN nanoparticles embedded in ZnO charge trapping layer

    SciTech Connect (OSTI)

    El-Atab, Nazek; Nayfeh, Ammar; Cimen, Furkan; Alkis, Sabri; Orta, Blend; Alevli, Mustafa; Dietz, Nikolaus; Okyay, Ali K.

    2014-06-23

    In this work, the fabrication of charge trapping memory cells with laser-synthesized indium-nitride nanoparticles (InN-NPs) embedded in ZnO charge trapping layer is demonstrated. Atomic layer deposited Al{sub 2}O{sub 3} layers are used as tunnel and blocking oxides. The gate contacts are sputtered using a shadow mask which eliminates the need for any lithography steps. High frequency C-V{sub gate} measurements show that a memory effect is observed, due to the charging of the InN-NPs. With a low operating voltage of 4?V, the memory shows a noticeable threshold voltage (V{sub t}) shift of 2?V, which indicates that InN-NPs act as charge trapping centers. Without InN-NPs, the observed memory hysteresis is negligible. At higher programming voltages of 10?V, a memory window of 5?V is achieved and the V{sub t} shift direction indicates that electrons tunnel from channel to charge storage layer.

  2. Bio-Derived Liquids to Hydrogen Distributed Reforming Working Group (BILIWG) Kick-Off Meeting Proceedings Hilton Garden Inn-BWI,Baltimore, MD October 24, 2006

    Broader source: Energy.gov [DOE]

    Proceedings from the October 24, 2006 Bio-Derived Liquids to Hydrogen Distributed Reforming Working Group Kick-Off Meeting.

  3. Differential expression of nanog1 and nanogp8 in colon cancer cells

    SciTech Connect (OSTI)

    Ishiguro, Tatsuya; Sato, Ai; Ohata, Hirokazu; Sakai, Hiroaki; Nakagama, Hitoshi; Okamoto, Koji

    2012-02-10

    Highlights: Black-Right-Pointing-Pointer Nanog is expressed in a majority of colon cancer cell lines examined. Black-Right-Pointing-Pointer Both nanog1 and nanogp8 are expressed in colon cancer cells with varying ratios. Black-Right-Pointing-Pointer Nanog mediates cell proliferation of colon cancer cells. Black-Right-Pointing-Pointer Nanog predominantly localizes in cytoplasm of colon cancer cells. -- Abstract: Nanog, a homeodomain transcription factor, is an essential regulator for promotion of self-renewal of embryonic stem cells and inhibition of their differentiation. It has been demonstrated that nanog1 as well as nanogp8, a retrogene of nanog1, is preferentially expressed in advanced stages of several types of cancer, suggesting their involvement during cancer progression. Here, we investigated the expression of Nanog in well-characterized colon cancer cell lines. Expression of Nanog was detectable in 5 (HCT116, HT29, RKO, SW48, SW620) out of seven cell lines examined. RNA expression analyses of nanog1 and nanogp8 indicated that, while nanog1 was a major form in SW620 as well as in teratoma cells Tera-2, nanogp8 was preferentially expressed in HT29 and HCT116. In accordance with this, shRNA-mediated knockdown of nanog1 caused the reduction of Nanog in SW620 but not in HT29. Inhibition of Nanog in SW620 cells negatively affected cell proliferation and tumor formation in mouse xenograft. Biochemical subcellular fractionation and immunostaining analyses revealed predominant localization of Nanog in cytoplasm in SW620 and HT29, while it was mainly localized in nucleus in Tera-2. Our data indicate that nanog1 and nanogp8 are differentially expressed in colon cancer cells, and suggest that their expression contributes to proliferation of colon cancer cells.

  4. Sex-based differences in gene expression in hippocampus following postnatal lead exposure

    SciTech Connect (OSTI)

    Schneider, J.S., E-mail: jay.schneider@jefferson.edu; Anderson, D.W.; Sonnenahalli, H.; Vadigepalli, R.

    2011-10-15

    The influence of sex as an effect modifier of childhood lead poisoning has received little systematic attention. Considering the paucity of information available concerning the interactive effects of lead and sex on the brain, the current study examined the interactive effects of lead and sex on gene expression patterns in the hippocampus, a structure involved in learning and memory. Male or female rats were fed either 1500 ppm lead-containing chow or control chow for 30 days beginning at weaning.Blood lead levels were 26.7 {+-} 2.1 {mu}g/dl and 27.1 {+-} 1.7 {mu}g/dl for females and males, respectively. The expression of 175 unique genes was differentially regulated between control male and female rats. A total of 167 unique genes were differentially expressed in response to lead in either males or females. Lead exposure had a significant effect without a significant difference between male and female responses in 77 of these genes. In another set of 71 genes, there were significant differences in male vs. female response. A third set of 30 genes was differentially expressed in opposite directions in males vs. females, with the majority of genes expressed at a lower level in females than in males. Highly differentially expressed genes in males and females following lead exposure were associated with diverse biological pathways and functions. These results show that a brief exposure to lead produced significant changes in expression of a variety of genes in the hippocampus and that the response of the brain to a given lead exposure may vary depending on sex. - Highlights: > Postnatal lead exposure has a significant effect on hippocampal gene expression patterns. > At least one set of genes was affected in opposite directions in males and females. > Differentially expressed genes were associated with diverse biological pathways.

  5. Redox Protein Expression Predicts Radiotherapeutic Response in Early-Stage Invasive Breast Cancer Patients

    SciTech Connect (OSTI)

    Woolston, Caroline M.; Al-Attar, Ahmad; Storr, Sarah J.; Ellis, Ian O.; Morgan, David A.L.; Martin, Stewart G.

    2011-04-01

    Purpose: Early-stage invasive breast cancer patients have commonly undergone breast-conserving surgery and radiotherapy. In a large majority of these patients, the treatment is effective; however, a proportion will develop local recurrence. Deregulated redox systems provide cancer cells protection from increased oxidative stress, such as that induced by ionizing radiation. Therefore, the expression of redox proteins was examined in tumor specimens from this defined cohort to determine whether such expression could predict response. Methods and Materials: The nuclear and cytoplasmic expression of nine redox proteins (glutathione, glutathione reductase, glutaredoxin, glutathione peroxidase 1, 3, and 4, and glutathione S-transferase-{theta}, -{pi}, and -{alpha}) was assessed using conventional immunohistochemistry on a tissue microarray of 224 tumors. Results: A high cytoplasmic expression of glutathione S-transferase-{theta} significantly correlated with a greater risk of local recurrence (p = .008) and, when combined with a low nuclear expression (p = .009), became an independent predictive factor (p = .002) for local recurrence. High cytoplasmic expression of glutathione S-transferase-{theta} also correlated with a worse overall survival (p = .009). Low nuclear and cytoplasmic expression of glutathione peroxidase 3 (p = .002) correlated with a greater risk of local recurrence and was an independent predictive factor (p = .005). These proteins did not correlate with tumor grade, suggesting their function might be specific to the regulation of oxidative stress rather than alterations of tumor phenotype. Only nuclear (p = .005) and cytoplasmic (p = .001) expression of glutathione peroxidase 4 correlated with the tumor grade. Conclusions: Our results support the use of redox protein expression, namely glutathione S-transferase-{theta} and glutathione peroxidase 3, to predict the response to radiotherapy in early-stage breast cancer patients. If incorporated into routine diagnostic tests, they have the potential to aid clinicians in their stratification of patients into more tailored treatment regimens. Future targeted therapies to these systems might improve the efficacy of reactive oxygen species-inducing therapies, such as radiotherapy.

  6. Weatherization and Intergovernmental Program Success Stories...

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    can improve the holidays by reducing energy and maintenance costs, thanks to its new LED holiday lights. Learn more. December 16, 2011 Cows like these in Skagit County,...

  7. Fermilab Office of General Counsel - Ethics Program - Ask the...

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Ask the Ethicist Questions & Answers Submit a Question Annual Reminders Reporting Fraud Holiday Parties Holidays and contractor gratuities From Fermilab Today, December 18,...

  8. RNA binding protein and binding site useful for expression of recombinant molecules

    DOE Patents [OSTI]

    Mayfield, Stephen

    2000-01-01

    The present invention relates to a gene expression system in eukaryotic and prokaryotic cells, preferably plant cells and intact plants. In particular, the invention relates to an expression system having a RB47 binding site upstream of a translation initiation site for regulation of translation mediated by binding of RB47 protein, a member of the poly(A) binding protein family. Regulation is further effected by RB60, a protein disulfide isomerase. The expression system is capable of functioning in the nuclear/cytoplasm of cells and in the chloroplast of plants. Translation regulation of a desired molecule is enhanced approximately 100 fold over that obtained without RB47 binding site activation.

  9. RNA binding protein and binding site useful for expression of recombinant molecules

    DOE Patents [OSTI]

    Mayfield, Stephen P.

    2006-10-17

    The present invention relates to a gene expression system in eukaryotic and prokaryotic cells, preferably plant cells and intact plants. In particular, the invention relates to an expression system having a RB47 binding site upstream of a translation initiation site for regulation of translation mediated by binding of RB47 protein, a member of the poly(A) binding protein family. Regulation is further effected by RB60, a protein disulfide isomerase. The expression system is capable of functioning in the nuclear/cytoplasm of cells and in the chloroplast of plants. Translation regulation of a desired molecule is enhanced approximately 100 fold over that obtained without RB47 binding site activation.

  10. Ethanol increases matrix metalloproteinase-12 expression via NADPH oxidase-dependent ROS production in macrophages

    SciTech Connect (OSTI)

    Kim, Mi Jin; Nepal, Saroj; Lee, Eung-Seok; Jeong, Tae Cheon; Kim, Sang-Hyun; Park, Pil-Hoon

    2013-11-15

    Matrix metalloproteinase-12 (MMP-12), an enzyme responsible for degradation of extracellular matrix, plays an important role in the progression of various diseases, including inflammation and fibrosis. Although most of those are pathogenic conditions induced by ethanol ingestion, the effect of ethanol on MMP-12 has not been explored. In the present study, we investigated the effect of ethanol on MMP-12 expression and its potential mechanisms in macrophages. Here, we demonstrated that ethanol treatment increased MMP-12 expression in primary murine peritoneal macrophages and RAW 264.7 macrophages at both mRNA and protein levels. Ethanol treatment also significantly increased the activity of nicotinamide adenine dinucleotide (NADPH) oxidase and the expression of NADPH oxidase-2 (Nox2). Pretreatment with an anti-oxidant (N-acetyl cysteine) or a selective inhibitor of NADPH oxidase (diphenyleneiodonium chloride (DPI)) prevented ethanol-induced MMP-12 expression. Furthermore, knockdown of Nox2 by small interfering RNA (siRNA) prevented ethanol-induced ROS production and MMP-12 expression in RAW 264.7 macrophages, indicating a critical role for Nox2 in ethanol-induced intracellular ROS production and MMP-12 expression in macrophages. We also showed that ethanol-induced Nox2 expression was suppressed by transient transfection with dominant negative I?B-? plasmid or pretreatment with Bay 11-7082, a selective inhibitor of NF-?B, in RAW 264.7 macrophages. In addition, ethanol-induced Nox2 expression was also attenuated by treatment with a selective inhibitor of p38 MAPK, suggesting involvement of p38 MAPK/NF-?B pathway in ethanol-induced Nox2 expression. Taken together, these results demonstrate that ethanol treatment elicited increase in MMP-12 expression via increase in ROS production derived from Nox2 in macrophages. - Highlights: Ethanol increases ROS production through up-regulation of Nox2 in macrophages. Enhanced oxidative stress contributes to ethanol-induced MMP-12 expression. p38 MAPK/NF-?B signaling pathway modulates ethanol-induced Nox2 expression.

  11. EERE Success Story-Chicago Solar Express Reduces Costs, Wait Times |

    Office of Environmental Management (EM)

    Department of Energy Chicago Solar Express Reduces Costs, Wait Times EERE Success Story-Chicago Solar Express Reduces Costs, Wait Times October 28, 2014 - 10:48am Addthis The Solar Express program in Chicago, Illinois-funded through a SunShot Initiative Rooftop Solar Challenge (RSC) I award of $750,000-is making it faster, easier, and cheaper for residents to go solar by cutting long wait times and fees for solar permits. Residents of Chicago can now acquire permits for their residential

  12. DOE Seeks Public-Private Sector Expressions of Interest for Global Nuclear Energy Partnership Initiative

    Broader source: Energy.gov [DOE]

    WASHINGTON, DC - U.S. Secretary of Energy Samuel Bodman today announced that the Department of Energy (DOE) is seeking expressions of interest from the public and private sectors by March 31, 2006,...

  13. Isolated yeast promoter sequence and a method of regulated heterologous expression

    DOE Patents [OSTI]

    Gao, Johnway; Skeen, Rodney S.; Hooker, Brian S.; Anderson, Daniel B.

    2005-05-31

    The present invention provides the promoter clone discovery of a glucoamylase gene of a starch utilizing yeast strain Schwanniomyces castellii. The isolated glucoamylase promoter is an inducible promoter, which can regulate strong gene expression in starch culture medium.

  14. U-116: IBM Tivoli Provisioning Manager Express for Software Distribution Multiple Vulnerabilities

    Broader source: Energy.gov [DOE]

    Multiple vulnerabilities have been reported in IBM Tivoli Provisioning Manager Express for Software Distribution, which can be exploited by malicious people to conduct SQL injection attacks and compromise a user's system

  15. DOE Seeks Public-Private Sector Expressions of Interest for Global...

    Office of Environmental Management (EM)

    Public-Private Sector Expressions of Interest for Global Nuclear Energy Partnership ... a safe, long-term, and environmentally clean energy supply for the U.S. and the world ...

  16. Uncertainty Estimation of Radiometric Data using a Guide to the Expression of Uncertainty in Measurement (GUM) Method

    SciTech Connect (OSTI)

    Habte, Aron

    2015-06-25

    This presentation summarizes uncertainty estimation of radiometric data using the Guide to the Expression of Uncertainty (GUM) method.

  17. Expression of the tumor suppressor gene, p53, during the development of

    Office of Scientific and Technical Information (OSTI)

    murine malignant mesotheliomas induced by asbestos fibers (Conference) | SciTech Connect Conference: Expression of the tumor suppressor gene, p53, during the development of murine malignant mesotheliomas induced by asbestos fibers Citation Details In-Document Search Title: Expression of the tumor suppressor gene, p53, during the development of murine malignant mesotheliomas induced by asbestos fibers Malignant mesotheliomas are rare tumors arising from the pleural or peritoneal lining after

  18. P70S6K 1 regulation of angiogenesis through VEGF and HIF-1{alpha} expression

    SciTech Connect (OSTI)

    Bian, Chuan-Xiu; Shi, Zhumei; Meng, Qiao; Jiang, Yue; Liu, Ling-Zhi; Jiang, Bing-Hua; Department of Pathology, Anatomy and Cell Biology, Thomas Jefferson University, Philadelphia, PA 19107

    2010-07-30

    Research highlights: {yields} P70S6K1 regulates VEGF expression; {yields} P70S6K1 induces transcriptional activation through HIF-1{alpha} binding site; {yields} P70S6K1 regulates HIF-1{alpha}, but not HIF-1{beta} protein expression; {yields} P70S6K1 mediates tumor growth and angiogenesis through HIF-1{alpha} and VEGF expression. -- Abstract: The 70 kDa ribosomal S6 kinase 1 (p70S6K1), a downstream target of phosphoinositide 3-kinase (PI3K) and ERK mitogen-activated protein kinase (MAPK), is an important regulator of cell cycle progression, and cell proliferation. Recent studies indicated an important role of p70S6K1 in PTEN-negative and AKT-overexpressing tumors. However, the mechanism of p70S6K1 in tumor angiogenesis remains to be elucidated. In this study, we specifically inhibited p70S6K1 activity in ovarian cancer cells using vector-based small interfering RNA (siRNA) against p70S6K1. We found that knockdown of p70S6K1 significantly decreased VEGF protein expression and VEGF transcriptional activation through the HIF-1{alpha} binding site at its enhancer region. The expression of p70S6K1 siRNA specifically inhibited HIF-1{alpha}, but not HIF-1{beta} protein expression. We also found that p70S6K1 down-regulation inhibited ovarian tumor growth and angiogenesis, and decreased cell proliferation and levels of VEGF and HIF-1{alpha} expression in tumor tissues. Our results suggest that p70S6K1 is required for tumor growth and angiogenesis through HIF-1{alpha} and VEGF expression, providing a molecular mechanism of human ovarian cancer mediated by p70S6K1 signaling.

  19. A Sorghum bicolor expression atlas reveals dynamic genotype-specific expression profiles for vegetative tissues of grain, sweet and bioenergy sorghums

    SciTech Connect (OSTI)

    Shakoor, N; Nair, R; Crasta, O; Morris, G; Feltus, A; Kresovich, S

    2014-01-23

    Background: Effective improvement in sorghum crop development necessitates a genomics-based approach to identify functional genes and QTLs. Sequenced in 2009, a comprehensive annotation of the sorghum genome and the development of functional genomics resources is key to enable the discovery and deployment of regulatory and metabolic genes and gene networks for crop improvement. Results: This study utilizes the first commercially available whole-transcriptome sorghum microarray (Sorgh-WTa520972F) to identify tissue and genotype-specific expression patterns for all identified Sorghum bicolor exons and UTRs. The genechip contains 1,026,373 probes covering 149,182 exons (27,577 genes) across the Sorghum bicolor nuclear, chloroplast, and mitochondrial genomes. Specific probesets were also included for putative non-coding RNAs that may play a role in gene regulation (e. g., microRNAs), and confirmed functional small RNAs in related species (maize and sugarcane) were also included in our array design. We generated expression data for 78 samples with a combination of four different tissue types (shoot, root, leaf and stem), two dissected stem tissues (pith and rind) and six diverse genotypes, which included 6 public sorghum lines (R159, Atlas, Fremont, PI152611, AR2400 and PI455230) representing grain, sweet, forage, and high biomass ideotypes. Conclusions: Here we present a summary of the microarray dataset, including analysis of tissue-specific gene expression profiles and associated expression profiles of relevant metabolic pathways. With an aim to enable identification and functional characterization of genes in sorghum, this expression atlas presents a new and valuable resource to the research community.

  20. ADAM15 expression is downregulated in melanoma metastasis compared to primary melanoma

    SciTech Connect (OSTI)

    Ungerer, Christopher; Doberstein, Kai; Boehm, Beate; Pfeilschifter, Josef; Mihic-Probst, Daniela; Gutwein, Paul

    2010-10-22

    Research highlights: {yields} Strong ADAM15 expression is found in normal melanocytes. {yields} ADAM15 expression is significantly downregulated in patients with melanoma metastasis. {yields} TGF-{beta} can downregulate ADAM15 expression in melanoma cells. {yields} Overexpression of ADAM15 in melanoma cells inhibits migration, proliferation and invasion of melanoma cells. {yields} Conclusion: ADAM15 represents an tumor suppressor protein in melanoma. -- Abstract: In a mouse melanoma metastasis model it has been recently shown that ADAM15 overexpression in melanoma cells significantly reduced the number of metastatic nodules on the lung. Unfortunately, the expression of ADAM15 in human melanoma tissue has not been determined so far. In our study, we characterized the expression of ADAM15 in tissue micro-arrays of patients with primary melanoma with melanoma metastasis. ADAM15 was expressed in melanocytes and endothelial cells of benign nevi and melanoma tissue. Importantly, ADAM15 was significantly downregulated in melanoma metastasis compared to primary melanoma. We further demonstrate that IFN-{gamma} and TGF-{beta} downregulate ADAM15 protein levels in melanoma cells. To investigate the role of ADAM15 in melanoma progression, we overexpressed ADAM15 in melanoma cells. Importantly, overexpression of ADAM15 in melanoma cells reduced the migration, invasion and the anchorage dependent and independent cell growth of melanoma cells. In summary, the downregulation of ADAM15 plays an important role in melanoma progression and ADAM15 act as a tumorsuppressor in melanoma.

  1. Knockdown of p53 suppresses Nanog expression in embryonic stem cells

    SciTech Connect (OSTI)

    Abdelalim, Essam Mohamed; Tooyama, Ikuo

    2014-01-10

    Highlights: We investigate the role of p53 in ESCs in the absence of DNA damage. p53 knockdown suppresses ESC proliferation. p53 knockdown downregulates Nanog expression. p53 is essential for mouse ESC self-renewal. -- Abstract: Mouse embryonic stem cells (ESCs) express high levels of cytoplasmic p53. Exposure of mouse ESCs to DNA damage leads to activation of p53, inducing Nanog suppression. In contrast to earlier studies, we recently reported that chemical inhibition of p53 suppresses ESC proliferation. Here, we confirm that p53 signaling is involved in the maintenance of mouse ESC self-renewal. RNA interference-mediated knockdown of p53 induced downregulation of p21 and defects in ESC proliferation. Furthermore, p53 knockdown resulted in a significant downregulation in Nanog expression at 24 and 48 h post-transfection. p53 knockdown also caused a reduction in Oct4 expression at 48 h post-transfection. Conversely, exposure of ESCs to DNA damage caused a higher reduction of Nanog expression in control siRNA-treated cells than in p53 siRNA-treated cells. These data show that in the absence of DNA damage, p53 is required for the maintenance of mouse ESC self-renewal by regulating Nanog expression.

  2. Enhanced human papillomavirus type 8 oncogene expression levels are crucial for skin tumorigenesis in transgenic mice

    SciTech Connect (OSTI)

    Hufbauer, M.; Lazic, D.; Akguel, B.; Brandsma, J.L.; Pfister, H.; Weissenborn, S.J.

    2010-08-01

    Human papillomavirus 8 (HPV8) is involved in skin cancer development in epidermodysplasia verruciformis patients. Transgenic mice expressing HPV8 early genes (HPV8-CER) developed papillomas, dysplasias and squamous cell carcinomas. UVA/B-irradiation and mechanical wounding of HPV8-CER mouse skin led to prompt papilloma induction in about 3 weeks. The aim of this study was to analyze the kinetics and level of transgene expression in response to skin irritations. Transgene expression was already enhanced 1 to 2 days after UVA/B-irradiation or tape-stripping and maintained during papilloma development. The enhanced transgene expression could be assigned to UVB and not to UVA. Papilloma development was thus always paralleled by an increased transgene expression irrespective of the type of skin irritation. A knock-down of E6 mRNA by tattooing HPV8-E6-specific siRNA led to a delay and a lower incidence of papilloma development. This indicates that the early increase of viral oncogene expression is crucial for induction of papillomatosis.

  3. Prominin-2 expression increases protrusions, decreases caveolae and inhibits Cdc42 dependent fluid phase endocytosis

    SciTech Connect (OSTI)

    Singh, Raman Deep Schroeder, Andreas S.; Scheffer, Luana; Holicky, Eileen L.; Wheatley, Christine L.; Marks, David L. Pagano, Richard E.

    2013-05-10

    Highlights: Prominin-2 expression induced protrusions that co-localized with lipid raft markers. Prominin-2 expression decreased caveolae, caveolar endocytosis and increased pCav1. Prominin-2 expression inhibited fluid phase endocytosis by inactivation of Cdc42. These endocytic effects can be reversed by adding exogenous cholesterol. Caveolin1 knockdown restored fluid phase endocytosis in Prominin2 expressing cells. -- Abstract: Background: Membrane protrusions play important roles in biological processes such as cell adhesion, wound healing, migration, and sensing of the external environment. Cell protrusions are a subtype of membrane microdomains composed of cholesterol and sphingolipids, and can be disrupted by cholesterol depletion. Prominins are pentaspan membrane proteins that bind cholesterol and localize to plasma membrane (PM) protrusions. Prominin-1 is of great interest as a marker for stem and cancer cells, while Prominin-2 (Prom2) is reportedly restricted to epithelial cells. Aim: To characterize the effects of Prom-2 expression on PM microdomain organization. Methods: Prom2-fluorescent protein was transfected in human skin fibroblasts (HSF) and Chinese hamster ovary (CHO) cells for PM raft and endocytic studies. Caveolae at PM were visualized using transmission electron microscopy. Cdc42 activation was measured and caveolin-1 knockdown was performed using siRNAs. Results: Prom2 expression in HSF and CHO cells caused extensive Prom2-positive protrusions that co-localized with lipid raft markers. Prom2 expression significantly decreased caveolae at the PM, reduced caveolar endocytosis and increased caveolin-1 phosphorylation. Prom2 expression also inhibited Cdc42-dependent fluid phase endocytosis via decreased Cdc42 activation. Effects on endocytosis were reversed by addition of cholesterol. Knockdown of caveolin-1 by siRNA restored Cdc42 dependent fluid phase endocytosis in Prom2-expressing cells. Conclusions: Prom2 protrusions primarily localize to lipid rafts and recruit cholesterol into protrusions and away from caveolae, leading to increased phosphorylation of caveolin-1, which inhibits Cdc42-dependent endocytosis. This study provides a new insight for the role for prominins in the regulation of PM lipid organization.

  4. Investigating the Correspondence Between Transcriptomic and Proteomic Expression Profiles Using Coupled Cluster Models.

    SciTech Connect (OSTI)

    Rogers, Simon; Girolami, Mark; Kolch, Walter; Waters, Katrina M.; Liu, Tao; Thrall, Brian D.; Wiley, H. S.

    2008-12-01

    Modern transcriptomics and proteomics enable us to survey the expression of RNAs and proteins at large scales. While these data are usually generated and analysed separately, there is an increasing interest in comparing and co-analysing transcriptome and proteome expression data. A major open question is whether transcriptome and proteome expression is linked and how it is coordinated. Results: Here we have developed a probabilistic clustering model that permits analysis of the links between transcriptomic and proteomic profiles in a sensible and flexible manner. Our coupled mixture model defines a prior probability distribution over the component to which a protein profile should be assigned conditioned on which component the associated mRNA profile belongs to. By providing probabilistic assignments this approach sits between the two extremes of concatenating the data on the assumption that mRNA and protein clusters would have a one-to-one relationship, and independent clustering where the mRNA profile provides no information on the protein profile and vice-versa. We apply this approach to a large dataset of quantitative transcriptomic and proteomic expression data obtained from a human breast epithelial cell line (HMEC) stimulated by epidermal growth factor (EGF) over a series of timepoints corresponding to one cell cycle. The results reveal a complex relationship between transcriptome and proteome with most mRNA clusters linked to at least two protein clusters, and vice versa. A more detailed analysis incorporating information on gene function from the gene ontology database shows that a high correlation of mRNA and protein expression is limited to the components of some molecular machines, such as the ribosome, cell adhesion complexes and the TCP-1 chaperonin involved in protein folding. Conclusions: The dynamic regulation of the transcriptome and proteome in mammalian cells in response to an acute mitogenic stimulus appears largely independent with very little correspondence between mRNA and protein expression. The exceptions involve a few selected multi-protein complexes that require the stoichiometric expression of components for correct function. This finding has wide ramifications regarding the understanding of gene and protein expression including its control and evolution. It also shows that transcriptomic and proteomic expression analysis are complementary and non-redundant.

  5. Catalog of gene expression in adult neural stem cells and their in vivo microenvironment

    SciTech Connect (OSTI)

    Williams, Cecilia; Wirta, Valtteri; Meletis, Konstantinos; Wikstroem, Lilian; Carlsson, Leif; Frisen, Jonas; Lundeberg, Joakim . E-mail: joakim.lundeberg@biotech.kth.se

    2006-06-10

    Stem cells generally reside in a stem cell microenvironment, where cues for self-renewal and differentiation are present. However, the genetic program underlying stem cell proliferation and multipotency is poorly understood. Transcriptome analysis of stem cells and their in vivo microenvironment is one way of uncovering the unique stemness properties and provides a framework for the elucidation of stem cell function. Here, we characterize the gene expression profile of the in vivo neural stem cell microenvironment in the lateral ventricle wall of adult mouse brain and of in vitro proliferating neural stem cells. We have also analyzed an Lhx2-expressing hematopoietic-stem-cell-like cell line in order to define the transcriptome of a well-characterized and pure cell population with stem cell characteristics. We report the generation, assembly and annotation of 50,792 high-quality 5'-end expressed sequence tag sequences. We further describe a shared expression of 1065 transcripts by all three stem cell libraries and a large overlap with previously published gene expression signatures for neural stem/progenitor cells and other multipotent stem cells. The sequences and cDNA clones obtained within this framework provide a comprehensive resource for the analysis of genes in adult stem cells that can accelerate future stem cell research.

  6. Knowledge-based analysis of microarray gene expression data by using support vector machines

    SciTech Connect (OSTI)

    William Grundy; Manuel Ares, Jr.; David Haussler

    2001-06-18

    The authors introduce a method of functionally classifying genes by using gene expression data from DNA microarray hybridization experiments. The method is based on the theory of support vector machines (SVMs). SVMs are considered a supervised computer learning method because they exploit prior knowledge of gene function to identify unknown genes of similar function from expression data. SVMs avoid several problems associated with unsupervised clustering methods, such as hierarchical clustering and self-organizing maps. SVMs have many mathematical features that make them attractive for gene expression analysis, including their flexibility in choosing a similarity function, sparseness of solution when dealing with large data sets, the ability to handle large feature spaces, and the ability to identify outliers. They test several SVMs that use different similarity metrics, as well as some other supervised learning methods, and find that the SVMs best identify sets of genes with a common function using expression data. Finally, they use SVMs to predict functional roles for uncharacterized yeast ORFs based on their expression data.

  7. Cancer Associated Fibroblasts express pro-inflammatory factors in human breast and ovarian tumors

    SciTech Connect (OSTI)

    Erez, Neta; Glanz, Sarah; Raz, Yael; Department of Obstetrics and Gynecology, LIS Maternity Hospital, Tel Aviv Sourasky Medical Center, affiliated with Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv ; Avivi, Camilla; Barshack, Iris; Department of Pathology, Sheba Medical Center, Tel Hashomer, affiliated with Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv

    2013-08-02

    Highlights: CAFs in human breast and ovarian tumors express pro-inflammatory factors. Expression of pro-inflammatory factors correlates with tumor invasiveness. Expression of pro-inflammatory factors is associated with NF-?b activation in CAFs. -- Abstract: Inflammation has been established in recent years as a hallmark of cancer. Cancer Associated Fibroblasts (CAFs) support tumorigenesis by stimulating angiogenesis, cancer cell proliferation and invasion. We previously demonstrated that CAFs also mediate tumor-enhancing inflammation in a mouse model of skin carcinoma. Breast and ovarian carcinomas are amongst the leading causes of cancer-related mortality in women and cancer-related inflammation is linked with both these tumor types. However, the role of CAFs in mediating inflammation in these malignancies remains obscure. Here we show that CAFs in human breast and ovarian tumors express high levels of the pro-inflammatory factors IL-6, COX-2 and CXCL1, previously identified to be part of a CAF pro-inflammatory gene signature. Moreover, we show that both pro-inflammatory signaling by CAFs and leukocyte infiltration of tumors are enhanced in invasive ductal carcinoma as compared with ductal carcinoma in situ. The pro-inflammatory genes expressed by CAFs are known NF-?B targets and we show that NF-?B is up-regulated in breast and ovarian CAFs. Our data imply that CAFs mediate tumor-promoting inflammation in human breast and ovarian tumors and thus may be an attractive target for stromal-directed therapeutics.

  8. Protocol for Uniformly Measuring and Expressing the Performance of Energy Storage Systems

    SciTech Connect (OSTI)

    Conover, David R.; Crawford, Aladsair J.; Viswanathan, Vilayanur V.; Ferreira, Summer; Schoenwald, David

    2014-06-01

    The Protocol for Uniformly Measuring and Expressing the Performance of Energy Storage Systems (PNNL-22010) was first issued in November 2012 as a first step toward providing a foundational basis for developing an initial standard for the uniform measurement and expression of energy storage system (ESS) performance. Its subsequent use in the field and review by the protocol working group and most importantly the users’ subgroup and the thermal subgroup has led to the fundamental modifications reflected in this update of the 2012 Protocol. As an update of the 2012 Protocol, this document (the June 2014 Protocol) is intended to supersede its predecessor and be used as the basis for measuring and expressing ESS performance. The foreword provides general and specific details about what additions, revisions, and enhancements have been made to the 2012 Protocol and the rationale for them in arriving at the June 2014 Protocol.

  9. Femtosecond spectroscopy probes the folding quality of antibody fragments expressed as GFP fusions in the cytoplasm

    SciTech Connect (OSTI)

    Didier, P. [Faculte de Pharmacie, UMR 7175, 74, route du Rhin, 67412 Illkirch (France); Weiss, E.; Sibler, A.-P. [Ecole Superieure de Biotechnologie de Strasbourg, UMR 7175, Boulevard Sebastien Brant, F-67412 Illkirch (France); Philibert, P.; Martineau, P. [Centre de recherche en cancerologie de Montpellier, UMR 5160, Val d'Aurelle-Paul Lamarque, 34298 Montpellier cedex 5 (France); Bigot, J.-Y. [Institut de Physique et Chimie des Materiaux de Strasbourg, UMR 7504, 23, rue du Loess, F-67037 Strasbourg (France); Guidoni, L. [Institut de Physique et Chimie des Materiaux de Strasbourg, UMR 7504, 23, rue du Loess, F-67037 Strasbourg (France); Laboratoire Materiaux et Phenomenes Quantiques, UMR 7162, Batiment Condorcet, 10 rue Alice Domon et Leonie Duquet, 75205 Paris cedex 13 (France)], E-mail: luca.guidoni@univ-paris-diderot.fr

    2008-02-22

    Time-resolved femtosecond spectroscopy can improve the application of green fluorescent proteins (GFPs) as protein-folding reporters. The study of ultrafast excited-state dynamics (ESD) of GFP fused to single chain variable fragment (scFv) antibody fragments, allowed us to define and measure an empirical parameter that only depends on the folding quality (FQ) of the fusion. This method has been applied to the analysis of genetic fusions expressed in the bacterial cytoplasm and allowed us to distinguish folded and thus functional antibody fragments (high FQ) with respect to misfolded antibody fragments. Moreover, these findings were strongly correlated to the behavior of the same scFvs expressed in animal cells. This method is based on the sensitivity of the ESD to the modifications in the tertiary structure of the GFP induced by the aggregation state of the fusion partner. This approach may be applicable to the study of the FQ of polypeptides over-expressed under reducing conditions.

  10. Cloning and expression of autogenes encoding RNA poly,erases of T7-like bacteriophages

    DOE Patents [OSTI]

    Studier, F. William (Stony Brook, NY); Dubendorff, John W. (Sound Beach, NY)

    1998-01-01

    This invention relates to the cloning and expression of autogenes encoding RNA polymerases of T7 and T7-like bacteriophages, in which the RNA polymerase gene is transcribed from a promoter which is recognized by the encoded RNA polymerase. Cloning of T7 autogenes was achieved by reducing the activity of the RNA polymerase sufficiently to permit host cell growth. T7 RNA polymerase activity was controlled by combining two independent methods: lac-repression of the recombinant lac operator-T7 promoter in the autogene and inhibition of the polymerase by T7 lysozyme. Expression systems for producing the RNA polymerases of T7 and other T7-like bacteriophages, and expression systems for producing selected gene products are described, as well as other related materials and methods.

  11. Cloning and expression of autogenes encoding RNA polymerases of T7-like bacteriophages

    DOE Patents [OSTI]

    Studier, F. William (Stony Brook, NY); Dubendorff, John W. (Sound Beach, NY)

    1998-01-01

    This invention relates to the cloning and expression of autogenes encoding RNA polymerases of T7 and T7-like bacteriophages, in which the RNA polymerase gene is transcribed from a promoter which is recognized by the encoded RNA polymerase. Cloning of T7 autogenes was achieved by reducing the activity of the RNA polymerase sufficiently to permit host cell growth. T7 RNA polymerase activity was controlled by combining two independent methods: lac-repression of the recombinant lac operator-T7 promoter in the autogene and inhibition of the polymerase by T7 lysozyme. Expression systems for producing the RNA polymerases of T7 and other T7-like bacteriophages, and expression systems for producing selected gene products are described, as well as other related materials and methods.

  12. Cloning and expression of autogenes encoding RNA polymerases of T7-like bacteriophages

    DOE Patents [OSTI]

    Studier, F.W.; Dubendorff, J.W.

    1998-10-20

    This invention relates to the cloning and expression of autogenes encoding RNA polymerases of T7 and T7-like bacteriophages, in which the RNA polymerase gene is transcribed from a promoter which is recognized by the encoded RNA polymerase. Cloning of T7 autogenes was achieved by reducing the activity of the RNA polymerase sufficiently to permit host cell growth. T7 RNA polymerase activity was controlled by combining two independent methods: lac-repression of the recombinant lac operator-T7 promoter in the autogene and inhibition of the polymerase by T7 lysozyme. Expression systems for producing the RNA polymerases of T7 and other T7-like bacteriophages, and expression systems for producing selected gene products are described, as well as other related materials and methods. 12 figs.

  13. Cloning and expression of autogenes encoding RNA polymerases of T7-like bacteriophages

    DOE Patents [OSTI]

    Studier, F.W.; Dubendorff, J.W.

    1998-11-03

    This invention relates to the cloning and expression of autogenes encoding RNA polymerases of T7 and T7-like bacteriophages, in which the RNA polymerase gene is transcribed from a promoter which is recognized by the encoded RNA polymerase. Cloning of T7 autogenes was achieved by reducing the activity of the RNA polymerase sufficiently to permit host cell growth. T7 RNA polymerase activity was controlled by combining two independent methods: lac-repression of the recombinant lac operator-T7 promoter in the autogene and inhibition of the polymerase by T7 lysozyme. Expression systems for producing the RNA polymerases of T7 and other T7-like bacteriophages, and expression systems for producing selected gene products are described, as well as other related materials and methods. 12 figs.

  14. Genetic analysis of the regulation of TCH gene expression, Final Report

    SciTech Connect (OSTI)

    Braam, Janet

    2008-10-28

    The Arabidopsis TCH genes, originally isolated as a consequence of their upregulation in response to the mechanical stimulus of touch, are also upregulated by a variety of seemingly disparate environmental and hormonal stimuli. To gain insight into the complexities of TCH gene regulation, a number of approaches were taken. Regulatory elements responsible for regulation were identified and characteristics of the regulation were evaluated. Reporter genes were used to monitor expression localization and dynamics. Microarray analyses of genome-wide expression behavior indicated that touch-inducible gene expression is more widespread than generally appreciated. Identification of all touch-regulated genes shed light on the types of cellular processes that may be altered in response to mechanical stress perturbations. Expression of the TCH2 gene, also called CML24, encoding a calmodulin (CaM)-like (CML) protein, was evaluated. CML24 shares over 40% amino acid sequence identity with CaM, has 4 EF hands and undergoes a Ca2+-dependent change in migration rate through denaturing gel electrophoresis, indicating that CML24 binds Ca2+ and, as a consequence, undergoes conformational changes. CML24 expression occurs in all major organs and is induced from 2- to 15-fold in plants subjected to touch, darkness, heat, cold, hydrogen peroxide, abscisic acid (ABA) and indole-3-acetic acid. The putative CML24 regulatory region confers reporter expression at sites of predicted mechanical stress, in regions undergoing growth, in vascular tissues and various floral organs and in stomata, trichomes and hydathodes. CML24 underexpressing transgenics are resistant to ABA inhibition of germination and seedling growth, defective in long-day induction of flowering, and have enhanced tolerance to CoCl2, molybdic acid, ZnSO4 and MgCl2. These data present evidence that CML24 encodes a potential Ca2+ sensor that may function to enable responses to ABA, day length and presence of various salts. Further investigation of CML24 function and regulation led to the finding that CML24 has a critical role in nitric oxide regulation. Distinct tilling mutant alleles demonstrated that CML24 can act as a switch in the response to day length perception. Because of potential redundancy with the related CML23 gene, CML23 T-DNA insertion mutants were identified and characterized. Together, CML23 and CML24 impact the autonomous regulatory pathway of the transition to flowering. Nitric oxide levels are elevated in cml23/cml24 double mutants. Therefore, CML23 and CML24 are potential calcium sensors regulate nitric oxide accumulation. In collaboration with Drs. McCann and Carpita, fourier transform infrared spectroscopy (FTIR) was used to assess, verify and classify wall architectural changes that occur as a result of single XTH insertion mutations. Thirty-four homozygous mutant lines of Arabidopsis representing 21 members of the xyloglucan endotransglucosylase/hydrolase gene family provided a set of mutants to characterize. Kohonen networks classified cell wall architectures of xth mutant lines and previously characterized cell wall mutants. The xth mutants were found to have chemical changes in their cell walls not detectable as phenotypic growth and development changes, consistent with the existence of feed-back loops that modify wall composition in response to a life-long deficiency of a cell wall enzyme. To gain insight into the potential physiological relevance of the distinct members of the XTH family, GUS reporter fusion genes were constructed, and plants expressing these transgenes were characterized to reveal spatial and temporal patterns of expression. In addition, Genevestigator sources were mined for comprehensive and comparative XTH expression regulation analysis. These data revealed that the Arabidopsis XTHs are likely expressed in every developmental stage from seed germination through flowering. All organs showed XTH::GUS expression and most, if not all, are found to express multiple XTH::GUS genes suggesting that XTHs may contribute to morphogenesis at every d

  15. Regulation of bcl-2 proto-oncogene expression during normal human lymphocyte proliferation

    SciTech Connect (OSTI)

    Reed, J.C.; Tsujimoto, Y.; Alpers, J.D.; Croce, C.M.; Nowell, P.C.

    1987-06-05

    The bcl-2 and c-myc proto-oncogenes are brought into juxtaposition with the immuno-globulin heavy chain locus in particular B-cell lymphomas, resulting in high levels of constitutive accumulation of their messenger RNAs. Precisely how the products of the bcl-2 and c-myc genes contribute to tumorigenesis is unknown, but observations that c-myc expression is rapidly induced in nonneoplastic lymphocytes upon stimulation of proliferation raise the possibility that this proto-oncogene is involved in the control of normal cellular growth. In addition to c-myc, the bcl-2 proto-oncogene also was expressed in normal human B and T lymphocytes after stimulation with appropriate mitogens. Comparison of the regulation of the expression of these proto-oncogenes demonstrated marked differences and provided evidence that, in contrast to c-myc, levels of bcl-2 messenger RNA are regulated primarily though transcriptional mechanisms. 10 references, 3 figures.

  16. Engineering of living cells for the expression of holo-phycobiliprotein-based constructs

    DOE Patents [OSTI]

    Glazer, Alexander N.; Tooley, Aaron J.; Cai, Yuping

    2004-05-25

    Recombinant cells which express a fluorescent holo-phycobiliprotein fusion protein and methods of use are described. The cells comprises a bilin, a recombinant bilin reductase, an apo-phycobiliprotein fusion protein precursor of the fusion protein comprising a corresponding apo-phycobiliprotein domain, and a recombinant phycobiliprotein domain-bilin lyase, which components react to form the holo-phycobiliprotein fusion protein. Also described are holo-phycobiliprotein based transcription reporter cells and assays, which cells conditionally express a heterologous-to-the-cell, fluorescent, first holo-phycobiliprotein domain.

  17. A Framework for Load Balancing of Tensor Contraction Expressions via Dynamic Task Partitioning

    SciTech Connect (OSTI)

    Lai, Pai-Wei; Stock, Kevin; Rajbhandari, Samyam; Krishnamoorthy, Sriram; Sadayappan, Ponnuswamy

    2013-11-17

    In this paper, we introduce the Dynamic Load-balanced Tensor Contractions (DLTC), a domain-specific library for efficient task parallel execution of tensor contraction expressions, a class of computation encountered in quantum chemistry and physics. Our framework decomposes each contraction into smaller unit of tasks, represented by an abstraction referred to as iterators. We exploit an extra level of parallelism by having tasks across independent contractions executed concurrently through a dynamic load balancing run- time. We demonstrate the improved performance, scalability, and flexibility for the computation of tensor contraction expressions on parallel computers using examples from coupled cluster methods.

