Science.gov

Sample records for 1-l liquid shipper

  1. Test and evaluation report for Westinghouse Hanford Company`s 1-L Liquid Shipper, Docket 95-41-7A, Type A container

    SciTech Connect

    Kelly, D.L.

    1995-11-01

    This report documents the U.S. Department of Transportation Specification 7A Type A (DOT-7A) compliance test results of the 1-L Liquid Shipper packaging. The 1-L Liquid Shipper consists of the 3M SafeSend polyethylene canister base and cap with integral sorbent material, a quad ring gasket, a plastic bag, and six foam inserts. A 1-L narrow-mouth, Teflon bottle with Tefzel lid holds contents. The inner bottle is placed inside the SafeSend container. The inner bottle content weight cannot exceed 2000 g (4.4 lb). Total bottle weight, including contents cannot exceed 2167.5 g (4.78 lb). The nominal gross weight of the assembled packaging is to be no more that 3830.5 g (8.4 lb). The approved packaging system is designed to ship Type A quantities of radioactive materials, normal form. Contents may be liquid or solid form. Liquid contents may have a specific gravity less than or equal to (<2). Solid materials are limited in weight, to include packaging, to the gross weight of the as-tested liquids and bottles.

  2. Scimitar-horned oryx (Oryx dammah) spermatozoa are functionally competent in a heterologous bovine in vitro fertilization system after cryopreservation on dry ice, in a dry shipper, or over liquid nitrogen vapor.

    PubMed

    Roth, T L; Bush, L M; Wildt, D E; Weiss, R B

    1999-02-01

    A heterologous bovine in vitro fertilization (IVF) system was used to study the functional competence of scimitar-horned oryx spermatozoa after cryopreservation. Four sperm-freezing methods were compared after dilution of ejaculates from six oryx with an equine semen extender: 1) dry ice, 2) dry shipper one-step, 3) dry shipper two-step, and 4) liquid nitrogen vapor. Post-thaw sperm motility, longevity, and acrosomal status were assessed and zona pellucida penetration, fertilization, and embryo cleavage were evaluated after coincubation of thawed oryx spermatozoa with in vitro-matured domestic cow oocytes. Sperm motility index (SMI) decreased (p < 0.05) over a 6-h period, but a high percentage (>/= 65%) of spermatozoa contained intact acrosomes in all treatments. Despite differences in sperm motility among methods, oocyte penetration, fertilization, and embryo cleavage did not differ (p >/= 0.05). However, cleavage success was < 50% across all treatments. There were positive correlations (p < 0.05; r = 0.81-0.97) between sample SMI at 3 and 6 h and fertilization, penetration, and cleavage, but no correlations (p >/= 0.05) between SMI at 0 or 1 h and IVF success. This study demonstrates that compatible heterologous gamete interaction allows thorough assessment of post-thaw sperm function in an endangered antelope. Scimitar-horned oryx spermatozoa appear relatively tolerant of varied cryopreservation methods, and preserved samples exhibit adequate post-thaw function to warrant use for assisted reproduction. PMID:9916019

  3. 7 CFR 905.161 - Repacking shipper.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... repacking shipper is a person who repacks and ships citrus fruit grown in the production area in Florida... repacked citrus fruit during a particular fiscal period of his or her intent to ship such citrus fruit... certified citrus fruit. (c) Any repacking shipper who handles citrus fruit shipped under a...

  4. 7 CFR 905.161 - Repacking shipper.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... repacking shipper is a person who repacks and ships citrus fruit grown in the production area in Florida... repacked citrus fruit during a particular fiscal period of his or her intent to ship such citrus fruit... certified citrus fruit. (c) Any repacking shipper who handles citrus fruit shipped under a...

  5. 7 CFR 905.161 - Repacking shipper.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... repacking shipper is a person who repacks and ships citrus fruit grown in the production area in Florida... repacked citrus fruit during a particular fiscal period of his or her intent to ship such citrus fruit... certified citrus fruit. (c) Any repacking shipper who handles citrus fruit shipped under a...

  6. 7 CFR 905.161 - Repacking shipper.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... repacking shipper is a person who repacks and ships citrus fruit grown in the production area in Florida... repacked citrus fruit during a particular fiscal period of his or her intent to ship such citrus fruit... certified citrus fruit. (c) Any repacking shipper who handles citrus fruit shipped under a...

  7. 7 CFR 905.161 - Repacking shipper.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... repacking shipper is a person who repacks and ships citrus fruit grown in the production area in Florida... repacked citrus fruit during a particular fiscal period of his or her intent to ship such citrus fruit... certified citrus fruit. (c) Any repacking shipper who handles citrus fruit shipped under a...

  8. 49 CFR 173.22 - Shipper's responsibility.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... SHIPMENTS AND PACKAGINGS Preparation of Hazardous Materials for Transportation § 173.22 Shipper's... offer a motor carrier any hazardous material specified in 49 CFR 385.403 unless that motor carrier holds... radioactive materials (see § 173.403), the shipper shall notify the consignee of the dates of shipment...

  9. 7 CFR 201.27 - Name of shipper or consignee.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 3 2010-01-01 2010-01-01 false Name of shipper or consignee. 201.27 Section 201.27... REGULATIONS Labeling Vegetable Seeds § 201.27 Name of shipper or consignee. The full name and address of either the shipper, or consignee, shall appear upon the label except that if the name and address of...

  10. 46 CFR 530.6 - Certification of shipper status.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 9 2011-10-01 2011-10-01 false Certification of shipper status. 530.6 Section 530.6 Shipping FEDERAL MARITIME COMMISSION REGULATIONS AFFECTING OCEAN SHIPPING IN FOREIGN COMMERCE SERVICE CONTRACTS General Provisions § 530.6 Certification of shipper status. (a) Certification. The shipper contract party shall sign and certify on...

  11. 49 CFR 377.203 - Extension of credit to shippers.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 49 Transportation 5 2013-10-01 2013-10-01 false Extension of credit to shippers. 377.203 Section... TRANSPORTATION CHARGES Extension of Credit to Shippers by Motor Common Carriers, Water Common Carriers, and Household Goods Freight Forwarders § 377.203 Extension of credit to shippers. (a) Authorization to...

  12. 49 CFR 1546.215 - Known shipper program.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 49 Transportation 9 2012-10-01 2012-10-01 false Known shipper program. 1546.215 Section 1546.215... Known shipper program. This section applies to each foreign air carrier operating a program under § 1546... carrier must have and carry out a known shipper program in accordance with its security program....

  13. 49 CFR 1544.239 - Known shipper program.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 49 Transportation 9 2012-10-01 2012-10-01 false Known shipper program. 1544.239 Section 1544.239... COMMERCIAL OPERATORS Operations § 1544.239 Known shipper program. This section applies to each aircraft... shipper program in accordance with its security program. The program must— (1) Determine the...

  14. 49 CFR 1546.215 - Known shipper program.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 49 Transportation 9 2010-10-01 2010-10-01 false Known shipper program. 1546.215 Section 1546.215... Known shipper program. This section applies to each foreign air carrier operating a program under § 1546... carrier must have and carry out a known shipper program in accordance with its security program....

  15. 49 CFR 1548.17 - Known shipper program.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 49 Transportation 9 2010-10-01 2010-10-01 false Known shipper program. 1548.17 Section 1548.17..., DEPARTMENT OF HOMELAND SECURITY CIVIL AVIATION SECURITY INDIRECT AIR CARRIER SECURITY § 1548.17 Known shipper..., each indirect air carrier must have and carry out a known shipper program in accordance with...

  16. 49 CFR 1548.17 - Known shipper program.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 49 Transportation 9 2012-10-01 2012-10-01 false Known shipper program. 1548.17 Section 1548.17..., DEPARTMENT OF HOMELAND SECURITY CIVIL AVIATION SECURITY INDIRECT AIR CARRIER SECURITY § 1548.17 Known shipper..., each indirect air carrier must have and carry out a known shipper program in accordance with...

  17. 49 CFR 1544.239 - Known shipper program.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 49 Transportation 9 2010-10-01 2010-10-01 false Known shipper program. 1544.239 Section 1544.239... COMMERCIAL OPERATORS Operations § 1544.239 Known shipper program. This section applies to each aircraft... shipper program in accordance with its security program. The program must— (1) Determine the...

  18. 49 CFR 173.22 - Shipper's responsibility.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... in 49 CFR 385.403 unless that motor carrier holds a safety permit issued by the Federal Motor Carrier... § 173.22, see the List of CFR Sections Affected which appears in the Finding Aids section of the printed... 49 Transportation 2 2010-10-01 2010-10-01 false Shipper's responsibility. 173.22 Section...

  19. 49 CFR 172.204 - Shipper's certification.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 49 Transportation 2 2011-10-01 2011-10-01 false Shipper's certification. 172.204 Section 172.204 Transportation Other Regulations Relating to Transportation PIPELINE AND HAZARDOUS MATERIALS SAFETY ADMINISTRATION, DEPARTMENT OF TRANSPORTATION HAZARDOUS MATERIALS REGULATIONS HAZARDOUS MATERIALS TABLE, SPECIAL PROVISIONS, HAZARDOUS...

  20. 7 CFR 917.115 - Nomination of shipper members for the Control Committee.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 8 2011-01-01 2011-01-01 false Nomination of shipper members for the Control... Nomination of shipper members for the Control Committee. (a) All shippers who, prior to February 1 of the... mail, of the time and place for a meeting of such shippers to elect nominees for shipper membership...

  1. 49 CFR 1546.215 - Known shipper program.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 49 Transportation 9 2014-10-01 2014-10-01 false Known shipper program. 1546.215 Section 1546.215 Transportation Other Regulations Relating to Transportation (Continued) TRANSPORTATION SECURITY ADMINISTRATION, DEPARTMENT OF HOMELAND SECURITY CIVIL AVIATION SECURITY FOREIGN AIR CARRIER SECURITY Operations § 1546.215 Known shipper program....

  2. 49 CFR 377.203 - Extension of credit to shippers.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ..., authorize discounts for early freight bill payments when credit is extended. (g)(1) Collection expense... 49 Transportation 5 2012-10-01 2012-10-01 false Extension of credit to shippers. 377.203 Section... TRANSPORTATION CHARGES Extension of Credit to Shippers by Motor Common Carriers, Water Common Carriers,...

  3. 49 CFR 377.203 - Extension of credit to shippers.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ..., authorize discounts for early freight bill payments when credit is extended. (g)(1) Collection expense... 49 Transportation 5 2011-10-01 2011-10-01 false Extension of credit to shippers. 377.203 Section... TRANSPORTATION CHARGES Extension of Credit to Shippers by Motor Common Carriers, Water Common Carriers,...

  4. 49 CFR 377.203 - Extension of credit to shippers.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ..., authorize discounts for early freight bill payments when credit is extended. (g)(1) Collection expense... 49 Transportation 5 2010-10-01 2010-10-01 false Extension of credit to shippers. 377.203 Section... TRANSPORTATION CHARGES Extension of Credit to Shippers by Motor Common Carriers, Water Common Carriers,...

  5. 49 CFR 377.203 - Extension of credit to shippers.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ..., authorize discounts for early freight bill payments when credit is extended. (g)(1) Collection expense... 49 Transportation 5 2014-10-01 2014-10-01 false Extension of credit to shippers. 377.203 Section... TRANSPORTATION CHARGES Extension of Credit to Shippers by Motor Common Carriers, Water Common Carriers,...

  6. 49 CFR 371.109 - Must I inform individual shippers which motor carriers I use?

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 49 Transportation 5 2013-10-01 2013-10-01 false Must I inform individual shippers which motor... individual shippers which motor carriers I use? (a) You must provide to each potential individual shipper who... must provide to each potential individual shipper who contacts you a statement indicating that you...

  7. 49 CFR 371.113 - May I provide individual shippers with a written estimate?

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... signed by the shipper before the shipment is loaded. (2) The household goods broker must explain the... 49 Transportation 5 2013-10-01 2013-10-01 false May I provide individual shippers with a written... shippers with a written estimate? (a) You may provide each individual shipper with an estimate...

  8. 7 CFR 201.23 - Name of shipper or consignee.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ..., Inspections, Marketing Practices), DEPARTMENT OF AGRICULTURE (CONTINUED) FEDERAL SEED ACT FEDERAL SEED ACT REGULATIONS Labeling Agricultural Seeds § 201.23 Name of shipper or consignee. The full name and address...

  9. 7 CFR 201.23 - Name of shipper or consignee.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ..., Inspections, Marketing Practices), DEPARTMENT OF AGRICULTURE (CONTINUED) FEDERAL SEED ACT FEDERAL SEED ACT REGULATIONS Labeling Agricultural Seeds § 201.23 Name of shipper or consignee. The full name and address...

  10. Experimental microbial contamination and disinfection of dry (vapour) shipper dewars designed for short-term storage and transportation of cryopreserved germplasm and other biological specimens.