  18. Matrigel Basement Membrane Matrix influences expression of microRNAs in cancer cell lines

    SciTech Connect (OSTI)

    Price, Karina J.; School of Medicine and Pharmacology, University of Western Australia, Nedlands, WA 6008 ; Tsykin, Anna; School of Molecular and Biomedical Science, University of Adelaide, Adelaide, SA 5005 ; Giles, Keith M.; Sladic, Rosemary T.; Epis, Michael R.; Ganss, Ruth; Goodall, Gregory J.; School of Molecular and Biomedical Science, University of Adelaide, Adelaide, SA 5005; Department of Medicine, University of Adelaide, Adelaide, SA 5005 ; Leedman, Peter J.

    2012-10-19

    Highlights: Black-Right-Pointing-Pointer Matrigel alters cancer cell line miRNA expression relative to culture on plastic. Black-Right-Pointing-Pointer Many identified Matrigel-regulated miRNAs are implicated in cancer. Black-Right-Pointing-Pointer miR-1290, -210, -32 and -29b represent a Matrigel-induced miRNA signature. Black-Right-Pointing-Pointer miR-32 down-regulates Integrin alpha 5 (ITGA5) mRNA. -- Abstract: Matrigel is a medium rich in extracellular matrix (ECM) components used for three-dimensional cell culture and is known to alter cellular phenotypes and gene expression. microRNAs (miRNAs) are small, non-coding RNAs that regulate gene expression and have roles in cancer. While miRNA profiles of numerous cell lines cultured on plastic have been reported, the influence of Matrigel-based culture on cancer cell miRNA expression is largely unknown. This study investigated the influence of Matrigel on the expression of miRNAs that might facilitate ECM-associated cancer cell growth. We performed miRNA profiling by microarray using two colon cancer cell lines (SW480 and SW620), identifying significant differential expression of miRNAs between cells cultured in Matrigel and on plastic. Many of these miRNAs have previously been implicated in cancer-related processes. A common Matrigel-induced miRNA signature comprised of up-regulated miR-1290 and miR-210 and down-regulated miR-29b and miR-32 was identified using RT-qPCR across five epithelial cancer cell lines (SW480, SW620, HT-29, A549 and MDA-MB-231). Experimental modulation of these miRNAs altered expression of their known target mRNAs involved in cell adhesion, proliferation and invasion, in colon cancer cell lines. Furthermore, ITGA5 was identified as a novel putative target of miR-32 that may facilitate cancer cell interactions with the ECM. We propose that culture of cancer cell lines in Matrigel more accurately recapitulates miRNA expression and function in cancer than culture on plastic and thus is a valuable approach to the in vitro study of miRNAs.

  19. FedEx Express Gasoline Hybrid Electric Delivery Truck Evaluation: 12-Month Report

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    FedEx Express Gasoline Hybrid Electric Delivery Truck Evaluation: 12-Month Report R. Barnitt Technical Report NREL/TP-5400-48896 January 2011 NREL is a national laboratory of the U.S. Department of Energy, Office of Energy Efficiency & Renewable Energy, operated by the Alliance for Sustainable Energy, LLC. National Renewable Energy Laboratory 1617 Cole Boulevard Golden, Colorado 80401 303-275-3000 * www.nrel.gov Contract No. DE-AC36-08GO28308 FedEx Express Gasoline Hybrid Electric Delivery

  20. Proceedings of the 1996 spring technical conference of the ASME Internal Combustion Engine Division. Volume 2: Engine design and engine systems; ICE-Volume 26-2

    SciTech Connect (OSTI)

    Uzkan, T.

    1996-12-31

    Although the cost of the petroleum crude has not increased much within the last decade, the drive to develop internal combustion engines is still continuing. The basic motivation of this drive is to reduce both emissions and costs. Recent developments in computer chip production and information management technology have opened up new applications in engine controls and monitoring. The development of new information is continuing at a rapid pace. Some of these research and development results were presented at the 1996 Spring Technical Conference of the ASME Internal Combustion Engine Division in Youngstown, Ohio, April 21--24, 1996. The papers presented covered various aspects of the design, development, and application of compression ignition and spark ignition engines. The conference was held at the Holiday Inn Metroplex Complex and hosted by Altronic Incorporated of Girard, Ohio. The written papers submitted to the conference have been published in three conference volumes. Volume 2 includes the papers on the topics of engine design, engine systems, and engine user experience.

  1. Use of CYP52A2A promoter to increase gene expression in yeast

    DOE Patents [OSTI]

    Craft, David L.; Wilson, C. Ron; Eirich, Dudley; Zhang, Yeyan

    2004-01-06

    A nucleic acid sequence including a CYP promoter operably linked to nucleic acid encoding a heterologous protein is provided to increase transcription of the nucleic acid. Expression vectors and host cells containing the nucleic acid sequence are also provided. The methods and compositions described herein are especially useful in the production of polycarboxylic acids by yeast cells.

  2. Analysis of differential protein expression in normal and neoplastic human breast epithelial cell lines

    SciTech Connect (OSTI)

    Williams, K.; Chubb, C.; Huberman, E.; Giometti, C.S.

    1997-07-01

    High resolution two dimensional get electrophoresis (2DE) and database analysis was used to establish protein expression patterns for cultured normal human mammary epithelial cells and thirteen breast cancer cell lines. The Human Breast Epithelial Cell database contains the 2DE protein patterns, including relative protein abundances, for each cell line, plus a composite pattern that contains all the common and specifically expressed proteins from all the cell lines. Significant differences in protein expression, both qualitative and quantitative, were observed not only between normal cells and tumor cells, but also among the tumor cell lines. Eight percent of the consistently detected proteins were found in significantly (P < 0.001) variable levels among the cell lines. Using a combination of immunostaining, comigration with purified protein, subcellular fractionation, and amino-terminal protein sequencing, we identified a subset of the differentially expressed proteins. These identified proteins include the cytoskeletal proteins actin, tubulin, vimentin, and cytokeratins. The cell lines can be classified into four distinct groups based on their intermediate filament protein profile. We also identified heat shock proteins; hsp27, hsp60, and hsp70 varied in abundance and in some cases in the relative phosphorylation levels among the cell lines. Finally, we identified IMP dehydrogenase in each of the cell lines, and found the levels of this enzyme in the tumor cell lines elevated 2- to 20-fold relative to the levels in normal cells.

  3. Systems for the expression of orthogonal translation components in eubacterial host cells

    DOE Patents [OSTI]

    Ryu, Youngha; Schultz, Peter G.

    2013-01-22

    The invention related to compositions and methods for the in vivo production of polypeptides comprising one or more unnatural amino acids. Specifically, the invention provides plasmid systems for the efficient eubacterial expression of polypeptides comprising one or more unnatural acids at genetically-programmed positions.

  4. Systems for the expression of orthogonal translation components in eubacterial host cells

    DOE Patents [OSTI]

    Ryu, Youngha; Schultz, Peter G.

    2011-06-14

    The invention relates to compositions and methods for the in vivo production of polypeptides comprising one or more unnatural amino acids. Specifically, the invention provides plasmid systems for the efficient eubacterial expression of polypeptides comprising one or more unnatural amino acids at genetically-programmed positions.

  5. Systems for the expression of orthogonal translation components eubacterial host cells

    DOE Patents [OSTI]

    Ryu, Youngha; Schultz, Peter G.

    2012-06-12

    The invention relates to compositions and methods for the in vivo production of polypeptides comprising one or more unnatural amino acids. Specifically, the invention provides plasmid systems for the efficient eubacterial expression of polypeptides comprising one or more unnatural amino acids at genetically-programmed positions.

  6. EIS-0447: Champlain Hudson Power Express Transmission Line Project, New York

    Broader source: Energy.gov [DOE]

    This EIS evaluated the potential environmental impacts of a DOE proposal to grant a Presidential permit to Champlain Hudson Power Express, Inc., to construct, operate, maintain, and connect a new 1000-megawatt (MW) electric transmission system across the U.S.-Canada border in northeastern New York State. The proposed transmission line would run from the Canadian Province of Quebec to New York City.

  7. Cellulase variants with improved expression, activity and stability, and use thereof

    DOE Patents [OSTI]

    Aehle, Wolfgang; Bott, Richard R; Bower, Benjamin; Caspi, Jonathan; Estell, David A; Goedegebuur, Frits; Hommes, Ronaldus W.J.; Kaper, Thijs; Kelemen, Bradley; Kralj, Slavko; Van Lieshout, Johan; Nikolaev, Igor; Van Stigt Thans, Sander; Wallace, Louise; Vogtentanz, Gudrun; Sandgren, Mats

    2014-03-25

    The present disclosure relates to cellulase variants. In particular the present disclosure relates to cellulase variants having improved expression, activity and/or stability. Also described are nucleic acids encoding the cellulase variants, compositions comprising the cellulase variants, and methods of use thereof.

  8. Cloning of a yeast alpha-amylase promoter and its regulated heterologous expression

    DOE Patents [OSTI]

    Gao, Johnway [Richland, WA; Skeen, Rodney S [Pendleton, OR; Hooker, Brian S [Kennewick, WA; Anderson, Daniel B [Pasco, WA

    2003-04-01

    The present invention provides the promoter clone discovery of an alpha-amylase gene of a starch utilizing yeast strain Schwanniomyces castellii. The isolated alpha-amylase promoter is an inducible promoter, which can regulate strong gene expression in starch culture medium.

  9. Liver X Receptor (LXR) activation negatively regulates visfatin expression in macrophages

    SciTech Connect (OSTI)

    Mayi, Therese Hervee; Rigamonti, Elena; INSERM UR1011, F-59000 Lille; UDSL, F-59000 Lille; Institut Pasteur de Lille, F-59019 Lille ; Pattou, Francois; Department of Endocrine Surgery, University Hospital, Lille; U859 Biotherapies for Diabetes, INSERM, Lille ; Staels, Bart; INSERM UR1011, F-59000 Lille; UDSL, F-59000 Lille; Institut Pasteur de Lille, F-59019 Lille ; Chinetti-Gbaguidi, Giulia; INSERM UR1011, F-59000 Lille; UDSL, F-59000 Lille; Institut Pasteur de Lille, F-59019 Lille

    2011-01-07

    Research highlights: {yields} Synthetic LXR ligands decreased visfatin expression in human macrophages. {yields} LXR activation leads to a modest and transient decrease of NAD{sup +} concentration. {yields} LXR activation decreased PPAR{gamma}-induced visfatin in human macrophages. -- Abstract: Adipose tissue macrophages (ATM) are the major source of visfatin, a visceral fat adipokine upregulated during obesity. Also known to play a role in B cell differentiation (pre-B cell colony-enhancing factor (PBEF)) and NAD biosynthesis (nicotinamide phosphoribosyl transferase (NAMPT)), visfatin has been suggested to play a role in inflammation. Liver X Receptor (LXR) and Peroxisome Proliferator-Activated Receptor (PPAR){gamma} are nuclear receptors expressed in macrophages controlling the inflammatory response. Recently, we reported visfatin as a PPAR{gamma} target gene in human macrophages. In this study, we examined whether LXR regulates macrophage visfatin expression. Synthetic LXR ligands decreased visfatin gene expression in a LXR-dependent manner in human and murine macrophages. The decrease of visfatin mRNA was paralleled by a decrease of protein secretion. Consequently, a modest and transient decrease of NAD{sup +} concentration was observed. Interestingly, LXR activation decreased the PPAR{gamma}-induced visfatin gene and protein secretion in human macrophages. Our results identify visfatin as a gene oppositely regulated by the LXR and PPAR{gamma} pathways in human macrophages.

  10. An EGF receptor inhibitor induces RAR-{beta} expression in breast and ovarian cancer cells

    SciTech Connect (OSTI)

    Grunt, Thomas W. . E-mail: thomas.grunt@meduniwien.ac.at; Puckmair, Klaudia; Tomek, Katharina; Kainz, Birgit; Gaiger, Alexander

    2005-04-22

    Inhibition of the epidermal growth factor (EGF)-receptor (EGFR) has become a promising anticancer treatment strategy. In addition, application of retinoids yields encouraging results for cancer prevention and therapy. Many tumors express no or low amounts of retinoic acid receptor-{beta}2 (RAR-{beta}2) due to epigenetic silencing via DNA hypermethylation. RAR-{beta}2 is the main mediator of the antiproliferative effect of retinoids. RAR-{beta}2 re-expression causes reversal of transformation, cell cycle arrest, and restoration of retinoid sensitivity. RAR-{beta}2 is thus a tumor suppressor. Western blotting, colorimetric in vitro cell proliferation assays, and reverse transcription-polymerase chain reaction demonstrated that the EGFR inhibitor PD153035 not only blocked activation of EGFR and inhibited cell growth, but also stimulated RAR-{beta} expression in MDA-MB-468 breast and OVCAR-3 ovarian carcinoma cells. Upregulation of RAR-{beta} by PD153035 was confirmed by real-time reverse transcription-polymerase chain reaction. In contrast, expression of other retinoid receptors and of estrogen receptor-{alpha} was not affected. PD153035-mediated re-induction of RAR-{beta} was associated with demethylation of the RAR-{beta}2 gene promoter P2 as demonstrated by methylation-specific polymerase chain reaction. These novel results on the ErbB/retinoid receptor cross-talk may be useful for designing future anticancer combination regimens.

  11. Expression of transforming growth factor alpha in plutonium-239-induced lung neoplasms in dogs: investigations of autocrine mechanisms of growth

    SciTech Connect (OSTI)

    Gillett, N.A.; Stegelmeier, B.L.; Chang, I.Y.; Kelly, G. )

    1991-06-01

    We have previously shown that 47% of radiation-induced lung neoplasms in dogs exhibit increased expression of epidermal growth factor receptor (EGFR). In this study, we investigated the expression of transforming growth factor alpha (TGF-alpha), a ligand for EGFR, to determine if an autocrine mechanism for growth stimulation was present in these tumors. As determined by immunohistochemistry, 59% (26/44) of the lung neoplasms examined had increased expression of TGF-alpha. Expression of TGF-alpha was not related to the etiology of the tumor, e.g., spontaneous or plutonium-induced; however, it was related to the phenotype of the tumor. Statistical analysis of the correlation of EGFR and TGF-alpha expression within the same tumor did not show a positive association; however, specific phenotypes did have statistically significant expression of EGFR or TGF-alpha, suggesting that overexpression of either the ligand or its receptor conferred a growth advantage to the neoplasm. Twenty-seven percent (32/117) of radiation-induced proliferative epithelial foci expressed TGF-alpha, and a portion of those foci (8/32) expressed both EGFR and TGF-alpha. This supports the hypothesis that these foci represent preneoplastic lesions, and suggests that those foci exhibiting increased expression of the growth factor or its receptor are at greater risk for progressing to neoplasia.

  12. Expression of a bacterial 3-dehydroshikimate dehydratase reduces lignin content and improves biomass saccharification efficiency

    SciTech Connect (OSTI)

    Eudes, Aymerick; Sathitsuksanoh, Noppadon; Baidoo, Edward E. K.; George, Anthe; Liang, Yan; Yang, Fan; Singh, Seema; Keasling, Jay D.; Simmons, Blake A.; Loqu, Dominique

    2015-01-13

    Lignin confers recalcitrance to plant biomass used as feedstocks in agro-processing industries or as source of renewable sugars for the production of bioproducts. The metabolic steps for the synthesis of lignin building blocks belong to the shikimate and phenylpropanoid pathways. Genetic engineering efforts to reduce lignin content typically employ gene knockout or gene silencing techniques to constitutively repress one of these metabolic pathways. Recently, new strategies have emerged offering better spatiotemporal control of lignin deposition, including the expression of enzymes that interfere with the normal process for cell wall lignification. In this study, we report that expression of a 3-dehydroshikimate dehydratase (QsuB from Corynebacterium glutamicum) reduces lignin deposition in Arabidopsis cell walls. QsuB was targeted to the plastids to convert 3-dehydroshikimate an intermediate of the shikimate pathway into protocatechuate. Compared to wild-type plants, lines expressing QsuB contain higher amounts of protocatechuate, p-coumarate, p-coumaraldehyde and p-coumaryl alcohol, and lower amounts of coniferaldehyde, coniferyl alcohol, sinapaldehyde and sinapyl alcohol. 2D-NMR spectroscopy and pyrolysis-gas chromatography/mass spectrometry (pyro-GC/MS) reveal an increase of p-hydroxyphenyl units and a reduction of guaiacyl units in the lignin of QsuB lines. Size-exclusion chromatography indicates a lower degree of lignin polymerization in the transgenic lines. Therefore, our data show that the expression of QsuB primarily affects the lignin biosynthetic pathway. Finally, biomass from these lines exhibits more than a twofold improvement in saccharification efficiency. We conclude that the expression of QsuB in plants, in combination with specific promoters, is a promising gain-of-function strategy for spatiotemporal reduction of lignin in plant biomass.

  13. Expression of a bacterial 3-dehydroshikimate dehydratase reduces lignin content and improves biomass saccharification efficiency

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Eudes, Aymerick; Sathitsuksanoh, Noppadon; Baidoo, Edward E. K.; George, Anthe; Liang, Yan; Yang, Fan; Singh, Seema; Keasling, Jay D.; Simmons, Blake A.; Loqué, Dominique

    2015-01-13

    Lignin confers recalcitrance to plant biomass used as feedstocks in agro-processing industries or as source of renewable sugars for the production of bioproducts. The metabolic steps for the synthesis of lignin building blocks belong to the shikimate and phenylpropanoid pathways. Genetic engineering efforts to reduce lignin content typically employ gene knockout or gene silencing techniques to constitutively repress one of these metabolic pathways. Recently, new strategies have emerged offering better spatiotemporal control of lignin deposition, including the expression of enzymes that interfere with the normal process for cell wall lignification. In this study, we report that expression of a 3-dehydroshikimatemore » dehydratase (QsuB from Corynebacterium glutamicum) reduces lignin deposition in Arabidopsis cell walls. QsuB was targeted to the plastids to convert 3-dehydroshikimate – an intermediate of the shikimate pathway – into protocatechuate. Compared to wild-type plants, lines expressing QsuB contain higher amounts of protocatechuate, p-coumarate, p-coumaraldehyde and p-coumaryl alcohol, and lower amounts of coniferaldehyde, coniferyl alcohol, sinapaldehyde and sinapyl alcohol. 2D-NMR spectroscopy and pyrolysis-gas chromatography/mass spectrometry (pyro-GC/MS) reveal an increase of p-hydroxyphenyl units and a reduction of guaiacyl units in the lignin of QsuB lines. Size-exclusion chromatography indicates a lower degree of lignin polymerization in the transgenic lines. Therefore, our data show that the expression of QsuB primarily affects the lignin biosynthetic pathway. Finally, biomass from these lines exhibits more than a twofold improvement in saccharification efficiency. We conclude that the expression of QsuB in plants, in combination with specific promoters, is a promising gain-of-function strategy for spatiotemporal reduction of lignin in plant biomass.« less

  14. Nucleic and amino acid sequences relating to a novel transketolase, and methods for the expression thereof

    DOE Patents [OSTI]

    Croteau, Rodney Bruce (Pullman, WA); Wildung, Mark Raymond (Colfax, WA); Lange, Bernd Markus (Pullman, WA); McCaskill, David G. (Pullman, WA)

    2001-01-01

    cDNAs encoding 1-deoxyxylulose-5-phosphate synthase from peppermint (Mentha piperita) have been isolated and sequenced, and the corresponding amino acid sequences have been determined. Accordingly, isolated DNA sequences (SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7) are provided which code for the expression of 1-deoxyxylulose-5-phosphate synthase from plants. In another aspect the present invention provides for isolated, recombinant DXPS proteins, such as the proteins having the sequences set forth in SEQ ID NO:4, SEQ ID NO:6 and SEQ ID NO:8. In other aspects, replicable recombinant cloning vehicles are provided which code for plant 1-deoxyxylulose-5-phosphate synthases, or for a base sequence sufficiently complementary to at least a portion of 1-deoxyxylulose-5-phosphate synthase DNA or RNA to enable hybridization therewith. In yet other aspects, modified host cells are provided that have been transformed, transfected, infected and/or injected with a recombinant cloning vehicle and/or DNA sequence encoding a plant 1-deoxyxylulose-5-phosphate synthase. Thus, systems and methods are provided for the recombinant expression of the aforementioned recombinant 1-deoxyxylulose-5-phosphate synthase that may be used to facilitate its production, isolation and purification in significant amounts. Recombinant 1-deoxyxylulose-5-phosphate synthase may be used to obtain expression or enhanced expression of 1-deoxyxylulose-5-phosphate synthase in plants in order to enhance the production of 1-deoxyxylulose-5-phosphate, or its derivatives such as isopentenyl diphosphate (BP), or may be otherwise employed for the regulation or expression of 1-deoxyxylulose-5-phosphate synthase, or the production of its products.

  15. Mixed lineage kinase 3 is required for matrix metalloproteinase expression and invasion in ovarian cancer cells

    SciTech Connect (OSTI)

    Zhan, Yu; Abi Saab, Widian F.; Modi, Nidhi; Stewart, Amanda M.; Liu, Jinsong; Chadee, Deborah N.

    2012-08-15

    Mixed lineage kinase 3 (MLK3) is a mitogen-activated protein kinase kinase kinase (MAP3K) that activates MAPK signaling pathways and regulates cellular responses such as proliferation, migration and apoptosis. Here we report high levels of total and phospho-MLK3 in ovarian cancer cell lines in comparison to immortalized nontumorigenic ovarian epithelial cell lines. Using small interfering RNA (siRNA)-mediated gene silencing, we determined that MLK3 is required for the invasion of SKOV3 and HEY1B ovarian cancer cells. Furthermore, mlk3 silencing substantially reduced matrix metalloproteinase (MMP)-1, -2, -9 and -12 gene expression and MMP-2 and -9 activities in SKOV3 and HEY1B ovarian cancer cells. MMP-1, -2, -9 and-12 expression, and MLK3-induced activation of MMP-2 and MMP-9 requires both extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK) activities. In addition, inhibition of activator protein-1 (AP-1) reduced MMP-1, MMP-9 and MMP-12 gene expression. Collectively, these findings establish MLK3 as an important regulator of MMP expression and invasion in ovarian cancer cells. -- Highlights: Black-Right-Pointing-Pointer Ovarian cancer cell lines have high levels of total and phosphorylated MLK3. Black-Right-Pointing-Pointer MLK3 is required for MMP expression and activity in ovarian cancer cells. Black-Right-Pointing-Pointer MLK3 is required for invasion of SKOV3 and HEY1B ovarian cancer cells. Black-Right-Pointing-Pointer MLK3-dependent regulation of MMP-2 and MMP-9 activities requires ERK and JNK.

  16. Modulation of keratinocyte expression of antioxidants by 4-hydroxynonenal, a lipid peroxidation end product

    SciTech Connect (OSTI)

    Zheng, Ruijin; Heck, Diane E.; Mishin, Vladimir; Black, Adrienne T.; Shakarjian, Michael P.; Kong, Ah-Ng Tony; Laskin, Debra L.; Laskin, Jeffrey D.

    2014-03-01

    4-Hydroxynonenal (4-HNE) is a lipid peroxidation end product generated in response to oxidative stress in the skin. Keratinocytes contain an array of antioxidant enzymes which protect against oxidative stress. In these studies, we characterized 4-HNE-induced changes in antioxidant expression in mouse keratinocytes. Treatment of primary mouse keratinocytes and PAM 212 keratinocytes with 4-HNE increased mRNA expression for heme oxygenase-1 (HO-1), catalase, NADPH:quinone oxidoreductase (NQO1) and glutathione S-transferase (GST) A1-2, GSTA3 and GSTA4. In both cell types, HO-1 was the most sensitive, increasing 8698 fold within 6 h. Further characterization of the effects of 4-HNE on HO-1 demonstrated concentration- and time-dependent increases in mRNA and protein expression which were maximum after 6 h with 30 ?M. 4-HNE stimulated keratinocyte Erk1/2, JNK and p38 MAP kinases, as well as PI3 kinase. Inhibition of these enzymes suppressed 4-HNE-induced HO-1 mRNA and protein expression. 4-HNE also activated Nrf2 by inducing its translocation to the nucleus. 4-HNE was markedly less effective in inducing HO-1 mRNA and protein in keratinocytes from Nrf2 ?/? mice, when compared to wild type mice, indicating that Nrf2 also regulates 4-HNE-induced signaling. Western blot analysis of caveolar membrane fractions isolated by sucrose density centrifugation demonstrated that 4-HNE-induced HO-1 is localized in keratinocyte caveolae. Treatment of the cells with methyl-?-cyclodextrin, which disrupts caveolar structure, suppressed 4-HNE-induced HO-1. These findings indicate that 4-HNE modulates expression of antioxidant enzymes in keratinocytes, and that this can occur by different mechanisms. Changes in expression of keratinocyte antioxidants may be important in protecting the skin from oxidative stress. - Highlights: Lipid peroxidation generates 4-hydroxynonenal, a reactive aldehyde. 4-HNE induces antioxidant proteins in mouse keratinocytes. Induction of antioxidant proteins is regulated via MAP kinases, Nrf2 and caveolae. 4-HNE is an effective signaling molecule in keratinocytes.

  17. DNA sequence and spatial expression pattern of a drought- and ABA-induced gene in tomato

    SciTech Connect (OSTI)

    Plant, A.L.; Cohen, A.; Moses, M.S.; Bray, E.A. )

    1991-05-01

    The genomic and cDNA sequence for the previously characterized drought- and ABA-induced gene pLE16 are presented. The single open reading frame contained within the gene has the capacity to encode a polypeptide of 12.7 kD with a predicted pI of 8.73. The amino-terminus is highly hydrophobic and is characteristic of signal sequences which target polypeptides for export from the cytoplasm. There is considerable homology (51.3% identity) between the amino-terminus of pLE16 and the amino-terminal domains of a group of proteins that comprise the phospholipid transfer proteins. Although this homology breaks down at the carboxy-terminal half of pLE16, the homology that exists suggests that pLE16 may be associated with membranes and may therefore play a role in maintaining membrane integrity during drought-stress. pLE16 is expressed in drought-stressed leaf, petiole and stem tissue and to a much lower extent in the seeds and pericarp of mature green tomato fruit. No expression was detected in the seeds or pericarp of red fruit or drought-stressed roots. Expression of pLE16 is induced in leaf tissue by a variety of other environmental stresses including PEG-mediated water deficit, salt, cold stress and heat stress. These stresses did not however induce expression of pLE16 in the roots. Examination of the 5{prime} flanking DNA sequences for this gene did not reveal the presence of the consensus ABA responsive element (ABRE), implicated in ABA induction of gene expression and so far common to the 5{prime} flanking DNA sequences of many genes that are ABA responsive. The expression of pLE16 in response to drought-stress and other environmental stresses in vegetative tissue, together with the lack of a consensus ABRE, suggests that the regulation of this gene by ABA may differ from those that are seed-specific.

  18. A Hybrid Approach to Protein Differential Expression in Mass Spectrometry-Based Proteomics

    SciTech Connect (OSTI)

    Wang, Xuan; Anderson, Gordon A.; Smith, Richard D.; Dabney, Alan R.

    2012-04-19

    Motivation: Quantitative mass spectrometry-based proteomics involves statistical inference on protein abundance, based on the intensities of each protein's associated spectral peaks. However, typical MS-based proteomics data sets have substantial proportions of missing observations, due at least in part to censoring of low intensities. This complicates intensity-based differential expression analysis. Results: We outline a statistical method for protein differential expression, based on a simple Binomial likelihood. By modeling peak intensities as binary, in terms of 'presence/ absence,' we enable the selection of proteins not typically amendable to quantitative analysis; e.g., 'one-state' proteins that are present in one condition but absent in another. In addition, we present an analysis protocol that combines quantitative and presence/ absence analysis of a given data set in a principled way, resulting in a single list of selected proteins with a single associated FDR.

  19. Process and genes for expression and overexpression of active [FeFe] hydrogenases

    DOE Patents [OSTI]

    Seibert, Michael; King, Paul W; Ghirardi, Maria Lucia; Posewitz, Matthew C; Smolinski, Sharon L

    2014-09-16

    A process for expression of active [FeFe]-hydrogenase in a host organism that does not contain either the structural gene(s) for [FeFe]-hydrogenases and/or homologues for the maturation genes HydE, HydF and HyG, comprising: cloning the structural hydrogenase gene(s) and/or the maturation genes HydE, HydF and HydG from an organisms that contains these genes into expression plasmids; transferring the plasmids into an organism that lacks a native [FeFe]-hydrogenase or that has a disrupted [FeFe]-hydrogenase and culturing it aerobically; and inducing anaerobiosis to provide [FeFe] hydrogenase biosynthesis and H?2#191 production.

  20. Explicit expressions for three-dimensional boundary integrals in linear elasticity

    SciTech Connect (OSTI)

    Nintcheu Fata, Sylvain

    2011-01-01

    On employing isoparametric, piecewise linear shape functions over a flat triangle, exact formulae are derived for all surface potentials involved in the numerical treatment of three-dimensional singular and hyper-singular boundary integral equations in linear elasticity. These formulae are valid for an arbitrary source point in space and are represented as analytical expressions along the edges of the integration triangle. They can be employed to solve integral equations defined on triangulated surfaces via a collocation method or may be utilized as analytical expressions for the inner integrals in a Galerkin technique. A numerical example involving a unit triangle and a source point located at various distances above it, as well as sample problems solved by a collocation boundary element method for the Lame equation are included to validate the proposed formulae.

  1. In silico method for modelling metabolism and gene product expression at genome scale

    SciTech Connect (OSTI)

    Lerman, Joshua A.; Hyduke, Daniel R.; Latif, Haythem; Portnoy, Vasiliy A.; Lewis, Nathan E.; Orth, Jeffrey D.; Rutledge, Alexandra C.; Smith, Richard D.; Adkins, Joshua N.; Zengler, Karsten; Palsson, Bernard O.

    2012-07-03

    Transcription and translation use raw materials and energy generated metabolically to create the macromolecular machinery responsible for all cellular functions, including metabolism. A biochemically accurate model of molecular biology and metabolism will facilitate comprehensive and quantitative computations of an organism's molecular constitution as a function of genetic and environmental parameters. Here we formulate a model of metabolism and macromolecular expression. Prototyping it using the simple microorganism Thermotoga maritima, we show our model accurately simulates variations in cellular composition and gene expression. Moreover, through in silico comparative transcriptomics, the model allows the discovery of new regulons and improving the genome and transcription unit annotations. Our method presents a framework for investigating molecular biology and cellular physiology in silico and may allow quantitative interpretation of multi-omics data sets in the context of an integrated biochemical description of an organism.

  2. DOE Environmental Impact Statement Public Scoping Meeting on Champlain Hudson Power Express Transmission Line Project

    Broader source: Energy.gov [DOE]

    The Department of Energy (DOE) is hosting seven meetings for public participation as part of its Environmental Impact Statement (EIS) preparation process pursuant to the National Environmental Policy Act (NEPA) to assess the potential environmental impacts from its proposed action of granting a Presidential permit to Champlain Hudson Power Express, Inc., to construct, operate, maintain, and connect a new electric transmission line across the U.S.- Canada border in northeastern New York.

  3. EIS-0450: TransWest Express Transmission Project; Wyoming, Colorado, Utah, and Nevada

    Broader source: Energy.gov [DOE]

    This EIS, prepared jointly by DOE's Western Area Power Administration and the Department of the Interior's Bureau of Land Management (Wyoming State Office), evaluates the potential environmental impacts of granting a right-of-way for the TransWest Express Transmission Project and amending a land use plan. The project consists of an overhead transmission line that would extend approximately 725 miles from south-central Wyoming, through Colorado and Utah. Western proposes to be a joint owner of the project.

  4. Microsoft Word - CafeExpressSecurityFormFinal June 2012 v2.doc

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Café Systems Express Access Form Version: 06/2012 Page 1 of 2 Applicant Information □ New User □ School or Department Transfer □ Inactivate *Date *NetID *Name (Last, First MI) *EMPLID Title NU Email Department Work Phone * indicates required field NUFinancials - Data Entry and Inquiry OPTIONAL: Model an existing user's access: Name: ____________________________ NetID: _________ □ Expense Entry and iBuyNU Shopper □ Budget Journal Entry □ CRT Entry □ CRT Inquiry □ Expense Entry,

  5. American Express OPEN has teamed up with Women Impacting Public Policy

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    American Express OPEN has teamed up with Women Impacting Public Policy (WIPP) and the U.S. Small Business Administration (SBA) to offer a ChallengeHER event with an OPEN for Government Contracts: Contract Connections session. This event is designed to help women-owned businesses build long-lasting, lucrative business relationships while assisting Government officials and Prime Contractors in meeting their small business goals. To learn more, please email: ContractConnections@govbizsolutions.com

  6. AG490 inhibits NFATc1 expression and STAT3 activation during RANKL induced osteoclastogenesis

    SciTech Connect (OSTI)

    Li, Chang-hong; Zhao, Jin-xia; Sun, Lin; Yao, Zhong-qiang; Deng, Xiao-li; Liu, Rui; Liu, Xiang-yuan

    2013-06-14

    Highlights: AG490 inhibits RANKL-induced osteoclastogenesis in RAW264.7 cells. AG490 affects cell proliferation and cell cycle distribution. AG490 reduces NFATc1 expression during RANKL-induced osteoclastogenesis. AG490 disrupts the activation of RANKL-mediated JAK2/STAT3 signaling pathway. STAT3 depletion partly mimics the effect of AG490 on RANKL-induced osteoclastogenesis. -- Abstract: Commonly, JAK/STAT relays cytokine signals for cell activation and proliferation, and recent studies have shown that the elevated expression of JAK/STAT is associated with the immune rejection of allografts and the inflammatory processes of autoimmune disease. However, the role which JAK2/STAT3 signaling plays in the receptor activator of nuclear factor-?B ligand (RANKL)-mediated osteoclastogenesis is unknown. In this study, we investigated the effects of AG490, specific JAK2 inhibitor, on osteoclast differentiation in vitro. AG490 significantly inhibited osteoclastogenesis in murine osteoclast precursor cell line RAW264.7 induced by RANKL. AG490 suppressed cell proliferation and delayed the G1 to S cell cycle transition. Furthermore, AG490 also suppressed the expression of nuclear factor of activated T cells (NFAT) c1 but not c-Fos in RAW264.7. Subsequently, we investigated various intracellular signaling components associated with osteoclastogenesis. AG490 had no effects on RANKL-induced activation of Akt, ERK1/2. Interestingly, AG490 partly inhibited RANKL-induced phosphorylation of Ser{sup 727} in STAT3. Additionally, down-regulation of STAT3 using siRNA resulted in suppression of TRAP, RANK and NFATc1 expression. In conclusion, we demonstrated that AG490 inhibited RANKL-induced osteoclastogenesis by suppressing NFATc1 production and cell proliferation via the STAT3 pathway. These results suggest that inhibition of JAK2 may be useful for the treatment of bone diseases characterized by excessive osteoclastogenesis.

  7. Microsoft Word - IARC_Expression of interest_Rev5_Nov2012.doc

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Rev.01/13/2012 Fermilab Technology Applications Program Expression of Interest Project/Proposal Name: Company/Institution Street Address Phone Company URL City, State, Zip Code Fax Contact Person Name Street Address Phone Title City, State, Zip Code Email Do you wish to have information in this EOI treated as proprietary? Yes No (please circle one) Provide additional comments if desired. Comments: Is your interest in the Fermilab Technology Applications Program "general" in nature or

  8. Expression analysis and prognostic significance of the SRA1 gene, in ovarian cancer

    SciTech Connect (OSTI)

    Leoutsakou, Theoni; Talieri, Maroulio; Scorilas, Andreas . E-mail: ascorilas@biol.uoa.gr

    2006-06-02

    The SR-related-CTD-associated-factors (SCAFs) have the ability to interact with the C-terminal domain of the RNA polymerase II, linking this way transcription to splicing. SRA1 (SR-A1) gene, encoding for a human high-molecular weight SCAF protein, is located on chromosome 19, between the IRF3 and the R-RAS oncogene and it has been demonstrated from members of our group that SRA1 is constitutively expressed in most of the human tissues, while it is overexpressed in a subset of ovarian tumors. In this study, we examine the expression of SRA1 gene in 111 ovarian malignant tissues and in the human ovarian carcinoma cell lines OVCAR-3, TOV21-G, and ES-2, using a semi-quantitative RT-PCR method. SRA1 gene was overexpressed in 61/111 (55%) of ovarian carcinomas. This higher expression was positively associated to the size of the tumor (p < 0.001), the grade and the stage of the disease (p = 0.003 and p = 0.006, respectively), and the debulking success (p < 0.001). Kaplan-Meier survival analysis revealed that lower SRA1 expression increases the probability of both the longer overall and the progression free survival of the patients. Multivariate Cox regression analysis revealed that SRA1 may be used as an independent prognostic biomarker in ovarian cancer. Our results suggest that SRA1 is associated with cancer progression and may possibly be characterized as a new marker of unfavorable prognosis for ovarian cancer.

  9. Gene expression profiling--Opening the black box of plant ecosystem responses to global change

    SciTech Connect (OSTI)

    Leakey, A.D.B.; Ainsworth, E.A.; Bernard, S.M.; Markelz, R.J.C.; Ort, D.R.; Placella, S.A.P.; Rogers, A.; Smith, M.D.; Sudderth, E.A.; Weston, D.J.; Wullschleger, S.D.; Yuan, S.

    2009-11-01

    The use of genomic techniques to address ecological questions is emerging as the field of genomic ecology. Experimentation under environmentally realistic conditions to investigate the molecular response of plants to meaningful changes in growth conditions and ecological interactions is the defining feature of genomic ecology. Since the impact of global change factors on plant performance are mediated by direct effects at the molecular, biochemical and physiological scales, gene expression analysis promises important advances in understanding factors that have previously been consigned to the 'black box' of unknown mechanism. Various tools and approaches are available for assessing gene expression in model and non-model species as part of global change biology studies. Each approach has its own unique advantages and constraints. A first generation of genomic ecology studies in managed ecosystems and mesocosms have provided a testbed for the approach and have begun to reveal how the experimental design and data analysis of gene expression studies can be tailored for use in an ecological context.

  10. Single nucleotide polymorphism in transcriptional regulatory regions and expression of environmentally responsive genes

    SciTech Connect (OSTI)

    Wang, Xuting; Tomso, Daniel J.; Liu Xuemei; Bell, Douglas A. . E-mail: BELL1@niehs.nih.gov

    2005-09-01

    Single nucleotide polymorphisms (SNPs) in the human genome are DNA sequence variations that can alter an individual's response to environmental exposure. SNPs in gene coding regions can lead to changes in the biological properties of the encoded protein. In contrast, SNPs in non-coding gene regulatory regions may affect gene expression levels in an allele-specific manner, and these functional polymorphisms represent an important but relatively unexplored class of genetic variation. The main challenge in analyzing these SNPs is a lack of robust computational and experimental methods. Here, we first outline mechanisms by which genetic variation can impact gene regulation, and review recent findings in this area; then, we describe a methodology for bioinformatic discovery and functional analysis of regulatory SNPs in cis-regulatory regions using the assembled human genome sequence and databases on sequence polymorphism and gene expression. Our method integrates SNP and gene databases and uses a set of computer programs that allow us to: (1) select SNPs, from among the >9 million human SNPs in the NCBI dbSNP database, that are similar to cis-regulatory element (RE) consensus sequences; (2) map the selected dbSNP entries to the human genome assembly in order to identify polymorphic REs near gene start sites; (3) prioritize the candidate polymorphic RE containing genes by searching the existing genotype and gene expression data sets. The applicability of this system has been demonstrated through studies on p53 responsive elements and is being extended to additional pathways and environmentally responsive genes.

  11. Influence of heart failure on nucleolar organization and protein expression in human hearts

    SciTech Connect (OSTI)

    Rosello-Lleti, Esther; Rivera, Miguel; Cortes, Raquel; Azorin, Inmaculada; Sirera, Rafael; Martinez-Dolz, Luis; Hove, Leif; Cinca, Juan; Lago, Francisca; Gonzalez-Juanatey, Jose R.; Salvador, Antonio; Portoles, Manuel

    2012-02-10

    Highlights: Black-Right-Pointing-Pointer Heart failure alters nucleolar morphology and organization. Black-Right-Pointing-Pointer Nucleolin expression is significant increased in ischemic and dilated cardiomyopathy. Black-Right-Pointing-Pointer Ventricular function of heart failure patients was related with nucleolin levels. -- Abstract: We investigate for the first time the influence of heart failure (HF) on nucleolar organization and proteins in patients with ischemic (ICM) or dilated cardiomyopathy (DCM). A total of 71 human hearts from ICM (n = 38) and DCM (n = 27) patients, undergoing heart transplantation and control donors (n = 6), were analysed by western-blotting, RT-PCR and cell biology methods. When we compared protein levels according to HF etiology, nucleolin was increased in both ICM (117%, p < 0.05) and DCM (141%, p < 0.01). Moreover, mRNA expression were also upregulated in ICM (1.46-fold, p < 0.05) and DCM (1.70-fold, p < 0.05. Immunofluorescence studies showed that the highest intensity of nucleolin was into nucleolus (p < 0.0001), and it was increased in pathological hearts (p < 0.0001). Ultrastructure analysis by electron microscopy showed an increase in the nucleus and nucleolus size in ICM (17%, p < 0.05 and 131%, p < 0.001) and DCM (56%, p < 0.01 and 69%, p < 0.01). Nucleolar organization was influenced by HF irrespective of etiology, increasing fibrillar centers (p < 0.001), perinucleolar chromatin (p < 0.01) and dense fibrillar components (p < 0.01). Finally, left ventricular function parameters were related with nucleolin levels in ischemic hearts (p < 0.0001). The present study demonstrates that HF influences on morphology and organization of nucleolar components, revealing changes in the expression and in the levels of nucleolin protein.

  12. The effects of deoxynivalenol on gene expression in the murine thymus

    SciTech Connect (OSTI)

    Kol, Sandra W.M. van; Hendriksen, Peter J.M.; Loveren, Henk van; Peijnenburg, Ad

    2011-02-01

    Deoxynivalenol (DON) is a mycotoxin produced by several Fusarium species and is often detected in grains. Because of its high abundance, there has been a large interest in the effects of DON in animals and humans. DON is known to be immunosuppressive at high concentrations and immunostimulatory at low concentrations. The present study aimed to acquire insight into the modes of action of DON. For this, C57Bl6 mice were orally exposed to 5, 10, or 25 mg/kg bw DON for 3, 6, or 24 h and thymuses were subjected to genome-wide expression microarray analysis. Gene set enrichment analysis (GSEA) demonstrated that DON downregulated genes involved in proliferation, mitochondria, protein synthesis, and ribosomal proteins. Furthermore, GSEA showed a selective downregulation of genes highly expressed at the early precursor thymocytes stage. This indicates that early precursor thymocytes, particularly at the double-positive CD4+CD8+ stage, are more vulnerable to DON than very early or late precursor thymocytes. There was a large overlap of genes upregulated by DON with genes previously reported to be either upregulated during T cell activation or upregulated during negative selection of thymocytes that recognize 'self-antigens'. This indicates that DON induces cellular events that also occur after activation of the T cell receptor, for example, release of calcium from the endoplasmatic reticulum. This T cell activation in the thymus then evokes negative selection and depletion of thymocytes, which provides a plausible explanation for the high sensitivity of the thymus for DON exposure. The expression patterns of four genes indicative for some of the processes that were affected after DON treatment were confirmed using real-time PCR. Immunocytological experiments with primary mouse thymocytes demonstrated the translocation of NFAT from the cytoplasm into the nucleus upon exposure top DON, thus providing further evidence for the involvement of T cell activation.

  13. Compositions and methods for the expression of selenoproteins in eukaryotic cells

    DOE Patents [OSTI]

    Gladyshev, Vadim (Lincoln, NE); Novoselov, Sergey (Puschino, RU)

    2012-09-25

    Recombinant nucleic acid constructs for the efficient expression of eukaryotic selenoproteins and related methods for production of recombinant selenoproteins are provided. The nucleic acid constructs comprise novel selenocysteine insertion sequence (SECIS) elements. Certain novel SECIS elements of the invention contain non-canonical quartet sequences. Other novel SECIS elements provided by the invention are chimeric SECIS elements comprising a canonical SECIS element that contains a non-canonical quartet sequence and chimeric SECIS elements comprising a non-canonical SECIS element that contains a canonical quartet sequence. The novel SECIS elements of the invention facilitate the insertion of selenocysteine residues into recombinant polypeptides.

  14. Suppression of Tla1 gene expression for improved solar conversion efficiency and photosynthetic productivity in plants and algae

    DOE Patents [OSTI]

    Melis, Anastasios; Mitra, Mautusi

    2010-06-29

    The invention provides method and compositions to minimize the chlorophyll antenna size of photosynthesis by decreasing TLA1 gene expression, thereby improving solar conversion efficiencies and photosynthetic productivity in plants, e.g., green microalgae, under bright sunlight conditions.

  15. Kinetics of expression of interleukin 2 receptors on class I and class II restricted murine T cell clones

    SciTech Connect (OSTI)

    Churilla, A.; Braciale, V.; Braciale, T.

    1986-03-05

    The kinetics of interleukin 2 receptor (IL-2R) expression has been examined on various class I and class II restricted, influenza specific murine T cell clones. Expression and relative levels of IL-2R were examined by Fluorescence Activated Cell Sorter analysis utilizing 3 anti-murine IL-2R monoclonal antibodies. Receptor expression was analyzed by scatchard analysis using radiolabeled recombinant human interleukin 2 to access the number of high and low affinity IL-2R per cell as well as the affinity of binding. The clones tested bound all 3 monoclonal antibodies and were inhibited in an IL-2 dependent proliferation assay by the addition of the antibodies to the culture. There was, however, differing degrees of inhibition ranging up to 99%, depending on the clone and the antibody used. IL-2R expression was detectable as early as 4-6 hours after antigenic stimulation of quiescent cells. After maximal levels of receptors were expressed, which was about 24 hours after stimulation, expression of IL-2R decreased with time on all clones examined (both class I and class II restricted). Differing rates of receptor loss is seen however, with some class II restricted clones retaining relatively high levels of receptors.

  16. Radiation-Induced Upregulation of Gene Expression From Adenoviral Vectors Mediated by DNA Damage Repair and Regulation

    SciTech Connect (OSTI)

    Nokisalmi, Petri; Rajecki, Maria; Pesonen, Sari; Escutenaire, Sophie; Soliymani, Rabah; Tenhunen, Mikko; Ahtiainen, Laura; Hemminki, Akseli

    2012-05-01

    Purpose: In the present study, we evaluated the combination of replication-deficient adenoviruses and radiotherapy in vitro. The purpose of the present study was to analyze the mechanism of radiation-mediated upregulation of adenoviral transgene expression. Methods and Materials: Adenoviral transgene expression (luciferase or green fluorescent protein) was studied with and without radiation in three cell lines: breast cancer M4A4-LM3, prostate cancer PC-3MM2, and lung cancer LNM35/enhanced green fluorescent protein. The effect of the radiation dose, modification of the viral capsid, and five different transgene promoters were studied. The cellular responses were studied using mass spectrometry and immunofluorescence analysis. Double strand break repair was modulated by inhibitors of heat shock protein 90, topoisomerase-I, and DNA protein kinase, and transgene expression was measured. Results: We found that a wide range of radiation doses increased adenoviral transgene expression regardless of the cell line, transgene, promoter, or viral capsid modification. Treatment with adenovirus, radiation, and double strand break repair inhibitors resulted in persistence of double strand breaks and subsequent increases in adenovirus transgene expression. Conclusions: Radiation-induced enhancement of adenoviral transgene expression is linked to DNA damage recognition and repair. Radiation induces a global cellular response that results in increased production of RNA and proteins, including adenoviral transgene products. This study provides a mechanistic rationale for combining radiation with adenoviral gene delivery.

  17. Methods for transforming and expression screening of filamentous fungal cells with a DNA library

    DOE Patents [OSTI]

    Teter, Sarah; Lamsa, Michael; Cherry, Joel; Ward, Connie

    2015-06-02

    The present invention relates to methods for expression screening of filamentous fungal transformants, comprising: (a) isolating single colony transformants of a DNA library introduced into E. coli; (b) preparing DNA from each of the single colony E. coli transformants; (c) introducing a sample of each of the DNA preparations of step (b) into separate suspensions of protoplasts of a filamentous fungus to obtain transformants thereof, wherein each transformant contains one or more copies of an individual polynucleotide from the DNA library; (d) growing the individual filamentous fungal transformants of step (c) on selective growth medium, thereby permitting growth of the filamentous fungal transformants, while suppressing growth of untransformed filamentous fungi; and (e) measuring activity or a property of each polypeptide encoded by the individual polynucleotides. The present invention also relates to isolated polynucleotides encoding polypeptides of interest obtained by such methods, to nucleic acid constructs, expression vectors, and recombinant host cells comprising the isolated polynucleotides, and to methods of producing the polypeptides encoded by the isolated polynucleotides.

  18. Changes in protein expression across laboratory and field experiments in Geobacter bemidjiensis

    SciTech Connect (OSTI)

    Merkley, Eric D.; Wrighton, Kelly C.; Castelle, Cindy; Anderson, Brian J.; Wilkins, Michael J.; Shah, Vega; Arbour, Tyler; Brown, Joseph N.; Singer, Steven W.; Smith, Richard D.; Lipton, Mary S.

    2015-03-06

    Bacterial extracellular metal respiration, as carried out by members of the genus Geobacter, is of interest for applications including microbial fuel cells and bioremediation. Geobacter bemidjiensis is the major species whose growth is stimulated during groundwater amendment with acetate. We have carried out label-free proteomics studies of Geobacter bemidjiensis grown with acetate as the electron donor and either fumarate, ferric citrate, or one of two hydrous ferric oxide mineral types as electron acceptor. The major class of proteins whose expression changes across these conditions is c-type cytochromes, many of which are known to be involved in extracellular metal reduction in other, better-characterized Geobacter species. Some proteins with multiple homologues in G. bemidjiensis (OmcS, OmcB) had different expression patterns than observed for their G. sulfurreducens homologues under similar growth conditions. We also compared the proteome from our study to a prior proteomics study of biomass recovered from an aquifer in Colorado, where the microbial community was dominated by strains closely-related to G. bemidjiensis. We detected an increased number of proteins with functions related to motility and chemotaxis in the Colorado field samples compared to the laboratory samples, suggesting the importance of motility for in situ extracellular metal respiration.

  19. Novel, highly expressed late nodulin gene (LjNOD16) from Lotus japonicus

    SciTech Connect (OSTI)

    Kapranov, P.; Bruijn, F.J. de; Szczyglowski, K.

    1997-04-01

    We have isolated a Lotus japonicus cDNA corresponding to a highly abundant, late nodule-specific RNA species that encodes a polypeptide with a predicted molecular mass of 15.6 kD. The protein and its corresponding gene were designated NIj16 and LjNOD16, respectively. LjNOD16 was found to be expressed only in the infected cells of L. japonicus nodules. Related DNA sequences could be identified in the genomes of both Glycine max and Medicago sativa. In the latter, a homologous mRNA species was detected in the nodules. Unlike LiNOD16, its alfalfa homologs appear to represent low-abundance mRNA species. However, the proteins corresponding to the LjNOD16 and its alfalfa homolog could be detected at similar levels in nodules but not in roots of both legume species. The predicted amino acid sequence analysis of nodulin NIj16 revealed the presence of a long {alpha}-helical region and a positively charged C terminus. The former domain has a very high propensity to form a coiled-coil type structure, indicating that nodulin NIj16 may interact with an as-yet-unidentified protein target(s) in the nodule-infected cells. Homology searches revealed no significant similarities to any known sequences in the databases, with the exception of two related, anonymous Arabidopsis expressed sequence tags.

  20. A replication-deficient rabies virus vaccine expressing Ebola virus glycoprotein is highly attenuated for neurovirulence

    SciTech Connect (OSTI)

    Papaneri, Amy B.; Wirblich, Christoph; Cann, Jennifer A.; Cooper, Kurt; Jahrling, Peter B.; Integrated Research Facility, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Fort Detrick MD, 21702 ; Schnell, Matthias J.; Blaney, Joseph E.

    2012-12-05

    We are developing inactivated and live-attenuated rabies virus (RABV) vaccines expressing Ebola virus (EBOV) glycoprotein for use in humans and endangered wildlife, respectively. Here, we further characterize the pathogenesis of the live-attenuated RABV/EBOV vaccine candidates in mice in an effort to define their growth properties and potential for safety. RABV vaccines expressing GP (RV-GP) or a replication-deficient derivative with a deletion of the RABV G gene (RV{Delta}G-GP) are both avirulent after intracerebral inoculation of adult mice. Furthermore, RV{Delta}G-GP is completely avirulent upon intracerebral inoculation of suckling mice unlike parental RABV vaccine or RV-GP. Analysis of RV{Delta}G-GP in the brain by quantitative PCR, determination of virus titer, and immunohistochemistry indicated greatly restricted virus replication. In summary, our findings indicate that RV-GP retains the attenuation phenotype of the live-attenuated RABV vaccine, and RV{Delta}G-GP would appear to be an even safer alternative for use in wildlife or consideration for human use.

  1. Transgenic plants expressing GLK1 and CCA1 having increased nitrogen assimilation capacity

    DOE Patents [OSTI]

    Coruzzi, Gloria (New York, NY); Gutierrez, Rodrigo A. (Santiago, CL); Nero, Damion C. (Woodside, NY)

    2012-04-10

    Provided herein are compositions and methods for producing transgenic plants. In specific embodiments, transgenic plants comprise a construct comprising a polynucleotide encoding CCA1, GLK1 or bZIP1, operably linked to a plant-specific promote, wherein the CCA1, GLK1 or bZIP1 is ectopically overexpressed in the transgenic plants, and wherein the promoter is optionally a constitutive or inducible promoter. In other embodiments, transgenic plants in which express a lower level of CCA1, GLK1 or bZIP1 are provided. Also provided herein are commercial products (e.g., pulp, paper, paper products, or lumber) derived from the transgenic plants (e.g., transgenic trees) produced using the methods provided herein.

  2. Analytical Expressions for the Hard-Scattering Production of Massive Partons

    SciTech Connect (OSTI)

    Wong, Cheuk-Yin

    2016-01-01

    We obtain explicit expressions for the two-particle differential cross section $E_c E_\\kappa d\\sigma (AB \\to c\\kappa X) /d\\bb c d \\bb \\kappa$ and the two-particle angular correlation function \\break $d\\sigma(AB$$ \\to$$ c\\kappa X)/d\\Delta \\phi \\, d\\Delta y$ in the hard-scattering production of massive partons in order to exhibit the ``ridge" structure on the away side in the hard-scattering process. The single-particle production cross section $d\\sigma(AB \\to cX) /dy_c c_T dc_T $ is also obtained and compared with the ALICE experimental data for charm production in $pp$ collisions at 7 TeV at LHC.

  3. High density growth of T7 expression strains with auto-induction option

    DOE Patents [OSTI]

    Studier, F. William (Stony Brook, NY)

    2010-07-20

    A bacterial growth medium for promoting auto-induction of transcription of cloned DNA in cultures of bacterial cells grown batchwise is disclosed. The transcription is under the control of a lac repressor. Also disclosed is a bacterial growth medium for improving the production of a selenomethionine-containing protein or polypeptide in a bacterial cell, the protein or polypeptide being produced by recombinant DNA techniques from a lac or T7lac promoter, the bacterial cell encoding a vitamin B12-dependent homocysteine methylase. Finally, disclosed is a bacterial growth medium for suppressing auto-induction of expression in cultures of bacterial cells grown batchwise, said transcription being under the control of lac repressor.

  4. Subacute effects of hexabromocyclododecane (HBCD) on hepatic gene expression profiles in rats

    SciTech Connect (OSTI)

    Canton, Rocio F. [Institute for Risk Assessment Sciences (IRAS), Utrecht University, P.O. Box 80.177, NL-3508 TD, Utrecht (Netherlands)], E-mail: rfcanton@gmail.com; Peijnenburg, Ad A.C.M.; Hoogenboom, Ron L.A.P. [RIKILT Institute of Food Safety, Wageningen University and Research Center, P.O. Box 230, 6700 AE Wageningen (Netherlands); Piersma, Aldert H.; Ven, Leo T.M. van der [National Institute of Public Health and the Environment (RIVM), Laboratory for Heath Protection Research, P.O. Box 1, 3720 BA Bilthoven (Netherlands); Berg, Martin van den [Institute for Risk Assessment Sciences (IRAS), Utrecht University, P.O. Box 80.177, NL-3508 TD, Utrecht (Netherlands); Heneweer, Marjoke [RIKILT Institute of Food Safety, Wageningen University and Research Center, P.O. Box 230, 6700 AE Wageningen (Netherlands)

    2008-09-01

    Hexabromoyclododecane (HBCD), used as flame retardant (FR) mainly in textile industry and in polystyrene foam manufacture, has been identified as a contaminant at levels comparable to other brominated FRs (BFRs). HBCD levels in biota are increasing slowly and seem to reflect the local market demand. The toxicological database of HBCD is too limited to perform at present a solid risk assessment, combining data from exposure and effect studies. In order to fill in some gaps, a 28-day HBCD repeated dose study (OECD407) was done in Wistar rats. In the present work liver tissues from these animals were used for gene expression profile analysis. Results show clear gender specificity with females having a higher number of regulated genes and therefore being more sensitive to HBCD than males. Several specific pathways were found to be affected by HBCD exposure, like PPAR-mediated regulation of lipid metabolism, triacylglycerol metabolism, cholesterol biosynthesis, and phase I and II pathways. These results were corroborated with quantitative RT-PCR analysis. Cholesterol biosynthesis and lipid metabolism were especially down-regulated in females. Genes involved in phase I and II metabolism were up-regulated predominantly in males, which could explain the observed lower HBCD hepatic disposition in male rats in this 28-day study. These sex-specific differences in gene expression profiles could also underlie sex-specific differences in toxicity (e.g. decreased thyroid hormone or increased serum cholesterol levels). To our knowledge, this is the fist study that describes the changes in rat hepatic gene profiles caused by this commonly used flame retardant.

  5. Azidothymidine and cisplatin increase p14ARF expression in OVCAR-3 ovarian cancer cell line

    SciTech Connect (OSTI)

    Vaskivuo, Liisa; Rysae, Jaana; Koivuperae, Johanna; Myllynen, Paeivi; Vaskivuo, Tommi; Chvalova, Katerina; Serpi, Raisa; Savolainen, Eeva-Riitta; Puistola, Ulla; Vaehaekangas, Kirsi . E-mail: kirsi.vahakangas@uku.fi

    2006-10-01

    p14{sup ARF} tumor suppressor protein regulates p53 by interfering with mdm2-p53 interaction. p14{sup ARF} is activated in response to oncogenic stimuli but little is known of the responses of endogenous p14{sup ARF} to different types of cellular stress or DNA damage. Azidothymidine (AZT) is being tested in several clinical trials as an enhancer of anticancer chemotherapy. However, the knowledge of the relationship between AZT and cellular pathways, e.g. p53 pathway, is very limited. In this study, we show that AZT, cisplatin (CDDP) and docetaxel (DTX) all induce unique molecular responses in OVCAR-3 ovarian carcinoma cells carrying a mutated p53, while in A2780, ovarian carcinoma and MCF-7 breast carcinoma cells with wild type p53, all of these drugs cause similar p53 responses. We found that endogenous p14{sup ARF} protein in OVCAR-3 cells is down-regulated by DTX but induced by AZT and a short CDDP pulse treatment. In HT-29 colon carcinoma cells with a mutated p53, all treatments down-regulated p14{sup ARF} protein. Both CDDP and AZT increased the expression of p14ARF mRNA in OVCAR-3 cells. Differences in cell death induced by these drugs did not explain the differences in protein and mRNA expressions. No increase in the level of either c-Myc or H-ras oncoproteins was seen in OVCAR-3 cells after AZT or CDDP-treatment. These results suggest that p14{sup ARF} can respond to DNA damage without oncogene activation in cell lines without functional p53.

  6. Flow cytometric analysis of expression of interleukin-2 receptor beta chain (p70-75) on various leukemic cells

    SciTech Connect (OSTI)

    Hoshino, S.; Oshimi, K.; Tsudo, M.; Miyasaka, M.; Teramura, M.; Masuda, M.; Motoji, T.; Mizoguchi, H. )

    1990-08-15

    We analyzed the expression of the interleukin-2 receptor (IL-2R) beta chain (p70-75) on various leukemic cells from 44 patients by flow cytometric analysis using the IL-2R beta chain-specific monoclonal antibody, designated Mik-beta 1. Flow cytometric analysis demonstrated the expression of the IL-2R beta chain on granular lymphocytes (GLs) from all eight patients with granular lymphocyte proliferative disorders (GLPDs), on adult T-cell leukemia (ATL) cells from all three patients with ATL, and on T-cell acute lymphoblastic leukemia (T-ALL) cells from one of three patients with T-ALL. Although GLs from all the GLPD patients expressed the IL-2R beta chain alone and not the IL-2R alpha chain (Tac-antigen: p55), ATL and T-ALL cells expressing the beta chain coexpressed the alpha chain. In two of seven patients with common ALL (cALL) and in both patients with B-cell chronic lymphocytic leukemia, the leukemic cells expressed the alpha chain alone. Neither the alpha chain nor the beta chain was expressed on leukemic cells from the remaining 28 patients, including all 18 patients with acute nonlymphocytic leukemia, five of seven patients with cALL, all three patients with multiple myeloma, and two of three patients with T-ALL. These results indicate that three different forms of IL-2R chain expression exist on leukemic cells: the alpha chain alone; the beta chain alone; and both the alpha and beta chains. To examine whether the results obtained by flow cytometric analysis actually reflect functional aspects of the expressed IL-2Rs, we studied the specific binding of 125I-labeled IL-2 (125I-IL-2) to leukemic cells in 18 of the 44 patients. In addition, we performed 125I-IL-2 crosslinking studies in seven patients. The results of IL-2R expression of both 125I-IL-2 binding assay and crosslinking studies were in agreement with those obtained by flow cytometric analysis.

  7. Transgenic expression of the dicotyledonous pattern recognition receptor EFR in rice leads to ligand-dependent activation of defense responses

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Schwessinger, Benjamin; Bahar, Ofir; Thomas, Nicolas; Holton, Nicolas; Nekrasov, Vladimir; Ruan, Deling; Canlas, Patrick E.; Daudi, Arsalan; Petzold, Christopher J.; Singan, Vasanth R.; et al

    2015-03-30

    Plant plasma membrane localized pattern recognition receptors (PRRs) detect extracellular pathogen-associated molecules. PRRs such as Arabidopsis EFR and rice XA21 are taxonomically restricted and are absent from most plant genomes. Here we show that rice plants expressing EFR or the chimeric receptor EFR::XA21, containing the EFR ectodomain and the XA21 intracellular domain, sense both Escherichia coli- and Xanthomonas oryzae pv. oryzae (Xoo)-derived elf18 peptides at sub-nanomolar concentrations. Treatment of EFR and EFR::XA21 rice leaf tissue with elf18 leads to MAP kinase activation, reactive oxygen production and defense gene expression. Although expression of EFR does not lead to robust enhanced resistancemore » to fully virulent Xoo isolates, it does lead to quantitatively enhanced resistance to weakly virulent Xoo isolates. EFR interacts with OsSERK2 and the XA21 binding protein 24 (XB24), two key components of the rice XA21-mediated immune response. Rice-EFR plants silenced for OsSERK2, or overexpressing rice XB24 are compromised in elf18-induced reactive oxygen production and defense gene expression indicating that these proteins are also important for EFR-mediated signaling in transgenic rice. Taken together, our results demonstrate the potential feasibility of enhancing disease resistance in rice and possibly other monocotyledonous crop species by expression of dicotyledonous PRRs. Our results also suggest that Arabidopsis EFR utilizes at least a subset of the known endogenous rice XA21 signaling components.« less

  8. Altered microRNA expression patterns in irradiated hematopoietic tissues suggest a sex-specific protective mechanism

    SciTech Connect (OSTI)

    Ilnytskyy, Yaroslav; Zemp, Franz J.; Koturbash, Igor [Department of Biological Sciences, University of Lethbridge, 4401 University Drive, Lethbridge, Alta., T1K 3M4 (Canada); Kovalchuk, Olga [Department of Biological Sciences, University of Lethbridge, 4401 University Drive, Lethbridge, Alta., T1K 3M4 (Canada)], E-mail: olga.kovalchuk@uleth.ca

    2008-12-05

    To investigate involvement of miRNAs in radiation responses we used microRNAome profiling to analyze the sex-specific response of radiation sensitive hematopoietic lymphoid tissues. We show that radiation exposure resulted in a significant and sex-specific deregulation of microRNA expression in murine spleen and thymus tissues. Among the regulated miRNAs, we found that changes in expression of miR-34a and miR-7 may be involved in important protective mechanisms counteracting radiation cytotoxicity. We observed a significant increase in the expression of tumor-suppressor miR-34a, paralleled by a decrease in the expression of its target oncogenes NOTCH1, MYC, E2F3 and cyclin D1. Additionally, we show that miR-7 targets the lymphoid-specific helicase LSH, a pivotal regulator of DNA methylation and genome stability. While miR-7 was significantly down-regulated LSH was significantly up-regulated. These cellular changes may constitute an attempt to counteract radiation-induced hypomethylation. Tissue specificity of miRNA responses and possible regulation of miRNA expression upon irradiation are discussed.

  9. Epidermal Growth Factor Receptor Expression As Prognostic Marker in Patients With Anal Carcinoma Treated With Concurrent Chemoradiation Therapy

    SciTech Connect (OSTI)

    Fraunholz, Ingeborg, E-mail: inge.fraunholz@kgu.de [Department of Radiotherapy and Oncology, Goethe University, Frankfurt/Main (Germany); Rdel, Franz; Kohler, Daniela [Department of Radiotherapy and Oncology, Goethe University, Frankfurt/Main (Germany); Diallo-Georgiopoulou, Margarita [Department of Radiotherapy and Oncology, Goethe University, Frankfurt/Main (Germany); Department of Radiation Oncology, Klinikum Offenbach, Offenbach/Main (Germany); Distel, Luitpold [Department of Radiation Oncology, Friedrich Alexander University, Erlangen (Germany); Falk, Stefan [Pathology Associates, Frankfurt/Main (Germany); Rdel, Claus [Department of Radiotherapy and Oncology, Goethe University, Frankfurt/Main (Germany)

    2013-08-01

    Purpose: To investigate the prognostic value of epidermal growth factor receptor (EGFR) expression in pretreatment tumor biopsy specimens of patients with anal cancer treated with concurrent 5-fluorouracil and mitomycin C-based chemoradiation therapy (CRT). Methods and Materials: Immunohistochemical staining for EGFR was performed in pretreatment biopsy specimens of 103 patients with anal carcinoma. EGFR expression was correlated with clinical and histopathologic characteristics and with clinical endpoints, including local failure-free survival (LFFS), colostomy-free survival (CFS), distant metastases-free survival (DMFS), cancer-specific survival (CSS), and overall survival (OS). Results: EGFR staining intensity was absent in 3%, weak in 23%, intermediate in 36% and intense in 38% of the patients. In univariate analysis, the level of EGFR staining was significantly correlated with CSS (absent/weak vs intermediate/intense expression: 5-year CSS, 70% vs 86%, P=.03). As a trend, this was also observed for DMFS (70% vs 86%, P=.06) and LFFS (70% vs 87%, P=.16). In multivariate analysis, N stage, tumor differentiation, and patients sex were independent prognostic factors for CSS, whereas EGFR expression only reached borderline significance (hazard ratio 2.75; P=.08). Conclusion: Our results suggest that elevated levels of pretreatment EGFR expression could be correlated with favorable clinical outcome in anal cancer patients treated with CRT. Further studies are warranted to elucidate how EGFR is involved in the response to CRT.

  10. CD26-mediated regulation of periostin expression contributes to migration and invasion of malignant pleural mesothelioma cells

    SciTech Connect (OSTI)

    Komiya, Eriko; Ohnuma, Kei; Yamazaki, Hiroto; Hatano, Ryo; Iwata, Satoshi; Okamoto, Toshihiro; Dang, Nam H.; Morimoto, Chikao

    2014-05-16

    Highlights: CD26-expressing MPM cells upregulate production of periostin. The intracytoplasmic region of CD26 mediates the upregulation of periostin. CD26 expression leads to nuclear translocation of Twist1 via phosphorylation of Src. Secreted periostin enhances migration and invasion of MPM cells. - Abstract: Malignant pleural mesothelioma (MPM) is an aggressive malignancy arising from mesothelial lining of pleura. It is generally associated with a history of asbestos exposure and has a very poor prognosis, partly due to the lack of a precise understanding of the molecular mechanisms associated with its malignant behavior. In the present study, we expanded on our previous studies on the enhanced motility and increased CD26 expression in MPM cells, with a particular focus on integrin adhesion molecules. We found that expression of CD26 upregulates periostin secretion by MPM cells, leading to enhanced MPM cell migratory and invasive activity. Moreover, we showed that upregulation of periostin expression results from the nuclear translocation of the basic helix-loop-helix transcription factor Twist1, a process that is mediated by CD26-associated activation of Src phosphorylation. While providing new and profound insights into the molecular mechanisms involved in MPM biology, these findings may also lead to the development of novel therapeutic strategies for MPM.

  11. Endothelial Cell Migration and Vascular Endothelial Growth Factor Expression Are the Result of Loss of Breast Tissue Polarity

    SciTech Connect (OSTI)

    Chen, Amy; Cuevas, Ileana; Kenny, Paraic A; Miyake, Hiroshi; Mace, Kimberley; Ghajar, Cyrus; Boudreau, Aaron; Bissell, Mina; Boudreau, Nancy

    2009-05-26

    Recruiting a new blood supply is a rate-limiting step in tumor progression. In a three-dimensional model of breast carcinogenesis, disorganized, proliferative transformed breast epithelial cells express significantly higher expression of angiogenic genes compared with their polarized, growth-arrested nonmalignant counterparts. Elevated vascular endothelial growth factor (VEGF) secretion by malignant cells enhanced recruitment of endothelial cells (EC) in heterotypic cocultures. Significantly, phenotypic reversion of malignant cells via reexpression of HoxD10, which is lost in malignant progression, significantly attenuated VEGF expression in a hypoxia-inducible factor 1{alpha}-independent fashion and reduced EC migration. This was due primarily to restoring polarity: forced proliferation of polarized, nonmalignant cells did not induce VEGF expression and EC recruitment, whereas disrupting the architecture of growth-arrested, reverted cells did. These data show that disrupting cytostructure activates the angiogenic switch even in the absence of proliferation and/or hypoxia and restoring organization of malignant clusters reduces VEGF expression and EC activation to levels found in quiescent nonmalignant epithelium. These data confirm the importance of tissue architecture and polarity in malignant progression.

  12. In vivo expression of the lacY gene in two segments leads to functional lac permease

    SciTech Connect (OSTI)

    Bibi, E.; Kaback, H.R. )

    1990-06-01

    The lacY gene of Escherichia coli was cut into two approximately equal-size fragments with Afl II and subcloned individually or together under separate lac operator/promoters in plasmid pT7-5. Under these conditions, lac permease is expressed in two portions: (i) the N-terminal portion (the N terminus, the first six putative transmembrane helices, and most of putative loop 7) and (ii) the C-terminal portion (the last six putative transmembrane helices and the C terminus). Cells harboring pT7-5 encoding both fragments transport lactose at about 30% the rate of cells expressing intact permease to a comparable steady-state level of accumulation. In contrast, cells expressing either half of the permease independently do not transport lactose. As judged by ({sup 35}S)methionine labeling and immunoblotting, intact permease in completely absent from the membrane of cells expressing lacY fragments either individually or together. Thus, transport activity must result from an association between independently synthesized pieces of lac permease. When the gene fragments are expressed individually, the N-terminal portion of the permease is observed inconsistently, and the C-terminal portion is not observed. When the gene fragments are expressed together, polypeptides identified as the N- and C-terminal moieties of the permease are found in the membrane. It is concluded that the N- or C-terminal halves of lac permease are proteolyzed when synthesized independently and that association between the two complementing polypeptides leads to a more stable, catalytically active complex.

  13. Fibroblast growth factor-2 up-regulates the expression of nestin through the RasRafERKSp1 signaling axis in C6 glioma cells

    SciTech Connect (OSTI)

    Chang, Kai-Wei; Huang, Yuan-Li; Wong, Zong-Ruei; Su, Peng-Han; Huang, Bu-Miin; Ju, Tsai-Kai; Technology Commons, College of Life Science, National Taiwan University, Taipei 106, Taiwan ; Yang, Hsi-Yuan

    2013-05-17

    Highlights: Nestin expression in C6 glioma cells is induced by FGF-2. Nestin expression is induced by FGF-2 via de novo RNA and protein synthesis. The FGFR inhibitor SU5402 blocks the FGF-2-induced nestin expression. The mRNA of FGFR1 and 3 are detected in C6 glioma cells. RasRafERKSp1 signaling pathway is responsibe for FGF-2-induced nestin expression. -- Abstract: Nestin is a 240-kDa intermediate filament protein expressed mainly in neural and myogenic stem cells. Although a substantial number of studies have focused on the expression of nestin during development of the central nervous system, little is known about the factors that induce and regulate its expression. Fibroblast growth factor-2 (FGF-2) is an effective mitogen and stimulates the proliferation and differentiation of a subset of nestin-expressing cells, including neural progenitor cells, glial precursor cells, and smooth muscle cells. To assess whether FGF-2 is a potent factor that induces the expression of nestin, C6 glioma cells were used. The results showed that nestin expression was up-regulated by FGF-2 via de novo RNA and protein synthesis. Our RT-PCR results showed that C6 glioma cells express FGFR1/3, and FGFRs is required for FGF-2-induced nestin expression. Further signaling analysis also revealed that FGF-2-induced nestin expression is mediated through FGFRMAPKERK signaling axis and the transcriptional factor Sp1. These findings provide new insight into the regulation of nestin in glial system and enable the further studies on the function of nestin in glial cells.

  14. Human metastatic melanoma cell lines express high levels of growth hormone receptor and respond to GH treatment

    SciTech Connect (OSTI)

    Sustarsic, Elahu G.; Department of Biological Sciences, Ohio University, Athens, OH ; Junnila, Riia K.; Kopchick, John J.

    2013-11-08

    Highlights: Most cancer types of the NCI60 have sub-sets of cell lines with high GHR expression. GHR is highly expressed in melanoma cell lines. GHR is elevated in advanced stage IV metastatic tumors vs. stage III. GH treatment of metastatic melanoma cell lines alters growth and cell signaling. -- Abstract: Accumulating evidence implicates the growth hormone receptor (GHR) in carcinogenesis. While multiple studies show evidence for expression of growth hormone (GH) and GHR mRNA in human cancer tissue, there is a lack of quantification and only a few cancer types have been investigated. The National Cancer Institutes NCI60 panel includes 60 cancer cell lines from nine types of human cancer: breast, CNS, colon, leukemia, melanoma, non-small cell lung, ovarian, prostate and renal. We utilized this panel to quantify expression of GHR, GH, prolactin receptor (PRLR) and prolactin (PRL) mRNA with real-time RT qPCR. Both GHR and PRLR show a broad range of expression within and among most cancer types. Strikingly, GHR expression is nearly 50-fold higher in melanoma than in the panel as a whole. Analysis of human metastatic melanoma biopsies confirmed GHR gene expression in melanoma tissue. In these human biopsies, the level of GHR mRNA is elevated in advanced stage IV tumor samples compared to stage III. Due to the novel finding of high GHR in melanoma, we examined the effect of GH treatment on three NCI60 melanoma lines (MDA-MB-435, UACC-62 and SK-MEL-5). GH increased proliferation in two out of three cell lines tested. Further analysis revealed GH-induced activation of STAT5 and mTOR in a cell line dependent manner. In conclusion, we have identified cell lines and cancer types that are ideal to study the role of GH and PRL in cancer, yet have been largely overlooked. Furthermore, we found that human metastatic melanoma tumors express GHR and cell lines possess active GHRs that can modulate multiple signaling pathways and alter cell proliferation. Based on this data, GH could be a new therapeutic target in melanoma.

  15. Equine cytochrome P450 2B6 Genomic identification, expression and functional characterization with ketamine

    SciTech Connect (OSTI)

    Peters, L.M.; Demmel, S.; Pusch, G.; Buters, J.T.M.; Zielinski, J.; Leeb, T.; Mevissen, M.; Schmitz, A.

    2013-01-01

    Ketamine is an anesthetic and analgesic regularly used in veterinary patients. As ketamine is almost always administered in combination with other drugs, interactions between ketamine and other drugs bear the risk of either adverse effects or diminished efficacy. Since cytochrome P450 enzymes (CYPs) play a pivotal role in the phase I metabolism of the majority of all marketed drugs, drugdrug interactions often occur at the active site of these enzymes. CYPs have been thoroughly examined in humans and laboratory animals, but little is known about equine CYPs. The characterization of equine CYPs is essential for a better understanding of drug metabolism in horses. We report annotation, cloning and heterologous expression of the equine CYP2B6 in V79 Chinese hamster fibroblasts. After computational annotation of all CYP2B genes, the coding sequence (CDS) of equine CYP2B6 was amplified by RT-PCR from horse liver total RNA and revealed an amino acid sequence identity of 77% and a similarity of 93.7% to its human ortholog. A non-synonymous variant c.226G>A in exon 2 of the equine CYP2B6 was detected in 97 horses. The mutant A-allele showed an allele frequency of 82%. Two further variants in exon 3 were detected in one and two horses of this group, respectively. Transfected V79 cells were incubated with racemic ketamine and norketamine as probe substrates to determine metabolic activity. The recombinant equine CYP2B6 N-demethylated ketamine to norketamine and produced metabolites of norketamine, such as hydroxylated norketamines and 5,6-dehydronorketamine. V{sub max} for S-/and R-norketamine formation was 0.49 and 0.45 nmol/h/mg cellular protein and K{sub m} was 3.41 and 2.66 ?M, respectively. The N-demethylation of S-/R-ketamine was inhibited concentration-dependently with clopidogrel showing an IC{sub 50} of 5.63 and 6.26 ?M, respectively. The functional importance of the recorded genetic variants remains to be explored. Equine CYP2B6 was determined to be a CYP enzyme involved in ketamine and norketamine metabolism, thus confirming results from inhibition studies with horse liver microsomes. Clopidogrel seems to be a feasible inhibitor for equine CYP2B6. The specificity still needs to be established with other single equine CYPs. Heterologous expression of single equine CYP enzymes opens new possibilities to substantially improve the understanding of drug metabolism and drug interactions in horses. -- Highlights: ? We annotate, express and functionally characterize equine CYP2B6. ? 3 genetic variants within this gene are described. ? Equine CYP2B6 N-demethylates ketamine and metabolizes norketamine. ? Equine CYP2B6 can be inhibited by clopidogrel.

  16. Expression of Autoactivated Stromelysin-1 in Mammary Glands of Transgenic Mice Leads to a Reactive Stroma During Early Development

    SciTech Connect (OSTI)

    Thomasset, N.; Lochter, A.; Sympson, C.J.; Lund, L.R.; Williams, D.R.; Behrendtsen, O.; Werb, Z.; Bissell, M.J.

    1998-04-24

    Extracellular matrix and extracellular matrix-degrading matrix metalloproteinases play a key role in interactions between the epithelium and the mesenchyme during mammary gland development and disease. In patients with breast cancer, the mammary mesenchyme undergoes a stromal reaction, the etiology of which is unknown. We previously showed that targeting of an autoactivating mutant of the matrix metalloproteinase stromelysin-1 to mammary epithelia of transgenic mice resulted in reduced mammary function during pregnancy and development of preneoplastic and neoplastic lesions. Here we examine the cascade of alterations before breast tumor formation in the mammary gland stroma once the expression of the stromelysin-1 transgene commences. Beginning in postpubertal virgin animals, low levels of transgene expression in mammary epithelia led to increased expression of endogenous stromelysin-1 in stromal fibroblasts and up-regulation of other matrix metalloproteinases, without basement membrane disruption. These changes were accompanied by the progressive development of a compensatory reactive stroma, characterized by increased collagen content and vascularization in glands from virgin mice. This remodeling of the gland affected epithelial-mesenchymal communication as indicated by inappropriate expression of tenascin-C starting by day 6 of pregnancy. This, together with increased transgene expression, led to basement membrane disruption starting by day 15 of pregnancy. We propose that the highly reactive stroma provides a prelude to breast epithelial tumors observed in these animals. Epithelial development depends on an exquisite series of inductive and instructive interactions between the differentiating epithelium and the mesenchymal (stromal) compartment. The epithelium, which consists of luminal and myoepithelial cells, is separated from the stroma by a basement membrane (BM), which plays a central role in mammary gland homeostasis and gene expression. In vivo, stromal cells produce fibronectin, collagens, proteoglycans, and some components of the BM, as well as a number of proteinases that can effectively degrade BM constituents. Stromal and epithelial cells of the mammary gland interact to regulate BM synthesis and degradation and, thus, mammary function. Matrix metalloproteinases (MMPs) are extracellular matrix (ECM)-degrading enzymes involved in mammary gland morphogenesis and involution. During late pregnancy and lactation, when the gland becomes fully functional, the expression of MMPs is low however, during involution, when the gland loses function and is remodeled, synthesis of ECM-degrading proteinases increases dramatically.11 Disturbance of the balance between MMPs and MMP inhibitors leads to either unscheduled involution or prolonged lactation. Mammary glands of virgin mice expressing an autoactivating stromelysin-1 (SL-1) transgene display supernumerary branches and precocious alveolar development, accompanied by the synthesis of {beta}-casein at levels found normally only during early pregnancy. During late pregnancy, increased expression of the SL-1 transgene leads to a reduction in expression of pregnancy-specific genes. Later in life, some SL-1 transgenic mice develop hyperplastic, dysplastic, and ductal carcinoma in situ-like lesions, as well as malignant tumors. Little is known about the sequence of changes that occurs before formation of an overt reactive stroma in breast cancer. In the present study, we address the question of whether and how the stromal compartment is altered as a consequence of inappropriate SL-1 transgene expression in the epithelium.

  17. Repression of miR-17-5p with elevated expression of E2F-1 and c-MYC in non-metastatic hepatocellular carcinoma and enhancement of cell growth upon reversing this expression pattern

    SciTech Connect (OSTI)

    El Tayebi, H.M.; Omar, K.; Hegy, S.; El Maghrabi, M.; El Brolosy, M.; Hosny, K.A.; Esmat, G.; Abdelaziz, A.I.

    2013-05-10

    Highlights: The oncogenic miR-17-5p is downregulated in non-metastatic hepatocellular carcinoma patients. E2F-1 and c-MYC transcripts are upregulated in non-metastatic HCC patients. miR-17-5p forced overexpression inhibited E2F-1 and c-MYC expression in HuH-7 cells. miR-17-5p mimicking increased HuH-7 cell growth, proliferation, migration and colony formation. miR-17-5p is responsible for HCC progression among the c-MYC/E2F-1/miR-17-5p triad members. -- Abstract: E2F-1, c-MYC, and miR-17-5p is a triad of two regulatory loops: a negative and a positive loop, where c-MYC induces the expression of E2F-1 that induces the expression of miR-17-5p which in turn reverses the expression of E2F-1 to close the loop. In this study, we investigated this triad for the first time in hepatocellular carcinoma (HCC), where miR-17-5p showed a significant down-regulation in 23 non-metastatic HCC biopsies compared to 10 healthy tissues; however, E2F-1 and c-MYC transcripts were markedly elevated. Forced over-expression of miR-17-5p in HuH-7 cells resulted in enhanced cell proliferation, growth, migration and clonogenicity with concomitant inhibition of E2F-1 and c-MYC transcripts expressions, while antagomirs of miR-17-5p reversed these events. In conclusion, this study revealed a unique pattern of expression for miR-17-5p in non-metastatic HCC patients in contrast to metastatic HCC patients. In addition we show that miR-17-5p is the key player among the triad that tumor growth and spread.

  18. Triangle Universities Nuclear Laboratory : 2011

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    every half-hour between Residence Inn by Marriott, Hilton Garden Inn, and the French Family Science Center (FFSC). Saturday, Nov. 7: 7:30 AM - 10 AM - Every half-hour...

  19. Protocol for Uniformly Measuring and Expressing the Performance of Energy Storage Systems

    SciTech Connect (OSTI)

    Bray, Kathryn L.; Conover, David R.; Kintner-Meyer, Michael CW; Viswanathan, Vijayganesh; Ferreira, Summer; Rose, David; Schoenwald, David

    2012-10-01

    The U.S. Department of Energys Energy Storage Systems (ESS) Program, through the support of Pacific Northwest National Laboratory (PNNL) and Sandia National Laboratories (SNL), facilitated the development of the protocol provided in this report. The focus of the protocol is to provide a uniform way of measuring, quantifying, and reporting the performance of EESs in various applications; something that does not exist today and, as such, is hampering the consideration and use of this technology in the market. The availability of an application-specific protocol for use in measuring and expressing performance-related metrics of ESSs will allow technology developers, power-grid operators and other end-users to evaluate the performance of energy storage technologies on a uniform and comparable basis. This will help differentiate technologies and products for specific application(s) and provide transparency in how performance is measured. It also will assist utilities and other consumers of ESSs make more informed decisions as they consider the potential application and use of ESSs, as well as form the basis for documentation that might be required to justify utility investment in such technologies.

  20. Protocol for uniformly measuring and expressing the performance of energy storage systems.

    SciTech Connect (OSTI)

    Ferreira, Summer Rhodes; Rose, David Martin; Schoenwald, David Alan; Bray, Kathy; Conover, David; Kintner-Meyer, Michael; Viswanathan, Vilayanur

    2013-08-01

    The U.S. Department of Energy's Energy Storage Systems (ESS) Program, through the support of Pacific Northwest National Laboratory (PNNL) and Sandia National Laboratories (SNL), facilitated the development of the protocol provided in this report. The focus of the protocol is to provide a uniform way of measuring, quantifying, and reporting the performance of ESSs in various applications; something that does not exist today and, as such, is hampering the consideration and use of this technology in the market. The availability of an application-specific protocol for use in measuring and expressing performance-related metrics of ESSs will allow technology developers, power-grid operators and other end-users to evaluate the performance of energy storage technologies on a uniform and comparable basis. This will help differentiate technologies and products for specific application(s) and provide transparency in how performance is measured. It also will assist utilities and other consumers of ESSs to make more informed decisions as they consider the potential application and use of ESSs, as well as form the basis for documentation that might be required to justify utility investment in such technologies.

  1. GIM3E: Condition-specific Models of Cellular Metabolism Developed from Metabolomics and Expression Data

    SciTech Connect (OSTI)

    Schmidt, Brian; Ebrahim, Ali; Metz, Thomas O.; Adkins, Joshua N.; Palsson, Bernard O.; Hyduke, Daniel R.