    PubMed

    Bielanski, A

    2005-04-15

    Cryopreservation, storage and transport of cryopreserved germplasm without the risk of disease transmission is of great concern to animal and human health authorities. Here we report on the efficacy of microbial decontamination of the liquid nitrogen (LN) dry (vapour) shippers used for short-term storage and transportation of germplasm and other biological specimens. Dry shippers containing either a hydrophobic or a non-hydrophobic LN absorbent were experimentally contaminated with high titers of cultures of Pseudomonas aeruginosa, Escherichia coli, Staphylococus aureus, bovine viral diarrhea virus (BVDV) and bovine herpesvirus-1 (BHV-1). Biocidals with broad spectrum antimicrobial activity and gas vapours of formalin and ethylene oxide were used for disinfection of the dewars. Among the biocidals used, treatment with sodium hypochlorite solution, the quaternary ammonium-based disinfectants and peracetic acid were the most effective and useful for dry shippers with a hydrophobic LN absorbent. None of the bacterial or viral microorganisms were detected in samples of semen and embryos stored in dry shippers following their disinfection with these biocides. An application of some other disinfectants, due to their foaming properties or to the permeability of the absorbent hydrophobic membrane (HM) was not effective or may have caused irreversible damage to the LN absorbent. Gas sterilization by ethylene oxide in contrast to formalin was fully effective for both types of dry shippers. PMID:15823351

  11. 75 FR 70080 - Notice of Railroad-Shipper Transportation Advisory Council Vacancies

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-11-16

    ... Surface Transportation Board Notice of Railroad-Shipper Transportation Advisory Council Vacancies AGENCY: Surface Transportation Board (Board), DOT. ACTION: Notice of vacancies on the Railroad-Shipper Transportation Advisory Council (RSTAC) and solicitation of nominations. SUMMARY: The Board hereby gives...

  12. 49 CFR 375.213 - What information must I provide to a prospective individual shipper?

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 49 Transportation 5 2010-10-01 2010-10-01 false What information must I provide to a prospective... I provide to a prospective individual shipper? (a) When you provide the written estimate to a prospective individual shipper, you must also provide the individual shipper with a copy of Department...

  13. 14 CFR 297.6 - Foreign cooperative shippers association as agent.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 14 Aeronautics and Space 4 2012-01-01 2012-01-01 false Foreign cooperative shippers association as... (AVIATION PROCEEDINGS) ECONOMIC REGULATIONS FOREIGN AIR FREIGHT FORWARDERS AND FOREIGN COOPERATIVE SHIPPERS ASSOCIATIONS General § 297.6 Foreign cooperative shippers association as agent. A foreign cooperative...

  14. 14 CFR 297.6 - Foreign cooperative shippers association as agent.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 14 Aeronautics and Space 4 2011-01-01 2011-01-01 false Foreign cooperative shippers association as... (AVIATION PROCEEDINGS) ECONOMIC REGULATIONS FOREIGN AIR FREIGHT FORWARDERS AND FOREIGN COOPERATIVE SHIPPERS ASSOCIATIONS General § 297.6 Foreign cooperative shippers association as agent. A foreign cooperative...

  15. 14 CFR 297.6 - Foreign cooperative shippers association as agent.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 14 Aeronautics and Space 4 2013-01-01 2013-01-01 false Foreign cooperative shippers association as... (AVIATION PROCEEDINGS) ECONOMIC REGULATIONS FOREIGN AIR FREIGHT FORWARDERS AND FOREIGN COOPERATIVE SHIPPERS ASSOCIATIONS General § 297.6 Foreign cooperative shippers association as agent. A foreign cooperative...

  16. 14 CFR 297.6 - Foreign cooperative shippers association as agent.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 14 Aeronautics and Space 4 2010-01-01 2010-01-01 false Foreign cooperative shippers association as... (AVIATION PROCEEDINGS) ECONOMIC REGULATIONS FOREIGN AIR FREIGHT FORWARDERS AND FOREIGN COOPERATIVE SHIPPERS ASSOCIATIONS General § 297.6 Foreign cooperative shippers association as agent. A foreign cooperative...

  17. 49 CFR 375.517 - May an individual shipper demand re-weighing?

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 49 Transportation 5 2011-10-01 2011-10-01 false May an individual shipper demand re-weighing? 375... of Shipments of Household Goods Weighing the Shipment § 375.517 May an individual shipper demand re... individual shipper may demand a re-weigh. You must base your freight bill charges upon the re-weigh weight....

  18. 49 CFR 375.517 - May an individual shipper demand re-weighing?

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 49 Transportation 5 2010-10-01 2010-10-01 false May an individual shipper demand re-weighing? 375... of Shipments of Household Goods Weighing the Shipment § 375.517 May an individual shipper demand re... individual shipper may demand a re-weigh. You must base your freight bill charges upon the re-weigh weight....

  19. 7 CFR 917.17 - Nomination of shipper members of the Control Committee.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 8 2011-01-01 2011-01-01 false Nomination of shipper members of the Control Committee....17 Nomination of shipper members of the Control Committee. Nominations for the 12 members of the Control Committee to represent shippers shall be made in the following manner: (a) By February 1 of...

  20. 49 CFR 375.219 - May I extend credit to shippers?

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 49 Transportation 5 2013-10-01 2013-10-01 false May I extend credit to shippers? 375.219 Section... Services to My Customers Collecting Transportation Charges § 375.219 May I extend credit to shippers? You may extend credit to shippers, but, if you do, it must be in accordance with § 375.807....

  1. 49 CFR 375.517 - May an individual shipper demand re-weighing?

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... of Shipments of Household Goods Weighing the Shipment § 375.517 May an individual shipper demand re-weighing? After you inform the individual shipper of the billing weight and total charges and before... 49 Transportation 5 2013-10-01 2013-10-01 false May an individual shipper demand re-weighing?...

  2. 49 CFR 375.213 - What information must I provide to a prospective individual shipper?

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... shipment of household goods, you must furnish to your prospective individual shipper all five of the... individual shipper? 375.213 Section 375.213 Transportation Other Regulations Relating to Transportation... I provide to a prospective individual shipper? (a) When you provide the written estimate to...

  3. 7 CFR 917.17 - Nomination of shipper members of the Control Committee.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 8 2010-01-01 2010-01-01 false Nomination of shipper members of the Control Committee....17 Nomination of shipper members of the Control Committee. Nominations for the 12 members of the Control Committee to represent shippers shall be made in the following manner: (a) By February 1 of...

  4. 76 FR 64426 - Notice of Railroad-Shipper Transportation Advisory Council Vacancy

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-10-18

    ... TRANSPORTATION Surface Transportation Board Notice of Railroad-Shipper Transportation Advisory Council Vacancy AGENCY: Surface Transportation Board (Board). ACTION: Notice of vacancies on the Railroad-Shipper... of 7 vacancies on RSTAC for: (1) Representatives of 3 small shippers; (2) representatives of 2...

  5. 7 CFR 917.115 - Nomination of shipper members for the Control Committee.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... Nomination of shipper members for the Control Committee. (a) All shippers who, prior to February 1 of the... 7 Agriculture 8 2010-01-01 2010-01-01 false Nomination of shipper members for the Control Committee. 917.115 Section 917.115 Agriculture Regulations of the Department of Agriculture...