    2013-11-15

    Motivation: Genome-scale metabolic models have been used extensively to investigate alterations in cellular metabolism. The accuracy of these models to represent cellular metabolism in specific conditions has been improved by constraining the model with omics data sources. However, few practical methods for integrating metabolomics data with other omics data sources into genome-scale models of metabolism have been reported. Results: GIMMME (Gene Inactivation Moderated by Metabolism, Metabolomics, and Expression) is an algorithm that enables the development of condition-specific models based on an objective function, transcriptomics, and intracellular metabolomics data. GIMMME establishes metabolite utilization requirements with metabolomics data, uses model-paired transcriptomics data to find experimentally supported solutions, and also provides calculations of the turnover (production / consumption) flux of metabolites. GIMMME was employed to investigate the effects of integrating additional omics datasets to create increasingly constrained solution spaces of Salmonella Typhimurium metabolism during growth in both rich and virulence media. This integration proved to be informative and resulted in a requirement of additional active reactions (12 in each case) or metabolites (26 or 29, respectively). The addition of constraints from transcriptomics also impacted the allowed solution space, and the cellular metabolites with turnover fluxes that were necessarily altered by the change in conditions increased from 118 to 271 of 1397. Availability: GIMMME has been implemented in Python and requires a COBRApy 0.2.x. The algorithm and sample data described here are freely available at: http://opencobra.sourceforge.net/

  2. Plant stimulation of soil microbial community succession: how sequential expression mediates soil carbon stabilization and turnover

    SciTech Connect (OSTI)

    Firestone, Mary

    2015-03-31

    It is now understood that most plant C is utilized or transformed by soil microorganisms en route to stabilization. Hence the composition of microbial communities that mediate decomposition and transformation of root C is critical, as are the metabolic capabilities of these communities. The change in composition and function of the C-transforming microbial communities over time in effect defines the biological component of soil C stabilization. Our research was designed to test 2 general hypotheses; the first two hypotheses are discussed first; H1: Root-exudate interactions with soil microbial populations results in the expression of enzymatic capacities for macromolecular, complex carbon decomposition; and H2: Microbial communities surrounding roots undergo taxonomic succession linked to functional gene activities as roots grow, mature, and decompose in soil. Over the term of the project we made significant progress in 1) quantifying the temporal pattern of root interactions with the soil decomposing community and 2) characterizing the role of root exudates in mediating these interactions.

  3. Receptor expression is essential for proliferation induced by dimerized Jak kinases

    SciTech Connect (OSTI)

    Fujii, Hodaka

    2008-06-13

    Two members of Jak kinases, Jak1 and Jak3, are associated with the cytoplasmic domains of the interleukin-2 (IL-2) receptor (IL-2R) {beta} chain (IL-2R{beta}) and the common cytokine receptor {gamma} chain ({gamma}c), respectively, and accumulating evidence indicates their functional importance in IL-2 signaling. Here, I showed that coumermycin-induced chemical heterodimerization of Jak1 and Jak3 but not homodimerization of Jak1 or Jak3 induces cell proliferation of an IL-2R-reconstituted cell line. In this regard, expression of IL-2R{beta} was essential for cell proliferation by chemical heterodimerization of Jak1 and Jak3, indicating that dimerized Jak1 and Jak3 induce heterodimerization of IL-2R{beta} and {gamma}c, which may activate receptor-bound signaling molecules. Previous reports using chemical dimerization suggest that dimerization of Jak kinases is sufficient to induce cell proliferation. The present study indicates that re-evaluation of this conclusion is necessary and that interpretation of functional analysis of signaling molecules using chemical dimerizers needs more careful assessment.

  4. ERK Oscillation-Dependent Gene Expression Patterns and Deregulation by Stress-Response

    SciTech Connect (OSTI)

    Waters, Katrina M.; Cummings, Brian S.; Shankaran, Harish; Scholpa, Natalie E.; Weber, Thomas J.

    2014-09-15

    Studies were undertaken to determine whether ERK oscillations regulate a unique subset of genes in human keratinocytes and subsequently, whether the p38 stress response inhibits ERK oscillations. A DNA microarray identified many genes that were unique to ERK oscillations, and network reconstruction predicted an important role for the mediator complex subunit 1 (MED1) node in mediating ERK oscillation-dependent gene expression. Increased ERK-dependent phosphorylation of MED1 was observed in oscillating cells compared to non-oscillating counterparts as validation. Treatment of keratinocytes with a p38 inhibitor (SB203580) increased ERK oscillation amplitudes and MED1 and phospho-MED1 protein levels. Bromate is a probable human carcinogen that activates p38. Bromate inhibited ERK oscillations in human keratinocytes and JB6 cells and induced an increase in phospho-p38 and decrease in phospho-MED1 protein levels. Treatment of normal rat kidney cells and primary salivary gland epithelial cells with bromate decreased phospho-MED1 levels in a reversible fashion upon treatment with p38 inhibitors (SB202190; SB203580). Our results indicate that oscillatory behavior in the ERK pathway alters homeostatic gene regulation patterns and that the cellular response to perturbation may manifest differently in oscillating vs non-oscillating cells.

  5. Analysis of Shewanella oneidensis Membrane Protein Expression in Response to Electron Acceptor Availability

    SciTech Connect (OSTI)

    Giometti, Carol S.; Khare, Tripti; Verberkmoes, Nathan; O'Loughlin, Ed; Lindberg, Carl; Thompson, Melissa; Hettich, Robert

    2006-04-05

    Shewanella oneidensis MR-1, a gram negative metal-reducing bacterium, can utilize a large number of electron acceptors. In the natural environment, S. oneidensis utilizes insoluble metal oxides as well as soluble terminal electron acceptors. The purpose of this ERSP project is to identify differentially expressed proteins associated with the membranes of S. oneidensis MR-1 cells grown with different electron acceptors, including insoluble metal oxides. We hypothesize that through the use of surface labeling, subcellular fractionation, and a combination of proteome analysis tools, proteins involved in the reduction of different terminal electron acceptors will be elucidated. We are comparing the protein profiles from cells grown with the soluble electron acceptors oxygen and fumarate and with those from cells grown with the insoluble iron oxides goethite, ferrihydrite and lepidocrocite. Comparison of the cell surface proteins isolated from cells grown with oxygen or anaerobically with fumarate revealed an increase in the abundance of over 25 proteins in anaerobic cells, including agglutination protein and flagellin proteins along with the several hypothetical proteins. In addition, the surface protein composition of cells grown with the insoluble iron oxides varies considerably from the protein composition observed with either soluble electron acceptor as well as between the different insoluble acceptors.

  6. PRAD1, a candidate BCL1 oncogene: Mapping and expression in centrocytic lymphoma

    SciTech Connect (OSTI)

    Rosenberg, C.L.; Arnold, A.; Harris, N.L. (Massachusetts General Hospital, Boston, MA (United States) Harvard Medical School, Boston, MA (United States)); Wong, E.; Petty, E.M.; Bale, A.E. (Yale Univ., New Haven, CT (United States)); Tsujimoto, Yoshihide (Wistar Inst., Philadelphia, PA (United States))

    1991-11-01

    Rearrangement of the BCL1 (B-cell lymphoma 1) region on chromosome 11q13 appears to be highly characteristic of centrocytic lymphoma and also is found infrequently in other B-cell neoplasma. Rearrangement is thought to deregulate a nearby protooncogene, but transcribed sequences in the immediate vicinity of BCL1 breakpoints had not been identified. PRAD1, previously designated D11S287E, was identified on 11q13 as a chromosomal breakpoint region rearranged with the parathyroid hormone gene in a subset of parathyroid adenomas; this highly conserved putative oncogene, which encodes a novel cyclin, has been linked to BCL1 and implicated also in subsets of breast and squamous cell neoplasms with 11q13 amplification. The authors report pulsed-field gel electrophoresis data showing BCL1 and PRAD1 to be no more than 130 kilobases apart. PRAD1 mRNA is abundantly expressed in seven of seven centrocytic lymphomas (Kiel classification), in contrast to 13 closely related but noncentrocytic lymphomas. Three of the seven centrocytic lymphomas had detectable BCL1 DNA rearrangement. Also, two unusual cases of CLL and BCL1 rearrangement overexpressed PRAD1, in contrast to five CLL controls. Thus, PRAD1 is an excellent candidate BCL1 oncogene. Its overexpression may be a key consequence of rearrangement of the BCL1 vicinity in B-cell neoplasms and a unifying pathogenetic feature in centrocytic lymphoma.

  7. Increased Production of Fatty Acids and Triglycerides in Aspergillus oryzae by Enhancing Expressions of Fatty Acid Synthesis-Related Genes

    SciTech Connect (OSTI)

    Tamano, Koichi; Bruno, Kenneth S.; Karagiosis, Sue A.; Culley, David E.; Deng, Shuang; Collett, James R.; Umemura, Myco; Koike, Hideaki; Baker, Scott E.; Machida, Masa

    2013-01-01

    Microbial production of fats and oils is being developedas a means of converting biomass to biofuels. Here we investigate enhancing expression of enzymes involved in the production of fatty acids and triglycerides as a means to increase production of these compounds in Aspergillusoryzae. Examination of the A.oryzaegenome demonstrates that it contains twofatty acid synthases and several other genes that are predicted to be part of this biosynthetic pathway. We enhancedthe expressionof fatty acid synthesis-related genes by replacing their promoters with thepromoter fromthe constitutively highly expressedgene tef1. We demonstrate that by simply increasing the expression of the fatty acid synthasegenes we successfullyincreasedtheproduction of fatty acids and triglyceridesby more than two fold. Enhancement of expression of the fatty acid pathway genes ATP-citrate lyase and palmitoyl-ACP thioesteraseincreasedproductivity to a lesser extent.Increasing expression ofacetyl-CoA carboxylase caused no detectable change in fatty acid levels. Increases in message level for each gene were monitored usingquantitative real-time RT-PCR. Our data demonstrates that a simple increase in the abundance of fatty acid synthase genes can increase the detectable amount of fatty acids.

  8. New methods for tightly regulated gene expression and highly efficient chromosomal integration of cloned genes for Methanosarcina species

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Guss, Adam M.; Rother, Michael; Zhang, Jun Kai; Kulkkarni, Gargi; Metcalf, William W.

    2008-01-01

    A highly efficient method for chromosomal integration of cloned DNA into Methanosarcina spp. was developed utilizing the site-specific recombination system from the Streptomyces phage φC31. Host strains expressing the φC31 integrase gene and carrying an appropriate recombination site can be transformed with non-replicating plasmids carrying the complementary recombination site at efficiencies similar to those obtained with self-replicating vectors. We have also constructed a series of hybrid promoters that combine the highly expressed M. barkeri P mcrB promoter with binding sites for the tetracycline-responsive, bacterial TetR protein. These promoters are tightly regulated by the presence or absence of tetracycline inmore » strains that express the tetR gene. The hybrid promoters can be used in genetic experiments to test gene essentiality by placing a gene of interest under their control. Thus, growth of strains with tetR -regulated essential genes becomes tetracycline-dependent. A series of plasmid vectors that utilize the site-specific recombination system for construction of reporter gene fusions and for tetracycline regulated expression of cloned genes are reported. These vectors were used to test the efficiency of translation at a variety of start codons. Fusions using an ATG start site were the most active, whereas those using GTG and TTG were approximately one half or one fourth as active, respectively. The CTG fusion was 95% less active than the ATG fusion.« less

  9. New methods for tightly regulated gene expression and highly efficient chromosomal integration of cloned genes forMethanosarcinaspecies

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Guss, Adam M.; Rother, Michael; Zhang, Jun Kai; Kulkkarni, Gargi; Metcalf, William W.

    2008-01-01

    A highly efficient method for chromosomal integration of cloned DNA intoMethanosarcina spp.was developed utilizing the site-specific recombination system from theStreptomycesphage ?C31. Host strains expressing the ?C31 integrase gene and carrying an appropriate recombination site can be transformed with non-replicating plasmids carrying the complementary recombination site at efficiencies similar to those obtained with self-replicating vectors. We have also constructed a series of hybrid promoters that combine the highly expressedM. barkeriPmcrBpromoter with binding sites for the tetracycline-responsive, bacterial TetR protein. These promoters are tightly regulated by the presence or absence of tetracycline in strains that express thetetRgene. The hybrid promoters can bemoreused in genetic experiments to test gene essentiality by placing a gene of interest under their control. Thus, growth of strains withtetR-regulated essential genes becomes tetracycline-dependent. A series of plasmid vectors that utilize the site-specific recombination system for construction of reporter gene fusions and for tetracycline regulated expression of cloned genes are reported. These vectors were used to test the efficiency of translation at a variety of start codons. Fusions using an ATG start site were the most active, whereas those using GTG and TTG were approximately one half or one fourth as active, respectively. The CTG fusion was 95% less active than the ATG fusion.less

  10. Association of brominated proteins and changes in protein expression in the rat kidney with subcarcinogenic to carcinogenic doses of bromate

    SciTech Connect (OSTI)

    Kolisetty, Narendrababu [Department of Pharmaceutical and Biomedical Sciences, College of Pharmacy, University of Georgia, Athens, GA 30602 (United States); Bull, Richard J. [MoBull Consulting, Richland, WA 99352 (United States); Muralidhara, Srinivasa; Costyn, Leah J. [Department of Pharmaceutical and Biomedical Sciences, College of Pharmacy, University of Georgia, Athens, GA 30602 (United States); Delker, Don A. [School of Medicine, University of Utah, Salt Lake City, UT 84132 (United States); Guo, Zhongxian [Water Quality Office, Public Utilities Board, 608576 (Singapore); Cotruvo, Joseph A. [Joseph Cotruvo and Associates, LLC, Washington, DC 20016 (United States); Fisher, Jeffrey W. [National Center for Toxicological Research, FDA, Jefferson, AR 72079 (United States); Cummings, Brian S., E-mail: bsc@rx.uga.edu [Department of Pharmaceutical and Biomedical Sciences, College of Pharmacy, University of Georgia, Athens, GA 30602 (United States)

    2013-10-15

    The water disinfection byproduct bromate (BrO{sub 3}{sup ?}) produces cytotoxic and carcinogenic effects in rat kidneys. Our previous studies demonstrated that BrO{sub 3}{sup ?} caused sex-dependent differences in renal gene and protein expression in rats and the elimination of brominated organic carbon in their urine. The present study examined changes in renal cell apoptosis and protein expression in male and female F344 rats treated with BrO{sub 3}{sup ?} and associated these changes with accumulation of 3-bromotyrosine (3-BT)-modified proteins. Rats were treated with 0, 11.5, 46 and 308 mg/L BrO{sub 3}{sup ?} in drinking water for 28 days and renal sections were prepared and examined for apoptosis (TUNEL-staining), 8-oxo-deoxyguanosine (8-oxoG), 3-BT, osteopontin, Kim-1, clusterin, and p-21 expression. TUNEL-staining in renal proximal tubules increased in a dose-related manner beginning at 11.5 mg BrO{sub 3}{sup ?}/L in female rats and 46 mg/L in males. Increased 8-oxoG staining was observed at doses as low as 46 mg/L. Osteopontin expression also increased in a dose-related manner after treatment with 46 mg/L, in males only. In contrast, Kim-1 expression increased in a dose-related manner in both sexes, although to a greater extent in females at the highest dose. Clusterin and p21 expression also increased in a dose-related manner in both sexes. The expression of 3-BT-modified proteins only increased in male rats, following a pattern previously reported for accumulation of ?-2{sub u}-globulin. Increases in apoptosis in renal proximal tubules of male and female rats at the lowest doses suggest a common mode of action for renal carcinogenesis for the two sexes that is independent of ?-2{sub u}-globulin nephropathy. - Highlights: Bromate induced nephrotoxicity in both male and female rats by similar mechanisms. Apoptosis was seen in both male and female rats at the lowest doses tested. Bromate-induced apoptosis correlated to 8-oxo-deoxyguanosine formation. Bromate increased the level of 3-bromotyrosine-modified proteins in male rats only. These data identify possible novel mechanisms for bromate-induced nephrotoxicity.

  11. Gender-specific reduction of hepatic Mrp2 expression by high-fat diet protects female mice from ANIT toxicity

    SciTech Connect (OSTI)

    Kong, Bo; Csanaky, Ivn L.; Aleksunes, Lauren M.; Patni, Meghan; Chen, Qi; Ma, Xiaochao; Jaeschke, Hartmut; Weir, Scott; Broward, Melinda; Klaassen, Curtis D.; University of Kansas Cancer Center, Kansas City, KS ; Guo, Grace L.

    2012-06-01

    Emerging evidence suggests that feeding a high-fat diet (HFD) to rodents affects the expression of genes involved in drug transport. However, gender-specific effects of HFD on drug transport are not known. The multidrug resistance-associated protein 2 (Mrp2, Abcc2) is a transporter highly expressed in the hepatocyte canalicular membrane and is important for biliary excretion of glutathione-conjugated chemicals. The current study showed that hepatic Mrp2 expression was reduced by HFD feeding only in female, but not male, C57BL/6J mice. In order to determine whether down-regulation of Mrp2 in female mice altered chemical disposition and toxicity, the biliary excretion and hepatotoxicity of the Mrp2 substrate, ?-naphthylisothiocyanate (ANIT), were assessed in male and female mice fed control diet or HFD for 4 weeks. ANIT-induced biliary injury is a commonly used model of experimental cholestasis and has been shown to be dependent upon Mrp2-mediated efflux of an ANIT glutathione conjugate that selectively injures biliary epithelial cells. Interestingly, HFD feeding significantly reduced early-phase biliary ANIT excretion in female mice and largely protected against ANIT-induced liver injury. In summary, the current study showed that, at least in mice, HFD feeding can differentially regulate Mrp2 expression and function and depending upon the chemical exposure may enhance or reduce susceptibility to toxicity. Taken together, these data provide a novel interaction between diet and gender in regulating hepatobiliary excretion and susceptibility to injury. -- Highlights: ? High-fat diet decreases hepatic Mrp2 expression only in female but not in male mice. ? HFD significantly reduces early-phase biliary ANIT excretion in female mice. ? HFD protects female mice against ANIT-induced liver injury.

  12. Transgenic expression of the dicotyledonous pattern recognition receptor EFR in rice leads to ligand-dependent activation of defense responses

    SciTech Connect (OSTI)

    Schwessinger, Benjamin; Bahar, Ofir; Thomas, Nicolas; Holton, Nicolas; Nekrasov, Vladimir; Ruan, Deling; Canlas, Patrick E.; Daudi, Arsalan; Petzold, Christopher J.; Singan, Vasanth R.; Kuo, Rita; Chovatia, Mansi; Daum, Christopher; Heazlewood, Joshua L.; Zipfel, Cyril; Ronald, Pamela C.

    2015-03-30

    Plant plasma membrane localized pattern recognition receptors (PRRs) detect extracellular pathogen-associated molecules. PRRs such as Arabidopsis EFR and rice XA21 are taxonomically restricted and are absent from most plant genomes. Here we show that rice plants expressing EFR or the chimeric receptor EFR::XA21, containing the EFR ectodomain and the XA21 intracellular domain, sense both Escherichia coli- and Xanthomonas oryzae pv. oryzae (Xoo)-derived elf18 peptides at sub-nanomolar concentrations. Treatment of EFR and EFR::XA21 rice leaf tissue with elf18 leads to MAP kinase activation, reactive oxygen production and defense gene expression. Although expression of EFR does not lead to robust enhanced resistance to fully virulent Xoo isolates, it does lead to quantitatively enhanced resistance to weakly virulent Xoo isolates. EFR interacts with OsSERK2 and the XA21 binding protein 24 (XB24), two key components of the rice XA21-mediated immune response. Rice-EFR plants silenced for OsSERK2, or overexpressing rice XB24 are compromised in elf18-induced reactive oxygen production and defense gene expression indicating that these proteins are also important for EFR-mediated signaling in transgenic rice. Taken together, our results demonstrate the potential feasibility of enhancing disease resistance in rice and possibly other monocotyledonous crop species by expression of dicotyledonous PRRs. Our results also suggest that Arabidopsis EFR utilizes at least a subset of the known endogenous rice XA21 signaling components.

  13. NNMCAB Board Minutes: September 2011 Taos

    Broader source: Energy.gov [DOE]

    Minutes of the September 28, 2011 Board meeting at Sagebrush Inn Conference Center Presentation DOE, Long Term Stewardship, Tom Longo

  14. Th Cell Gene Expression and Function in Response to Low Dose and Acute Radiation

    SciTech Connect (OSTI)

    Daila S. Gridley, PhD

    2012-03-30

    FINAL TECHNICAL REPORT Supported by the Low Dose Radiation Research Program, Office of Science U.S. Department of Energy Grant No. DE-FG02-07ER64345 Project ID: 0012965 Award Register#: ER64345 Project Manager: Noelle F. Metting, Sc.D. Phone: 301-903-8309 Division SC-23.2 noelle.metting@science.doe.gov Submitted March 2012 To: https://www.osti.gov/elink/241.3.jsp Title: Th Cell Gene Expression and Function in Response to Low Dose and Acute Radiation PI: Daila S. Gridley, Ph.D. Human low dose radiation data have been derived primarily from studies of space and airline flight personnel, nuclear plant workers and others exposed occupationally, as well as victims in the vicinity of atomic bomb explosions. The findings remain inconclusive due to population inconsistencies and complex interactions among total dose, dose rate, radiation quality and age at exposure. Thus, safe limits for low dose occupational irradiation are currently based on data obtained with doses far exceeding the levels expected for the general population and health risks have been largely extrapolated using the linear-nonthreshold dose-response model. The overall working hypothesis of the present study is that priming with low dose, low-linear energy transfer (LET) radiation can ameliorate the response to acute high-dose radiation exposure. We also propose that the efficacy of low-dose induced protection will be dependent upon the form and regimen of the high-dose exposure: photons versus protons versus simulated solar particle event protons (sSPE). The emphasis has been on gene expression and function of CD4+ T helper (Th) lymphocytes harvested from spleens of whole-body irradiated C57BL/6 mice, a strain that provides the genetic background for many genetically engineered strains. Evaluations of the responses of other selected cells, tissues such as skin, and organs such as lung, liver and brain were also initiated (partially funded by other sources). The long-term goal is to provide information that will be useful in estimating human health risks due to radiation that may occur during exposures in the work environment, nuclear/radiological catastrophes, as well as radiotherapy. Several papers have been published, accepted for publication or are in preparation. A number of poster and oral presentations have been made at scientific conferences and workshops. Archived tissues of various types will continue to be evaluated via funding from other sources (the DoE Low Dose Radiation Research Program, Office of Science and this specific grant will be appropriately included in the Acknowledgements of all subsequent publications/presentations). A post-doc and several students have participated in this study. More detailed description of the accomplishments is described in attached file.

  15. Myocardial regeneration in adriamycin cardiomyopathy by nuclear expression of GLP1 using ultrasound targeted microbubble destruction

    SciTech Connect (OSTI)

    Chen, Shuyuan; Chen, Jiaxi; Huang, Pintong; Meng, Xing-Li; Clayton, Sandra; Shen, Jin-Song; Grayburn, Paul A.

    2015-03-20

    Recently GLP-1 was found to have cardioprotective effects independent of those attributable to tight glycemic control. Methods and results: We employed ultrasound targeted microbubble destruction (UTMD) to deliver piggybac transposon plasmids encoding the GLP-1 gene with a nuclear localizing signal to rat hearts with adriamycin cardiomyopathy. After a single UTMD treatment, overexpression of transgenic GLP-1 was found in nuclei of rat heart cells with evidence that transfected cardiac cells had undergone proliferation. UTMD-GLP-1 gene therapy restored LV mass, fractional shortening index, and LV posterior wall diameter to nearly normal. Nuclear overexpression of GLP-1 by inducing phosphorylation of FoxO1-S256 and translocation of FoxO1 from the nucleus to the cytoplasm significantly inactivated FoxO1 and activated the expression of cyclin D1 in nuclei of cardiac muscle cells. Reversal of adriamycin cardiomyopathy appeared to be mediated by dedifferentiation and proliferation of nuclear FoxO1-positive cardiac muscle cells with evidence of embryonic stem cell markers (OCT4, Nanog, SOX2 and c-kit), cardiac early differentiation markers (NKX2.5 and ISL-1) and cellular proliferation markers (BrdU and PHH3) after UTMD with GLP-1 gene therapy. Conclusions: Intranuclear myocardial delivery of the GLP-1gene can reverse established adriamycin cardiomyopathy by stimulating myocardial regeneration. - Highlights: • The activation of nuclear FoxO1 in cardiac muscle cells associated with adriamycin cardiomyopathy. • Myocardial nuclear GLP-1 stimulates myocardial regeneration and reverses adriamycin cardiomyopathy. • The process of myocardial regeneration associated with dedifferentiation and proliferation.

  16. Concentration-dependent gene expression responses to flusilazole in embryonic stem cell differentiation cultures

    SciTech Connect (OSTI)

    Dartel, Dorien A.M. van; Pennings, Jeroen L.A.; Fonteyne, Liset J.J. de la; Brauers, Karen J.J.; Claessen, Sandra; Delft, Joost H. van; Kleinjans, Jos C.S.; Piersma, Aldert H.

    2011-03-01

    The murine embryonic stem cell test (EST) is designed to evaluate developmental toxicity based on compound-induced inhibition of embryonic stem cell (ESC) differentiation into cardiomyocytes. The addition of transcriptomic evaluation within the EST may result in enhanced predictability and improved characterization of the applicability domain, therefore improving usage of the EST for regulatory testing strategies. Transcriptomic analyses assessing factors critical for risk assessment (i.e. dose) are needed to determine the value of transcriptomic evaluation in the EST. Here, using the developmentally toxic compound, flusilazole, we investigated the effect of compound concentration on gene expression regulation and toxicity prediction in ESC differentiation cultures. Cultures were exposed for 24 h to multiple concentrations of flusilazole (0.54-54 {mu}M) and RNA was isolated. In addition, we sampled control cultures 0, 24, and 48 h to evaluate the transcriptomic status of the cultures across differentiation. Transcriptomic profiling identified a higher sensitivity of development-related processes as compared to cell division-related processes in flusilazole-exposed differentiation cultures. Furthermore, the sterol synthesis-related mode of action of flusilazole toxicity was detected. Principal component analysis using gene sets related to normal ESC differentiation was used to describe the dynamics of ESC differentiation, defined as the 'differentiation track'. The concentration-dependent effects on development were reflected in the significance of deviation of flusilazole-exposed cultures from this transcriptomic-based differentiation track. Thus, the detection of developmental toxicity in EST using transcriptomics was shown to be compound concentration-dependent. This study provides further insight into the possible application of transcriptomics in the EST as an improved alternative model system for developmental toxicity testing.

  17. Gestational exposure to diethylstilbestrol alters cardiac structure/function, protein expression and DNA methylation in adult male mice progeny

    SciTech Connect (OSTI)

    Haddad, Rami, E-mail: rami.haddad@mail.mcgill.ca [Lady Davis Institute for Medical Research, Jewish General Hospital, 3755 chemin Cote Ste Catherine, Montral, Qubec, Canada H3T 1E2 (Canada) [Lady Davis Institute for Medical Research, Jewish General Hospital, 3755 chemin Cote Ste Catherine, Montral, Qubec, Canada H3T 1E2 (Canada); Division of Experimental Medicine, Department of Medicine, McGill University, 850 Sherbrooke Street, Montral, Qubec, Canada H3A 1A2 (Canada); Kasneci, Amanda, E-mail: amanda.kasneci@mail.mcgill.ca [Lady Davis Institute for Medical Research, Jewish General Hospital, 3755 chemin Cote Ste Catherine, Montral, Qubec, Canada H3T 1E2 (Canada)] [Lady Davis Institute for Medical Research, Jewish General Hospital, 3755 chemin Cote Ste Catherine, Montral, Qubec, Canada H3T 1E2 (Canada); Mepham, Kathryn, E-mail: katherine.mepham@mail.mcgill.ca [Lady Davis Institute for Medical Research, Jewish General Hospital, 3755 chemin Cote Ste Catherine, Montral, Qubec, Canada H3T 1E2 (Canada) [Lady Davis Institute for Medical Research, Jewish General Hospital, 3755 chemin Cote Ste Catherine, Montral, Qubec, Canada H3T 1E2 (Canada); Division of Experimental Medicine, Department of Medicine, McGill University, 850 Sherbrooke Street, Montral, Qubec, Canada H3A 1A2 (Canada); Sebag, Igal A., E-mail: igal.sebag@mcgill.ca [Division of Cardiology, Jewish General Hospital, 3755 chemin Cote Ste Catherine, Montral, Qubec, Canada H3T 1E2 (Canada); and others

    2013-01-01

    Pregnant women, and thus their fetuses, are exposed to many endocrine disruptor compounds (EDCs). Fetal cardiomyocytes express sex hormone receptors making them potentially susceptible to re-programming by estrogenizing EDCs. Diethylstilbestrol (DES) is a proto-typical, non-steroidal estrogen. We hypothesized that changes in adult cardiac structure/function after gestational exposure to the test compound DES would be a proof in principle for the possibility of estrogenizing environmental EDCs to also alter the fetal heart. Vehicle (peanut oil) or DES (0.1, 1.0 and 10.0 ?g/kg/da.) was orally delivered to pregnant C57bl/6n dams on gestation days 11.514.5. At 3 months, male progeny were left sedentary or were swim trained for 4 weeks. Echocardiography of isoflurane anesthetized mice revealed similar cardiac structure/function in all sedentary mice, but evidence of systolic dysfunction and increased diastolic relaxation after swim training at higher DES doses. The calcium homeostasis proteins, SERCA2a, phospholamban, phospho-serine 16 phospholamban and calsequestrin 2, are important for cardiac contraction and relaxation. Immunoblot analyses of ventricle homogenates showed increased expression of SERCA2a and calsequestrin 2 in DES mice and greater molecular remodeling of these proteins and phospho-serine 16 phospholamban in swim trained DES mice. DES increased cardiac DNA methyltransferase 3a expression and DNA methylation in the CpG island within the calsequestrin 2 promoter in heart. Thus, gestational DES epigenetically altered ventricular DNA, altered cardiac function and expression, and reduced the ability of adult progeny to cardiac remodel when physically challenged. We conclude that gestational exposure to estrogenizing EDCs may impact cardiac structure/function in adult males. -- Highlights: ? Gestational DES changes cardiac SERCA2a and CASQ2 expression. ? Echocardiography identified systolic dysfunction and increased diastolic relaxation. ? DES increased DNMT3a expression and increased CpG DNA methylation. ? DES impacts fetal heart reducing cardiac reserve on challenge in adulthood. ? Fetal heart can be re-programmed by a non-steroidal estrogen.

  18. Twenty-first workshop on geothermal reservoir engineering: Proceedings

    SciTech Connect (OSTI)

    1996-01-26

    PREFACE The Twenty-First Workshop on Geothermal Reservoir Engineering was held at the Holiday Inn, Palo Alto on January 22-24, 1996. There were one-hundred fifty-five registered participants. Participants came from twenty foreign countries: Argentina, Austria, Canada, Costa Rica, El Salvador, France, Iceland, Indonesia, Italy, Japan, Mexico, The Netherlands, New Zealand, Nicaragua, the Philippines, Romania, Russia, Switzerland, Turkey and the UK. The performance of many geothermal reservoirs outside the United States was described in several of the papers. Professor Roland N. Horne opened the meeting and welcomed visitors. The key note speaker was Marshall Reed, who gave a brief overview of the Department of Energy's current plan. Sixty-six papers were presented in the technical sessions of the workshop. Technical papers were organized into twenty sessions concerning: reservoir assessment, modeling, geology/geochemistry, fracture modeling hot dry rock, geoscience, low enthalpy, injection, well testing, drilling, adsorption and stimulation. Session chairmen were major contributors to the workshop, and we thank: Ben Barker, Bobbie Bishop-Gollan, Tom Box, Jim Combs, John Counsil, Sabodh Garg, Malcolm Grant, Marcel0 Lippmann, Jim Lovekin, John Pritchett, Marshall Reed, Joel Renner, Subir Sanyal, Mike Shook, Alfred Truesdell and Ken Williamson. Jim Lovekin gave the post-dinner speech at the banquet and highlighted the exciting developments in the geothermal field which are taking place worldwide. The Workshop was organized by the Stanford Geothermal Program faculty, staff, and graduate students. We wish to thank our students who operated the audiovisual equipment. Shaun D. Fitzgerald Program Manager.

  19. Natural Gas Weekly Update

    Annual Energy Outlook [U.S. Energy Information Administration (EIA)]

    Holiday Notice: Due to the federal holiday in observance of Martin Luther King Day on Monday, January 21, 2002, the next issue of the Natural Gas Weekly Update will be published on...

  20. Natural Gas Weekly Update, Printer-Friendly Version

    Annual Energy Outlook [U.S. Energy Information Administration (EIA)]

    Holiday Notice: Due to the federal holiday in observance of Martin Luther King Day on Monday, January 21, 2002, the next issue of the Natural Gas Weekly Update will be published on...

  1. Natural Gas Weekly Update, Printer-Friendly Version

    Annual Energy Outlook [U.S. Energy Information Administration (EIA)]

    2.852 2.813 Holiday Closed January Delivery 2.996 3.041 2.991 Holiday Closed Source: Reuters Information Service Storage: Net injections into working gas storage were 15 billion...

  2. Blog | Department of Energy

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    your tips for saving energy and money during the holidays. December 19, 2012 Using LED lights for your holiday decorations can save you energy and money. | Photo courtesy of...

  3. South Carolina Community Lights Up the Season with Energy-Efficient...

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    South Carolina's new LED holiday light display. | Photo courtesy of Richland County, S.C. Carolers sing in front of Forest Acres, South Carolina's new LED holiday light display. ...

  4. Wreaths Across America

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    wreath-laying ceremonies at over 1,000 locations worldwide, including 12 locations in New Mexico. G ving Employee Giving Campaign Holiday Food Drive Holiday Gift Drive LANL Laces...

  5. Blog | Department of Energy

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Photo courtesy of Chris Gunn, NREL. Energy Saving Holiday Kitchen Trivia Test your energy-saving knowledge with our holiday kitchen trivia. November 19, 2012 Save time and money on...

  6. January Events

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    for New Year's Holiday Bradbury Science Museum - 1350 Central Ave, Los Alamos, NM 87544, USA The Bradbury Science Museum will be CLOSED for the New Year's holiday. Jan 9 Sat 11:00...

  7. Aluminium induced oxidative stress results in decreased mitochondrial biogenesis via modulation of PGC-1? expression

    SciTech Connect (OSTI)

    Sharma, Deep Raj; Sunkaria, Aditya; Wani, Willayat Yousuf; Sharma, Reeta Kumari; Kandimalla, Ramesh J.L.; Bal, Amanjit; Gill, Kiran Dip

    2013-12-01

    The present investigation was carried out to elucidate a possible molecular mechanism related to the effects of aluminium-induced oxidative stress on various mitochondrial respiratory complex subunits with special emphasis on the role of Peroxisome proliferator activated receptor gamma co-activator 1? (PGC-1?) and its downstream targets i.e. Nuclear respiratory factor-1(NRF-1), Nuclear respiratory factor-2(NRF-2) and Mitochondrial transcription factor A (Tfam) in mitochondrial biogenesis. Aluminium lactate (10 mg/kg b.wt./day) was administered intragastrically to rats for 12 weeks. After 12 weeks of exposure, we found an increase in ROS levels, mitochondrial DNA oxidation and decrease in citrate synthase activity in the Hippocampus (HC) and Corpus striatum (CS) regions of rat brain. On the other hand, there was a decrease in the mRNA levels of the mitochondrial encoded subunitsNADH dehydrogenase (ND) subunits i.e. ND1, ND2, ND3, Cytochrome b (Cytb), Cytochrome oxidase (COX) subunits i.e. COX1, COX3, ATP synthase (ATPase) subunit 6 along with reduced expression of nuclear encoded subunits COX4, COX5A, COX5B of Electron transport chain (ETC). Besides, a decrease in mitochondrial DNA copy number and mitochondrial content in both regions of rat brain was observed. The PGC-1? was down-regulated in aluminium treated rats along with NRF-1, NRF-2 and Tfam, which act downstream from PGC-1? in aluminium treated rats. Electron microscopy results revealed a significant increase in the mitochondrial swelling, loss of cristae, chromatin condensation and decreases in mitochondrial number in case of aluminium treated rats as compared to control. So, PGC-1? seems to be a potent target for aluminium neurotoxicity, which makes it an almost ideal target to control or limit the damage that has been associated with the defective mitochondrial function seen in neurodegenerative diseases. - Highlights: Aluminium decreases the mRNA levels of mitochondrial and nuclear encoded subunits. It decreases the mtDNA copy number and mitochondrial content in rat brain. It down-regulates the mRNA and protein levels of PGC-1?, NRF-1, NRF-2 and Tfam. It also disturbs the mitochondrial or nuclear architecture of neurons. Finally it also decreases mitochondrial number in HC and CS regions of rat brain.

  8. o-p?-DDT-mediated uterotrophy and gene expression in immature C57BL/6 mice and SpragueDawley rats

    SciTech Connect (OSTI)

    Kwekel, Joshua C.; Forgacs, Agnes L.; Williams, Kurt J.; Zacharewski, Timothy R.

    2013-12-15

    1,1,1-Trichloro-2,2-bis(2-chlorophenyl-4-chlorophenyl)ethane (o,p?-DDT) is an organochlorine pesticide and endocrine disruptor known to activate the estrogen receptor. Comprehensive ligand- and species-comparative dose- and time-dependent studies were conducted to systematically assess the uterine physiological, morphological and gene expression responses elicited by o,p?-DDT and ethynyl estradiol (EE) in immature ovariectomized C57BL/6 mice and SpragueDawley rats. Custom cDNA microarrays were used to identify conserved and divergent differential gene expression responses. A total of 1256 genes were differentially expressed by both ligands in both species, 559 of which exhibited similar temporal expression profiles suggesting that o,p?-DDT elicits estrogenic effects at high doses when compared to EE. However, 51 genes exhibited species-specific uterine expression elicited by o,p?-DDT. For example, carbonic anhydrase 2 exhibited species- and ligand-divergent expression as confirmed by quantitative real-time PCR. The identification of comparable temporal phenotypic responses linked to gene expression demonstrates that systematic comparative gene expression assessments are valuable for elucidating conserved and divergent estrogen signaling mechanisms in rodent uterotrophy. - Highlights: o,p?-DDT and enthynyl estradiol (EE) both elicit uterotrophy in mice and rats. o,p?-DDT and EE have different kinetics in uterine wet weight induction. o,p?-DDT elicited stromal hypertrophy in rats but myometrial hypertrophy in mice. 1256 genes were differentially expressed by both ligands in both species. Only 51 genes had species-specific uterine expression.

  9. Remember the Batteries and Maybe a Charger?

    Broader source: Energy.gov [DOE]

    For the holiday gift-giving season take a look at the ENERGY STAR list of certified rechargeable batteries.

  10. Deck Those Halls!

    Broader source: Energy.gov [DOE]

    Preparing to decorate your home for the holiday season? Consider LED lighting as an energy-efficient option.

  11. Black Friday Savings All Year ’Round

    Broader source: Energy.gov [DOE]

    On Friday, shoppers across the country will flock to stores and retailers seeking the best bargains of the holiday season.

  12. Cloning and expression of soluble truncated variants of Borrelia OspA, OspB and Vmp7

    DOE Patents [OSTI]

    Dunn, J.J.; Barbour, A.G.

    1996-11-05

    A method is provided for preparing soluble recombinant variations of Borrelia lipoproteins such as Borrelia burgdorferi outer surface protein A (OspA) and outer surface protein B (OspB), and B. hermsii variable major protein 7 (Vmp7). The method includes synthesizing a set of oligonucleotide primers, amplifying the template DNA utilizing the PCR, purifying the amplification products, cloning the amplification products into a suitable expression vector, transforming a suitable host utilizing the cloned expression vector, cultivating the transformed host for protein production and subsequently isolating and purifying the resulting protein. Also provided are soluble, recombinant variations of Borrelia burgdorferi outer surface protein A (OspA), outer surface protein B (OspB), and B. hermsii variable major protein 7 (Vmp7). The expression vectors harboring DNA encoding the recombinant variations, pET9-OspA, pET9-OspB and pET9-Vmp7, as well as the E. coli host BL21(DE3)/pLysS transformed with each of these vectors, are also disclosed. 38 figs.

  13. Cloning and expression of soluble truncated variants of Borrelia OspA, OspB and Vmp7

    DOE Patents [OSTI]

    Dunn, John J.; Barbour, Alan G.

    1996-11-05

    A method is provided herein for preparing soluble recombinant variations of Borrelia lipoproteins such as Borrelia burgdorferi outer surface protein A (OspA) and outer surface protein B (OspB), and B. hermsii variable major protein 7 (Vmp7). The method includes synthesizing a set of oligonucleotide primers, amplifying the template DNA utilizing the PCR, purifying the amplification products, cloning the amplification products into a suitable expression vector, transforming a suitable host utilizing the cloned expression vector, cultivating the transformed host for protein production and subsequently isolating and purifying the resulting protein. Also provided are soluble, recombinant variations of Borrelia burgdorferi outer surface protein A (OspA), outer surface protein B (OspB), and B. hermsii variable major protein 7 (Vmp7). The expression vectors harboring DNA encoding the recombinant variations, pET9-OspA, pET9-OspB and pET9-Vmp7, as well as the E. coli host BL21(DE3)/pLysS transformed with each of these vectors, are also disclosed.

  14. Assessment of ERCC1 and XPF Protein Expression Using Quantitative Immunohistochemistry in Nasopharyngeal Carcinoma Patients Undergoing Curative Intent Treatment

    SciTech Connect (OSTI)

    Jagdis, Amanda; Phan, Tien; Faculty of Medicine, University of Calgary, Calgary, Alberta ; Klimowicz, Alexander C.; Faculty of Medicine, University of Calgary, Calgary, Alberta ; Laskin, Janessa J.; Faculty of Medicine, University of British Columbia, Vancouver, British Columbia ; Lau, Harold Y.; Faculty of Medicine, University of Calgary, Calgary, Alberta ; Petrillo, Stephanie K.; Siever, Jodi E.; Thomson, Thomas A.; Faculty of Medicine, University of British Columbia, Vancouver, British Columbia ; Magliocco, Anthony M.; Faculty of Medicine, University of Calgary, Calgary, Alberta ; Hao, Desire; Faculty of Medicine, University of Calgary, Calgary, Alberta

    2013-04-01

    Purpose: We sought to evaluate the prognostic/predictive value of ERCC1 and XPF in patients with nonmetastatic nasopharyngeal carcinoma (NPC) treated with curative intent. Methods and Materials: ERCC1 and XPF protein expression was evaluated by immunofluorescence combined with automated quantitative analysis (AQUA) using the FL297 and 3F2 antibodies, respectively. ERCC1 and XPF protein expression levels were correlated with clinical outcomes. Results: Patient characteristics were as follows: mean age 52 years (range, 18-85 years), 67% male, 72% Karnofsky performance status (KPS) ?90%, World Health Organization (WHO) type 1/2/3 = 12%/28%/60%, stage III/IV 65%. With a median follow-up time of 50 months (range, 2.9 to 120 months), the 5-year overall survival (OS) was 70.8%. Median standardized nuclear AQUA scores were used as cutpoints for ERCC1 (n=138) and XPF (n=130) protein expression. Agreement between dichotomized ERCC1 and XPF scores was high at 79.4% (kappa = 0.587, P<.001). Neither biomarker predicted locoregional recurrence, DFS, or OS after adjustment for age and KPS, irrespective of stratification by stage, WHO type, or treatment. Conclusions: Neither ERCC1 nor XPF, analyzed by quantitative immunohistochemistry using the FL297 and 3F2 antibodies, was prognostic or predictive in this cohort of NPC patients.

  15. Estrogen induced concentration dependent differential gene expression in human breast cancer (MCF7) cells: Role of transcription factors

    SciTech Connect (OSTI)

    Chandrasekharan, Sabarinath; Kandasamy, Krishna Kumar; Dayalan, Pavithra; Ramamurthy, Viraragavan

    2013-08-02

    Highlights: Estradiol (E2) at low dose induced cell proliferation in breast cancer cells. E2 at high concentration induced cell stress in breast cancer cells. Estrogen receptor physically interacts only with a few transcription factors. Differential expression of genes with Oct-1 binding sites increased under stress. Transcription factor binding sites showed distinct spatial distribution on genes. -- Abstract: Background: Breast cancer cells respond to estrogen in a concentration dependent fashion, resulting in proliferation or apoptosis. The mechanism of this concentration dependent differential outcome is not well understood yet. Methodology: Meta-analysis of the expression data of MCF7 cells treated with low (1 nM) or high (100 nM) dose of estradiol (E2) was performed. We identified genes differentially expressed at the low or the high dose, and examined the nature of regulatory elements in the vicinity of these genes. Specifically, we looked for the difference in the presence, abundance and spatial distribution of binding sites for estrogen receptor (ER) and selected transcription factors (TFs) in the genomic region up to 25 kb upstream and downstream from the transcription start site (TSS) of these genes. Results: It was observed that at high dose E2 induced the expression of stress responsive genes, while at low dose, genes involved in cell cycle were induced. We found that the occurrence of transcription factor binding regions (TFBRs) for certain factors such as Sp1 and SREBP1 were higher on regulatory regions of genes expressed at low dose. At high concentration of E2, genes with a higher frequency of Oct-1 binding regions were predominantly involved. In addition, there were differences in the spatial distribution pattern of the TFBRs in the genomic regions among the two sets of genes. Discussion: E2 induced predominantly proliferative/metabolic response at low concentrations; but at high concentration, stressrescue responses were induced. At high E2 concentration, classical genomic pathway involving ER binding to the regulatory regions was reduced, and alternate or indirect activation of genes through Oct-1 became more prominent.

  16. Urine acidification has no effect on peroxisome proliferator-activated receptor (PPAR) signaling or epidermal growth factor (EGF) expression in rat urinary bladder urothelium

    SciTech Connect (OSTI)

    Achanzar, William E. Moyer, Carolyn F.; Marthaler, Laura T.; Gullo, Russell; Chen, Shen-Jue; French, Michele H.; Watson, Linda M.; Rhodes, James W.; Kozlosky, John C.; White, Melvin R.; Foster, William R.; Burgun, James J.; Car, Bruce D.; Cosma, Gregory N.; Dominick, Mark A.

    2007-09-15

    We previously reported prevention of urolithiasis and associated rat urinary bladder tumors by urine acidification (via diet acidification) in male rats treated with the dual peroxisome proliferator-activated receptor (PPAR){alpha}/{gamma} agonist muraglitazar. Because urine acidification could potentially alter PPAR signaling and/or cellular proliferation in urothelium, we evaluated urothelial cell PPAR{alpha}, PPAR{delta}, PPAR{gamma}, and epidermal growth factor receptor (EGFR) expression, PPAR signaling, and urothelial cell proliferation in rats fed either a normal or an acidified diet for 5, 18, or 33 days. A subset of rats in the 18-day study also received 63 mg/kg of the PPAR{gamma} agonist pioglitazone daily for the final 3 days to directly assess the effects of diet acidification on responsiveness to PPAR{gamma} agonism. Urothelial cell PPAR{alpha} and {gamma} expression and signaling were evaluated in the 18- and 33-day studies by immunohistochemical assessment of PPAR protein (33-day study only) and quantitative real-time polymerase chain reaction (qRT-PCR) measurement of PPAR-regulated gene expression. In the 5-day study, EGFR expression and phosphorylation status were evaluated by immunohistochemical staining and egfr and akt2 mRNA levels were assessed by qRT-PCR. Diet acidification did not alter PPAR{alpha}, {delta}, or {gamma} mRNA or protein expression, PPAR{alpha}- or {gamma}-regulated gene expression, total or phosphorylated EGFR protein, egfr or akt2 gene expression, or proliferation in urothelium. Moreover, diet acidification had no effect on pioglitazone-induced changes in urothelial PPAR{gamma}-regulated gene expression. These results support the contention that urine acidification does not prevent PPAR{gamma} agonist-induced bladder tumors by altering PPAR{alpha}, {gamma}, or EGFR expression or PPAR signaling in rat bladder urothelium.

  17. 01-02253B_OR_Knox_map.ai

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Knoxville Nashville Oak Ridge via State Route 162 North OAK RIDGE INN & SUITES THE RIDGE INN and THE SUPER 8 MOTEL ORNL GRAPHITE REACTOR NATIONAL TRANSPORTATION RESEARCH CENTER ETTP OBSERVATION CENTER MELTON HILL DAM HAMPTON INN BULL RUN STEAM PLANT (TVA) COMFORT INN JAMESON INN BENEFITS OFFICE OF TECH. TRANSFER SNS PROJECT OFFICE COMMERCE PARK OAK RIDGE/KNOXVILLE ROUTE MAP A B C D E F G H I J K L M N O P Q R S T U V W ORAU / ORISE / POLLARD AUDITORIUM U.S. POST OFFICE METHODIST MEDICAL

  18. Activation of nuclear receptor NR5A2 increases Glut4 expression and glucose metabolism in muscle cells

    SciTech Connect (OSTI)

    Bolado-Carrancio, A.; Riancho, J.A.; Sainz, J.; Rodrguez-Rey, J.C.

    2014-04-04

    Highlights: NR5A2 expression in C2C12 is associated with myotube differentiation. DLPC induces an increase in GLUT4 levels and glucose uptake in C2C12 myotubes. In high glucose conditions the activation of NR5A2 inhibits fatty acids oxidation. - Abstract: NR5A2 is a nuclear receptor which regulates the expression of genes involved in cholesterol metabolism, pluripotency maintenance and cell differentiation. It has been recently shown that DLPC, a NR5A2 ligand, prevents liver steatosis and improves insulin sensitivity in mouse models of insulin resistance, an effect that has been associated with changes in glucose and fatty acids metabolism in liver. Because skeletal muscle is a major tissue in clearing glucose from blood, we studied the effect of the activation of NR5A2 on muscle metabolism by using cultures of C2C12, a mouse-derived cell line widely used as a model of skeletal muscle. Treatment of C2C12 with DLPC resulted in increased levels of expression of GLUT4 and also of several genes related to glycolysis and glycogen metabolism. These changes were accompanied by an increased glucose uptake. In addition, the activation of NR5A2 produced a reduction in the oxidation of fatty acids, an effect which disappeared in low-glucose conditions. Our results suggest that NR5A2, mostly by enhancing glucose uptake, switches muscle cells into a state of glucose preference. The increased use of glucose by muscle might constitute another mechanism by which NR5A2 improves blood glucose levels and restores insulin sensitivity.

  19. Functional Toll-like receptor 4 expressed in lactotrophs mediates LPS-induced proliferation in experimental pituitary hyperplasia

    SciTech Connect (OSTI)

    Sabatino, Mara Eugenia; Sosa, Liliana del Valle; Petiti, Juan Pablo; Mukdsi, Jorge Humberto; Mascanfroni, Ivn Daro; Pellizas, Claudia Gabriela; Gutirrez, Silvina; Torres, Alicia Ins; De Paul, Ana Luca

    2013-11-15

    Toll like receptor 4 (TLR4) has been characterized for its ability to recognize bacterial endotoxin lipopolysaccharide (LPS). Considering that infections or inflammatory processes might contribute to the progression of pituitary tumors, we analyzed the TLR4 functional role by evaluating the LPS effect on lactotroph proliferation in primary cultures from experimental pituitary tumors, and examined the involvement of PI3K-Akt and NF-?B activation in this effect. In addition, the role of 17?-estradiol as a possible modulator of LPS-induced PRL cell proliferation was further investigated. In estrogen-induced hyperplasic pituitaries, LPS triggered lactotroph cell proliferation. However, endotoxin failed to increase the number of lactotrophs taking up BrdU in normal pituitaries. Moreover, incubation with anti-TLR4 antibody significantly reduced LPS-induced lactotroph proliferation, suggesting a functional role of this receptor. As a sign of TLR4 activation, an LPS challenge increased IL-6 release in normal and tumoral cells. By flow cytometry, TLR4 baseline expression was revealed at the plasma membrane of tumoral lactotrophs, without changes noted in the percentage of double PRL/TLR4 positive cells after LPS stimulus. Increases in TLR4 intracellular expression were detected as well as rises in CD14, p-Akt and NF-?B after an LPS challenge, as assessed by western blotting. The TLR4/PRL and PRL/NF-?B co-localization was also corroborated by immunofluorescence and the involvement of PI3K/Akt signaling in lactotroph proliferation and IL-6 release was revealed through the PI3K inhibitor Ly-294002. In addition, 17?-estradiol attenuated the LPS-evoked increase in tumoral lactotroph proliferation and IL-6 release. Collectively these results demonstrate the presence of functional TLR4 in lactotrophs from estrogen-induced hyperplasic pituitaries, which responded to the proliferative stimulation and IL-6 release induced by LPS through TLR4/CD14, with a contribution of the PI3K-Akt and NF-?B signaling pathways. - Highlights: In hyperplastic pituitaries, LPS triggered the lactotroph cell proliferation and IL-6 release. Functional Toll-like receptor 4 (TLR4) is expressed at the plasma membrane of tumoral lactotrophs. Increases in TLR4 and CD14 intracellular expression levels were detected after an LPS challenge. The proliferative stimulation and IL-6 release involved the PI3K-Akt pathway and NF-?B activation. 17?-estradiol attenuated the LPS-evoked tumoral lactotroph proliferation and IL-6 secretion.

  20. Kinase Expression and Chromosomal Rearrangements in Papillary Thyroid Cancer Tissues: Investigations at the Molecular and Microscopic Levels

    SciTech Connect (OSTI)

    Weier, Heinz-Ulrich; Kwan, Johnson; Lu, Chun-Mei; Ito, Yuko; Wang, Mei; Baumgartner, Adolf; Hayward, Simon W.; Weier, Jingly F.; Zitzelsberger, Horst F.

    2009-07-07

    Structural chromosome aberrations are known hallmarks of many solid tumors. In the papillary form of thyroid cancer (PTC), for example, activation of the receptor tyrosine kinase (RTK) genes, ret or the neurotrophic tyrosine kinase receptor type I (NTRK1) by intra- or interchromosomal rearrangements have been suggested as a cause of the disease. The 1986 accident at the nuclear power plant in Chernobyl, USSR, led to the uncontrolled release of high levels of radioisotopes. Ten years later, the incidence of childhood papillary thyroid cancer (chPTC) near Chernobyl had risen by two orders of magnitude. Tumors removed from some of these patients showed aberrant expression of the ret RTK gene due to a ret/PTC1 or ret/PTC3 rearrangement involving chromosome 10. However, many cultured chPTC cells show a normal G-banded karyotype and no ret rearrangement. We hypothesize that the 'ret-negative' tumors inappropriately express a different oncogene or have lost function of a tumor suppressor as a result of chromosomal rearrangements, and decided to apply molecular and cytogenetic methods to search for potentially oncogenic chromosomal rearrangements in Chernobyl chPTC cases. Knowledge of the kind of genetic alterations may facilitate the early detection and staging of chPTC as well as provide guidance for therapeutic intervention.

  1. Molecular cloning of amphioxus uncoupling protein and assessment of its uncoupling activity using a yeast heterologous expression system

    SciTech Connect (OSTI)

    Chen, Kun; Sun, Guoxun; Lv, Zhiyuan; Wang, Chen; Jiang, Xueyuan; Li, Donghai; Zhang, Chenyu

    2010-10-01

    Research highlights: {yields} Invertebrates, for example amphioxus, do express uncoupling proteins. {yields} Both the sequence and the uncoupling activity of amphioxus UCP resemble UCP2. {yields} UCP1 is the only UCP that can form dimer on yeast mitochondria. -- Abstract: The present study describes the molecular cloning of a novel cDNA fragment from amphioxus (Branchiostoma belcheri) encoding a 343-amino acid protein that is highly homologous to human uncoupling proteins (UCP), this protein is therefore named amphioxus UCP. This amphioxus UCP shares more homology with and is phylogenetically more related to mammalian UCP2 as compared with UCP1. To further assess the functional similarity of amphioxus UCP to mammalian UCP1 and -2, the amphioxus UCP, rat UCP1, and human UCP2 were separately expressed in Saccharomyces cerevisiae, and the recombinant yeast mitochondria were isolated and assayed for the state 4 respiration rate and proton leak, using pYES2 empty vector as the control. UCP1 increased the state 4 respiration rate by 2.8-fold, and the uncoupling activity was strongly inhibited by GDP, while UCP2 and amphioxus UCP only increased the state 4 respiration rate by 1.5-fold and 1.7-fold in a GDP-insensitive manner, moreover, the proton leak kinetics of amphioxus UCP was very similar to UCP2, but much different from UCP1. In conclusion, the amphioxus UCP has a mild, unregulated uncoupling activity in the yeast system, which resembles mammalian UCP2, but not UCP1.

  2. Over-expression of human endosulfatase-1 exacerbates cadmium-induced injury to transformed human lung cells in vitro

    SciTech Connect (OSTI)

    Zhang, Huiying; Department of Environmental and Molecular Toxicology, College of Agriculture and Life Sciences, NC State University, Raleigh, NC 27695 ; Newman, Donna R.; Bonner, James C.; Sannes, Philip L.

    2012-11-15

    Environmental exposure to cadmium is known to cause damage to alveolar epithelial cells of the lung, impair their capacity to repair, and result in permanent structural alterations. Cell surface heparan sulfate proteoglycans (HSPGs) can modulate cell responses to injury through their interactions with soluble effector molecules. These interactions are often sulfate specific, and the removal of sulfate groups from HS side chains could be expected to influence cellular injury, such as that caused by exposure to cadmium. The goal of this study was to define the role 6-O-sulfate plays in cellular responses to cadmium exposure in two pulmonary epithelial cancer cell lines (H292 and A549) and in normal human primary alveolar type II (hAT2) cells. Sulfate levels were modified by transduced transient over-expression of 6-O-endosulfatase (HSulf-1), a membrane-bound enzyme which specifically removes 6-O-sulfate groups from HSPG side chains. Results showed that cadmium decreased cell viability and activated apoptosis pathways at low concentrations in hAT2 cells but not in the cancer cells. HSulf-1 over-expression, on the contrary, decreased cell viability and activated apoptosis pathways in H292 and A549 cells but not in hAT2 cells. When combined with cadmium, HSulf-1 over-expression further decreased cell viability and exacerbated the activation of apoptosis pathways in the transformed cells but did not add to the toxicity in hAT2 cells. The finding that HSulf-1 sensitizes these cancer cells and intensifies the injury induced by cadmium suggests that 6-O-sulfate groups on HSPGs may play important roles in protection against certain environmental toxicants, such as heavy metals. -- Highlights: ? Primary human lung alveolar type 2 (hAT2) cells and H292 and A549 cells were used. ? Cadmium induced apoptosis in hAT2 cells but not in H292 or A549 cells. ? HSulf-1exacerbates apoptosis induced by cadmium in H292 and A549 but not hAT2 cells.

  3. The expression level of HJURP has an independent prognostic impact and predicts the sensitivity to radiotherapy in breast cancer

    SciTech Connect (OSTI)

    Hu, Zhi; Huang, Ge; Sadanandam, Anguraj; Gu, Shenda; Lenburg, Marc E; Pai, Melody; Bayani, Nora; Blakely, Eleanor A; Gray, Joe W; Mao, Jian-Hua

    2010-06-25

    Introduction: HJURP (Holliday Junction Recognition Protein) is a newly discovered gene reported to function at centromeres and to interact with CENPA. However its role in tumor development remains largely unknown. The goal of this study was to investigate the clinical significance of HJURP in breast cancer and its correlation with radiotherapeutic outcome. Methods: We measured HJURP expression level in human breast cancer cell lines and primary breast cancers by Western blot and/or by Affymetrix Microarray; and determined its associations with clinical variables using standard statistical methods. Validation was performed with the use of published microarray data. We assessed cell growth and apoptosis of breast cancer cells after radiation using high-content image analysis. Results: HJURP was expressed at higher level in breast cancer than in normal breast tissue. HJURP mRNA levels were significantly associated with estrogen receptor (ER), progesterone receptor (PR), Scarff-Bloom-Richardson (SBR) grade, age and Ki67 proliferation indices, but not with pathologic stage, ERBB2, tumor size, or lymph node status. Higher HJURP mRNA levels significantly decreased disease-free and overall survival. HJURP mRNA levels predicted the prognosis better than Ki67 proliferation indices. In a multivariate Cox proportional-hazard regression, including clinical variables as covariates, HJURP mRNA levels remained an independent prognostic factor for disease-free and overall survival. In addition HJURP mRNA levels were an independent prognostic factor over molecular subtypes (normal like, luminal, Erbb2 and basal). Poor clinical outcomes among patients with high HJURP expression werevalidated in five additional breast cancer cohorts. Furthermore, the patients with high HJURP levels were much more sensitive to radiotherapy. In vitro studies in breast cancer cell lines showed that cells with high HJURP levels were more sensitive to radiation treatment and had a higher rate of apoptosis than those with low levels. Knock down of HJURP in human breast cancer cells using shRNA reduced the sensitivity to radiation treatment. HJURP mRNA levels were significantly correlated with CENPA mRNA levels. Conclusions: HJURP mRNA level is a prognostic factor for disease-free and overall survival in patients with breast cancer and is a predictive biomarker for sensitivity to radiotherapy.

  4. Determination of gene expression patterns using high-throughput RNA in situ hybridizaion to whole-mount Drosophila embryos

    SciTech Connect (OSTI)

    Weiszmann, R.; Hammonds, A.S.; Celniker, S.E.

    2009-04-09

    We describe a high-throughput protocol for RNA in situ hybridization (ISH) to Drosophila embryos in a 96-well format. cDNA or genomic DNA templates are amplified by PCR and then digoxigenin-labeled ribonucleotides are incorporated into antisense RNA probes by in vitro transcription. The quality of each probe is evaluated before ISH using a RNA probe quantification (dot blot) assay. RNA probes are hybridized to fixed, mixed-staged Drosophila embryos in 96-well plates. The resulting stained embryos can be examined and photographed immediately or stored at 4oC for later analysis. Starting with fixed, staged embryos, the protocol takes 6 d from probe template production through hybridization. Preparation of fixed embryos requires a minimum of 2 weeks to collect embryos representing all stages. The method has been used to determine the expression patterns of over 6,000 genes throughout embryogenesis.

  5. Ran GTPase protein promotes human pancreatic cancer proliferation by deregulating the expression of Survivin and cell cycle proteins

    SciTech Connect (OSTI)

    Deng, Lin; Department of Oncology, Tangdu Hospital, Fourth Military Medical University, Xian, Shaanxi 710038 ; Lu, Yuanyuan; Zhao, Xiaodi; Sun, Yi; Shi, Yongquan; Fan, Hongwei; Liu, Changhao; Zhou, Jinfeng; Nie, Yongzhan; Wu, Kaichun; Fan, Daiming; Guo, Xuegang

    2013-10-18

    Highlights: Overexpression of Ran in pancreatic cancer was correlated with histological grade. Downregulation of Ran could induce cell apoptosis and inhibit cell proliferation. The effects were mediated by cell cycle proteins, Survivin and cleaved Caspase-3. -- Abstract: Ran, a member of the Ras GTPase family, has important roles in nucleocytoplasmic transport. Herein, we detected Ran expression in pancreatic cancer and explored its potential role on tumour progression. Overexpressed Ran in pancreatic cancer tissues was found highly correlated with the histological grade. Downregulation of Ran led to significant suppression of cell proliferation, cell cycle arrest at the G1/S phase and induction of apoptosis. In vivo studies also validated that result. Further studies revealed that those effects were at least partly mediated by the downregulation of Cyclin A, Cyclin D1, Cyclin E, CDK2, CDK4, phospho-Rb and Survivin proteins and up regulation of cleaved Caspase-3.

  6. The effects of 5-fluorouracil and doxorubicin on expression of human immunodeficiency virus type 1 long terminal repeat

    SciTech Connect (OSTI)

    Panozzo, J.; Akan, E.; Griffiths, T.D.; Woloschak, G.E.

    1996-03-01

    Previous work by many groups has documented induction of the HIV-LTR following exposure of cells to ultraviolet light and other DNA damaging agents. Our experiments set out to determine the relative activation or repression of the HIV-LTR in response to two classes of chemotherapeutic agents: Doxorubicin is a DNA-damage inducing agent, and 5-fluorouracil has an antimetabolic mode of action. Using HeLa cells stably transfected with a construct in which HIV-LTR drives expression of the chloramphenicol acetyl transferase reporter gene, we demonstrated an up to 10-fold induction following doxorubicin treatment in 24 h post-treatment. This induction was repressed by treatment with salicylic acid, suggesting a role for prostaglandin/cyclo-oxygenase pathways and/or NFKB in the inductive response. Induction by 5-fluorouracil, in contrast, was more modest (two-fold at most) though it was consistently elevated over controls.

  7. Enhanced expression of Nrf2 in mice attenuates the fatty liver produced by a methionine- and choline-deficient diet

    SciTech Connect (OSTI)

    Zhang, Yu-Kun Jennifer; Yeager, Ronnie L.; Tanaka, Yuji; Klaassen, Curtis D.

    2010-06-15

    Oxidative stress has been proposed as an important promoter of the progression of fatty liver diseases. The current study investigates the potential functions of the Nrf2-Keap1 signaling pathway, an important hepatic oxidative stress sensor, in a rodent fatty liver model. Mice with no (Nrf2-null), normal (wild type, WT), and enhanced (Keap1 knockdown, K1-kd) expression of Nrf2 were fed a methionine- and choline-deficient (MCD) diet or a control diet for 5 days. Compared to WT mice, the MCD diet-caused hepatosteatosis was more severe in the Nrf2-null mice and less in the K1-kd mice. The Nrf2-null mice had lower hepatic glutathione and exhibited more lipid peroxidation, whereas the K1-kd mice had the highest amount of glutathione in the liver and developed the least lipid peroxidation among the three genotypes fed the MCD diet. The Nrf2 signaling pathway was activated by the MCD diet, and the Nrf2-targeted cytoprotective genes Nqo1 and Gst{alpha}1/2 were induced in WT and even more in K1-kd mice. In addition, Nrf2-null mice on both control and MCD diets exhibited altered expression profiles of fatty acid metabolism genes, indicating Nrf2 may influence lipid metabolism in liver. For example, mRNA levels of long chain fatty acid translocase CD36 and the endocrine hormone Fgf21 were higher in livers of Nrf2-null mice and lower in the K1-kd mice than WT mice fed the MCD diet. Taken together, these observations indicate that Nrf2 could decelerate the onset of fatty livers caused by the MCD diet by increasing hepatic antioxidant and detoxification capabilities.

  8. RNA Helicase DDX5 Regulates MicroRNA Expression and Contributes to Cytoskeletal Reorganization in Basal Breast Cancer Cells

    SciTech Connect (OSTI)

    Wang, Daojing; Huang, Jing; Hu, Zhi

    2011-11-15

    RNA helicase DDX5 (also p68) is involved in all aspects of RNA metabolism and serves as a transcriptional co-regulator, but its functional role in breast cancer remains elusive. Here, we report an integrative biology study of DDX5 in breast cancer, encompassing quantitative proteomics, global MicroRNA profiling, and detailed biochemical characterization of cell lines and human tissues. We showed that protein expression of DDX5 increased progressively from the luminal to basal breast cancer cell lines, and correlated positively with that of CD44 in the basal subtypes. Through immunohistochemistry analyses of tissue microarrays containing over 200 invasive human ductal carcinomas, we observed that DDX5 was upregulated in the majority of malignant tissues, and its expression correlated strongly with those of Ki67 and EGFR in the triple-negative tumors. We demonstrated that DDX5 regulated a subset of MicroRNAs including miR-21 and miR-182 in basal breast cancer cells. Knockdown of DDX5 resulted in reorganization of actin cytoskeleton and reduction of cellular proliferation. The effects were accompanied by upregulation of tumor suppressor PDCD4 (a known miR-21 target); as well as upregulation of cofilin and profilin, two key proteins involved in actin polymerization and cytoskeleton maintenance, as a consequence of miR-182 downregulation. Treatment with miR-182 inhibitors resulted in morphologic phenotypes resembling those induced by DDX5 knockdown. Using bioinformatics tools for pathway and network analyses, we confirmed that the network for regulation of actin cytoskeleton was predominantly enriched for the predicted downstream targets of miR-182. Our results reveal a new functional role of DDX5 in breast cancer via the DDX5?miR-182?actin cytoskeleton pathway, and suggest the potential clinical utility of DDX5 and its downstream MicroRNAs in the theranostics of breast cancer.

  9. Decreased expression of RNA interference machinery, Dicer and Drosha, is associated with poor outcome in ovarian cancer patients

    SciTech Connect (OSTI)

    Merritt, William M.; Lin, Yvonne G.; Han, Liz Y.; Kamat, Aparna A.; Spannuth, Whitney A.; Schmandt, Rosemarie; Urbauer, Diana; Pennacchio, Len A.; Cheng, Jan-Fang; Zeidan, Alexandra; Wang, Hua; Mueller, Peter; Lenburg, Marc E.; Gray, Joe W.; Mok, Samuel; Birrer, Michael J.; Lopez-Berestein, Gabriel; Coleman, Robert L.; Bar-Eli, Menashe; Sood, Anil K.

    2008-05-06

    The clinical and functional significance of RNA interference (RNAi) machinery, Dicer and Drosha, in ovarian cancer is not known and was examined. Dicer and Drosha expression was measured in ovarian cancer cell lines (n=8) and invasive epithelial ovarian cancer specimens (n=111) and correlated with clinical outcome. Validation was performed with previously published cohorts of ovarian, breast, and lung cancer patients. Anti-Galectin-3 siRNA and shRNA transfections were used for in vitro functional studies. Dicer and Drosha mRNA and protein levels were decreased in 37% to 63% of ovarian cancer cell lines and in 60% and 51% of human ovarian cancer specimens, respectively. Low Dicer was significantly associated with advanced tumor stage (p=0.007), and low Drosha with suboptimal surgical cytoreduction (p=0.02). Tumors with both high Dicer and Drosha were associated with increased median patient survival (>11 years vs. 2.66 years for other groups; p<0.001). In multivariate analysis, high Dicer (HR=0.48; p=0.02), high-grade histology (HR=2.46; p=0.03), and poor chemoresponse (HR=3.95; p<0.001) were identified as independent predictors of disease-specific survival. Findings of poor clinical outcome with low Dicer expression were validated in separate cohorts of cancer patients. Galectin-3 silencing with siRNA transfection was superior to shRNA in cell lines with low Dicer (78-95% vs. 4-8% compared to non-targeting sequences), and similar in cell lines with high Dicer. Our findings demonstrate the clinical and functional impact of RNAi machinery alterations in ovarian carcinoma and support the use of siRNA constructs that do not require endogenous Dicer and Drosha for therapeutic applications.

  10. Nuclear factor-?B is a common upstream signal for growth differentiation factor-5 expression in brown adipocytes exposed to pro-inflammatory cytokines and palmitate

    SciTech Connect (OSTI)

    Hinoi, Eiichi; Iezaki, Takashi; Ozaki, Kakeru; Yoneda, Yukio

    2014-10-03

    Highlights: GDF5 expression is up-regulated by IL-1?, TNF-? and palmitate in brown pre-adipocytes. NF-?B stimulates promoter activity and expression of GDF5 in brown pre-adipocytes. Recruitment of NF-?B to the GDF5 promoter is facilitated in BAT from ob/ob mice. An NF-?B inhibitor prevents upregulation of GDF5 expression in brown pre-adipocytes. - Abstract: We have previously demonstrated that genetic and acquired obesity similarly led to drastic upregulation in brown adipose tissue (BAT), rather than white adipose tissue, of expression of both mRNA and corresponding protein for the bone morphogenic protein/growth differentiation factor (GDF) member GDF5 capable of promoting brown adipogenesis. In this study, we evaluated expression profiles of GDF5 in cultured murine brown pre-adipocytes exposed to pro-inflammatory cytokines and free fatty acids (FFAs), which are all shown to play a role in the pathogenesis of obesity. Both interleukin-1? (IL-1?) and tumor necrosis factor-? (TNF-?) were effective in up-regulating GDF5 expression in a concentration-dependent manner, while similar upregulation was seen in cells exposed to the saturated FFA palmitate, but not to the unsaturated FFA oleate. In silico analysis revealed existence of the putative nuclear factor-?B (NF-?B) binding site in the 5?-flanking region of mouse GDF5, whereas introduction of NF-?B subunits drastically facilitated both promoter activity and expression of GDF5 in brown pre-adipocytes. Chromatin immunoprecipitation analysis confirmed significant facilitation of the recruitment of NF-?B to the GDF5 promoter in lysed extracts of BAT from leptin-deficient ob/ob obese mice. Upregulation o GDF5 expression was invariably inhibited by an NF-?B inhibitor in cultured brown pre-adipocytes exposed to IL-1?, TNF-? and palmitate. These results suggest that obesity leads to upregulation of GDF5 expression responsible for the promotion of brown adipogenesis through a mechanism relevant to activation of the NF-?B pathway in response to particular pro-inflammatory cytokines and/or saturated FFAs in BAT.

  11. MicroRNA-101 mediates the suppressive effect of laminar shear stress on mTOR expression in vascular endothelial cells

    SciTech Connect (OSTI)

    Chen, Kui; Fan, Wendong; Wang, Xing; Ke, Xiao [Division of Cardiology, First Affiliated Hospital of Sun Yat-sen University, Guangzhou 510080 (China)] [Division of Cardiology, First Affiliated Hospital of Sun Yat-sen University, Guangzhou 510080 (China); Wu, Guifu, E-mail: eecpchina@yahoo.com.cn [Key Laboratory on Assisted Circulation, Ministry of Health, Guangzhou 510080 (China)] [Key Laboratory on Assisted Circulation, Ministry of Health, Guangzhou 510080 (China); Hu, Chengheng, E-mail: huchenghengpci@yahoo.com.cn [Division of Cardiology, First Affiliated Hospital of Sun Yat-sen University, Guangzhou 510080 (China)] [Division of Cardiology, First Affiliated Hospital of Sun Yat-sen University, Guangzhou 510080 (China)

    2012-10-12

    Highlights: Black-Right-Pointing-Pointer Laminar shear stress upregulates miR-101 expression in vascular endothelial cells. Black-Right-Pointing-Pointer miR-101 represses mTOR expression through a specific 3 Prime UTR binding site. Black-Right-Pointing-Pointer Overexpression of miR-101 inhibits G1/S transition and endothelial cell proliferation. Black-Right-Pointing-Pointer Blockade of miR-101 attenuates the suppressive effect of laminar flow on mTOR expression. -- Abstract: Shear stress associated with blood flow plays an important role in regulating gene expression and cell function in endothelial cells (ECs). MicroRNAs (miRNAs) are highly conserved, small non-coding RNAs that negatively regulate the expression of target genes by binding to the mRNA 3 Prime -untranslated region (3 Prime UTR) at the posttranscriptional level involved in diverse cellular processes. This study demonstrates that microRNA-101 in response to laminar shear stress (LSS) is involved in the flow regulation of gene expression in ECs. qRT-PCR analysis showed that miR-101 expression was significantly upregulated in human umbilical vein endothelial cells (HUVECs) exposed to 12 dyn/cm{sup 2} laminar shear stress for 12 h. We found that transfection of miR-101 significantly decreased the luciferase activity of plasmid reporter containing the 3 Prime UTR of mammalian target of rapamycin (mTOR) gene. Western analysis revealed that the protein level of mTOR was significantly reduced in ECs transfected with miR-101. Furthermore, miR-101 overexpression induced cell cycle arrest at the G1/S transition and suppressed endothelial cell proliferation. Finally, transfection of miR-101 inhibitors attenuated the suppressive effects of LSS on mTOR expression, which identified the efficacy of loss-of-function of miR-101 in laminar flow-treated ECs. In conclusion, we have demonstrated that upregulation of miR-101 in response to LSS contributes to the suppressive effects of LSS on mTOR expression and EC proliferation. These studies advance our understanding of the posttranscriptional mechanisms by which shear stress modulates endothelial homeostasis.

  12. Role for DNA methylation in the regulation of miR-200c and miR-141 expression in normal and cancer cells

    SciTech Connect (OSTI)

    Vrba, Lukas; Jensen, Taylor J.; Garbe, James C.; Heimark, Ronald L.; Cress, Anne E.; Dickinson, Sally; Stampfer, Martha R.; Futscher, Bernard W.

    2009-12-23

    BACKGROUND: The microRNA-200 family participates in the maintenance of an epithelial phenotype and loss of its expression can result in epithelial to mesenchymal transition (EMT). Furthermore, the loss of expression of miR-200 family members is linked to an aggressive cancer phenotype. Regulation of the miR-200 family expression in normal and cancer cells is not fully understood. METHODOLOGY/ PRINCIPAL FINDINGS: Epigenetic mechanisms participate in the control of miR-200c and miR-141 expression in both normal and cancer cells. A CpG island near the predicted mir-200c/mir-141 transcription start site shows a striking correlation between miR-200c and miR-141 expression and DNA methylation in both normal and cancer cells, as determined by MassARRAY technology. The CpG island is unmethylated in human miR-200/miR-141 expressing epithelial cells and in miR-200c/miR-141 positive tumor cells. The CpG island is heavily methylated in human miR-200c/miR-141 negative fibroblasts and miR-200c/miR-141 negative tumor cells. Mouse cells show a similar inverse correlation between DNA methylation and miR-200c expression. Enrichment of permissive histone modifications, H3 acetylation and H3K4 trimethylation, is seen in normal miR-200c/miR-141-positive epithelial cells, as determined by chromatin immunoprecipitation coupled to real-time PCR. In contrast, repressive H3K9 dimethylation marks are present in normal miR-200c/miR-141-negative fibroblasts and miR-200c/miR-141 negative cancer cells and the permissive histone modifications are absent. The epigenetic modifier drug, 5-aza-2'-deoxycytidine, reactivates miR-200c/miR-141 expression showing that epigenetic mechanisms play a functional role in their transcriptional control. CONCLUSIONS/ SIGNIFICANCE: We report that DNA methylation plays a role in the normal cell type-specific expression of miR-200c and miR-141 and this role appears evolutionarily conserved, since similar results were obtained in mouse. Aberrant DNA methylation of the miR-200c/141 CpG island is closely linked to their inappropriate silencing in cancer cells. Since the miR-200c cluster plays a significant role in EMT, our results suggest an important role for DNA methylation in the control of phenotypic conversions in normal cells.

  13. Isolation and bacterial expression of a sesquiterpene synthase CDNA clone from peppermint(mentha .chi. piperita, L.) that produces the aphid alarm pheromone (E)-.beta.-farnesene

    DOE Patents [OSTI]

    Croteau, Rodney Bruce; Wildung, Mark Raymond; Crock, John E.

    1999-01-01

    A cDNA encoding (E)-.beta.-farnesene synthase from peppermint (Mentha piperita) has been isolated and sequenced, and the corresponding amino acid sequence has been determined. Accordingly, an isolated DNA sequence (SEQ ID NO:1) is provided which codes for the expression of (E)-.beta.-farnesene synthase (SEQ ID NO:2), from peppermint (Mentha piperita). In other aspects, replicable recombinant cloning vehicles are provided which code for (E)-.beta.-farnesene synthase, or for a base sequence sufficiently complementary to at least a portion of (E)-.beta.-farnesene synthase DNA or RNA to enable hybridization therewith. In yet other aspects, modified host cells are provided that have been transformed, transfected, infected and/or injected with a recombinant cloning vehicle and/or DNA sequence encoding (E)-.beta.-farnesene synthase. Thus, systems and methods are provided for the recombinant expression of the aforementioned recombinant (E)-.beta.-farnesene synthase that may be used to facilitate its production, isolation and purification in significant amounts. Recombinant (E)-.beta.-farnesene synthase may be used to obtain expression or enhanced expression of (E)-.beta.-farnesene synthase in plants in order to enhance the production of (E)-.beta.-farnesene, or may be otherwise employed for the regulation or expression of (E)-.beta.-farnesene synthase, or the production of its product.

  14. Isolation and bacterial expression of a sesquiterpene synthase cDNA clone from peppermint (Mentha x piperita, L.) that produces the aphid alarm pheromone (E)-.beta.-farnesene

    DOE Patents [OSTI]

    Croteau, Rodney Bruce; Crock, John E.

    2005-01-25

    A cDNA encoding (E)-.beta.-farnesene synthase from peppermint (Mentha piperita) has been isolated and sequenced, and the corresponding amino acid sequence has been determined. Accordingly, an isolated DNA sequence (SEQ ID NO:1) is provided which codes for the expression of (E)-.beta.-farnesene synthase (SEQ ID NO:2), from peppermint (Mentha piperita). In other aspects, replicable recombinant cloning vehicles are provided which code for (E)-.beta.-farnesene synthase, or for a base sequence sufficiently complementary to at least a portion of (E)-.beta.-farnesene synthase DNA or RNA to enable hybridization therewith. In yet other aspects, modified host cells are provided that have been transformed, transfected, infected and/or injected with a recombinant cloning vehicle and/or DNA sequence encoding (E)-.beta.-farnesene synthase. Thus, systems and methods are provided for the recombinant expression of the aforementioned recombinant (E)-.beta.-famesene synthase that may be used to facilitate its production, isolation and purification in significant amounts. Recombinant (E)-.beta.-farnesene synthase may be used to obtain expression or enhanced expression of (E)-.beta.-famesene synthase in plants in order to enhance the production of (E)-.beta.-farnesene, or may be otherwise employed for the regulation or expression of (E)-.beta.-farnesene synthase, or the production of its product.

  15. Analysis of the Salmonella regulatory network suggests involvement of SsrB and H-NS in σE-regulated SPI-2 gene expression

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Li, Jie; Overall, Christopher C.; Nakayasu, Ernesto S.; Kidwai, Afshan S.; Jones, Marcus B.; Johnson, Rudd; Nguyen, Nhu T.; McDermott, Jason E.; Ansong, Charles; Heffron, Fred; et al

    2015-02-10

    The extracytoplasmic functioning sigma factor ?E is known to play an essential role for Salmonella enterica serovar Typhimurium to survive and proliferate in macrophages and mice. However, its regulatory network is not well characterized, especially during infection. Here we used microarray to identify genes regulated by ?E in Salmonella grown in three conditions: a nutrient-rich condition and two others that mimic early and late intracellular infection. We found that in each condition ?E regulated different sets of genes, and notably, several global regulators. When comparing nutrient-rich and infection-like conditions, large changes were observed in the expression of genes involved inmoreSalmonella pathogenesis island (SPI)-1 type-three secretion system (TTSS), SPI-2 TTSS, protein synthesis, and stress responses. In total, the expression of 58% of Salmonella genes was affected by ?E in at least one of the three conditions. An important finding is that ?E up-regulates SPI-2 genes, which are essential for Salmonella intracellular survival, by up-regulating SPI-2 activator ssrB expression at the early stage of infection and down-regulating SPI-2 repressor hns expression at a later stage. Moreover, ?E is capable of countering the silencing of H-NS, releasing the expression of SPI-2 genes. This connection between ?E and SPI-2 genes, combined with the global regulatory effect of ?E, may account for the lethality of rpoE-deficient Salmonella in murine infection.less

  16. JBP485 improves gentamicin-induced acute renal failure by regulating the expression and function of Oat1 and Oat3 in rats

    SciTech Connect (OSTI)

    Guo, Xinjin [Department of Clinical Pharmacology, College of Pharmacy, Dalian Medical University, Dalian (China); Meng, Qiang; Liu, Qi; Wang, Changyuan; Sun, Huijun; Peng, Jinyong; Ma, Xiaochi [Department of Clinical Pharmacology, College of Pharmacy, Dalian Medical University, Dalian (China); Provincial Key Laboratory for Pharmacokinetics and Transport, Dalian Medical University, Liaoning (China); Kaku, Taiichi [Japan Bioproducts Industry Co. Ltd., Tomigaya, Shibuya-ku, Tokyo (Japan); Liu, Kexin, E-mail: kexinliu@dlmedu.edu.cn [Department of Clinical Pharmacology, College of Pharmacy, Dalian Medical University, Dalian (China); Provincial Key Laboratory for Pharmacokinetics and Transport, Dalian Medical University, Liaoning (China)

    2013-09-01

    We investigated the effects of JBP485 (an anti-inflammatory dipeptide and a substrate of OAT) on regulation of the expression and function of renal Oat1 and Oat3, which can accelerate the excretion of accumulated uremic toxins (e.g. indoxyl sulfate) in the kidney to improve gentamicin-induced ARF in rats. JBP485 caused a significant decrease in the accumulation of endogenous substances (creatinine, blood urea nitrogen and indoxyl sulfate) in vivo, an increase in the excretion of exogenous compounds (lisinopril and inulin) into urine, and up-regulation of the expressions of renal Oat1 and Oat3 in the kidney tissues and slices via substrate induction. To determine the effect of JBP485 on the accelerated excretion of uremic toxins mediated by Oat1 and Oat3, the mRNA and protein expression levels of renal basolateral Oats were assessed by quantitative real-time PCR, western blot, immunohistochemical analysis and an immunofluorescence method. Gentamicin down-regulated the expression of Oats mRNA and protein in rat kidney, and these effects were reversed after administration of JBP485. In addition, JBP485 caused a significant decrease in MPO and MDA levels in the kidney, and improved the pathological condition of rat kidney. These results indicated that JBP485 improved acute renal failure by increasing the expression and function of Oat1 and Oat3, and by decreasing overoxidation of the kidney in gentamicin-induced ARF rats. - Highlights: JBP485 could up-regulate function and expression of Oat1 and Oat3 in kidney. Effects of JBP485 on ARF are mediated by stimulating excretion of uremic toxins. JBP485 protected against gentamicin-induced ARF by decreasing MPO and MDA.

  17. No Effect of the Transforming Growth Factor {beta}1 Promoter Polymorphism C-509T on TGFB1 Gene Expression, Protein Secretion, or Cellular Radiosensitivity

    SciTech Connect (OSTI)

    Reuther, Sebastian; Metzke, Elisabeth; Bonin, Michael; Petersen, Cordula; Dikomey, Ekkehard; Raabe, Annette

    2013-02-01

    Purpose: To study whether the promoter polymorphism (C-509T) affects transforming growth factor {beta}1 gene (TGFB1) expression, protein secretion, and/or cellular radiosensitivity for both human lymphocytes and fibroblasts. Methods and Materials: Experiments were performed with lymphocytes taken either from 124 breast cancer patients or 59 pairs of normal monozygotic twins. We used 15 normal human primary fibroblast strains as controls. The C-509T genotype was determined by polymerase chain reaction-restriction fragment length polymorphism or TaqMan single nucleotide polymorphism (SNP) genotyping assay. The cellular radiosensitivity of lymphocytes was measured by G0/1 assay and that of fibroblasts by colony assay. The amount of extracellular TGFB1 protein was determined by enzyme-linked immunosorbent assay, and TGFB1 expression was assessed via microarray analysis or reverse transcription-polymerase chain reaction. Results: The C-509T genotype was found not to be associated with cellular radiosensitivity, neither for lymphocytes (breast cancer patients, P=.811; healthy donors, P=.181) nor for fibroblasts (P=.589). Both TGFB1 expression and TGFB1 protein secretion showed considerable variation, which, however, did not depend on the C-509T genotype (protein secretion: P=.879; gene expression: lymphocytes, P=.134, fibroblasts, P=.605). There was also no general correlation between TGFB1 expression and cellular radiosensitivity (lymphocytes, P=.632; fibroblasts, P=.573). Conclusion: Our data indicate that any association between the SNP C-509T of TGFB1 and risk of normal tissue toxicity cannot be ascribed to a functional consequence of this SNP, either on the level of gene expression, protein secretion, or cellular radiosensitivity.