  6. 49 CFR 1544.239 - Known shipper program.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 49 Transportation 9 2014-10-01 2014-10-01 false Known shipper program. 1544.239 Section 1544.239 Transportation Other Regulations Relating to Transportation (Continued) TRANSPORTATION SECURITY ADMINISTRATION, DEPARTMENT OF HOMELAND SECURITY CIVIL AVIATION SECURITY AIRCRAFT OPERATOR SECURITY: AIR CARRIERS AND COMMERCIAL OPERATORS Operations §...

  7. 49 CFR 1544.239 - Known shipper program.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 49 Transportation 9 2011-10-01 2011-10-01 false Known shipper program. 1544.239 Section 1544.239 Transportation Other Regulations Relating to Transportation (Continued) TRANSPORTATION SECURITY ADMINISTRATION, DEPARTMENT OF HOMELAND SECURITY CIVIL AVIATION SECURITY AIRCRAFT OPERATOR SECURITY: AIR CARRIERS...

  8. 7 CFR 46.31 - Duties of shippers.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Duties of shippers. 46.31 Section 46.31 Agriculture Regulations of the Department of Agriculture AGRICULTURAL MARKETING SERVICE (Standards, Inspections, Marketing Practices), DEPARTMENT OF AGRICULTURE MARKETING OF PERISHABLE AGRICULTURAL COMMODITIES REGULATIONS...

  9. 75 FR 343 - Fresh Garlic From the People's Republic of China: Initiation of New Shipper Reviews

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-01-05

    ...: Fresh Garlic from the People's Republic of China, 59 FR 59209 (November 16, 1994) (Order). On November... International Trade Administration Fresh Garlic From the People's Republic of China: Initiation of New Shipper... shipper review of the antidumping duty order on fresh garlic from the People's Republic of China...

  10. 18 CFR 284.223 - Transportation by interstate pipelines on behalf of shippers.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... interstate pipelines on behalf of shippers. 284.223 Section 284.223 Conservation of Power and Water Resources... by Interstate Pipelines on Behalf of Others and Services by Local Distribution Companies § 284.223 Transportation by interstate pipelines on behalf of shippers. Subject to the provisions of this subpart and...

  11. 18 CFR 284.223 - Transportation by interstate pipelines on behalf of shippers.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... interstate pipelines on behalf of shippers. 284.223 Section 284.223 Conservation of Power and Water Resources... by Interstate Pipelines on Behalf of Others and Services by Local Distribution Companies § 284.223 Transportation by interstate pipelines on behalf of shippers. Subject to the provisions of this subpart and...

  12. 18 CFR 284.223 - Transportation by interstate pipelines on behalf of shippers.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... interstate pipelines on behalf of shippers. 284.223 Section 284.223 Conservation of Power and Water Resources... by Interstate Pipelines on Behalf of Others and Services by Local Distribution Companies § 284.223 Transportation by interstate pipelines on behalf of shippers. Subject to the provisions of this subpart and...

  13. 76 FR 74044 - Honey From Argentina: Final Results of Antidumping Duty New Shipper Review

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-11-30

    ... Honey From Argentina: Preliminary Results of Antidumping Duty New Shipper Review, 76 FR 54202 (August 31... Antidumping and Countervailing Duty Proceedings: Assessment of Antidumping Duties, 68 FR 23954 (May 6, 2003... International Trade Administration Honey From Argentina: Final Results of Antidumping Duty New Shipper...

  14. 18 CFR 284.223 - Transportation by interstate pipelines on behalf of shippers.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... interstate pipelines on behalf of shippers. 284.223 Section 284.223 Conservation of Power and Water Resources... by Interstate Pipelines on Behalf of Others and Services by Local Distribution Companies § 284.223 Transportation by interstate pipelines on behalf of shippers. Subject to the provisions of this subpart and...

  15. 18 CFR 284.223 - Transportation by interstate pipelines on behalf of shippers.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... interstate pipelines on behalf of shippers. 284.223 Section 284.223 Conservation of Power and Water Resources... by Interstate Pipelines on Behalf of Others and Services by Local Distribution Companies § 284.223 Transportation by interstate pipelines on behalf of shippers. Subject to the provisions of this subpart and...

  16. 14 CFR 297.6 - Foreign cooperative shippers association as agent.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 14 Aeronautics and Space 4 2014-01-01 2014-01-01 false Foreign cooperative shippers association as agent. 297.6 Section 297.6 Aeronautics and Space OFFICE OF THE SECRETARY, DEPARTMENT OF TRANSPORTATION (AVIATION PROCEEDINGS) ECONOMIC REGULATIONS FOREIGN AIR FREIGHT FORWARDERS AND FOREIGN COOPERATIVE SHIPPERS ASSOCIATIONS General § 297.6...

  17. 19 CFR 4.63 - Outward cargo declaration; shippers' export declarations.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... of the section in the Census Regulations (see 15 CFR 30.39, 30.50 through 30.57) where the particular... 19 Customs Duties 1 2010-04-01 2010-04-01 false Outward cargo declaration; shippers' export... Outward cargo declaration; shippers' export declarations. (a) No vessel shall be cleared directly for...

  18. 49 CFR 375.605 - How must I notify an individual shipper of any service delays?

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... REGULATIONS Transportation of Shipments § 375.605 How must I notify an individual shipper of any service delays? (a) When you are unable to perform either the pickup or delivery of a shipment on the dates or... 49 Transportation 5 2013-10-01 2013-10-01 false How must I notify an individual shipper of...

  19. 77 FR 5773 - Chlorinated Isocyanurates From the People's Republic of China: Initiation of New Shipper Review

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-02-06

    ... Isocyanurates From the People's Republic of China: Initiation of New Shipper Review, 76 FR 6399 (February 4... Isocyanurates from the People's Republic of China, 70 FR 36561 (June 24, 2005). On December 30, 2011, the... threshold requirements for initiation of a new shipper review for shipments of chlorinated isos from the...

  20. 15 CFR 732.5 - Steps regarding Shipper's Export Declaration or Automated Export System record, Destination...

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... Declaration or Automated Export System record, Destination Control Statements, and recordkeeping. 732.5... THE EAR § 732.5 Steps regarding Shipper's Export Declaration or Automated Export System record... Automated Export System (AES) record. Exporters or agents authorized to complete the Shipper's...

  1. 15 CFR 732.5 - Steps regarding Shipper's Export Declaration or Automated Export System record, Destination...

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... Declaration or Automated Export System record, Destination Control Statements, and recordkeeping. 732.5... THE EAR § 732.5 Steps regarding Shipper's Export Declaration or Automated Export System record... Automated Export System (AES) record. Exporters or agents authorized to complete the Shipper's...