  18. Arsenic augments the uptake of oxidized LDL by upregulating the expression of lectin-like oxidized LDL receptor in mouse aortic endothelial cells

    SciTech Connect (OSTI)

    Hossain, Ekhtear; Ota, Akinobu; Karnan, Sivasundaram; Damdindorj, Lkhagvasuren; Takahashi, Miyuki; Konishi, Yuko; Konishi, Hiroyuki; Hosokawa, Yoshitaka

    2013-12-15

    Although chronic arsenic exposure is a well-known risk factor for cardiovascular diseases, including atherosclerosis, the molecular mechanism underlying arsenic-induced atherosclerosis remains obscure. Therefore, this study aimed to elucidate this molecular mechanism. We examined changes in the mRNA level of the lectin-like oxidized LDL (oxLDL) receptor (LOX-1) in a mouse aortic endothelial cell line, END-D, after sodium arsenite (SA) treatment. SA treatment significantly upregulated LOX-1 mRNA expression; this finding was also verified at the protein expression level. Flow cytometry and fluorescence microscopy analyses showed that the cellular uptake of fluorescence (Dil)-labeled oxLDL was significantly augmented with SA treatment. In addition, an anti-LOX-1 antibody completely abrogated the augmented uptake of Dil-oxLDL. We observed that SA increased the levels of the phosphorylated forms of nuclear factor of kappa light polypeptide gene enhancer in B cells (NF-?B)/p65. SA-induced upregulation of LOX-1 protein expression was clearly prevented by treatment with an antioxidant, N-acetylcysteine (NAC), or an NF-?B inhibitor, caffeic acid phenethylester (CAPE). Furthermore, SA-augmented uptake of Dil-oxLDL was also prevented by treatment with NAC or CAPE. Taken together, our results indicate that arsenic upregulates LOX-1 expression through the reactive oxygen species-mediated NF-?B signaling pathway, followed by augmented cellular oxLDL uptake, thus highlighting a critical role of the aberrant LOX-1 signaling pathway in the pathogenesis of arsenic-induced atherosclerosis. - Highlights: Sodium arsenite (SA) increases LOX-1 expression in mouse aortic endothelial cells. SA enhances cellular uptake of oxidized LDL in dose-dependent manner. SA-induced ROS generation enhances phosphorylation of NF-?B. SA upregulates LOX-1 expression through ROS-activated NF-?B signaling pathway.

  19. Phylogenetic and comparative gene expression analysis of barley (Hordeum vulgare)WRKY transcription factor family reveals putatively retained functions betweenmonocots and dicots

    SciTech Connect (OSTI)

    Mangelsen, Elke; Kilian, Joachim; Berendzen, Kenneth W.; Kolukisaoglu, Uner; Harter, Klaus; Jansson, Christer; Wanke, Dierk

    2008-02-01

    WRKY proteins belong to the WRKY-GCM1 superfamily of zinc finger transcription factors that have been subject to a large plant-specific diversification. For the cereal crop barley (Hordeum vulgare), three different WRKY proteins have been characterized so far, as regulators in sucrose signaling, in pathogen defense, and in response to cold and drought, respectively. However, their phylogenetic relationship remained unresolved. In this study, we used the available sequence information to identify a minimum number of 45 barley WRKY transcription factor (HvWRKY) genes. According to their structural features the HvWRKY factors were classified into the previously defined polyphyletic WRKY subgroups 1 to 3. Furthermore, we could assign putative orthologs of the HvWRKY proteins in Arabidopsis and rice. While in most cases clades of orthologous proteins were formed within each group or subgroup, other clades were composed of paralogous proteins for the grasses and Arabidopsis only, which is indicative of specific gene radiation events. To gain insight into their putative functions, we examined expression profiles of WRKY genes from publicly available microarray data resources and found group specific expression patterns. While putative orthologs of the HvWRKY transcription factors have been inferred from phylogenetic sequence analysis, we performed a comparative expression analysis of WRKY genes in Arabidopsis and barley. Indeed, highly correlative expression profiles were found between some of the putative orthologs. HvWRKY genes have not only undergone radiation in monocot or dicot species, but exhibit evolutionary traits specific to grasses. HvWRKY proteins exhibited not only sequence similarities between orthologs with Arabidopsis, but also relatedness in their expression patterns. This correlative expression is indicative for a putative conserved function of related WRKY proteins in mono- and dicot species.

  20. Enhanced citric acid production in aspergillus with inactivated asparagine-linked glycosylation protein 3 (ALG3), and/or increased laeA expression

    DOE Patents [OSTI]

    Dai, Ziyu; Baker, Scott E.

    2015-12-08

    Provided herein are fungi, such as Aspergillus niger, having a dolichyl-P-Man:Man(5)GlcNAc(2)-PP-dolichyl mannosyltransferase (Alg3) gene genetic inactivation, increased expression of a loss of aflR expression A (Lae), or both. In some examples, such mutants have several phenotypes, including an increased production of citric acid relative to the parental strain. Methods of using the disclosed fungi to make citric acid are also provided, as are compositions and kits including the disclosed fungi.

  1. Low p53 Binding Protein 1 (53BP1) Expression Is Associated With Increased Local Recurrence in Breast Cancer Patients Treated With Breast-Conserving Surgery and Radiotherapy

    SciTech Connect (OSTI)

    Neboori, Hanmanth J.R.; Haffty, Bruce G.; Wu Hao; Yang Qifeng; Aly, Amal; Goyal, Sharad; Schiff, Devora; Moran, Meena S.; Golhar, Ryan; Chen Chunxia; Moore, Dirk; and others

    2012-08-01

    Purpose: To investigate whether the expression of p53 binding protein 1 (53BP1) has prognostic significance in a cohort of early-stage breast cancer patients treated with breast-conserving surgery and radiotherapy (BCS+RT). Methods and Materials: A tissue microarray of early-stage breast cancer treated with BCS+RT from a cohort of 514 women was assayed for 53BP1, estrogen receptor, progesterone receptor, and HER2 expression by immunohistochemistry. Through log-rank tests and univariate and multivariate models, the staining profile of each tumor was correlated with clinical endpoints, including ipsilateral breast recurrence-free survival (IBRFS), distant metastasis-free survival (DMFS), cause-specific survival (CSS), recurrence-free survival (RFS), and overall survival (OS). Results: Of the 477 (93%) evaluable tumors, 63 (13%) were scored as low. Low expression of 53BP1 was associated with worse outcomes for all endpoints studied, including 10-year IBRFS (76.8% vs. 90.5%; P=.01), OS (66.4% vs. 81.7%; P=.02), CSS (66.0% vs. 87.4%; P<.01), DMFS (55.9% vs. 87.0%; P<.01), and RFS (45.2% vs. 80.6%; P<.01). Multivariate analysis incorporating various clinico-pathologic markers and 53BP1 expression found that 53BP1 expression was again an independent predictor of all endpoints (IBRFS: P=.0254; OS: P=.0094; CSS: P=.0033; DMFS: P=.0006; RFS: P=.0002). Low 53BP1 expression was also found to correlate with triple-negative (TN) phenotype (P<.01). Furthermore, in subset analysis of all TN breast cancer, negative 53BP1 expression trended for lower IBRFS (72.3% vs. 93.9%; P=.0361) and was significant for worse DMFS (48.2% vs. 86.8%; P=.0035) and RFS (37.8% vs. 83.7%; P=.0014). Conclusion: Our data indicate that low 53BP1 expression is an independent prognostic indicator for local relapse among other endpoints in early-stage breast cancer and TN breast cancer patients treated with BCS+RT. These results should be verified in larger cohorts of patients to validate their clinical significance.

  2. Case study on the utility of hepatic global gene expression profiling in the risk assessment of the carcinogen furan

    SciTech Connect (OSTI)

    Jackson, Anna Francina; Williams, Andrew; Recio, Leslie; Waters, Michael D.; Lambert, Iain B.; Yauk, Carole L.

    2014-01-01

    Furan is a chemical hepatocarcinogen in mice and rats. Its previously postulated cancer mode of action (MOA) is chronic cytotoxicity followed by sustained regenerative proliferation; however, its molecular basis is unknown. To this end, we conducted toxicogenomic analysis of B3C6F1 mouse livers following three week exposures to non-carcinogenic (0, 1, 2 mg/kg bw) or carcinogenic (4 and 8 mg/kg bw) doses of furan. We saw enrichment for pathways responsible for cytotoxicity: stress-activated protein kinase (SAPK) and death receptor (DR5 and TNF-alpha) signaling, and proliferation: extracellular signal-regulated kinases (ERKs) and TNF-alpha. We also noted the involvement of NF-kappaB and c-Jun in response to furan, which are genes that are known to be required for liver regeneration. Furan metabolism by CYP2E1 produces cis-2-butene-1,4-dial (BDA), which is required for ensuing cytotoxicity and oxidative stress. NRF2 is a master regulator of gene expression during oxidative stress and we suggest that chronic NFR2 activity and chronic inflammation may represent critical transition events between the adaptive (regeneration) and adverse (cancer) outcomes. Another objective of this study was to demonstrate the applicability of toxicogenomics data in quantitative risk assessment. We modeled benchmark doses for our transcriptional data and previously published cancer data, and observed consistency between the two. Margin of exposure values for both transcriptional and cancer endpoints were also similar. In conclusion, using furan as a case study we have demonstrated the value of toxicogenomics data in elucidating dose-dependent MOA transitions and in quantitative risk assessment. - Highlights: Global gene expression changes in furan-exposed mouse livers were analyzed. A molecular mode of action for furan-induced hepatocarcinogenesis is proposed. Key pathways include NRF2, SAPK, ERK and death receptor signaling. Important roles for TNF-alpha, c-Jun, and NF-?B in tumorigenesis are proposed. BMD and MoE values from transcriptional and apical data are compared.

  3. SPINE: SParse eIgengene NEtwork linking gene expression clusters in Dehalococcoides mccartyi to perturbations in experimental conditions

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Mansfeldt, Cresten B.; Logsdon, Benjamin A.; Debs, Garrett E.; Richardson, Ruth E.; Mande, Shekhar C.

    2015-02-25

    We present a statistical model designed to identify the effect of experimental perturbations on the aggregate behavior of the transcriptome expressed by the bacterium Dehalococcoides mccartyi strain 195. Strains of Dehalococcoides are used in sub-surface bioremediation applications because they organohalorespire tetrachloroethene and trichloroethene (common chlorinated solvents that contaminate the environment) to non-toxic ethene. However, the biochemical mechanism of this process remains incompletely described. Additionally, the response of Dehalococcoides to stress-inducing conditions that may be encountered at field-sites is not well understood. The constructed statistical model captured the aggregate behavior of gene expression phenotypes by modeling the distinct eigengenes of 100more » transcript clusters, determining stable relationships among these clusters of gene transcripts with a sparse network-inference algorithm, and directly modeling the effect of changes in experimental conditions by constructing networks conditioned on the experimental state. Based on the model predictions, we discovered new response mechanisms for DMC, notably when the bacterium is exposed to solvent toxicity. The network identified a cluster containing thirteen gene transcripts directly connected to the solvent toxicity condition. Transcripts in this cluster include an iron-dependent regulator (DET0096-97) and a methylglyoxal synthase (DET0137). To validate these predictions, additional experiments were performed. Continuously fed cultures were exposed to saturating levels of tetrachloethene, thereby causing solvent toxicity, and transcripts that were predicted to be linked to solvent toxicity were monitored by quantitative reverse-transcription polymerase chain reaction. Twelve hours after being shocked with saturating levels of tetrachloroethene, the control transcripts (encoding for a key hydrogenase and the 16S rRNA) did not significantly change. By contrast, transcripts for DET0137 and DET0097 displayed a 46.8±11.5 and 14.6±9.3 fold up-regulation, respectively, supporting the model. This is the first study to identify transcripts in Dehalococcoides that potentially respond to tetrachloroethene solvent-toxicity conditions that may be encountered near contamination source zones in sub-surface environments.« less

  4. MNK1 expression increases during cellular senescence and modulates the subcellular localization of hnRNP A1

    SciTech Connect (OSTI)

    Ziaei, Samira; The Graduate School and University Center of CUNY, New York, NY ; Shimada, Naoko; Kucharavy, Herman; Hubbard, Karen; The Graduate School and University Center of CUNY, New York, NY

    2012-03-10

    Heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1) is an RNA-binding protein that modulates splice site usage, polyadenylation, and cleavage efficiency. This protein has also been implicated in mRNA stability and transport from the nucleus. We have previously demonstrated that hnRNP A1 had diminished protein levels and showed cytoplasmic accumulation in senescent human diploid fibroblasts. Furthermore, we have shown that inhibition of p38 MAPK, a key regulator of cellular senescence, elevated hnRNP A1 protein levels and inhibited hnRNP A1 cytoplasmic localization. In this study, we have explored the possible involvement of MNK1, one of the downstream effector of p38 MAPK, in the regulation of hnRNP A1. We have demonstrated that pharmacological inhibition of MNK1 by CGP 57380 decreased the phosphorylation levels of hnRNP A1 in young and senescent fibroblast cells and blocked the cytoplasmic accumulation of hnRNP A1 in senescent cells. In addition, MNK1 formed a complex with hnRNP A1 in vivo. The expression levels of MNK1, phospho-MNK1, and phospho-eIF4E proteins were found to be elevated in senescent cells. These data suggest that MNK1 regulates the phosphorylation and the subcellular distribution of hnRNP A1 and that MNK1 may play a role in the induction of senescence. -- Highlights: Black-Right-Pointing-Pointer MNK1 and not MAPKAPK2 phosphorylates hnRNP A1. Black-Right-Pointing-Pointer MNK1 has elevated levels in senescent cells, this has not been reported previously. Black-Right-Pointing-Pointer MNK1 activity induces cytoplasmic accumulation of hnRNP A1 in senescent cells. Black-Right-Pointing-Pointer Altered cytoplasmic localization of hnRNP A1 may alter gene expression patterns. Black-Right-Pointing-Pointer Our studies may increase our understanding of RNA metabolism during cellular aging.

  5. CD147 and AGR2 expression promote cellular proliferation and metastasis of head and neck squamous cell carcinoma

    SciTech Connect (OSTI)

    Sweeny, Larissa; Liu, Zhiyong; Bush, Benjamin D.; Hartman, Yolanda; Zhou, Tong; Rosenthal, Eben L.

    2012-08-15

    The signaling pathways facilitating metastasis of head and neck squamous cell carcinoma (HNSCC) cells are not fully understood. CD147 is a transmembrane glycoprotein known to induce cell migration and invasion. AGR2 is a secreted peptide also known to promote cell metastasis. Here we describe their importance in the migration and invasion of HNSCC cells (FADU and OSC-19) in vitro and in vivo. In vitro, knockdown of CD147 or AGR2 decreased cellular proliferation, migration and invasion. In vivo, knockdown of CD147 or AGR2 expression decreased primary tumor growth as well as regional and distant metastasis. -- Highlights: Black-Right-Pointing-Pointer We investigated AGR2 in head and neck squamous cell carcinoma for the first time. Black-Right-Pointing-Pointer We explored the relationship between AGR2 and CD147 for the first time. Black-Right-Pointing-Pointer AGR2 and CD147 appear to co-localize in head and squamous cell carcinoma samples. Black-Right-Pointing-Pointer Knockdown of both AGR2 and CD147 reduced migration and invasion in vitro. Black-Right-Pointing-Pointer Knockdown of both AGR2 and CD147 decreased metastasis in vivo.

  6. Calcitriol inhibits Ether-a go-go potassium channel expression and cell proliferation in human breast cancer cells

    SciTech Connect (OSTI)

    Garcia-Becerra, Rocio; Diaz, Lorenza; Camacho, Javier; Barrera, David; Ordaz-Rosado, David; Morales, Angelica; Ortiz, Cindy Sharon; Avila, Euclides; Bargallo, Enrique; Arrecillas, Myrna; Halhali, Ali; Larrea, Fernando

    2010-02-01

    Antiproliferative actions of calcitriol have been shown to occur in many cell types; however, little is known regarding the molecular basis of this process in breast carcinoma. Ether-a-go-go (Eag1) potassium channels promote oncogenesis and are implicated in breast cancer cell proliferation. Since calcitriol displays antineoplastic effects while Eag1 promotes tumorigenesis, and both factors antagonically regulate cell cycle progression, we investigated a possible regulatory effect of calcitriol upon Eag1 as a mean to uncover new molecular events involved in the antiproliferative activity of this hormone in human breast tumor-derived cells. RT real-time PCR and immunocytochemistry showed that calcitriol suppressed Eag1 expression by a vitamin D receptor (VDR)-dependent mechanism. This effect was accompanied by inhibition of cell proliferation, which was potentiated by astemizole, a nonspecific Eag1 inhibitor. Immunohistochemistry and Western blot demonstrated that Eag1 and VDR abundance was higher in invasive-ductal carcinoma than in fibroadenoma, and immunoreactivity of both proteins was located in ductal epithelial cells. Our results provide evidence of a novel mechanism involved in the antiproliferative effects of calcitriol and highlight VDR as a cancer therapeutic target for breast cancer treatment and prevention.

  7. The MGGB equation-of-state for multifield applications: a numerical recipe for analytic expression of sesame EOS data

    SciTech Connect (OSTI)

    Kashiwa, B. A.

    2010-12-01

    Abstract A thermodynamically consistent and fully general equationof state (EOS) for multifield applications is described. EOS functions are derived from a Helmholtz free energy expressed as the sum of thermal (fluctuational) and collisional (condensedphase) contributions; thus the free energy is of the MieGruneisen1 form. The phasecoexistence region is defined using a parameterized saturation curve by extending the form introduced by Guggenheim,2 which scales the curve relative to conditions at the critical point. We use the zerotemperature condensedphase contribution developed by Barnes,3 which extends the ThomasFermiDirac equation to zero pressure. Thus, the functional form of the EOS could be called MGGB (for Mie GruneisenGuggenheimBarnes). Substancespecific parameters are obtained by fitting the lowdensity energy to data from the Sesame4 library; fitting the zerotemperature pressure to the Sesame cold curve; and fitting the saturation curve and latent heat to laboratory data,5 if available. When suitable coexistence data, or Sesame data, are not available, then we apply the Principle of Corresponding States.2 Thus MGGB can be thought of as a numerical recipe for rendering the tabular Sesame EOS data in an analytic form that includes a proper coexistence region, and which permits the accurate calculation of derivatives associated with compressibility, expansivity, Joule coefficient, and specific heat, all of which are required for multifield applications. 1

  8. Cloning and heterologous expression of two aryl-aldehyde dehydrogenases from the white-rot basidiomycete Phanerochaete chrysosporium

    SciTech Connect (OSTI)

    Nakamura, Tomofumi; Fukuoka Institute of Health and Environmental Sciences, 39 Mukaizano, Dazaifu-shi, Fukuoka 818-0135 ; Ichinose, Hirofumi; Wariishi, Hiroyuki; Bio-Architecture Center, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812-8581; Innovation Center for Medical Redox Navigation, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka 812-8581

    2010-04-09

    We identified two aryl-aldehyde dehydrogenase proteins (PcALDH1 and PcALDH2) from the white-rot basidiomycete Phanerochaete chrysosporium. Both PcALDHs were translationally up-regulated in response to exogenous addition of vanillin, one of the key aromatic compounds in the pathway of lignin degradation by basidiomycetes. To clarify the catalytic functions of PcALDHs, we isolated full-length cDNAs encoding these proteins and heterologously expressed the recombinant enzymes using a pET/Escherichia coli system. The open reading frames of both PcALDH1 and PcALDH2 consisted of 1503 nucleotides. The deduced amino acid sequences of both proteins showed high homologies with aryl-aldehyde dehydrogenases from other organisms and contained ten conserved domains of ALDHs. Moreover, a novel glycine-rich motif 'GxGxxxG' was located at the NAD{sup +}-binding site. The recombinant PcALDHs catalyzed dehydrogenation reactions of several aryl-aldehyde compounds, including vanillin, to their corresponding aromatic acids. These results strongly suggested that PcALDHs metabolize aryl-aldehyde compounds generated during fungal degradation of lignin and various aromatic xenobiotics.

  9. A reduced mechanism for methane and one-step rate expressions for fuel-lean catalytic combustion of small alkanes on noble metals

    SciTech Connect (OSTI)

    Deshmukh, S.R.; Vlachos, D.G.

    2007-06-15

    A reduced mechanism and a one-step rate expression for fuel-lean methane/air catalytic combustion on an Rh catalyst are proposed. These are developed from a detailed microkinetic model using a computer-aided model reduction strategy that employs reaction path analysis, sensitivity analysis, partial equilibrium analysis, and simple algebra to deduce the most abundant reaction intermediate and the rate-determining step. The mechanism and the one-step rate expression are then tested on Pt catalyst. It is found that the reaction proceeds effectively via the same mechanistic pathway on both noble metals, but the effective reaction orders differ due to the difference in the adsorption strength of oxygen. Based on the homologous series idea, the rate expression is extended to small alkanes (ethane and propane; butane is also briefly discussed) and is found to reasonably describe experimental data. Estimation of the relevant parameters in the rate expression for various fuels and catalysts using the semiempirical bond-order conservation theory, quantum mechanical density functional theory, and/or simple experiments is discussed. Finally, it is proposed that detailed microkinetic models with coverage-dependent parameters can assist in rationalizing the apparent discrepancies between experimental data from various research groups. (author)

  10. In Situ Expression of Acidic and Thermophilic Carbohydrate Active Enzymes by Filamentous Fungi (JGI Seventh Annual User Meeting 2012: Genomics of Energy and Environment)

    ScienceCinema (OSTI)

    Mosier, Annika [Stanford University

    2013-01-22

    Annika Mosier, graduate student from Stanford University presents a talk titled "In Situ Expression of Acidic and Thermophilic Carbohydrate Active Enzymes by Filamentous Fungi" at the JGI User 7th Annual Genomics of Energy & Environment Meeting on March 22, 2012 in Walnut Creek, Calif

  11. In Situ Expression of Acidic and Thermophilic Carbohydrate Active Enzymes by Filamentous Fungi (JGI Seventh Annual User Meeting 2012: Genomics of Energy and Environment)

    SciTech Connect (OSTI)

    Mosier, Annika [Stanford University] [Stanford University

    2012-03-22

    Annika Mosier, graduate student from Stanford University presents a talk titled "In Situ Expression of Acidic and Thermophilic Carbohydrate Active Enzymes by Filamentous Fungi" at the JGI User 7th Annual Genomics of Energy & Environment Meeting on March 22, 2012 in Walnut Creek, Calif

  12. The candidate tumor suppressor CST6 alters the gene expression profile of human breast carcinoma cells: Down-regulation of the potent mitogenic, motogenic, and angiogenic factor autotaxin

    SciTech Connect (OSTI)

    Song Jin; Jie Chunfa; Polk, Paula; Shridhar, Ravi; Clair, Timothy; Zhang, Jun; Yin, Lijia; Keppler, Daniel . E-mail: dkeppl@lsuhsc.edu

    2006-02-03

    We recently coined CST6 as a novel candidate tumor suppressor gene for breast cancer. CST6 indeed is expressed in the normal human breast epithelium, but little or not at all in breast carcinomas and breast cancer cell lines. Moreover, ectopic expression of CST6 in human breast cancer cells suppressed cell proliferation, migration, invasion, and orthotopic tumor growth. To obtain insights into the molecular mechanism by which CST6 exhibits its pleiotropic effects on tumor cells, we compared global gene expression profiles in mock- and CST6-transfected human MDA-MB-435S cells. Out of 12,625 transcript species, 61 showed altered expression. These included genes for extracellular matrix components, cytokines, kinases, and phosphatases, as well as several key transcription factors. TaqMan PCR assays were used to confirm the microarray data for 7 out of 11 genes. One down-regulated gene product, secreted autotaxin/lyso-phospholipase D, was of particular interest because its down-regulation by CST6 could explain most of CST6's effect on the breast cancer cells. This study thus provides First evidence that CST6 plays a role in the modulation of genes, particularly, genes that are highly relevant to breast cancer progression.

  13. Promoter sequence of 3-phosphoglycerate kinase gene 2 of lactic acid-producing fungus rhizopus oryzae and a method of expressing a gene of interest in fungal species

    DOE Patents [OSTI]

    Gao, Johnway [Richland, WA; Skeen, Rodney S [Pendleton, OR

    2003-03-04

    The present invention provides the promoter clone discovery of phosphoglycerate kinase gene 2 of a lactic acid-producing filamentous fungal strain, Rhizopus oryzae. The isolated promoter can constitutively regulate gene expression under various carbohydrate conditions. In addition, the present invention also provides a design of an integration vector for the transformation of a foreign gene in Rhizopus oryzae.

  14. Promoter sequence of 3-phosphoglycerate kinase gene 1 of lactic acid-producing fungus rhizopus oryzae and a method of expressing a gene of interest in fungal species

    DOE Patents [OSTI]

    Gao, Johnway [Richland, WA; Skeen, Rodney S [Pendleton, OR

    2002-10-15

    The present invention provides the promoter clone discovery of phosphoglycerate kinase gene 1 of a lactic acid-producing filamentous fungal strain, Rhizopus oryzae. The isolated promoter can constitutively regulate gene expression under various carbohydrate conditions. In addition, the present invention also provides a design of an integration vector for the transformation of a foreign gene in Rhizopus oryzae.

  15. Ghrelin inhibits proliferation and increases T-type Ca{sup 2+} channel expression in PC-3 human prostate carcinoma cells

    SciTech Connect (OSTI)

    Diaz-Lezama, Nundehui; Hernandez-Elvira, Mariana; Sandoval, Alejandro; Monroy, Alma; Felix, Ricardo; Monjaraz, Eduardo

    2010-12-03

    Research highlights: {yields} Ghrelin decreases prostate carcinoma PC-3 cells proliferation. {yields} Ghrelin favors apoptosis in PC-3 cells. {yields} Ghrelin increase in intracellular free Ca{sup 2+} levels in PC-3 cells. {yields} Grelin up-regulates expression of T-type Ca{sup 2+} channels in PC-3 cells. {yields} PC-3 cells express T-channels of the Ca{sub V}3.1 and Ca{sub V}3.2 subtype. -- Abstract: Ghrelin is a multifunctional peptide hormone with roles in growth hormone release, food intake and cell proliferation. With ghrelin now recognized as important in neoplastic processes, the aim of this report is to present findings from a series of in vitro studies evaluating the cellular mechanisms involved in ghrelin regulation of proliferation in the PC-3 human prostate carcinoma cells. The results showed that ghrelin significantly decreased proliferation and induced apoptosis. Consistent with a role in apoptosis, an increase in intracellular free Ca{sup 2+} levels was observed in the ghrelin-treated cells, which was accompanied by up-regulated expression of T-type voltage-gated Ca{sup 2+} channels. Interestingly, T-channel antagonists were able to prevent the effects of ghrelin on cell proliferation. These results suggest that ghrelin inhibits proliferation and may promote apoptosis by regulating T-type Ca{sup 2+} channel expression.

  16. Bisphenol A at a low concentration boosts mouse spermatogonial cell proliferation by inducing the G protein-coupled receptor 30 expression

    SciTech Connect (OSTI)

    Sheng, Zhi-Guo; Huang, Wei; Liu, Yu-Xiang; Zhu, Ben-Zhan

    2013-02-15

    Bisphenol A (BPA) is one of the most prevalent chemicals in daily-use materials, therefore, human exposure to BPA is ubiquitous. We found that low concentrations of BPA stimulate the spermatogonial GC-1 cells proliferation by G protein-coupled receptor 30 (GPR30)-mediated epidermal growth factor receptor (EGFR)-extracellular regulated kinase (ERK)-c-Fos pathway. However, through the same pathway GPR30 expression has been shown to be induced by EGF, an EGFR ligand. Thus, we want to know if low concentrations of BPA are able to induce the GPR30 expression and the possible mechanism(s) in GC-1 cells. By transient transfection with expression plasmids, 10{sup ?9} M BPA significantly transactivates the Gpr30-5?-flanking region through activating the GPR30, cGMP-dependent protein kinase (PKG), estrogen receptor-? (ER-?), and EFGR-ERK pathways. Furthermore, an activator protein-1 (AP-1) site located within this region is found to be responsible for the transactivation of BPA. Expectedly, through the same pathways, BPA significantly induces the gene and protein expression of GPR30. c-Fos is further observed to be strongly recruited to the AP-1 site in a chromatin immunoprecipitation assay and its dysfunction on the AP-1 site markedly suppresses the expression of GPR30, p-ERK1/2, p-Ser118-ER-? and cell proliferation by BPA. Our results demonstrate that a low-concentration BPA induces GPR30 expression through the GPR30-EFGR-ERK-c-Fos, ER-?, and PKG pathways, presumably boosting the cells proliferation via a regulatory loop. The present study provides a novel insight into the potential role of GPR30 in the initiation and progression of male germ cell cancer induced by environmentally relevant BPA. - Highlights: ? Low concentrations of BPA activate the PKG and GPR30-EFGR-ERK-ER-? pathways. ? Low concentrations of BPA activate the AP-1 site of Gpr30-5?-flanking region. ? Low concentrations of BPA induce the expression of GPR30 gene and protein. ? Low concentrations of BPA boost GC-1 cells proliferation via a regulatory loop.

  17. Expression of low-, intermediate-, and high-affinity IL-2 receptors on B cell lines derived from patients with undifferentiated lymphoma of Burkitt's and non-Burkitt's types

    SciTech Connect (OSTI)

    Benjamin, D.; Rosolen, A.; Wormsley, S.B.; DeBault, L.E.; Colamonici, O.R. )

    1990-08-01

    IL-2 receptors on T cells exist in at least three forms which differ in their ligand-binding affinity. The low-affinity IL-2 receptor (IL-2R) consists of the 55-kDa Tac protein (p55 alpha), the intermediate-affinity site corresponds to the 70-kDa molecule (p70 beta), and the high-affinity IL-2R consists of a noncovalent heterodimeric structure involving both p55 alpha and p70 beta. We studied 24 B cell lines (8 EBV-negative and 16 EBV-positive) for IL-2R expression in the presence or absence of the tumor promoter, teleocidin. 125I-IL-2 radioreceptor binding assays and crosslinking studies demonstrated the sole expression of p55 alpha in EBV-negative cell lines only, whereas p55 alpha present in EBV-positive cell lines was always associated with p70 beta to construct high-affinity IL-2R. p70 beta was not detected in any of the EBV-negative cell lines, but was expressed on most of the EBV-positive cell lines (13 of 16). Our data also indicate that the expression of p55 alpha and p70 beta by radiolabeling correlates with their expression in flow cytometry, and that a large excess of p55 alpha is required to construct high-affinity IL-2R. Coexpression of p55 alpha and p70 beta on human B cells contributed to constructing high-affinity IL-2R hybrid complex as shown by rapid association rate contributed by p55 alpha and slow dissociation rate by p70 beta; teleocidin's ability to induce p55 alpha on cell lines which express p70 beta only, resulting in appearance of high-affinity IL-2R; and blocking p55 alpha by anti-Tac mAb in cell lines which constitutively express high-affinity IL-2R eliminated both high- and low-affinity components. The existence of low, intermediate, and high IL-2R on human B cells bears important future implications for understanding the mechanism of IL-2 signaling and the role of IL-2 in B cell activation, proliferation, and differentiation.

  18. Context dependent reversion of tumor phenotype by connexin-43 expression in MDA-MB231 cells and MCF-7 cells: Role of ?-catenin/connexin43 association

    SciTech Connect (OSTI)

    Talhouk, Rabih S.; Fares, Mohamed-Bilal; Rahme, Gilbert J.; Hariri, Hanaa H.; Rayess, Tina; Dbouk, Hashem A.; Bazzoun, Dana; Al-Labban, Dania; El-Sabban, Marwan E.

    2013-12-10

    Connexins (Cx), gap junction (GJ) proteins, are regarded as tumor suppressors, and Cx43 expression is often down regulated in breast tumors. We assessed the effect of Cx43 over-expression in 2D and 3D cultures of two breast adenocarcinoma cell lines: MCF-7 and MDA-MB-231. While Cx43 over-expression decreased proliferation of 2D and 3D cultures of MCF-7 by 56% and 80% respectively, MDA-MB-231 growth was not altered in 2D cultures, but exhibited 35% reduction in 3D cultures. C-terminus truncated Cx43 did not alter proliferation. Untransfected MCF-7 cells formed spherical aggregates in 3D cultures, and MDA-MB-231 cells formed stellar aggregates. However, MCF-7 cells over-expressing Cx43 formed smaller sized clusters and Cx43 expressing MDA-MB-231 cells lost their stellar morphology. Extravasation ability of both MCF-7 and MDA-MB-231 cells was reduced by 60% and 30% respectively. On the other hand, silencing Cx43 in MCF10A cells, nonneoplastic human mammary cell line, increased proliferation in both 2D and 3D cultures, and disrupted acinar morphology. Although Cx43 over-expression did not affect total levels of ?-catenin, ?-catenin and ZO-2, it decreased nuclear levels of ?-catenin in 2D and 3D cultures of MCF-7 cells, and in 3D cultures of MDA-MB-231 cells. Cx43 associated at the membrane with ?-catenin, ?-catenin and ZO-2 in 2D and 3D cultures of MCF-7 cells, and only in 3D conditions in MDA-MB-231 cells. This study suggests that Cx43 exerts tumor suppressive effects in a context-dependent manner where GJ assembly with ?-catenin, ?-catenin and ZO-2 may be implicated in reducing growth rate, invasiveness, and, malignant phenotype of 2D and 3D cultures of MCF-7 cells, and 3D cultures of MDA-MB-231 cells, by sequestering ?-catenin away from nucleus. - Highlights: Cx43 over-expressing MCF-7 and MDA-MB-231 were grown in 2D and 3D cultures. Proliferation and growth morphology were affected in a context dependent manner. Extravasation ability of both MCF-7 and MDA-MB-231 cells was reduced. Cx43-mediated gap junction complex assembly correlated with observed changes. We propose that membranous Cx43 sequesters ?-catenin away from the nucleus.

  19. Aberrant, ectopic expression of VEGF and VEGF receptors 1 and 2 in malignant colonic epithelial cells. Implications for these cells growth via an autocrine mechanism

    SciTech Connect (OSTI)

    Ahluwalia, Amrita [Veterans Affairs Long Beach Healthcare System, Long Beach, CA (United States)] [Veterans Affairs Long Beach Healthcare System, Long Beach, CA (United States); Jones, Michael K. [Veterans Affairs Long Beach Healthcare System, Long Beach, CA (United States) [Veterans Affairs Long Beach Healthcare System, Long Beach, CA (United States); Department of Medicine, University of California, Irvine, CA (United States); Szabo, Sandor [Veterans Affairs Long Beach Healthcare System, Long Beach, CA (United States) [Veterans Affairs Long Beach Healthcare System, Long Beach, CA (United States); Department of Pathology, University of California, Irvine, CA (United States); Tarnawski, Andrzej S., E-mail: amrita.ahluwalia@va.gov [Veterans Affairs Long Beach Healthcare System, Long Beach, CA (United States); Department of Medicine, University of California, Irvine, CA (United States)

    2013-08-09

    Highlights: Malignant colonic epithelial cells express VEGF and its receptors. Cultured colon cancer cells secrete VEGF into the medium. Inhibition of VEGF receptor significantly decreases colon cancer cell proliferation. VEGF is critical for colon cancer cell growth. -- Abstract: Vascular endothelial growth factor A (referred to as VEGF) is implicated in colon cancer growth. Currently, the main accepted mechanism by which VEGF promotes colon cancer growth is via the stimulation of angiogenesis, which was originally postulated by late Judah Folkman. However, the cellular source of VEGF in colon cancer tissue; and, the expression of VEGF and its receptors VEGF-R1 and VEGF-R2 in colon cancer cells are not fully known and are subjects of controversy. Material and methods: We examined and quantified expression of VEGF, VEGF-R1 and VEGF-R2 in three different human colonic tissue arrays containing sections of adenocarcinoma (n = 43) and normal mucosa (n = 41). In human colon cancer cell lines HCT116 and HT29 and normal colon cell lines NCM356 and NCM460, we examined expression of VEGF, VEGF-R1 and VEGF-R2 mRNA and protein, VEGF production and secretion into the culture medium; and, the effect of a potent, selective inhibitor of VEGF receptors, AL-993, on cell proliferation. Results: Human colorectal cancer specimens had strong expression of VEGF in cancer cells and also expressed VEGF-R1 and VEGF-R2.In vitro studies showed that human colon cancer cell lines, HCT116 and HT29, but not normal colonic cell lines, express VEGF, VEGF-R1 and VEGF-R2 and secrete VEGF into the medium up to a concentration 2000 pg/ml within 48 h. Furthermore, we showed that inhibition of VEGF receptors using a specific VEGF-R inhibitor significantly reduced proliferation (by >50%) of cultured colon cancer cell lines. Conclusions: Our findings support the contention that VEGF generated by colon cancer cells stimulates their growth directly through an autocrine mechanism that is independent of its primary function in the induction of angiogenesis.

  20. Curcumin blocks interleukin (IL)-2 signaling in T-lymphocytes by inhibiting IL-2 synthesis, CD25 expression, and IL-2 receptor signaling

    SciTech Connect (OSTI)

    Forward, Nicholas A.; Conrad, David M.; Power Coombs, Melanie R.; Doucette, Carolyn D.; Furlong, Suzanne J.; Lin, Tong-Jun; Department of Pediatrics, Dalhousie University, Halifax, Nova Scotia ; Hoskin, David W.

    2011-04-22

    Highlights: {yields} Curcumin inhibits CD4{sup +} T-lymphocyte proliferation. {yields} Curcumin inhibits interleukin-2 (IL-2) synthesis and CD25 expression by CD4{sup +} T-lymphocytes. {yields} Curcumin interferes with IL-2 receptor signaling by inhibiting JAK3 and STAT5 phosphorylation. {yields} IL-2-dependent regulatory T-lymphocyte function and Foxp3 expression is downregulated by curcumin. -- Abstract: Curcumin (diferulomethane) is the principal curcuminoid in the spice tumeric and a potent inhibitor of activation-induced T-lymphocyte proliferation; however, the molecular basis of this immunosuppressive effect has not been well studied. Here we show that micromolar concentrations of curcumin inhibited DNA synthesis by mouse CD4{sup +} T-lymphocytes, as well as interleukin-2 (IL-2) and CD25 ({alpha} chain of the high affinity IL-2 receptor) expression in response to antibody-mediated cross-linking of CD3 and CD28. Curcumin acted downstream of protein kinase C activation and intracellular Ca{sup 2+} release to inhibit I{kappa}B phosphorylation, which is required for nuclear translocation of the transcription factor NF{kappa}B. In addition, IL-2-dependent DNA synthesis by mouse CTLL-2 cells, but not constitutive CD25 expression, was impaired in the presence of curcumin, which demonstrated an inhibitory effect on IL-2 receptor (IL-2R) signaling. IL-2-induced phosphorylation of STAT5A and JAK3, but not JAK1, was diminished in the presence of curcumin, indicating inhibition of critical proximal events in IL-2R signaling. In line with the inhibitory action of curcumin on IL-2R signaling, pretreatment of CD4{sup +}CD25{sup +} regulatory T-cells with curcumin downregulated suppressor function, as well as forkhead box p3 (Foxp3) expression. We conclude that curcumin inhibits IL-2 signaling by reducing available IL-2 and high affinity IL-2R, as well as interfering with IL-2R signaling.

  1. Analysis of the Salmonella regulatory network suggests involvement of SsrB and H-NS in σE-regulated SPI-2 gene expression

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Li, Jie; Overall, Christopher C.; Nakayasu, Ernesto S.; Kidwai, Afshan S.; Jones, Marcus B.; Johnson, Rudd; Nguyen, Nhu T.; McDermott, Jason E.; Ansong, Charles; Heffron, Fred; et al

    2015-02-10

    The extracytoplasmic functioning sigma factor σE is known to play an essential role for Salmonella enterica serovar Typhimurium to survive and proliferate in macrophages and mice. However, its regulatory network is not well characterized, especially during infection. Here we used microarray to identify genes regulated by σE in Salmonella grown in three conditions: a nutrient-rich condition and two others that mimic early and late intracellular infection. We found that in each condition σE regulated different sets of genes, and notably, several global regulators. When comparing nutrient-rich and infection-like conditions, large changes were observed in the expression of genes involved inmore » Salmonella pathogenesis island (SPI)-1 type-three secretion system (TTSS), SPI-2 TTSS, protein synthesis, and stress responses. In total, the expression of 58% of Salmonella genes was affected by σE in at least one of the three conditions. An important finding is that σE up-regulates SPI-2 genes, which are essential for Salmonella intracellular survival, by up-regulating SPI-2 activator ssrB expression at the early stage of infection and down-regulating SPI-2 repressor hns expression at a later stage. Moreover, σE is capable of countering the silencing of H-NS, releasing the expression of SPI-2 genes. This connection between σE and SPI-2 genes, combined with the global regulatory effect of σE, may account for the lethality of rpoE-deficient Salmonella in murine infection.« less

  2. Expression, purification, characterization and crystallization of non- and phosphorylated states of JAK2 and JAK3 kinase domain

    SciTech Connect (OSTI)

    Hall, Troii; Emmons, Thomas L.; Chrencik, Jill E.; Gormley, Jennifer A.; Weinberg, Robin A.; Leone, Joseph W.; Hirsch, Jeffrey L.; Saabye, Matthew J.; Schindler, John F.; Day, Jacqueline E.; Williams, Jennifer M.; Kiefer, James R.; Lightle, Sandra A.; Harris, Melissa S.; Guru, Siradanahalli; Fischer, H. David; Tomasselli, Alfredo G. (Pfizer)

    2012-05-29

    Janus-associated kinases (JAKs) play critical roles in cytokine signaling, and have emerged as viable therapeutic targets in inflammation and oncology related diseases. To date, targeting JAK proteins with highly selective inhibitor compounds have remained elusive. We have expressed the active kinase domains for both JAK2 and JAK3 and devised purification protocols to resolve the non-, mono- (Y1007) and diphosphorylated (Y1007 and Y1008) states of JAK2 and non- and monophosphorylated states of JAK3 (Y980). An optimal purified protein yield of 20, 29 and 69 mg per 20 L cell culture was obtained for the three JAK2 forms, respectively, and 12.2 and 2.3 mg per 10 L fermentation for the two JAK3 forms allowing detailed biochemical and biophysical studies. To monitor the purification process we developed a novel HPLC activity assay where a sequential order of phosphorylation was observed whereby the first tyrosine residue was completely phosphorylated prior to phosphorylation of the tandem tyrosine residue. A Caliper-based microfluidics assay was used to determine the kinetic parameters (K{sub m} and k{sub cat}) for each phosphorylated state, showing that monophosphorylated (Y1007) JAK2 enzyme activity increased 9-fold over that of the nonphosphorylated species, and increased an additional 6-fold for the diphosphorylated (Y1007/Y1008) species, while phosphorylation of JAK3 resulted in a negligible increase in activity. Moreover, crystal structures have been generated for each isolated state of JAK2 and JAK3 with resolutions better than 2.4 {angstrom}. The generation of these reagents has enabled kinetic and structural characterization to inform the design of potent and selective inhibitors of the JAK family.