  2. 15 CFR 732.5 - Steps regarding Shipper's Export Declaration or Automated Export System record, Destination...

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... Declaration or Automated Export System record, Destination Control Statements, and recordkeeping. 732.5... THE EAR § 732.5 Steps regarding Shipper's Export Declaration or Automated Export System record... Automated Export System (AES) record. Exporters or agents authorized to complete the Shipper's...

  3. 15 CFR 732.5 - Steps regarding Shipper's Export Declaration or Automated Export System record, Destination...

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... Declaration or Automated Export System record, Destination Control Statements, and recordkeeping. 732.5... THE EAR § 732.5 Steps regarding Shipper's Export Declaration or Automated Export System record... Automated Export System (AES) record. Exporters or agents authorized to complete the Shipper's...

  4. 15 CFR 732.5 - Steps regarding Shipper's Export Declaration or Automated Export System record, Destination...

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... Declaration or Automated Export System record, Destination Control Statements, and recordkeeping. 732.5... THE EAR § 732.5 Steps regarding Shipper's Export Declaration or Automated Export System record... Automated Export System (AES) record. Exporters or agents authorized to complete the Shipper's...

  5. Shippers and freight operators perceptions of sustainable initiatives.

    PubMed

    Vieira, José Geraldo Vidal; Mendes, Juliana Veiga; Suyama, Suzi Sanae

    2016-02-01

    The purpose of this paper is to document the extent to which Brazilian companies in retail channels have committed to adopting environmental sustainability measures, goals and strategies for waste disposal in the evaluation and planning of sustainable transportation. This paper also aims to examine the different viewpoints of sustainability issues and the preventive actions taken by companies in terms of controlling carbon dioxide emissions and proper disposal of tires, lubricant oils, accessories and spare parts. Finally, taking the perspective of these companies, this paper examines their difficulties in meeting environmental regulations. The research involved a survey completed by 185 representatives of different types of companies, including shippers (represented by manufacturers), LSPs (logistics service providers) and carriers. The non-linear canonical correlation was calculated to verify the opinions of these representatives from different companies regarding issues that impact on the environment, the preventive actions they adopt to reduce their environmental impact and their difficulties in meeting environmental regulations. The results show that shippers and LSPs have the same perceptions regarding these sustainability issues and preventive actions. In addition, the companies perceive high costs and the lack of training for their partners as the major challenges experienced in addressing these issues and undertaking preventive actions. Therefore they need to plan their transportation activities. PMID:26227446

  6. 75 FR 22742 - New Pneumatic Off-the-Road Tires From the People's Republic of China: Rescission of New Shipper...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-04-30

    ... New Shipper Review, 74 FR 56575 (``Initiation Notice''). On March 22, 2009, OGF withdrew its request... Antidumping Duty New Shipper Review, 74 FR 31911 (July 6, 2009) (rescinding new shipper review after 60-day... International Trade Administration New Pneumatic Off-the-Road Tires From the People's Republic of...

  7. 49 CFR 371.117 - Must I provide individual shippers with my policies concerning cancellation, deposits, and refunds?

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... individual shipper's request to cancel a shipment and the disposition of each request for a period of three... 49 Transportation 5 2013-10-01 2013-10-01 false Must I provide individual shippers with my... Brokers § 371.117 Must I provide individual shippers with my policies concerning cancellation,...

  8. 49 CFR 375.203 - What actions of an individual shipper may limit or reduce my normal liability?

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 49 Transportation 5 2013-10-01 2013-10-01 false What actions of an individual shipper may limit or... actions of an individual shipper may limit or reduce my normal liability? (a) If an individual shipper includes perishable, dangerous, or hazardous articles in the shipment without your knowledge, you need...

  9. 49 CFR 375.609 - What must I do for shippers who store household goods in transit?

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... REGULATIONS Transportation of Shipments § 375.609 What must I do for shippers who store household goods in... 49 Transportation 5 2013-10-01 2013-10-01 false What must I do for shippers who store household... expire, you must comply with this section. (b) You must notify the individual shipper, in writing of...

  10. 75 FR 36635 - Certain Frozen Warmwater Shrimp From the Socialist Republic of Vietnam: Rescission of New Shipper...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-06-28

    ... Socialist Republic of Vietnam, 70 FR 5152 (February 1, 2005) (``Shrimp Order''). We announced our... Shipper Review, 75 FR 3446 (January 21, 2010) (``Preliminary Rescission''). We have analyzed the comments... of Antidumping Duty New Shipper Review, 75 FR 20563 (April 20, 2010). On May 18, 2010, the...

  11. 9 CFR 201.39 - Payment to be made to consignor or shipper by market agencies; exceptions.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... shipper by market agencies; exceptions. 201.39 Section 201.39 Animals and Animal Products GRAIN INSPECTION... shipper by market agencies; exceptions. (a) No market agency shall, except as provided in paragraph (b) of... from the Secretary or a court of competent jurisdiction, unless (1) such market agency has reason...

  12. 75 FR 38080 - Certain Steel Nails From the People's Republic of China: Rescission of New Shipper Review

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-07-01

    ... People's Republic of China: Initiation of Antidumping Duty New Shipper Review, 74 FR 48907 (September 25... Republic of China: Notice of Rescission of Antidumping Duty New Shipper Review, 74 FR 31911 (July 6, 2009.... DATES: Effective Date: July 1, 2010. SUMMARY: In response to a request from Maanshan Leader...

  13. 75 FR 34425 - Certain Steel Nails from the People's Republic of China: Final Results of the First New Shipper...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-06-17

    ...''). See Notice of Antidumping Duty Order: Certain Steel Nails From the People's Republic of China, 73 FR... from the People's Republic of China: Notice of Preliminary Results of the New Shipper Review, 75 FR... Final Results of the First New Shipper Review, 75 FR 14423 (March 25, 2010). On January 13, 2010,...

  14. 75 FR 5764 - Honey from the People's Republic of China: Initiation of New Shipper Antidumping Duty Reviews

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-02-04

    ... at Less Than Fair Value and Antidumping Duty Order; Honey from the People's Republic of China, 66 FR... International Trade Administration Honey from the People's Republic of China: Initiation of New Shipper... determined that two requests for new shipper reviews (``NSRs'') of the antidumping duty order on honey...

  15. 75 FR 55307 - Honey From the People's Republic of China: Preliminary Intent To Rescind New Shipper Reviews

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-09-10

    ...: Initiation of New Shipper Antidumping Duty Reviews, 75 FR 5764 (February 4, 2010). On February 4, 2010, the... Results for New Shipper Review, 75 FR 38980 (July 7, 2010). Expansion of the POR When the sale of the... 19 CFR 351.214(f)(2)(ii). \\4\\ See Antidumping Duties; Countervailing Duties; Final Rule, 62 FR...

  16. 75 FR 61425 - Raw Flexible Magnets from the People's Republic of China: Rescission of New Shipper Review

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-10-05

    ... Antidumping Duty New Shipper Review, 75 FR 22740 (April 30, 2010). On August 27, 2010, Jingzhou Meihou... Nails From the People's Republic of China: Rescission of New Shipper Review, 75 FR 38080 (July 1, 2010... International Trade Administration Raw Flexible Magnets from the People's Republic of China: Rescission of...

  17. 49 CFR 375.513 - Must I give the individual shipper an opportunity to observe the weighing?

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... of the shipment. You must advise the individual shipper, or any other person entitled to observe the... 49 Transportation 5 2013-10-01 2013-10-01 false Must I give the individual shipper an opportunity... PROTECTION REGULATIONS Pick Up of Shipments of Household Goods Weighing the Shipment § 375.513 Must I...

  18. 76 FR 24857 - Fresh Garlic From the People's Republic of China: Preliminary Intent To Rescind New Shipper Reviews

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-05-03

    ... From the People's Republic of China: Initiation of New Shipper Reviews, 75 FR 38986 (July 7, 2010... of Antidumping Duty New Shipper Reviews, 75 FR 74002 (November 30, 2010). On February 11, 2011, the... flexibility to extend the POR. See Antidumping Duties; Countervailing Duties; Final Rule, 62 FR 27296,...

  19. 9 CFR 201.39 - Payment to be made to consignor or shipper by market agencies; exceptions.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... shipper by market agencies; exceptions. 201.39 Section 201.39 Animals and Animal Products GRAIN INSPECTION... shipper by market agencies; exceptions. (a) No market agency shall, except as provided in paragraph (b) of... from the Secretary or a court of competent jurisdiction, unless (1) such market agency has reason...

  20. 77 FR 266 - Fresh Garlic From the People's Republic of China: Initiation of New Shipper Reviews

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-01-04

    ..., 1994. See Antidumping Duty Order: Fresh Garlic From the People's Republic of China, 59 FR 59209... International Trade Administration Fresh Garlic From the People's Republic of China: Initiation of New Shipper...) under the antidumping duty order on fresh garlic from the People's Republic of China (PRC), meet...

  1. 77 FR 65670 - Honey From Argentina: Final Rescission of Antidumping Duty New Shipper Review

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-10-30

    ... Shipper Review, 77 FR 45334 (July 31, 2012) (Preliminary Rescission). FOR FURTHER INFORMATION CONTACT... and Revocation of Antidumping Duty and Countervailing Duty Orders, 77 FR 58524 (September 21, 2012... did not receive comments from any party. As discussed below, based on our analysis of the record,...

  2. 77 FR 45334 - Honey From Argentina: Preliminary Rescission of Antidumping Duty New Shipper Review

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-07-31

    ... initiated this NSR. See Honey from Argentina: Notice of Initiation of Antidumping New Shipper Review, 77 FR... of Antidumping Duty Order: Honey From Argentina, 66 FR 63672 (December 10, 2001). \\2\\ We note that... doing business as Ap cola Danangie (``Danangie'') did not satisfy the regulatory requirements for an...

  3. 15 CFR 758.1 - The Shipper's Export Declaration (SED) or Automated Export System (AES) record.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ...) or Automated Export System (AES) record. 758.1 Section 758.1 Commerce and Foreign Trade Regulations... (SED) or Automated Export System (AES) record. (a) The Shipper's Export Declaration (SED) or Automated Export System (AES) record. The SED (Form 7525-V, Form 7525-V-Alt, or Automated Export System record)...

  4. 15 CFR 758.1 - The Shipper's Export Declaration (SED) or Automated Export System (AES) record.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ...) or Automated Export System (AES) record. 758.1 Section 758.1 Commerce and Foreign Trade Regulations... (SED) or Automated Export System (AES) record. (a) The Shipper's Export Declaration (SED) or Automated Export System (AES) record. The SED (Form 7525-V, Form 7525-V-Alt, or Automated Export System record)...

  5. 15 CFR 758.1 - The Shipper's Export Declaration (SED) or Automated Export System (AES) record.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ...) or Automated Export System (AES) record. 758.1 Section 758.1 Commerce and Foreign Trade Regulations... (SED) or Automated Export System (AES) record. (a) The Shipper's Export Declaration (SED) or Automated Export System (AES) record. The SED (Form 7525-V, Form 7525-V-Alt, or Automated Export System record)...

  6. 15 CFR 758.1 - The Shipper's Export Declaration (SED) or Automated Export System (AES) record.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ...) or Automated Export System (AES) record. 758.1 Section 758.1 Commerce and Foreign Trade Regulations... (SED) or Automated Export System (AES) record. (a) The Shipper's Export Declaration (SED) or Automated Export System (AES) record. The SED (Form 7525-V, Form 7525-V-Alt, or Automated Export System record)...

  7. 19 CFR 351.214 - New shipper reviews under section 751(a)(2)(B) of the Act.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... shipper reviews under section 751(a)(2)(B) of the Act. (a) Introduction. The URAA established a new...) In an antidumping proceeding involving imports from a nonmarket economy country, a certification...

  8. 75 FR 61130 - Fresh Garlic From the People's Republic of China: Final Results of New Shipper Review

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-10-04

    ... Results of New Shipper Review, 75 FR 24578 (May 5, 2010) (Preliminary Results). We are continuing to find... Duty Administrative, 75 FR 34976, (June 21, 2010) (14th AR). Therefore, the Department will determine... or separated into constituent cloves, whether or not peeled, fresh, chilled, frozen,...

  9. 76 FR 52315 - Fresh Garlic From the People's Republic of China: Final Rescission of New Shipper Reviews of...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-08-22

    ...: Preliminary Intent To Rescind New Shipper Reviews, 76 FR 24857 (May 3, 2011) (Preliminary Intent to Rescind.... See Preliminary Intent to Rescind, 76 FR at 24858. After analyzing Yantai Jinyan's case brief and... and Countervailing Duty Administrative Reviews and Request for Revocation in Part, 75 FR 81565,...

  10. 75 FR 15412 - Silicon Metal From the People's Republic of China: Notice of Amended Final Results of New Shipper...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-03-29

    ... (``the Department'') determination of the appropriate surrogate value for silica fume in Silicon Metal From the People's Republic of China: Notice of Final Results of 2005/2006 New Shipper Reviews, 72 FR 58...-product silica fume in the Department's normal value calculation. FOR FURTHER INFORMATION CONTACT:...