  3. Proteomic and Functional Analysis of the Cellulase System Expressed by Postia placenta during Brown Rot of Solid Wood

    SciTech Connect (OSTI)

    Ryu, Jae San; Shary, Semarjit; Houtman, Carl J.; Panisko, Ellen A.; Korripally, Premsagar; St John, Franz J.; Crooks, Casey; Siika-aho, Matti; Magnuson, Jon K.; Hammel, Ken

    2011-11-01

    Abstract Brown rot basidiomycetes have an important ecological role in lignocellulose recycling and are notable for their rapid degradation of wood polymers via oxidative and hydrolytic mechanisms. However, most of these fungi apparently lack processive (exo-acting) cellulases, such as cellobiohydrolases, which are generally required for efficient cellulolysis. The recent sequencing of the Postia placenta genome now permits a proteomic approach to this longstanding conundrum. We grew P. placenta on solid aspen wood, extracted proteins from the biodegrading substrate, and analyzed tryptic digests by shotgun liquid chromatography-tandem mass spectrometry. Comparison of the data with the predicted P. placenta proteome revealed the presence of 34 likely glycoside hydrolases, but only four of these-two in glycoside hydrolase family 5, one in family 10, and one in family 12-have sequences that suggested possible activity on cellulose. We expressed these enzymes heterologously and determined that they all exhibited endoglucanase activity on phosphoric acid-swollen cellulose. They also slowly hydrolyzed filter paper, a more crystalline substrate, but the soluble/insoluble reducing sugar ratios they produced classify them as nonprocessive. Computer simulations indicated that these enzymes produced soluble/insoluble ratios on reduced phosphoric acid-swollen cellulose that were higher than expected for random hydrolysis, which suggests that they could possess limited exo activity, but they are at best 10-fold less processive than cellobiohydrolases. It appears likely that P. placenta employs a combination of oxidative mechanisms and endo-acting cellulases to degrade cellulose efficiently in the absence of a significant processive component.

  4. Increased expression of bHLH transcription factor E2A (TCF3) in prostate cancer promotes proliferation and confers resistance to doxorubicin induced apoptosis

    SciTech Connect (OSTI)

    Patel, Divya; Chaudhary, Jaideep; Dept. of Biological Sciences, Clark Atlanta University, 223 James P. Brawley Dr. SW, Atlanta, GA 30314

    2012-05-25

    Highlights: Black-Right-Pointing-Pointer E2A, considered as a tumor suppressor is highly expressed in prostate cancer. Black-Right-Pointing-Pointer Silencing of E2A attenuates cell proliferation and promotes apoptosis. Black-Right-Pointing-Pointer E2A regulates c-myc, Id1, Id3 and CDKN1A expression. Black-Right-Pointing-Pointer Loss of E2A promotes doxorubicin dependent apoptosis in prostate cancer cells. Black-Right-Pointing-Pointer Results suggest that E2A acts as a tumor promoter at least in prostate cancer. -- Abstract: E2A (TCF3) is a multifunctional basic helix loop helix (bHLH), transcription factor. E2A regulates transcription of target genes by homo- or heterodimerization with cell specific bHLH proteins. In general, E2A promotes cell differentiation, acts as a negative regulator of cell proliferation in normal cells and cancer cell lines and is required for normal B-cell development. Given the diverse biological pathways regulated/influenced by E2A little is known about its expression in cancer. In this study we investigated the expression of E2A in prostate cancer. Unexpectedly, E2A immuno-histochemistry demonstrated increased E2A expression in prostate cancer as compared to normal prostate. Silencing of E2A in prostate cancer cells DU145 and PC3 led to a significant reduction in proliferation due to G1 arrest that was in part mediated by increased CDKN1A(p21) and decreased Id1, Id3 and c-myc. E2A silencing in prostate cancer cell lines also resulted in increased apoptosis due to increased mitochondrial permeability and caspase 3/7 activation. Moreover, silencing of E2A increased sensitivity to doxorubicin induced apoptosis. Based on our results, we propose that E2A could be an upstream regulator of Id1 and c-Myc which are highly expressed in prostate cancer. These results for the first time demonstrate that E2A could in fact acts as a tumor promoter at least in prostate cancer.

  5. Mutant HNF-1{alpha} and mutant HNF-1{beta} identified in MODY3 and MODY5 downregulate DPP-IV gene expression in Caco-2 cells

    SciTech Connect (OSTI)

    Gu Ning [Laboratory of Metabolism, Graduate School of Human and Environmental Studies, Kyoto University, Kyoto (Japan); Laboratory of Neurochemistry, Graduate School of Human and Environmental Studies, Kyoto University, Kyoto (Japan); Adachi, Tetsuya [Department of Genomic Drug Discovery Science, Graduate School of Pharmaceutical Sciences, Kyoto University, Kyoto (Japan); Matsunaga, Tetsuro [Laboratory of Metabolism, Graduate School of Human and Environmental Studies, Kyoto University, Kyoto (Japan); Takeda, Jun [Department of Endocrinology Diabetes and Rheumatology, Graduate School of Medicine, Gifu University School of Medicine, Gifu (Japan); Tsujimoto, Gozoh [Department of Genomic Drug Discovery Science, Graduate School of Pharmaceutical Sciences, Kyoto University, Kyoto (Japan); Ishihara, Akihiko [Laboratory of Neurochemistry, Graduate School of Human and Environmental Studies, Kyoto University, Kyoto (Japan); Yasuda, Koichiro [Laboratory of Metabolism, Graduate School of Human and Environmental Studies, Kyoto University, Kyoto (Japan); Diabetic Center, Tsunashimakai-Kosei Hospital, Himeji (Japan); Tsuda, Kinsuke [Laboratory of Metabolism, Graduate School of Human and Environmental Studies, Kyoto University, Kyoto (Japan)]. E-mail: jinkan@tom.life.h.kyoto-u.ac.jp

    2006-08-04

    Dipeptidylpeptidase IV (DPP-IV) is a well-documented drug target for the treatment of type 2 diabetes. Hepatocyte nuclear factors (HNF)-1{alpha} and HNF-1{beta}, known as the causal genes of MODY3 and MODY5, respectively, have been reported to be involved in regulation of DPP-IV gene expression. But, it is not completely clear (i) that they play roles in regulation of DPP-IV gene expression, and (ii) whether DPP-IV gene activity is changed by mutant HNF-1{alpha} and mutant HNF-1{beta} in MODY3 and MODY5. To explore these questions, we investigated transactivation effects of wild HNF-1{alpha} and 13 mutant HNF-1{alpha}, as well as wild HNF-1{beta} and 2 mutant HNF-1{beta}, on DPP-IV promoter luciferase gene in Caco-2 cells by means of a transient experiment. Both wild HNF-1{alpha} and wild HNF-1{beta} significantly transactivated DPP-IV promoter, but mutant HNF-1{alpha} and mutant HNF-1{beta} exhibited low transactivation activity. Moreover, to study whether mutant HNF-1{alpha} and mutant HNF-1{beta} change endogenous DPP-IV enzyme activity, we produced four stable cell lines from Caco-2 cells, in which wild HNF-1{alpha} or wild HNF-1{beta}, or else respective dominant-negative mutant HNF-1{alpha}T539fsdelC or dominant-negative mutant HNF-1{beta}R177X, was stably expressed. We found that DPP-IV gene expression and enzyme activity were significantly increased in wild HNF-1{alpha} cells and wild HNF-1{beta} cells, whereas they decreased in HNF-1{alpha}T539fsdelC cells and HNF-1{beta}R177X cells, compared with DPP-IV gene expression and enzyme activity in Caco-2 cells. These results suggest that both wild HNF-1{alpha} and wild HNF-1{beta} have a stimulatory effect on DPP-IV gene expression, but that mutant HNF-1{alpha} and mutant HNF-1{beta} attenuate the stimulatory effect.

  6. A Method to Estimate Uncertainty in Radiometric Measurement Using the Guide to the Expression of Uncertainty in Measurement (GUM) Method; NREL (National Renewable Energy Laboratory)

    SciTech Connect (OSTI)

    Habte, A.; Sengupta, M.; Reda, I.

    2015-03-01

    Radiometric data with known and traceable uncertainty is essential for climate change studies to better understand cloud radiation interactions and the earth radiation budget. Further, adopting a known and traceable method of estimating uncertainty with respect to SI ensures that the uncertainty quoted for radiometric measurements can be compared based on documented methods of derivation.Therefore, statements about the overall measurement uncertainty can only be made on an individual basis, taking all relevant factors into account. This poster provides guidelines and recommended procedures for estimating the uncertainty in calibrations and measurements from radiometers. The approach follows the Guide to the Expression of Uncertainty in Measurement (GUM). derivation.Therefore, statements about the overall measurement uncertainty can only be made on an individual basis, taking all relevant factors into account. This poster provides guidelines and recommended procedures for estimating the uncertainty in calibrations and measurements from radiometers. The approach follows the Guide to the Expression of Uncertainty in Measurement (GUM).

  7. Compatibility issues of potential payloads for the USA/9904/B(U)F-85 RTG transportation system (RTGTS) for the 'Pluto Express' mission

    SciTech Connect (OSTI)

    Miller, Roger G.; Barklay, Chadwick D.; Howell, Edwin I.; Frazier, Timothy A.

    1997-01-10

    The specific electric power system for the 'Pluto Express' mission has yet to be specified. However, electric power will be provided by either radioisotopic thermoelectric generators (RTG), radioisotope thermophotovoltaic systems (RTPV), alkali metal thermal to electrical conversion (AMTEC) systems, radioisotope Stirling systems, or a combination of these. The selected radioisotopic power system will also be transported using the USA/9904/B(U)F-85, Radioisotope Thermoelectric Generator (RTG) Transportation System (RTGTS). As a result, all of the potential payloads present uniquely different environmental and physical configuration requirements. This paper presents the major compatibility issues of the potential payloads for the USA/9904/B(U)F-85 RTG Transportation System for the 'Pluto Express' mission.

  8. Lack of anti-tumor activity with the ?-catenin expression inhibitor EZN-3892 in the C57BL/6J Min/+ model of intestinal carcinogenesis

    SciTech Connect (OSTI)

    Hasson, Rian M.; Briggs, Alexandra; Rizvi, Hira; Carothers, Adelaide M.; Davids, Jennifer S.; Bertagnolli, Monica M.; Cho, Nancy L.

    2014-02-14

    Highlights: Wnt/?-catenin signaling is aberrantly activated in most colorectal cancers. Locked nucleic acid (LNA)-based antisense is a novel tool for cancer therapy. ?-Catenin inhibition was observed in mature intestinal tissue of LNA-treated mice. Further investigation of Wnt/?-catenin targeted therapies is warranted. - Abstract: Background: Previously, we showed that short-term inhibition of ?-catenin expression and reversal of aberrant ?-catenin subcellular localization by the selective COX-2 inhibitor celecoxib is associated with adenoma regression in the C57BL/6J Min/+ mouse. Conversly, long-term administration resulted in tumor resistance, leading us to investigate alternative methods for selective ?-catenin chemoprevention. In this study, we hypothesized that disruption of ?-catenin expression by EZN-3892, a selective locked nucleic acid (LNA)-based ?-catenin inhibitor, would counteract the tumorigenic effect of Apc loss in Min/+ adenomas while preserving normal intestinal function. Materials and methods: C57BL/6J Apc{sup +/+} wild-type (WT) and Min/+ mice were treated with the maximum tolerated dose (MTD) of EZN-3892 (30 mg/kg). Drug effect on tumor numbers, ?-catenin protein expression, and nuclear ?-catenin localization were determined. Results: Although the tumor phenotype and ?-catenin nuclear localization in Min/+ mice did not change following drug administration, we observed a decrease in ?-catenin expression levels in the mature intestinal tissue of treated Min/+ and WT mice, providing proof of principle regarding successful delivery of the LNA-based antisense vehicle. Higher doses of EZN-3892 resulted in fatal outcomes in Min/+ mice, likely due to ?-catenin ablation in the intestinal tissue and loss of function. Conclusions: Our data support the critical role of Wnt/?-catenin signaling in maintaining intestinal homeostasis and highlight the challenges of effective drug delivery to target disease without permanent toxicity to normal cellular function.

  9. Piperine activates human pregnane X receptor to induce the expression of cytochrome P450 3A4 and multidrug resistance protein 1

    SciTech Connect (OSTI)

    Wang, Yue-Ming; Lin, Wenwei; Chai, Sergio C.; Wu, Jing; Ong, Su Sien; Schuetz, Erin G.; Chen, Taosheng

    2013-10-01

    Activation of the pregnane X receptor (PXR) and subsequently its target genes, including those encoding drug transporters and metabolizing enzymes, while playing substantial roles in xenobiotic detoxification, might cause undesired drug-drug interactions. Recently, an increased awareness has been given to dietary components for potential induction of dietdrug interactions through activation of PXR. Here, we studied, whether piperine (PIP), a major component extracted from the widely-used daily spice black pepper, could induce PXR-mediated expression of cytochrome P450 3A4 (CYP3A4) and multidrug resistance protein 1 (MDR1). Our results showed that PIP activated human PXR (hPXR)-mediated CYP3A4 and MDR1 expression in human hepatocytes, intestine cells, and a mouse model; PIP activated hPXR by recruiting its coactivator SRC-1 in both cellular and cell-free systems; PIP bound to the hPXR ligand binding domain in a competitive ligand binding assay in vitro. The dichotomous effects of PIP on induction of CYP3A4 and MDR1 expression observed here and inhibition of their activity reported elsewhere challenges the potential use of PIP as a bioavailability enhancer and suggests that caution should be taken in PIP consumption during drug treatment in patients, particularly those who favor daily pepper spice or rely on certain pepper remedies. - Highlights: Piperine induces PXR-mediated CYP3A4 and MDR1 expression. Piperine activates PXR by binding to PXR and recruiting coactivator SRC-1. Piperine induces PXR activation in vivo. Caution should be taken in piperine consumption during drug treatment.

  10. Fetal and neonatal exposure to nicotine leads to augmented hepatic and circulating triglycerides in adult male offspring due to increased expression of fatty acid synthase

    SciTech Connect (OSTI)

    Ma, Noelle; Nicholson, Catherine J.; Wong, Michael; Holloway, Alison C.; Hardy, Daniel B.

    2014-02-15

    While nicotine replacement therapy is assumed to be a safer alternative to smoking during pregnancy, the long-term consequences for the offspring remain elusive. Animal studies now suggest that maternal nicotine exposure during perinatal life leads to a wide range of adverse outcomes for the offspring including increased adiposity. The focus of this study was to investigate if nicotine exposure during pregnancy and lactation leads to alterations in hepatic triglyceride synthesis. Female Wistar rats were randomly assigned to receive daily subcutaneous injections of saline (vehicle) or nicotine bitartrate (1 mg/kg/day) for two weeks prior to mating until weaning. At postnatal day 180 (PND 180), nicotine exposed offspring exhibited significantly elevated levels of circulating and hepatic triglycerides in the male offspring. This was concomitant with increased expression of fatty acid synthase (FAS), the critical hepatic enzyme in de novo triglyceride synthesis. Given that FAS is regulated by the nuclear receptor Liver X receptor (LXR?), we measured LXR? expression in both control and nicotine-exposed offspring. Nicotine exposure during pregnancy and lactation led to an increase in hepatic LXR? protein expression and enriched binding to the putative LXRE element on the FAS promoter in PND 180 male offspring. This was also associated with significantly enhanced acetylation of histone H3 [K9,14] surrounding the FAS promoter, a hallmark of chromatin activation. Collectively, these findings suggest that nicotine exposure during pregnancy and lactation leads to an increase in circulating and hepatic triglycerides long-term via changes in the transcriptional and epigenetic regulation of the hepatic lipogenic pathway. - Highlights: Our data reveals the links nicotine exposure in utero and long-term hypertriglyceridemia. It is due to nicotine-induced augmented expression of hepatic FAS and LXR? activity. Moreover, this involves nicotine-induced enhanced acetylation of histone H3 [K9,14]. This provides a mechanism for developmental origins of health and disease (DOHaD)

  11. Tris(1,3-dichloro-2-propyl) phosphate perturbs the expression of genes involved in immune response and lipid and steroid metabolism in chicken embryos

    SciTech Connect (OSTI)

    Farhat, Amani; Buick, Julie K.; Williams, Andrew; Yauk, Carole L.; O'Brien, Jason M.; Crump, Doug; Williams, Kim L.; Chiu, Suzanne; Kennedy, Sean W.

    2014-03-01

    We previously demonstrated that in ovo exposure to the flame retardant tris(1,3-dichloro-2-propyl) phosphate (TDCPP) decreased plasma thyroxine levels, reduced growth parameters, and decreased gallbladder size in chicken embryos. In the current study DNA microarrays were used to evaluate global mRNA expression in liver tissue of male chicken embryos that exhibited the above mentioned effects. Injected doses were dimethyl sulfoxide vehicle control, 7.6 or 45 ?g TDCPP/g egg. TDCPP caused significant changes in the expression of five genes at the low dose and 47 genes at the high dose (False Discovery Rate p ? 0.1, fold change ? 1.5). The gene expression analysis suggested a compromised immune function, a state of cholestatic liver/biliary fibrosis, and disrupted lipid and steroid metabolism. Circulating bile acid levels were elevated, which is an indication of liver dysfunction, and plasma cholesterol levels were reduced; however, hepatic bile acid and cholesterol levels were unaltered. Interactome analyses identified apolipoprotein E, hepatocyte nuclear factor 4 alpha, and peroxisome proliferator-activated receptor alpha as key regulatory molecules involved in the effects of TDCPP. Our results demonstrate a targeted effect of TDCPP toxicity on lipid metabolism, including cholesterol, that helps explain the aforementioned phenotypic effects, as chicken embryos are highly dependent on yolk lipids for growth and maintenance throughout development. Finally, our results are in concordance with the literature that describes TDCPP as a cancer-causing agent, since the majority of dysregulated genes were involved in cancer pathways. - Highlights: TDCPP dysregulates genes involved in immune function and lipid metabolism. A targeted effect of TDCPP toxicity on cholesterol metabolism is apparent. A state of cholestatic liver fibrosis is suggested by the expression profile. Elevated plasma bile acids suggest that TDCPP causes liver dysfunction.

  12. Apigenin suppresses migration and invasion of transformed cells through down-regulation of C-X-C chemokine receptor 4 expression

    SciTech Connect (OSTI)

    Wang, Lei; Kuang, Lisha; Hitron, John Andrew; Son, Young-Ok; Wang, Xin; Budhraja, Amit; Lee, Jeong-Chae; Pratheeshkumar, Poyil; Chen, Gang; Zhang, Zhuo; Luo, Jia; Shi, Xianglin

    2013-10-01

    Environmental exposure to arsenic is known to cause various cancers. There are some potential relationships between cell malignant transformation and C-X-C chemokine receptor type 4 (CXCR4) expressions. Metastasis, one of the major characteristics of malignantly transformed cells, contributes to the high mortality of cells. CXCR4 and its natural chemokine ligand C-X-C motif ligand 12 (CXCL12) play a critical role in metastasis. Therefore, identification of nutritional factors which are able to inhibit CXCR4 is important for protection from environmental arsenic-induced carcinogenesis and for abolishing metastasis of malignantly transformed cells. The present study demonstrates that apigenin (4?,5,7-trihydroxyflavone), a natural dietary flavonoid, suppressed CXCR4 expression in arsenic-transformed Beas-2B cells (B-AsT) and several other types of transformed/cancer cells in a dose- and time-dependent manner. Neither proteasome nor lysosome inhibitor had any effect in reducing the apigenin-induced down-regulation of CXCR4, indicating that apigenin-induced down-regulation of CXCR4 is not due to proteolytic degradation. The down-regulation of CXCR4 is mainly due to the inhibition of nuclear factor ?B (NF-?B) transcriptional activity. Apigenin also abolished migration and invasion of transformed cells induced by CXCL12. In a xenograft mouse model, apigenin down-regulated CXCR4 expression and suppressed tumor growth. Taken together, our results show that apigenin is a novel inhibitor of CXCR4 expression. This dietary flavonoid has the potential to suppress migration and invasion of transformed cells and prevent environmental arsenic-induced carcinogenesis. - Highlights: Apigenin has a potential in preventing environmental arsenic induced carcinogenesis. Apigenin suppresses CXCR4 in malignant transformed cells in vitro and in vivo. The down-regulation of CXCR4 is mainly due to inhibition of NF-?B activity.

  13. Effects of charge and surface ligand properties of nanoparticles on oxidative stress and gene expression within the gut of Daphnia magna

    SciTech Connect (OSTI)

    Dominguez, Gustavo A.; Lohse, Samuel E.; Torelli, Marco; Murphy, Catherine; Hamers, Robert J.; Orr, Galya; Klaper, Rebecca D.

    2015-05-01

    Concern has been raised regarding the current and future release of engineered nanomaterials into aquatic environments from industry and other sources. However, not all nanomaterials may cause an environ-mental impact and identifying which nanomaterials may be of greatest concern has been difficult. It is thought that the surface groups of a functionalized nanoparticles (NPs) may play a significant role in determining their interactions with aquatic organisms, but the way in which surface properties of NPs impact their toxicity in whole organisms has been minimally explored. A major point of interaction of NPs with aquatic organisms is in the gastrointestinal tract as they ingest particulates from the water column or from the sediment. The main goal of this study was to use model gold NP (AuNPs) to evaluate the potential effects of the different surfaces groups on NPs on the gut of an aquatic model organism, Daphnia magna. In this study, we exposed daphnids to a range of AuNPs concentrations and assessed the impact of AuNP exposure in the daphnid gut by measuring reactive oxygen species (ROS) production and expression of genes associated with oxidative stress and general cellular stress: glutathione S-transferase(gst), catalase (cat), heat shock protein 70 (hsp70), and metallothionein1 (mt1). We found ROS formation and gene expression were impacted by both charge and the specific surface ligand used. We detected some degree of ROS production in all NP exposures, but positively charged AuNPs induced a greater ROS response. Similarly, we observed that, compared to controls, both positively charged AuNPs and only one negatively AuNP impacted expression of genes associated with cellular stress. Finally, ligand-AuNP exposures showed a different toxicity and gene expression profile than the ligand alone, indicating a NP specific effect.

  14. Three Human Cell Types Respond to Multi-Walled Carbon Nanotubes and Titanium Dioxide Nanobelts with Cell-Specific Transcriptomic and Proteomic Expression Patterns.

    SciTech Connect (OSTI)

    Tilton, Susan C.; Karin, Norman J.; Tolic, Ana; Xie, Yumei; Lai, Xianyin; Hamilton, Raymond F.; Waters, Katrina M.; Holian, Andrij; Witzmann, Frank A.; Orr, Galya

    2014-08-01

    The growing use of engineered nanoparticles (NPs) in commercial and medical applications raises the urgent need for tools that can predict NP toxicity. Global transcriptome and proteome analyses were conducted on three human cell types, exposed to two high aspect ratio NP types, to identify patterns of expression that might indicate high versus low NP toxicity. Three cell types representing the most common routes of human exposure to NPs, including macrophage-like (THP-1), small airway epithelial and intestinal (Caco-2/HT29-MTX) cells, were exposed to TiO2 nanobelts (TiO2-NB; high toxicity) and multi-walled carbon nanotubes (MWCNT; low toxicity) at low (10 g/mL) and high (100 g/mL) concentrations for 1 and 24 h. Unique patterns of gene and protein expressions were identified for each cell type, with no differentially expressed (p < 0.05, 1.5-fold change) genes or proteins overlapping across all three cell types. While unique to each cell type, the early response was primarily independent of NP type, showing similar expression patterns in response to both TiO2-NB and MWCNT. The early response might, therefore, indicate a general response to insult. In contrast, the 24 h response was unique to each NP type. The most significantly (p < 0.05) enriched biological processes in THP-1 cells indicated TiO2-NB regulation of pathways associated with inflammation, apoptosis, cell cycle arrest, DNA replication stress and genomic instability, while MWCNT-regulated pathways indicated increased cell proliferation, DNA repair and anti-apoptosis. These two distinct sets of biological pathways might, therefore, underlie cellular responses to high and low NP toxicity, respectively.

  15. Low doses of ochratoxin A upregulate the protein expression of organic anion transporters Oat1, Oat2, Oat3 and Oat5 in rat kidney cortex

    SciTech Connect (OSTI)

    Zlender, Vilim; Breljak, Davorka; Ljubojevic, Marija; Flajs, Dubravka; Balen, Daniela; Brzica, Hrvoje; Domijan, Ana-Marija; Peraica, Maja; Fuchs, Radovan; Anzai, Naohiko; Sabolic, Ivan

    2009-09-15

    Mycotoxin ochratoxin A (OTA) is nephrotoxic in various animal species. In rodents, OTA intoxication impairs various proximal tubule (PT) functions, including secretion of p-aminohippurate (PAH), possibly via affecting the renal organic anion (OA) transporters (Oat). However, an effect of OTA on the activity/expression of specific Oats in the mammalian kidney has not been reported. In this work, male rats were gavaged various doses of OTA every 2nd day for 10 days, and in their kidneys we studied: tubule integrity by microscopy, abundance of basolateral (rOat1, rOat3) and brush-border (rOat2, rOat5) rOat proteins by immunochemical methods, and expression of rOats mRNA by RT-PCR. The OTA treatment caused: a) dose-dependent damage of the cells in S3 segments of medullary rays, b) dual effect upon rOats in PT: low doses (50-250 {mu}g OTA/kg b.m.) upregulated the abundance of all rOats, while a high dose (500 {mu}g OTA/kg b.m.) downregulated the abundance of rOat1, and c) unchanged mRNA expression for all rOats at low OTA doses, and its downregulation at high OTA dose. Changes in the expression of renal Oats were associated with enhanced OTA accumulation in tissue and excretion in urine, whereas the indicators of oxidative stress either remained unchanged (malondialdehyde, glutathione, 8-hydroxydeoxyguanosine) or became deranged (microtubules). While OTA accumulation and downregulation of rOats in the kidney are consistent with the previously reported impaired renal PAH secretion in rodents intoxicated with high OTA doses, the post-transcriptional upregulation of Oats at low OTA doses may contribute to OTA accumulation and development of nephrotoxicity.

  16. Activation/proliferation and apoptosis of bystander goat lymphocytes induced by a macrophage-tropic chimeric caprine arthritis encephalitis virus expressing SIV Nef

    SciTech Connect (OSTI)

    Bouzar, Baya Amel; Rea, Angela; Hoc-Villet, Stephanie; Garnier, Celine; Guiguen, Francois; Jin Yuhuai; Narayan, Opendra; Chebloune, Yahia . E-mail: ychebloune@kumc.edu

    2007-08-01

    Caprine arthritis encephalitis virus (CAEV) is the natural lentivirus of goats, well known for its tropism for macrophages and its inability to cause infection in lymphocytes. The viral genome lacks nef, tat, vpu and vpx coding sequences. To test the hypothesis that when nef is expressed by the viral genome, the virus became toxic for lymphocytes during replication in macrophages, we inserted the SIVsmm PBj14 nef coding sequences into the genome of CAEV thereby generating CAEV-nef. This recombinant virus is not infectious for lymphocytes but is fully replication competent in goat macrophages in which it constitutively expresses the SIV Nef. We found that goat lymphocytes cocultured with CAEV-nef-infected macrophages became activated, showing increased expression of the interleukin-2 receptor (IL-2R). Activation correlated with increased proliferation of the cells. Interestingly, a dual effect in terms of apoptosis regulation was observed in exposed goat lymphocytes. Nef was found first to induce a protection of lymphocytes from apoptosis during the first few days following exposure to infected macrophages, but later it induced increased apoptosis in the activated lymphocytes. This new recombinant virus provides a model to study the functions of Nef in the context of infection of macrophages, but in absence of infection of T lymphocytes and brings new insights into the biological effects of Nef on lymphocytes.

  17. Simbach Braunau GEPP | Open Energy Information

    Open Energy Info (EERE)

    Home Simbach Braunau GEPP General Information Name Simbach Braunau GEPP Facility Power Plant Sector Geothermal energy Location Information Location Braunau am Inn; Austria...

  18. Triangle Universities Nuclear Laboratory : 2011

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    into what is now downtown Durhams entertainment district. Located within 20 minutes walking distance or a 5 minuted drive from the Marriott Residence Inn, a stroll through the...

  19. Laurens County, South Carolina: Energy Resources | Open Energy...

    Open Energy Info (EERE)

    Clinton, South Carolina Cross Hill, South Carolina Fountain Inn, South Carolina Gray Court, South Carolina Joanna, South Carolina Laurens, South Carolina Mountville, South...

  20. NNMCAB Board Minutes: September 2003 Taos

    Broader source: Energy.gov [DOE]

    Minutes of the September 17, 2003 Board meeting at Sagebrush Inn Conference Center Presentation LANL, 2002 Environmental Surveillance Report, Lars Soholt

  1. NNMCAB Board Minutes: May 2004 Taos

    Broader source: Energy.gov [DOE]

    Minutes of the May 22, 2004 Board meeting at Sagebrush Inn Conference Center Presentation DOE, Report on the NNMCAB, Ed Wilmot

  2. Webster Parish, Louisiana: Energy Resources | Open Energy Information

    Open Energy Info (EERE)

    Climate Zone Number 3 Climate Zone Subtype A. Places in Webster Parish, Louisiana Cotton Valley, Louisiana Cullen, Louisiana Dixie Inn, Louisiana Doyline, Louisiana Dubberly,...

  3. Unlocking Plant Metabolic Diversity

    SciTech Connect (OSTI)

    Osbourn, Anne [John Innes Centre

    2014-03-19

    Anne Osbourn, John Innes Centre, UK, at the 9th Annual Genomics of Energy & Environment Meeting on March 19, 2014 in Walnut Creek, Calif.

  4. Verde Biofuels Inc | Open Energy Information

    Open Energy Info (EERE)

    Biofuels Inc Jump to: navigation, search Name: Verde Biofuels Inc Place: Fountain Inn, South Carolina Product: The company is a biodiesel producer and distributor. References:...

  5. NNMCAB Board Minutes: May 2006 Taos

    Broader source: Energy.gov [DOE]

    Minutes of the May 20, 2006 Board meeting at Sagebrush Inn Conference Center Consideration and Action on EM SSAB Chairs Letters

  6. Pegasus Energietechnik AG | Open Energy Information

    Open Energy Info (EERE)

    AG Place: Mhldorf am Inn, Germany Zip: 84453 Sector: Renewable Energy Product: Germany-based renewable energy developer assisting with projects in Europe. References:...

  7. Award Laureates | U.S. DOE Office of Science (SC)

    Office of Science (SC) Website

    McInnes Bernard Matthew Poelker Barry F. Smith Ceremony Recap 2013 Adam P. Arkin ... Moore Gustavus J. Simmons James L. Smith 1987 James W. Gordon Miklos Gyulassy ...

  8. Wreaths Across America

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Giving » Wreaths Across America Wreaths Across America Our employees can sponsor a holiday wreath for a military gravesite at the Santa Fe National Cemetery or Carlsbad Cemetery. LANS employees helped purchase 2,147 wreaths for the 2015 event. October 30, 2015 LANL employees holding holiday wreaths at cemetery Laboratory employees and community partners purchase holiday wreaths for placement on military graves at the Santa Fe National Cemetery and Carlsbad Cemetery during the annual December

  9. Gasoline and Diesel Fuel Update

    Gasoline and Diesel Fuel Update (EIA)

    Holiday Release Schedule The prices are published around 5:00 p.m. Monday (Eastern time), except on government holidays, when the data are released on Tuesday (but still represent Monday's price). Data for: Alternate Release Date Release Day Holiday October 12, 2015 October 13, 2015 Tuesday Columbus January 18, 2016 January 19, 2016 Tuesday Martin Luther King Jr. February 15, 2016 February 16, 2016 Tuesday President's May 30, 2016 May 31, 2016 Tuesday Memorial July 4, 2016 July 5, 2016 Tuesday

  10. Education Homepage | Department of Energy

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    Services » Education Homepage Education Homepage NEW! Energy Literacy Video Series NEW! Energy Literacy Video Series Read more BioenergizeME Infographic Challenge: Cellulosic Ethanol BioenergizeME Infographic Challenge: Cellulosic Ethanol Read more Find Resources for Planning Your Clean Energy Career Find Resources for Planning Your Clean Energy Career Read more How Do Holiday Lights Work? How Do Holiday Lights Work? Learn the science behind the lights that brighten the holiday season. Read

  11. Changes in expression of renal Oat1, Oat3 and Mrp2 in cisplatin-induced acute renal failure after treatment of JBP485 in rats

    SciTech Connect (OSTI)

    Liu, Tao, E-mail: liutaomedical@qq.com [Department of Clinical Pharmacology, College of Pharmacy, Dalian Medical University, 9 West Section, Lvshun South Road, Lvshunkou District, Dalian 116044 (China)] [Department of Clinical Pharmacology, College of Pharmacy, Dalian Medical University, 9 West Section, Lvshun South Road, Lvshunkou District, Dalian 116044 (China); Meng, Qiang, E-mail: mengq531@yahoo.cn [Department of Clinical Pharmacology, College of Pharmacy, Dalian Medical University, 9 West Section, Lvshun South Road, Lvshunkou District, Dalian 116044 (China) [Department of Clinical Pharmacology, College of Pharmacy, Dalian Medical University, 9 West Section, Lvshun South Road, Lvshunkou District, Dalian 116044 (China); Provincial Key Laboratory for Pharmacokinetics and Transport, Liaoning, Dalian Medical University (China); Wang, Changyuan, E-mail: wangcyuan@163.com [Department of Clinical Pharmacology, College of Pharmacy, Dalian Medical University, 9 West Section, Lvshun South Road, Lvshunkou District, Dalian 116044 (China) [Department of Clinical Pharmacology, College of Pharmacy, Dalian Medical University, 9 West Section, Lvshun South Road, Lvshunkou District, Dalian 116044 (China); Provincial Key Laboratory for Pharmacokinetics and Transport, Liaoning, Dalian Medical University (China); Liu, Qi, E-mail: llaqii@yahoo.com.cn [Department of Clinical Pharmacology, College of Pharmacy, Dalian Medical University, 9 West Section, Lvshun South Road, Lvshunkou District, Dalian 116044 (China) [Department of Clinical Pharmacology, College of Pharmacy, Dalian Medical University, 9 West Section, Lvshun South Road, Lvshunkou District, Dalian 116044 (China); Provincial Key Laboratory for Pharmacokinetics and Transport, Liaoning, Dalian Medical University (China); Guo, Xinjin, E-mail: guo.xinjin@163.com [Department of Clinical Pharmacology, College of Pharmacy, Dalian Medical University, 9 West Section, Lvshun South Road, Lvshunkou District, Dalian 116044 (China)] [Department of Clinical Pharmacology, College of Pharmacy, Dalian Medical University, 9 West Section, Lvshun South Road, Lvshunkou District, Dalian 116044 (China); Sun, Huijun, E-mail: sunhuijun@hotmail.com [Department of Clinical Pharmacology, College of Pharmacy, Dalian Medical University, 9 West Section, Lvshun South Road, Lvshunkou District, Dalian 116044 (China) [Department of Clinical Pharmacology, College of Pharmacy, Dalian Medical University, 9 West Section, Lvshun South Road, Lvshunkou District, Dalian 116044 (China); Provincial Key Laboratory for Pharmacokinetics and Transport, Liaoning, Dalian Medical University (China); Peng, Jinyong, E-mail: jinyongpeng2005@163.com [Department of Clinical Pharmacology, College of Pharmacy, Dalian Medical University, 9 West Section, Lvshun South Road, Lvshunkou District, Dalian 116044 (China) [Department of Clinical Pharmacology, College of Pharmacy, Dalian Medical University, 9 West Section, Lvshun South Road, Lvshunkou District, Dalian 116044 (China); Provincial Key Laboratory for Pharmacokinetics and Transport, Liaoning, Dalian Medical University (China); and others

    2012-11-01

    The purpose of this study is to investigate whether the effect of cyclo-trans-4-L-hydroxyprolyl-L-serine (JBP485) on acute renal failure (ARF) induced by cisplatin is related to change in expression of renal Oat1, Oat3 and Mrp2 in rats. JBP485 reduced creatinine, blood urea nitrogen (BUN) and indoxyl sulfate (IS) in plasma and malondialdehyde (MDA) in kidney, and recovered the glomerular filtration rate (GFR) and the activity of superoxide dismutase (SOD) in cisplatin-treated rats. The plasma concentration of PAH (para-aminohippurate) determined by LCMS/MS was increased markedly after intravenous administration of cisplatin, whereas cumulative urinary excretion of PAH and the uptake of PAH in kidney slices were significantly decreased. qRT-PCR and Western-blot showed a decrease in mRNA and protein of Oat1 and Oat3, an increase in mRNA and protein of Mrp2 in cisplatin-treated rats, and an increase in IS (a uremic toxin) after co-treatment with JBP485. It indicated that JBP485 promoted urinary excretion of toxins by upregulating renal Mrp2. This therefore gives in part the explanation about the mechanism by which JBP485 improves ARF induced by cisplatin in rats. -- Highlights: ? Cisplatin induces acute renal failure (ARF). ? The expression of Oat1, Oat3 and Mrp2 were changed during ARF. ? The regulated expression of Oat1, Oat3 and Mrp2 is an adaptive protected response. ? JBP485 could facilitate the adaptive protective action.

  12. Reciprocal Regulation of Hypoxia-Inducible Factor 2α and GLI1 Expression Associated With the Radioresistance of Renal Cell Carcinoma

    SciTech Connect (OSTI)

    Zhou, Jiancheng; Wu, Kaijie; Gao, Dexuan; Zhu, Guodong; Wu, Dapeng; Wang, Xinyang; Chen, Yule; Du, Yuefeng; Song, Wenbin; Ma, Zhenkun; Authement, Craig; Saha, Debabrata; Hsieh, Jer-Tsong; He, Dalin

    2014-11-15

    Purpose: Renal cell carcinoma (RCC) is often considered a radioresistant tumor, but the molecular mechanism underlying its radioresistance is poorly understood. This study explored the roles of hypoxia-inducible factor 2α (HIF2α) and sonic hedgehog (SHH)-GLI1 signaling in mediating the radioresistance of RCC cells and to unveil the interaction between these 2 signaling pathways. Methods and Materials: The activities of SHH-GLI1 signaling pathway under normoxia and hypoxia in RCC cells were examined by real-time polymerase chain reaction, Western blot, and luciferase reporter assay. The expression of HIF2α and GLI1 in RCC patients was examined by immunohistochemistry, and their correlation was analyzed. Furthermore, RCC cells were treated with HIF2α-specific shRNA (sh-HIF2α), GLI1 inhibitor GANT61, or a combination to determine the effect of ionizing radiation (IR) on RCC cells based on clonogenic assay and double-strand break repair assay. Results: RCC cells exhibited elevated SHH-GLI1 activities under hypoxia, which was mediated by HIF2α. Hypoxia induced GLI1 activation through SMO-independent pathways that could be ablated by PI3K inhibitor or MEK inhibitor. Remarkably, the SHH-GLI1 pathway also upregulated HIF2α expression in normoxia. Apparently, there was a positive correlation between HIF2α and GLI1 expression in RCC patients. The combination of sh-HIF2α and GLI1 inhibitor significantly sensitized RCC cells to IR. Conclusions: Cross-talk between the HIF2α and SHH-GLI1 pathways was demonstrated in RCC. Cotargeting these 2 pathways, significantly sensitizing RCC cells to IR, provides a novel strategy for RCC treatment.

  13. Nuclear NF-?B Expression Correlates With Outcome Among Patients With Head and Neck Squamous Cell Carcinoma Treated With Primary Chemoradiation Therapy

    SciTech Connect (OSTI)

    Balermpas, Panagiotis [Department of Radiation Therapy and Oncology, J. W. Goethe University Frankfurt am Main, Frankfurt (Germany)] [Department of Radiation Therapy and Oncology, J. W. Goethe University Frankfurt am Main, Frankfurt (Germany); Michel, Yvonne [Senckenberg Institute of Pathology, J. W. Goethe University Frankfurt am Main, Frankfurt (Germany)] [Senckenberg Institute of Pathology, J. W. Goethe University Frankfurt am Main, Frankfurt (Germany); Wagenblast, Jens [Department of Otorhinolaryngology, J. W. Goethe University Frankfurt am Main, Frankfurt (Germany)] [Department of Otorhinolaryngology, J. W. Goethe University Frankfurt am Main, Frankfurt (Germany); Seitz, Oliver [Department of Maxillofacial Surgery, J. W. Goethe University Frankfurt am Main, Frankfurt (Germany)] [Department of Maxillofacial Surgery, J. W. Goethe University Frankfurt am Main, Frankfurt (Germany); Sipek, Florian; Rdel, Franz; Rdel, Claus [Department of Radiation Therapy and Oncology, J. W. Goethe University Frankfurt am Main, Frankfurt (Germany)] [Department of Radiation Therapy and Oncology, J. W. Goethe University Frankfurt am Main, Frankfurt (Germany); Fokas, Emmanouil, E-mail: emmanouil.fokas@kgu.de [Department of Radiation Therapy and Oncology, J. W. Goethe University Frankfurt am Main, Frankfurt (Germany)] [Department of Radiation Therapy and Oncology, J. W. Goethe University Frankfurt am Main, Frankfurt (Germany)

    2013-07-15

    Background: To examine whether nuclear NF-?B expression correlates with outcome in patients with head and neck squamous cell carcinoma (HNSCC) treated with primary chemoradiation therapy (CRT). Methods and Materials: Between 2007 and 2010, 101 patients with locally advanced primary HNSCC were treated with definitive simultaneous CRT. Pretreatment biopsy specimens were analyzed for NF-?B p65 (RelA) nuclear immunoreactivity. A sample was assigned to be positive with more than 5% positive nuclear expression. The predictive relevance of NF-?B and clinicopathologic factors for overall survival (OS), progression-free survival (PFS), local progression-free survival (LPFS), and metastasis-free survival (DMFS) was examined by univariate and multivariate analysis. Results: No significant differences between the groups were observed with regard to age, sex, total radiation dose, fractionation mode, total chemotherapy applied, T stage or grading. Patients with p65 nuclear positive biopsy specimens showed significantly a higher rate of lymph node metastasis (cN2c or cN3 status, P=.034). Within a mean follow-up time of 25 months (range, 2.33-62.96 months) OS, PFS, and DMFS were significantly poorer in the p65 nuclear positive group (P=.008, P=.027, and P=.008, respectively). These correlations remained significant in multivariate analysis. Conclusion: NF-?B/p65 nuclear expression is associated with increased lymphatic and hematogenous tumor dissemination and decreased survival in HNSCC patients treated with primary CRT. Our results may foster further investigation of a predictive relevance of NF-?B/p65 and its role as a suitable target for a molecular-based targeted therapy in HNSCC cancer.

  14. This Week In Petroleum Printer-Friendly Version

    Annual Energy Outlook [U.S. Energy Information Administration (EIA)]

    America to a new winter holiday Festivus. The Airing of Grievances, one of the core traditions of Festivus, involved friends and family telling each other about all the...

  15. CASR and personnel features highlight Insider | The Ames Laboratory

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    carcinogens damage DNA. Forsling was featured for her second career as a musician and piano teacher. The issue also contained stories on Ames Laboratory holiday traditions and...

  16. Tax Credits, Rebates & Savings | Department of Energy

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    MeasuresWhole Building ENERGY STAR Sales Tax Holiday for Energy-Efficient Products Although the eligibility of some products is limited according to their sale price,...

  17. wehearthat | Jefferson Lab

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    homepage. Note: some restrictions apply, coupons do not stack on all special holiday or Drone offers. Thank you in advance for sharing these offers with your colleagues, friends...

  18. Collaborative Military Vehicle Design | GE Global Research

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Research: Holiday Shopping & Electric Vehicles IMG0475 Innovation 247: We're Always Open primusenginefeaturedimage3 GE Innovation and Manufacturing in Europe ...