  11. 77 FR 59178 - Seamless Refined Copper Pipe and Tube From Mexico: Final Results of Antidumping Duty New Shipper...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-09-26

    ... Copper Pipe and Tube From Mexico: Preliminary Results of Antidumping Duty New Shipper Review, 77 FR 25136... Sn--Tin 0.8 Te--Tellurium 0.8 Zn--Zinc 1.0 Zr--Zirconium 0.3 Other elements (each) 0.3 Excluded...

  12. 49 CFR 375.203 - What actions of an individual shipper may limit or reduce my normal liability?

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... reduce my normal liability? 375.203 Section 375.203 Transportation Other Regulations Relating to... PROTECTION REGULATIONS Before Offering Services to My Customers Liability Considerations § 375.203 What actions of an individual shipper may limit or reduce my normal liability? (a) If an individual...

  13. 49 CFR 375.203 - What actions of an individual shipper may limit or reduce my normal liability?

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... reduce my normal liability? 375.203 Section 375.203 Transportation Other Regulations Relating to... PROTECTION REGULATIONS Before Offering Services to My Customers Liability Considerations § 375.203 What actions of an individual shipper may limit or reduce my normal liability? (a) If an individual...

  14. 49 CFR 375.203 - What actions of an individual shipper may limit or reduce my normal liability?

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... reduce my normal liability? 375.203 Section 375.203 Transportation Other Regulations Relating to... PROTECTION REGULATIONS Before Offering Services to My Customers Liability Considerations § 375.203 What actions of an individual shipper may limit or reduce my normal liability? (a) If an individual...

  15. 76 FR 45775 - Fourth New Shipper Review of Certain Frozen Warmwater Shrimp From the Socialist Republic of...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-08-01

    ... Republic of Vietnam: Preliminary Results of Antidumping Duty New Shipper Review, 76 FR 20627 (April 13...) and peeled shrimp; (2) to which a ``dusting'' layer of rice or wheat flour of at least 95 percent... Economies: Valuing the Factor of Production: Labor, Request for Comment, 76 FR 9544 (Feb. 18, 2011). On...

  16. A LARGE-SCALE SIMULATION OF INTERNATIONAL MARITIME CONTAINER SHIPPING CONSIDERING OPTIMAL BEHAVIOR OF SHIPPERS AND OCEANGOING CARRIERS

    NASA Astrophysics Data System (ADS)

    Shibasaki, Ryuichi; Watanabe, Tomihiro; Ieda, Hitoshi

    This paper develops a large-scale simulation model of international maritime container shipping industry considering optimal behaviors of both shippers and oceangoing carriers, in order to measure impact of port and international logistics policies for each country including Japan. Concretely, the authors develop a short-term model (income maximization model of carriers) including shippers' choice of carrier when maritime cargo shipping demand between ports are given and a mid-term model (Nash equilibrium model of shippers and carriers) including shippers' choice of import/export port and route of hinterland transport and carriers' profit maximization behavior when cargo shipping demand between regions are given. The developed model is applied to the actual large-scale international maritime container shipping network in Eastern Asia. From a trial calculation based on the actual cargo shipping demand, the performance of the model is validated in terms of convergency and reproducibility. Also, the sensitivity of the model output taking the actual port policies into account is confirmed.

  17. 78 FR 88 - Fresh Garlic From the People's Republic of China: Initiation of Antidumping Duty New Shipper...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-01-02

    ... Republic of China, 59 FR 59209 (November 16, 1994). Pursuant to the requirements set forth in 19 CFR 351... International Trade Administration Fresh Garlic From the People's Republic of China: Initiation of Antidumping... new shipper review (NSR) under the antidumping duty order on fresh garlic from the People's...

  18. 78 FR 40428 - Fresh Garlic from the People's Republic of China: Initiation of Antidumping Duty New Shipper...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-07-05

    ... the People's Republic of China, 59 FR 59209 (November 16, 1994). Qingshui and Merry each certified... International Trade Administration Fresh Garlic from the People's Republic of China: Initiation of Antidumping... shipper reviews (NSRs) of the antidumping duty order on fresh garlic from the People's Republic of...

  19. 75 FR 24578 - Fresh Garlic from the People's Republic of China: Preliminary Results of New Shipper Review

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-05-05

    ... China, 68 FR 47538 (August 11, 2003), and accompanying Issues and Decision Memorandum at Comment 1 (PVA... Review, 74 FR 31241 (June 30, 2009). On October 29, 2009, the Department placed a copy of the CBP data... of China: Extension of Time Limit for the Preliminary Results of the New Shipper Review, 74 FR...

  20. 75 FR 69415 - Fresh Garlic From the People's Republic of China: Preliminary Results of New Shipper Reviews and...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-11-12

    ...The Department of Commerce (Department) is conducting new shipper reviews (NSRs) of Jinxiang Chengda Imp & Exp Co., Ltd. (Chengda), Jinxiang Yuanxin Imp & Exp Co., Ltd. (Yuanxin), and Zhengzhou Huachao Industrial Co., Ltd. (Huachao) under the antidumping duty order on fresh garlic from the People's Republic of China (PRC) covering the period of review (POR) of November 1, 2008 through October......

  1. 78 FR 73837 - Glycine From the People's Republic of China: Final Rescission of Antidumping Duty New Shipper...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-12-09

    ... of China: Preliminary Rescission of Antidumping Duty New Shipper Review; 2012, 78 FR 52501 (August 23... Antidumping Duty Order: Glycine From the People's Republic of China, 60 FR 16116 (March 29, 1995). Tolling... March 1, 2012, through August 31, 2012, for Hebei Donghua Jiheng Fine Chemical Company, Ltd....

  2. 77 FR 65669 - Glycine From the People's Republic of China: Notice of Initiation of Antidumping Duty New Shipper...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-10-30

    ... Antidumping Duty Order: Glycine From the People's Republic of China, 60 FR 16115 (March 29, 1995) (Order). In... bond only for entries of subject merchandise which Donghua Fine Chemical exported. Interested parties...), we are initiating an antidumping duty new shipper review of Hebei Donghua Jiheng Fine Chemical...

  3. 46 CFR 545.1 - Interpretation of Shipping Act of 1984-Refusal to negotiate with shippers' associations.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 46 Shipping 9 2012-10-01 2012-10-01 false Interpretation of Shipping Act of 1984-Refusal to negotiate with shippers' associations. 545.1 Section 545.1 Shipping FEDERAL MARITIME COMMISSION REGULATIONS AFFECTING OCEAN SHIPPING IN FOREIGN COMMERCE INTERPRETATIONS AND STATEMENTS OF POLICY § 545.1 Interpretation of Shipping Act of 1984—Refusal...