  19. Blog | Department of Energy

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    and other workers at the Hanford Site donated bicycles, helmets, toys, clothes, books, diapers, and other items to 107 children in need over the holidays. January 13, 2016...

  20. This Week In Petroleum Printer-Friendly Version

    Gasoline and Diesel Fuel Update (EIA)

    seasons that were routinely plagued with holiday price spikes andor elevated prices stemming from infrastructure problems, such as occasional refinery outages. Between Memorial...

  1. Natural Gas Weekly Update

    Annual Energy Outlook [U.S. Energy Information Administration (EIA)]

    20, when traders returned to colder-than-expected weather following the Martin Luther King, Jr. holiday weekend. The upward pressure on spot prices continued through yesterday's...

  2. Natural Gas Weekly Update, Printer-Friendly Version

    Gasoline and Diesel Fuel Update (EIA)

    20, when traders returned to colder-than-expected weather following the Martin Luther King, Jr. holiday weekend. The upward pressure on spot prices continued through yesterday's...

  3. Natural Gas Weekly Update

    Annual Energy Outlook [U.S. Energy Information Administration (EIA)]

    of Tuesday's (January 21) trading session coming out of the 3-day Martin Luther King, Jr. holiday weekend, spot prices rose robustly at most market locations with...

  4. Natural Gas Weekly Update, Printer-Friendly Version

    Gasoline and Diesel Fuel Update (EIA)

    of Tuesday's (January 21) trading session coming out of the 3-day Martin Luther King, Jr. holiday weekend, spot prices rose robustly at most market locations with...

  5. Training | Advanced Photon Source

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Delivery Monday - Friday (except holidays) User's Name co APS Contact Person's Name CAT name (if applicable) APS Sector , Building (e.g., "APS Sector 16, Building 434")...

  6. Vendor Information

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    regarding holiday gifts LANL Support and Commitment to Small Businesses NSSB building Subcontractor welding for the Strategic Computing Complex cooling tower construction....

  7. Energy Savings You Can Savor

    ScienceCinema (OSTI)

    Hill, Lauria

    2013-05-29

    Lauria Hill, an Administrative Support Specialist with the office of Energy Efficiency and Renewable Energy, walks us through the intricacies of a very special holiday treat.

  8. Sandia National Laboratories: Careers: Students & Postdocs: Internship...

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Are interns compensated for employee holidays? Housing Does Sandia manage the housing process for interns? What are my transportation options for getting to Sandia from my summer...

  9. Making 3D Printed Christmas Ornaments | GE Global Research

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    high-tech manufacturing to Christmas tree decorations to ring in the holiday season using additive manufacturing techniques. An error occurred. Try watching this video on...

  10. Solara AG | Open Energy Information

    Open Energy Info (EERE)

    Place: Hamburg, Germany Zip: D-22765 Product: Distributerinstaller of small-scale photovoltaics and turbines for boats, campervans and holiday homes, as well as grid-connected...

  11. SSRL HEADLINES December 2003

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Holiday Greetings from the Director SLAC Scientific Policy Committee Fall Meeting LCLS Update Second SPPS Experimental Run Underway 2004 Workshop on Ultrafast X-ray Science...

  12. This Week In Petroleum Printer-Friendly Version

    Gasoline and Diesel Fuel Update (EIA)

    or early spring. With peak summer gasoline demand occurring between the Memorial Day and Labor Day holidays, retail gasoline prices would correspondingly peak at some point during...

  13. 3D Printed Toy | GE Global Research

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Scientists at GE Global Research get into the holiday spirit by bringing high-tech additive manufacturing techniques to Christmas tree ornaments. You Might Also Like...

  14. "Season of Giving" launches

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    to make a monetary donation, contribute non-perishable food items, or donate a frozen turkey to complete holiday meals for families in our communities. The Laboratory's annual...

  15. 2014 - 11 | Jefferson Lab

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    November 2014 Fri, 11142014 - 4:33pm Maximum Vacation Carryover Balance & Upcoming Holiday Shutdown Reminder Thu, 11132014 - 3:58pm Pressure Systems Awareness Training Offered...

  16. Demonstration Home Program-San Diego | Department of Energy

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    and lessons learned from its Home Tour event in San Diego. PDF icon California Center for Sustainable Energy presentation More Documents & Publications Leveraging Holidays and...

  17. Brazoria County, Texas: Energy Resources | Open Energy Information

    Open Energy Info (EERE)

    Texas Hillcrest, Texas Holiday Lakes, Texas Iowa Colony, Texas Jones Creek, Texas Lake Jackson, Texas Liverpool, Texas Manvel, Texas Oyster Creek, Texas Pearland, Texas Quintana,...

  18. Energy Efficiency Can Be at the Top of Your Shopping List

    Broader source: Energy.gov [DOE]

    I hope your holidays are filled with cool ways to fine-tune your life that will get you closer to the cutting edge of energy efficiency.

  19. Search | National Nuclear Security Administration

    National Nuclear Security Administration (NNSA)

    Facility history holiday honors and awards house of representatives hpc hr hrp HSEMC hydrogen i-rapter iaea icf imaging impc IND infrastructure Infrastructure & Sustainability...

  20. Natural Gas Weekly Update, Printer-Friendly Version

    Gasoline and Diesel Fuel Update (EIA)

    days during the holiday period as cold weather blanketed much of the area. .(See Temperature Map) (See Deviation Map) (Temperature map) (Temperature Deviation Map). Prices in...

  1. City of Lompoc Utilities - Residential Energy Efficient Appliance...

    Broader source: Energy.gov (indexed) [DOE]

    Rebate Amount Clothes Washer: 120 Dishwasher: 50 LED Holiday Lights: 4 - 8 Refrigerator Replacement Rebate: 144 Refrigerator Buy-Back Program: 35 Custom: 0.15watt...

  2. EERE Blog | Department of Energy

    Office of Energy Efficiency and Renewable Energy (EERE) Indexed Site

    appliances lower compared to past holiday seasons. December 16, 2015 New Hydropower, Hidden in Plain Sight The federal government's new Section 242 Hydroelectric Production...

  3. Approved for Public Release; Further Dissemination Unlimited

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    ... 1.8 billion gallons treated. The Decommissioning, Waste, Fuels, and Remediation ... the Holiday o Globally Harmonized System Status Update o Perform 360 o Safety Check o ...

  4. SSRL HEADLINES December 2002

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Wafers Using Synchrotron Radiation Holiday Greetings from the SSRL Director SPEAR3 Lehman Review Biannual Scientific Policy Committee Meeting SSRLUO-EC Meeting Latest SRI 2003...

  5. Volunteer Program

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    National VolunteerMatch Retired and Senior Volunteer Program United Way of Northern New Mexico United Way of Santa Fe County Giving Employee Giving Campaign Holiday Food Drive...

  6. Students are Hitting the Books – Instead of the Mall

    Broader source: Energy.gov [DOE]

    The Science Bowl encourages students to take up a new holiday tradition of opening their presents - and then opening their books.

  7. Appliances and Electronics | Department of Energy

    Broader source: Energy.gov (indexed) [DOE]

    appliances Home office and electronics. Follow Us followontwitter.png followonfacebook.png Appliance & Electronics Blogs Save Energy on Appliances this Holiday Season...

  8. Fermilab Legal Office - Ethics Program - Annual Reminders

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Documents FRA Contract Modification No. M009 Modification No. M024 Ethics Program Ask the Ethicist Submit a Question Contact Us Annual Reminders Reporting Fraud Holiday Parties...

  9. Fermilab Office of General Counsel - Ethics Program

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Ask the Ethicist Questions & Answers Submit a Question Annual Reminders Reporting Fraud Holiday Parties FRA CODE OF BUSINESS ETHICS AND CONDUCT PROGRAM Fermi Research...

  10. Fermilab Office of General Counsel - Ethics Program - Submit...

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Ask the Ethicist Questions & Answers Submit a Question Annual Reminders Reporting Fraud Holiday Parties Fermilab employees can complete the form below to submit a question to...

  11. BPA-2011-01733-FOIA Response

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    Sue Holden-Baker Kammy Rogers-Holiday Dennis Oster Team Members Mark Jackson Sean Egusa Doug Johnson Chuck Combs Len Morales Robert Shier ...

  12. CLIMATE OF LAS VEGAS, NEVADA ANDREW S. GORELOW

    National Nuclear Security Administration (NNSA)

    3 MISCELLANEOUS DATA HEATING/COOLING DEGREE DAYS...............................37 SNOW......................................................38 SUNSHINE...............................................39-40 CLOUDIEST MONTHS AND YEARS................................41 RELATIVE HUMIDITY.........................................42 FOG.......................................................43 PRESSURE..................................................44 HOLIDAY

  13. Transgenic mice that express the human multidrug-resistance gene in bone marrow enable a rapid identification of agents that reverse drug resistance

    SciTech Connect (OSTI)

    Mickisch, G.H.; Merlino, G.T.; Galski, H.; Gottesman, M.M.; Pastan, I. )

    1991-01-15

    The development of preclinical models for the rapid testing of agents that circumvent multidrug resistance in cancer is a high priority of research on drug resistance. A common form of multidrug resistance in human cancer results from expression of the MDR1 gene, which encodes a M{sub r} 170,000 glycoprotein that functions as a plasma membrane energy-dependent multidrug efflux pump. The authors have engineered transgenic mice that express this multidrug transporter in their bone marrow and demonstrated that these animals are resistant to leukopenia by a panel of anticancer drugs including anthracyclines, vinca alkaloids, etoposide, taxol, and actinomycin D. Differential leukocyte counts indicate that both neutrophils and lympohcytes are pretected. Drugs such as cisplatin, methotrexate, and 5-fluorouracil, which are not handled by the multidrug transporter, produce bone marrow suppression in both normal and transgenic mice. The resistance conferred by the MDR1 gene can be circumvented in a dose-dependent manner by simultaneous administration of agents previously shown to be inhibitors of the multidrug transporter in vitro, including verapamil isomers, quinidine, and quinine. They conclude that MDR1-transgenic mice provide a rapid and reliable system to determine the bioactivity of agents that reverse multidrug resistance in animals.

  14. Gradient Theory simulations of pure fluid interfaces using a generalized expression for influence parameters and a Helmholtz energy equation of state for fundamentally consistent two-phase calculations

    DOE Public Access Gateway for Energy & Science Beta (PAGES Beta)

    Dahms, Rainer N.

    2014-12-31

    The fidelity of Gradient Theory simulations depends on the accuracy of saturation properties and influence parameters, and require equations of state (EoS) which exhibit a fundamentally consistent behavior in the two-phase regime. Widely applied multi-parameter EoS, however, are generally invalid inside this region. Hence, they may not be fully suitable for application in concert with Gradient Theory despite their ability to accurately predict saturation properties. The commonly assumed temperature-dependence of pure component influence parameters usually restricts their validity to subcritical temperature regimes. This may distort predictions for general multi-component interfaces where temperatures often exceed the critical temperature of vapor phasemore » components. Then, the calculation of influence parameters is not well defined. In this paper, one of the first studies is presented in which Gradient Theory is combined with a next-generation Helmholtz energy EoS which facilitates fundamentally consistent calculations over the entire two-phase regime. Illustrated on pentafluoroethane as an example, reference simulations using this method are performed. They demonstrate the significance of such high-accuracy and fundamentally consistent calculations for the computation of interfacial properties. These reference simulations are compared to corresponding results from cubic PR EoS, widely-applied in combination with Gradient Theory, and mBWR EoS. The analysis reveals that neither of those two methods succeeds to consistently capture the qualitative distribution of obtained key thermodynamic properties in Gradient Theory. Furthermore, a generalized expression of the pure component influence parameter is presented. This development is informed by its fundamental definition based on the direct correlation function of the homogeneous fluid and by presented high-fidelity simulations of interfacial density profiles. As a result, the new model preserves the accuracy of previous temperature-dependent expressions, remains well-defined at supercritical temperatures, and is fully suitable for calculations of general multi-component two-phase interfaces.« less

  15. Genome Clone Libraries and Data from the Integrated Molecular Analysis of Genomes and their Expression (I.M.A.G.E.) Consortium

    DOE Data Explorer [Office of Scientific and Technical Information (OSTI)]

    The I.M.A.G.E. Consortium was initiated in 1993 by four academic groups on a collaborative basis after informal discussions led to a common vision of how to achieve an important goal in the study of the human genome: the Integrated Molecular Analysis of Genomes and their Expression Consortium's primary goal is to create arrayed cDNA libraries and associated bioinformatics tools, and make them publicly available to the research community. The primary organisms of interest include intensively studied mammalian species, including human, mouse, rat and non-human primate species. The Consortium has also focused on several commonly studied model organisms; as part of this effort it has arrayed cDNAs from zebrafish, and Fugu (pufferfish) as well as Xenopus laevis and X. tropicalis (frog). Utilizing high speed robotics, over nine million individual cDNA clones have been arrayed into 384-well microtiter plates, and sufficient replicas have been created to distribute copies both to sequencing centers and to a network of five distributors located worldwide. The I.M.A.G.E. Consortium represents the world's largest public cDNA collection, and works closely with the National Institutes of Health's Mammalian Gene Collection(MGC) to help it achieve its goal of creating a full-length cDNA clone for every human and mouse gene. I.M.A.G.E. is also a member of the ORFeome Collaboration, working to generate a complete set of expression-ready open reading frame clones representing each human gene. Custom informatics tools have been developed in support of these projects to better allow the research community to select clones of interest and track and collect all data deposited into public databases about those clones and their related sequences. I.M.A.G.E. clones are publicly available, free of any royalties, and may be used by anyone agreeing with the Consortium's guidelines.

  16. Gradient Theory simulations of pure fluid interfaces using a generalized expression for influence parameters and a Helmholtz energy equation of state for fundamentally consistent two-phase calculations

    SciTech Connect (OSTI)

    Dahms, Rainer N.

    2014-12-31

    The fidelity of Gradient Theory simulations depends on the accuracy of saturation properties and influence parameters, and require equations of state (EoS) which exhibit a fundamentally consistent behavior in the two-phase regime. Widely applied multi-parameter EoS, however, are generally invalid inside this region. Hence, they may not be fully suitable for application in concert with Gradient Theory despite their ability to accurately predict saturation properties. The commonly assumed temperature-dependence of pure component influence parameters usually restricts their validity to subcritical temperature regimes. This may distort predictions for general multi-component interfaces where temperatures often exceed the critical temperature of vapor phase components. Then, the calculation of influence parameters is not well defined. In this paper, one of the first studies is presented in which Gradient Theory is combined with a next-generation Helmholtz energy EoS which facilitates fundamentally consistent calculations over the entire two-phase regime. Illustrated on pentafluoroethane as an example, reference simulations using this method are performed. They demonstrate the significance of such high-accuracy and fundamentally consistent calculations for the computation of interfacial properties. These reference simulations are compared to corresponding results from cubic PR EoS, widely-applied in combination with Gradient Theory, and mBWR EoS. The analysis reveals that neither of those two methods succeeds to consistently capture the qualitative distribution of obtained key thermodynamic properties in Gradient Theory. Furthermore, a generalized expression of the pure component influence parameter is presented. This development is informed by its fundamental definition based on the direct correlation function of the homogeneous fluid and by presented high-fidelity simulations of interfacial density profiles. As a result, the new model preserves the accuracy of previous temperature-dependent expressions, remains well-defined at supercritical temperatures, and is fully suitable for calculations of general multi-component two-phase interfaces.

  17. Modifications in Dynamic Contrast-Enhanced Magnetic Resonance Imaging Parameters After ?-Particle-Emitting {sup 227}Th-trastuzumab Therapy of HER2-Expressing Ovarian Cancer Xenografts

    SciTech Connect (OSTI)

    Heyerdahl, Helen; Re, Kathrine; Brevik, Ellen Mengshoel; Dahle, Jostein

    2013-09-01

    Purpose: The purpose of this study was to investigate the effect of ?-particle-emitting {sup 227}Th-trastuzumab radioimmunotherapy on tumor vasculature to increase the knowledge about the mechanisms of action of {sup 227}Th-trastuzumab. Methods and Materials: Human HER2-expressing SKOV-3 ovarian cancer xenografts were grown bilaterally in athymic nude mice. Mice with tumor volumes 253 36 mm{sup 3} (mean SEM) were treated with a single injection of either {sup 227}Th-trastuzumab at a dose of 1000 kBq/kg body weight (treated group, n=14 tumors) or 0.9% NaCl (control group, n=10 tumors). Dynamic T1-weighted contrast-enhanced magnetic resonance imaging (DCEMRI) was used to study the effect of {sup 227}Th-trastuzumab on tumor vasculature. DCEMRI was performed before treatment and 1, 2, and 3 weeks after therapy. Tumor contrast-enhancement curves were extracted voxel by voxel and fitted to the Brix pharmacokinetic model. Pharmacokinetic parameters for the tumors that underwent radioimmunotherapy were compared with the corresponding parameters of control tumors. Results: Significant increases of k{sub ep}, the rate constant of diffusion from the extravascular extracellular space to the plasma (P<.05), and k{sub el,} the rate of clearance of contrast agent from the plasma (P<.01), were seen in the radioimmunotherapy group 2 and 3 weeks after injection, compared with the control group. The product of k{sub ep} and the amplitude parameter A, associated with increased vessel permeability and perfusion, was also significantly increased in the radioimmunotherapy group 2 and 3 weeks after injection (P<.01). Conclusions: Pharmacokinetic modeling of MRI contrast-enhancement curves evidenced significant alterations in parameters associated with increased tumor vessel permeability and tumor perfusion after {sup 227}Th-trastuzumab treatment of HER2-expressing ovarian cancer xenografts.

  18. CX-007991: Categorical Exclusion Determination

    Broader source: Energy.gov [DOE]

    Wood Pole Replacements Along the Drummond-Macks Inn, Macks Inn-Madison, and Targhee Tap 115-Kilovolt Transmission Line Right-of-ways CX(s) Applied: B1.3 Date: 02/10/2012 Location(s): Idaho Offices(s): Bonneville Power Administration

  19. CX-009709: Categorical Exclusion Determination

    Broader source: Energy.gov [DOE]

    Wood Pole Replacements Along the Drummond-Macks Inn, Macks Inn-Madison, and Swan Valley-Teton 115-kilovolt Transmission Line Rights-of-Way CX(s) Applied: B1.3 Date: 11/29/2012 Location(s): Idaho, Idaho Offices(s): Bonneville Power Administration

  20. International shipment of light weight radioisotopic heater units (LWRHU) using the USA/9516/B(U)F Mound 1 kW shipping package in support of the {open_quotes}Pluto Express{close_quotes} mission

    SciTech Connect (OSTI)

    Barklay, C.D.; Merten, C.W.

    1997-01-01

    Radioisotopes have provided heat that has been used to maintain specific operating environments within remote satellites and spacecraft. For the {open_quotes}Pluto Express{close_quotes} mission the {sup 238}PuO{sub 2} fueled light weight radioisotopic heater unit (LWRHU) will be used within the spacecraft. Since the current plan for the {open_quotes}Pluto Express{close_quotes} mission incorporates the use of a Russian launch platform for the spacecraft, the LWRHUs must be transported in an internationally certified shipping container. An internationally certified shipping package that is versatile enough to be reconfigured to transport the LWRHUs that will be required to support the {open_quotes}Pluto Express{close_quotes} mission is the Mound USA/9516/B(U)F. {copyright} {ital 1997 American Institute of Physics.}

  1. Epigenetic involvement of Alien/ESET complex in thyroid hormone-mediated repression of E2F1 gene expression and cell proliferation

    SciTech Connect (OSTI)

    Hong, Wei; College of Basic Medicine, Tianjin Medical University, 300070 Tianjin ; Li, Jinru; Wang, Bo; Chen, Linfeng; Niu, Wenyan; Yao, Zhi; Baniahmad, Aria

    2011-12-02

    Highlights: Black-Right-Pointing-Pointer Corepressor Alien interacts with histone methyltransferase ESET in vivo. Black-Right-Pointing-Pointer Alien/ESET complex is recruited to nTRE of T3-responsive gene by liganded TR{beta}1. Black-Right-Pointing-Pointer ESET-mediated H3K9 methylation is required for liganded TR{beta}1-repressed transcription. Black-Right-Pointing-Pointer ESET is involved in T3-repressed G1/S phase transition and proliferation. -- Abstract: The ligand-bound thyroid hormone receptor (TR) is known to repress via a negative TRE (nTRE) the expression of E2F1, a key transcription factor that controls the G1/S phase transition. Alien has been identified as a novel interacting factor of E2F1 and acts as a corepressor of E2F1. The detailed molecular mechanism by which Alien inhibits E2F1 gene expression remains unclear. Here, we report that the histone H3 lysine 9 (H3K9) methyltransferase (HMT) ESET is an integral component of the corepressor Alien complex and the Alien/ESET complex is recruited to both sites, the E2F1 and the nTRE site of the E2F1 gene while the recruitment to the negative thyroid hormone response element (nTRE) is induced by the ligand-bound TR{beta}1 within the E2F1 gene promoter. We show that, overexpression of ESET promotes, whereas knockdown of ESET releases, the inhibition of TR{beta}1-regulated gene transcription upon T3 stimulation; and H3K9 methylation is required for TR{beta}1-repressed transcription. Furthermore, depletion of ESET impairs thyroid hormone-repressed proliferation as well as the G1/S transition of the cell cycle. Taken together, our data indicate that ESET is involved in TR{beta}1-mediated transcription repression and provide a molecular basis of thyroid hormone-induced repression of proliferation.

  2. Employees from Throughout the Nuclear Security Enterprise Give Back this

    National Nuclear Security Administration (NNSA)

    Holiday Season | National Nuclear Security Administration Employees from Throughout the Nuclear Security Enterprise Give Back this Holiday Season | National Nuclear Security Administration Facebook Twitter Youtube Flickr RSS People Mission Managing the Stockpile Preventing Proliferation Powering the Nuclear Navy Emergency Response Recapitalizing Our Infrastructure Countering Nuclear Terrorism About Our Programs Our History Who We Are Our Leadership Our Locations Budget Our Operations Library

  3. Lux expression in eukaryotic cells

    DOE Patents [OSTI]

    Gupta, Rakesh K.; Patterson, Stacy S.; Sayler, Gary S.; Ripp, Steven A.

    2007-11-27

    The luxA, B, C, D, and E genes from Photorhabdus luminescens have been introduced into Saccharomyces cerevisiae bioluminescent yeast cells.

  4. Federal Express | Open Energy Information

    Open Energy Info (EERE)

    Type Other Relationship Partnering Center within NREL National Center for Photovoltaics Partnership Year 1996 Link to project description http:www.nrel.govnewspress...

  5. Transwest Express | Open Energy Information

    Open Energy Info (EERE)

    released; no ROD issued. Data Completion Notes Link to document library (last updated 052014): http:www.blm.govwysteninfoNEPAdocumentshddtranswestdocs.html Documents...

  6. Cloning, expression, crystallization and preliminary X-ray analysis of a putative multiple antibiotic resistance repressor protein (MarR) from Xanthomonas campestris

    SciTech Connect (OSTI)

    Tu, Zhi-Le; Li, Juo-Ning; Chin, Ko-Hsin; Chou, Chia-Cheng; Lee, Cheng-Chung; Shr, Hui-Lin; Lyu, Ping-Chiang; Gao, Fei Philip; Wang, Andrew H.-J.; Chou, Shan-Ho

    2005-07-01

    A putative repressor for the multiple antibiotic resistance operon from a plant pathogen X. campestris pv. campestris has been overexpressed in E. coli, purified and crystallized. The crystals diffracted to 2.3 with good quality. The multiple antibiotic resistance operon (marRAB) is a member of the multidrug-resistance system. When induced, this operon enhances resistance of bacteria to a variety of medically important antibiotics, causing a serious global health problem. MarR is a marR-encoded protein that represses the transcription of the marRAB operon. Through binding with salicylate and certain antibiotics, however, MarR can derepress and activate the marRAB operon. In this report, the cloning, expression, crystallization and preliminary X-ray analysis of XC1739, a putative MarR repressor protein present in the Xanthomonas campestris pv. campestris, a Gram-negative bacterium causing major worldwide disease of cruciferous crops, are described. The XC1739 crystals diffracted to a resolution of at least 1.8 . They are orthorhombic and belong to space group P2{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 39.5, b = 54.2 and c = 139.5 , respectively. They contain two molecules in the asymmetric unit from calculation of the self-rotation function.

  7. Expression, purification, crystallization and preliminary X-ray crystallographic analysis of L-lactate dehydrogenase and its H171C mutant from Bacillus subtilis

    SciTech Connect (OSTI)

    Zhang, Yanfeng; Gao, Xiaoli

    2012-08-31

    L-Lactate dehydrogenase (LDH) is an important enzyme involved in the last step of glycolysis that catalyzes the reversible conversion of pyruvate to L-lactate with the simultaneous oxidation of NADH to NAD{sup +}. In this study, wild-type LDH from Bacillus subtilis (BsLDH-WT) and the H171C mutant (BsLDH-H171C) were expressed in Escherichia coli and purified to near-homogeneity. BsLDH-WT was crystallized in the presence of fructose 1,6-bisphosphate (FBP) and NAD{sup +} and the crystal diffracted to 2.38 {angstrom} resolution. The crystal belonged to space group P3, with unit-cell parameters a = b = 171.04, c = 96.27 {angstrom}. BsLDH-H171C was also crystallized as the apoenzyme and in complex with NAD{sup +}, and data sets were collected to 2.20 and 2.49 {angstrom} resolution, respectively. Both BsLDH-H171C crystals belonged to space group P3, with unit-cell parameters a = b = 133.41, c = 99.34 {angstrom} and a = b = 133.43, c = 99.09 {angstrom}, respectively. Tetramers were observed in the asymmetric units of all three crystals.

  8. Assembly of 500,000 inter-specific catfish expressed sequence tags and large scale gene-associated marker development for whole genome association studies

    SciTech Connect (OSTI)

    Catfish Genome Consortium; Wang, Shaolin; Peatman, Eric; Abernathy, Jason; Waldbieser, Geoff; Lindquist, Erika; Richardson, Paul; Lucas, Susan; Wang, Mei; Li, Ping; Thimmapuram, Jyothi; Liu, Lei; Vullaganti, Deepika; Kucuktas, Huseyin; Murdock, Christopher; Small, Brian C; Wilson, Melanie; Liu, Hong; Jiang, Yanliang; Lee, Yoona; Chen, Fei; Lu, Jianguo; Wang, Wenqi; Xu, Peng; Somridhivej, Benjaporn; Baoprasertkul, Puttharat; Quilang, Jonas; Sha, Zhenxia; Bao, Baolong; Wang, Yaping; Wang, Qun; Takano, Tomokazu; Nandi, Samiran; Liu, Shikai; Wong, Lilian; Kaltenboeck, Ludmilla; Quiniou, Sylvie; Bengten, Eva; Miller, Norman; Trant, John; Rokhsar, Daniel; Liu, Zhanjiang

    2010-03-23

    Background-Through the Community Sequencing Program, a catfish EST sequencing project was carried out through a collaboration between the catfish research community and the Department of Energy's Joint Genome Institute. Prior to this project, only a limited EST resource from catfish was available for the purpose of SNP identification. Results-A total of 438,321 quality ESTs were generated from 8 channel catfish (Ictalurus punctatus) and 4 blue catfish (Ictalurus furcatus) libraries, bringing the number of catfish ESTs to nearly 500,000. Assembly of all catfish ESTs resulted in 45,306 contigs and 66,272 singletons. Over 35percent of the unique sequences had significant similarities to known genes, allowing the identification of 14,776 unique genes in catfish. Over 300,000 putative SNPs have been identified, of which approximately 48,000 are high-quality SNPs identified from contigs with at least four sequences and the minor allele presence of at least two sequences in the contig. The EST resource should be valuable for identification of microsatellites, genome annotation, large-scale expression analysis, and comparative genome analysis. Conclusions-This project generated a large EST resource for catfish that captured the majority of the catfish transcriptome. The parallel analysis of ESTs from two closely related Ictalurid catfishes should also provide powerful means for the evaluation of ancient and recent gene duplications, and for the development of high-density microarrays in catfish. The inter- and intra-specific SNPs identified from all catfish EST dataset assembly will greatly benefit the catfish introgression breeding program and whole genome association studies.

  9. Novel inherited mutations and variable expressivity of BRCA1 alleles, including the founder mutation 185delAG in Ashkenazi Jewish families

    SciTech Connect (OSTI)

    Friedman, L.S.; Szabo, C.I.; Ostermeyer, E.A.

    1995-12-01

    Thirty-seven families with four or more cases of breast cancer or breast and ovarian cancer were analyzed for mutations in BRCA1. Twelve different germ-line mutations, four novel and eight previously observed, were detected in 16 families. Five families of Ashkenazi Jewish descent carried the 185delAG mutation and shared the same haplotype at eight polymorphic markers spanning {approximately}850 kb at BRCA1. Expressivity of 185delAG in these families varied, from early-onset bilateral breast cancer and ovarian cancer to late-onset breast cancer without ovarian cancer. Mutation 4184delTCAA occurred independently in two families. In one family, penetrance was complete, with females developing early-onset breast cancer or ovarian cancer and the male carrier developing prostatic cancer, whereas, in the other family, penetrance was incomplete and only breast cancer occurred, diagnosed at ages 38-81 years. Two novel nonsense mutations led to the loss of mutant BRCA1 transcript in families with 10 and 6 cases of early-onset breast cancer and ovarian cancer. A 665-nt segment of the BRCA1 3{prime}-UTR and 1.3 kb of genomic sequence including the putative promoter region were invariant by single-strand conformation analysis in 13 families without coding-sequence mutations. Overall in our series, BRCA1 mutations have been detected in 26 families: 16 with positive BRCA1 lod scores, 7 with negative lod scores (reflecting multiple sporadic breast cancers), and 3 not tested for linkage. Three other families have positive lod scores for linkage to BRCA2, but 13 families without detected BRCA1 mutations have negative lod scores for both BRCA1 and BRCA2. 57 refs., 5 figs., 3 tabs.

  10. Treatment of HER2-Expressing Breast Cancer and Ovarian Cancer Cells With Alpha Particle-Emitting {sup 227}Th-Trastuzumab

    SciTech Connect (OSTI)

    Heyerdahl, Helen; Krogh, Cecilie; Borrebaek, Jorgen; Larsen, Asmund; Dahle, Jostein

    2011-02-01

    Purpose: To evaluate the cytotoxic effects of low-dose-rate alpha particle-emitting radioimmunoconjugate {sup 227}Th-p-isothiocyanato-benzyl-DOTA-trastuzumab ({sup 227}Th-trastuzumab [where DOTA is 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid]) internalized by breast and ovarian cancer cell lines in order to assess the potential of {sup 227}Th-trastuzumab as a therapeutic agent against metastatic cancers that overexpress the HER2 oncogene. Methods and Materials: Clonogenic survival and cell growth rates of breast cancer cells treated with {sup 227}Th-trastuzumab were compared with rates of cells treated with nonbinding {sup 227}Th-rituximab, cold trastuzumab, and X-radiation. Cell growth experiments were also performed with ovarian cancer cells. Cell-associated radioactivity was measured at several time points, and the mean radiation dose to cells was calculated. Results: SKBR-3 cells got 50% of the mean absorbed radiation dose from internalized activity and 50% from cell surface-bound activity, while BT-474 and SKOV-3 cells got 75% radiation dose from internalized activity and 25% from cell surface-bound activity. Incubation of breast cancer cells with 2.5 kBq/ml {sup 227}Th-trastuzumab for 1 h at 4{sup o}C, followed by washing, resulted in mean absorbed radiation doses of 2 to 2.5 Gy. A dose-dependent inhibition of cell growth and an increase in apoptosis were induced in all cell lines. Conclusions: Clinically relevant activity concentrations of {sup 227}Th-trastuzumab induced a specific cytotoxic effect in three HER2-expressing cell lines. The cytotoxic effect of {sup 227}Th-trastuzumab was higher than that of single-dose X-radiation (relative biological effectiveness = 1.2). These results warrant further studies of treatment of breast cancer and ovarian cancer with {sup 227}Th-trastuzumab.

  11. CYP2E1-dependent and leptin-mediated hepatic CD57 expression on CD8 + T cells aid progression of environment-linked nonalcoholic steatohepatitis

    SciTech Connect (OSTI)

    Seth, Ratanesh Kumar; Das, Suvarthi; Kumar, Ashutosh; Chanda, Anindya; Kadiiska, Maria B.; Michelotti, Gregory; Manautou, Jose; Diehl, Anna Mae; Chatterjee, Saurabh

    2014-01-01

    Environmental toxins induce a novel CYP2E1/leptin signaling axis in liver. This in turn activates a poorly characterized innate immune response that contributes to nonalcoholic steatohepatitis (NASH) progression. To identify the relevant subsets of T-lymphocytes in CYP2E1-dependent, environment-linked NASH, we utilized a model of diet induced obese (DIO) mice that are chronically exposed to bromodichloromethane. Mice deficient in CYP2E1, leptin (ob/ob mice), or both T and B cells (Pfp/Rag2 double knockout (KO) mice) were used to delineate the role of each of these factors in metabolic oxidative stress-induced T cell activation. Results revealed that elevated levels of lipid peroxidation, tyrosyl radical formation, mitochondrial tyrosine nitration and hepatic leptin as a consequence of metabolic oxidative stress caused increased levels of hepatic CD57, a marker of peripheral blood lymphocytes including NKT cells. CD8 + CD57 + cytotoxic T cells but not CD4 + CD57 + cells were significantly decreased in mice lacking CYP2E1 and leptin. There was a significant increase in the levels of T cell cytokines IL-2, IL-1?, and IFN-? in bromodichloromethane exposed DIO mice but not in mice that lacked CYP2E1, leptin or T and B cells. Apoptosis as evidenced by TUNEL assay and levels of cleaved caspase-3 was significantly lower in leptin and Pfp/Rag2 KO mice and highly correlated with protection from NASH. The results described above suggest that higher levels of oxidative stress-induced leptin mediated CD8 + CD57 + T cells play an important role in the development of NASH. It also provides a novel insight of immune dysregulation and may be a key biomarker in NASH. - Highlights: Metabolic oxidative stress caused increased levels of hepatic CD57 expression. CD8+ CD57+ cytotoxic T cells were decreased in mice lacking CYP2E1 and leptin. There was a significant increase in T cell cytokines in toxin-treated mice. Apoptosis was significantly lower in leptin and Pfp/Rag2 KO mice. Leptin mediated CD8+CD57+ T cells play an important role in NASH.

  12. SU-E-T-121: Dosimetric Characterization of Gafchromic Film EBT3 Using Vidar DosimetryPro Advantage RED and EPSON Expression 10000XL Scanners

    SciTech Connect (OSTI)

    Medina, L; Adrada, A; Filipuzzi, M; Garrigo, E; Venencia, C

    2014-06-01

    Purpose: The purpose of this paper is to characterize EBT3 using two types of scanner, analyzing the factors of influence of each dosimetry system. Methods: The film used in this study was GAFCHROMIC EBT3, the films were exposed at a dose range between 0Gy a 9Gy in a solid water phantom, SSD=100cm, 5cm depth and perpendicularly to the 6MV photon beam generated by a Novalis TX linear accelerator equipped with an HDMLC. A Farmer type ion chamber TN30013 (PTW) was used to determine the dose delivered to the film. The films were digitized with a scanner EPSON expression 10000XL and the VIDAR DosimetryPro Adventage RED. Software RIT113v6.1 was used for construction of the calibration curve and analysis. The film characteristics investigated were: response at different dose levels, sensitivity to orientation and side and resolution through the results of the spatial response function by analyzing a step pattern. Additionally, 20 IMRT treatment fields were measured with both scanner and compared with calculated dose using gamma index analysis (3%-3mm). Results: The OD obtained for dose level 2Gy in the orientation portrait of the film on the scanner EPSON is (0,2220,19) and for Vidar RED (0,2520,10) and landscape is for EPSON (0,2110,25) and for Vidar RED (0,2500,11) . The orientation dependence with respect to film side is about 0,09% for EPSON and about 0.03% for VIDAR. The spatial response function increase in response to the Gaussian function FWHM EPSON scanner (0.18mm) compared with VIDAR scanner function (less than 0.06mm) was observed. We analyzed 20 total plan dose distributions the number of pixels with gamma>1 (3%-3mm) was 0.7%1.2 [0.1%; 2.82%] for EBT3-VIDAR y 2%2.9 [0.2%; 3.5%] for EBT3-EPSON. Conclusion: VIDAR scanner shows better sensitivity. EBT3 film shows a different response between portrait and landscape orientation. Step pattern is better reproduce by VIDAR scanner.

  13. Compatibility issues of potential payloads for the USA/9904/B(U)F-85 RTG transportation system (RTGTS) for the {open_quotes}Pluto Express{close_quotes} mission

    SciTech Connect (OSTI)

    Miller, R.G.; Barklay, C.D.; Howell, E.I.; Frazier, T.A.

    1997-01-01

    The specific electric power system for the {open_quotes}Pluto Express{close_quotes} mission has yet to be specified. However, electric power will be provided by either radioisotopic thermoelectric generators (RTG), radioisotope thermophotovoltaic systems (RTPV), alkali metal thermal to electrical conversion (AMTEC) systems, radioisotope Stirling systems, or a combination of these. The selected radioisotopic power system will also be transported using the USA/9904/B(U)F-85, Radioisotope Thermoelectric Generator (RTG) Transportation System (RTGTS). As a result, all of the potential payloads present uniquely different environmental and physical configuration requirements. This paper presents the major compatibility issues of the potential payloads for the USA/9904/B(U)F-85 RTG Transportation System for the {open_quotes}Pluto Express{close_quotes} mission. {copyright} {ital 1997 American Institute of Physics.}

  14. Crystallographic analysis of human hemoglobin elucidates the structural basis of the potent and dual antisickling activity of pyridyl derivatives of vanillin

    SciTech Connect (OSTI)

    Abdulmalik, Osheiza; Ghatge, Mohini S.; Musayev, Faik N.; Parikh, Apurvasena; Chen, Qiukan; Yang, Jisheng; Nnamani, Ijeoma; Danso-Danquah, Richmond; Eseonu, Dorothy N.; Asakura, Toshio; Abraham, Donald J.; Venitz, Jurgen; Safo, Martin K.

    2011-11-01

    Pyridyl derivatives of vanillin increase the fraction of the more soluble oxygenated sickle hemoglobin and/or directly increase the solubility of deoxygenated sickle hemoglobin. Crystallographic analysis reveals the structural basis of the potent and dual antisickling activity of these derivatives. Vanillin has previously been studied clinically as an antisickling agent to treat sickle-cell disease. In vitro investigations with pyridyl derivatives of vanillin, including INN-312 and INN-298, showed as much as a 90-fold increase in antisickling activity compared with vanillin. The compounds preferentially bind to and modify sickle hemoglobin (Hb S) to increase the affinity of Hb for oxygen. INN-312 also led to a considerable increase in the solubility of deoxygenated Hb S under completely deoxygenated conditions. Crystallographic studies of normal human Hb with INN-312 and INN-298 showed that the compounds form Schiff-base adducts with the N-terminus of the ?-subunits to constrain the liganded (or relaxed-state) Hb conformation relative to the unliganded (or tense-state) Hb conformation. Interestingly, while INN-298 binds and directs its meta-positioned pyridine-methoxy moiety (relative to the aldehyde moiety) further down the central water cavity of the protein, that of INN-312, which is ortho to the aldehyde, extends towards the surface of the protein. These studies suggest that these compounds may act to prevent sickling of SS cells by increasing the fraction of the soluble high-affinity Hb S and/or by stereospecific inhibition of deoxygenated Hb S polymerization.

  15. In other news

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    In other news Community Connections: Your link to news and opportunities from Los Alamos National Laboratory Latest Issue:Mar. 2016 all issues All Issues » submit In other news Holiday Gift Drive; Electric Car Challenge success; national Wreaths Across America December 1, 2014 The holiday season is a good time to make a difference. The holiday season is a good time to make a difference. Contact Community Programs Director Kurt Steinhaus Email Editor Ute Haker Email Please click on the title(s)

  16. Arsenite evokes IL-6 secretion, autocrine regulation of STAT3 signaling, and miR-21 expression, processes involved in the EMT and malignant transformation of human bronchial epithelial cells

    SciTech Connect (OSTI)

    Luo, Fei; Xu, Yuan; Ling, Min; Zhao, Yue; Xu, Wenchao; Liang, Xiao; Jiang, Rongrong; Wang, Bairu; Bian, Qian; Liu, Qizhan

    2013-11-15

    Arsenite is an established human carcinogen, and arsenite-induced inflammation contributes to malignant transformation of cells, but the molecular mechanisms by which cancers are produced remain to be established. The present results showed that, evoked by arsenite, secretion of interleukin-6 (IL-6), a pro-inflammatory cytokine, led to the activation of STAT3, a transcription activator, and to increased levels of a microRNA, miR-21. Blocking IL-6 with anti-IL-6 antibody and inhibiting STAT3 activation reduced miR-21 expression. For human bronchial epithelial cells, cultured in the presence of anti-IL-6 antibody for 3 days, the arsenite-induced EMT and malignant transformation were reversed. Thus, IL-6, acting on STAT3 signaling, which up-regulates miR-21in an autocrine manner, contributes to the EMT induced by arsenite. These data define a link from inflammation to EMT in the arsenite-induced malignant transformation of HBE cells. This link, mediated through miRNAs, establishes a mechanism for arsenite-induced lung carcinogenesis. - Highlights: Arsenite evokes IL-6 secretion. IL-6 autocrine mediates STAT3 signaling and up-regulates miR-21expression. Inflammation is involved in arsenite-induced EMT.

  17. Activation of the PI3K-Akt pathway by human T cell leukemia virus type 1 (HTLV-1) oncoprotein Tax increases Bcl3 expression, which is associated with enhanced growth of HTLV-1-infected T cells

    SciTech Connect (OSTI)

    Saito, Kousuke; Saito, Mineki; Taniura, Naoko; Okuwa, Takako; Ohara, Yoshiro

    2010-08-01

    Bcl3 is a member of the I{kappa}B family that regulates genes involved in cell proliferation and apoptosis. Recent reports indicated that Bcl3 is overexpressed in HTLV-1-infected T cells via Tax-mediated transactivation, and acts as a negative regulator of viral transcription. However, the role of Bcl3 in cellular signal transduction and the growth of HTLV-1-infected T cells have not been reported. In this study, we showed that the knockdown of Bcl3 by short hairpin RNA inhibited the growth of HTLV-1-infected T cells. Although phosphatidylinositol-3 kinase (PI3K) inhibitor reduced Bcl3 expression, inactivation of glycogen synthase kinase 3 (GSK3), an effector kinase of the PI3K/Akt signaling pathway, restored Bcl3 expression in Tax-negative but not in Tax-positive T cells. Our results indicate that the overexpression of Bcl3 in HTLV-1-infected T cells is regulated not only by transcriptional but also by post-transcriptional mechanisms, and is involved in overgrowth of HTLV-1-infected T cells.

  18. HERO Ski Trip to Mt. Hood Meadows February

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    If there is enough interest, we may be able to charter a bus to drive us up and back. Stay at the Best Western Plus Hood River Inn which is just 30 miles from Mt. Hood's largest...

  19. A D O E En

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    power to enhance clean energy production A D O E En e rg y Inn ov ati o n H ub Nuclear Energy Cover photo: 3D visualizations allow a physical walk-through of the top 20% of...

  20. Center for Nanophase Materials Sciences - Newsletter

    Broader source: All U.S. Department of Energy (DOE) Office Webpages (Extended Search)

    alcohol-free facility. The Guest House is a 3 floor, 47 room, 71 bed inn (23 rooms with King beds and 24 rooms with 2 ex-long double beds). All rooms have a mini fridge and...