  4. 46 CFR 545.1 - Interpretation of Shipping Act of 1984-Refusal to negotiate with shippers' associations.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 46 Shipping 9 2010-10-01 2010-10-01 false Interpretation of Shipping Act of 1984-Refusal to negotiate with shippers' associations. 545.1 Section 545.1 Shipping FEDERAL MARITIME COMMISSION REGULATIONS AFFECTING OCEAN SHIPPING IN FOREIGN COMMERCE INTERPRETATIONS AND STATEMENTS OF POLICY § 545.1 Interpretation of Shipping Act of 1984—Refusal...

  5. 46 CFR 545.1 - Interpretation of Shipping Act of 1984-Refusal to negotiate with shippers' associations.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 46 Shipping 9 2014-10-01 2014-10-01 false Interpretation of Shipping Act of 1984-Refusal to negotiate with shippers' associations. 545.1 Section 545.1 Shipping FEDERAL MARITIME COMMISSION REGULATIONS AFFECTING OCEAN SHIPPING IN FOREIGN COMMERCE INTERPRETATIONS AND STATEMENTS OF POLICY § 545.1 Interpretation of Shipping Act of 1984—Refusal...

  6. 46 CFR 545.1 - Interpretation of Shipping Act of 1984-Refusal to negotiate with shippers' associations.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 46 Shipping 9 2011-10-01 2011-10-01 false Interpretation of Shipping Act of 1984-Refusal to negotiate with shippers' associations. 545.1 Section 545.1 Shipping FEDERAL MARITIME COMMISSION REGULATIONS AFFECTING OCEAN SHIPPING IN FOREIGN COMMERCE INTERPRETATIONS AND STATEMENTS OF POLICY § 545.1 Interpretation of Shipping Act of 1984—Refusal...

  7. 46 CFR 545.1 - Interpretation of Shipping Act of 1984-Refusal to negotiate with shippers' associations.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 46 Shipping 9 2013-10-01 2013-10-01 false Interpretation of Shipping Act of 1984-Refusal to negotiate with shippers' associations. 545.1 Section 545.1 Shipping FEDERAL MARITIME COMMISSION REGULATIONS AFFECTING OCEAN SHIPPING IN FOREIGN COMMERCE INTERPRETATIONS AND STATEMENTS OF POLICY § 545.1 Interpretation of Shipping Act of 1984—Refusal...

  8. 49 CFR 375.203 - What actions of an individual shipper may limit or reduce my normal liability?

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... agrees to ship household goods released at a value greater than 60 cents per pound ($1.32 per kilogram... shipper will be entitled to full recovery up to the declared value of the article or articles, not to exceed the declared value of the entire shipment. General Responsibilities...

  9. Radcalc: An Analytical Tool for Shippers of Radioactive Material and Waste

    SciTech Connect

    Kapoor, A.K.; Stuhl, L.A.

    2008-07-01

    The U.S. Department of Energy (DOE) ships radioactive materials in support of its research and development, environmental restoration, and national defense activities. The Radcalc software program assists personnel working on behalf of DOE in packaging and transportation determinations (e.g., isotopic decay, decay heat, regulatory classification, and gas generation) for shipment of radioactive materials and waste. Radcalc performs: - The U.S. Department of Transportation determinations and classifications (i.e., activity concentration for exempt material Type A or B, effective A1/A2, limited quantity, low specific activity, highway route controlled quantity, fissile quantity, fissile excepted, reportable quantity, list of isotopes required on shipping papers) - DOE calculations (i.e., transuranic waste, Pu-239 equivalent curies, fissile-gram equivalents) - The U.S. Nuclear Regulatory Commission packaging category (i.e., Category I, II, or III) - Dose-equivalent curie calculations - Radioactive decay calculations using a novel decay methodology and a decay data library of 1,867 isotopes typical of the range of materials encountered in DOE laboratory environments - Hydrogen and helium gas calculations - Pressure calculations. Radcalc is a validated and cost-effective tool to provide consistency, accuracy, reproducibility, timeliness, quality, compliance, and appropriate documentation to shippers of radioactive materials and waste at DOE facilities nationwide. Hundreds of shippers and engineers throughout the DOE Complex routinely use this software to automate various determinations and to validate compliance with the regulations. The effective use of software by DOE sites contributes toward minimizing risk involved in radioactive waste shipments and assuring the safety of workers and the public. (authors)

  10. Secure transmission of shipper/receiver confirmatory measurements. [Gillaroo Encrypted Secure Communication

    SciTech Connect

    Tisinger, R.M.; Brumfield, T.

    1988-01-01

    Shipper/receiver confirmatory measurements will be performed at Y-12 using equipment designed by Y-12 and the Safeguards Assay Group at Los Alamos. Equipment consists of three stationary NaI(Tl) gamma-ray counter arrays and a Digital Equipment Corporation Micro-11 processor for processing data. In the case of outgoing shipments, the counter data will be incorporated in a shipping file with other necessary documentation and transmitted to the intended recipient. In the case of incoming shipments, this station will receive a similar file from the sending organization. Protection of all data at the S/RD level is required. Transmission of the data files can be made point-to-point over telephone lines with proper encryption. Two commercial devices used in conjunction with a personal computer are being evaluated to provide secure communication endorsed by the National Security Agency (NSA). One device is the Gillaroo from P. E. Systems, Inc., and the other is the STU-III, which is available as a PC board or a standalone unit, whereas the STU-III is a Secure Telephone Unit with an RS-232 port for connection to a computer. In both cases, file encryption is performed in hardware using keys supplied by NSA. The prime advantage of this approach is a simplicity. Aside from key management, the whole operation can be automatically controlled by command files on the computer without involving additional interfaces with secure networks or mail systems. Additional benefits are rapid response, immediate confirmation of receipt of message, a communication channel independent of the shipment itself, and low cost.

  11. Novel roles for LIX1L in promoting cancer cell proliferation through ROS1-mediated LIX1L phosphorylation

    PubMed Central

    Nakamura, Satoki; Kahyo, Tomoaki; Tao, Hong; Shibata, Kiyoshi; Kurabe, Nobuya; Yamada, Hidetaka; Shinmura, Kazuya; Ohnishi, Kazunori; Sugimura, Haruhiko

    2015-01-01

    Herein, we report the characterization of Limb expression 1-like, (LIX1L), a putative RNA-binding protein (RBP) containing a double-stranded RNA binding motif, which is highly expressed in various cancer tissues. Analysis of MALDI-TOF/TOF mass spectrometry and RNA immunoprecipitation-sequencing of interacting proteins and the microRNAs (miRNAs) bound to LIX1L revealed that LIX1L interacts with proteins (RIOK1, nucleolin and PABPC4) and miRNAs (has-miRNA-520a-5p, −300, −216b, −326, −190a, −548b-3p, −7–5p and −1296) in HEK-293 cells. Moreover, the reduction of phosphorylated Tyr136 (pTyr136) in LIX1L through the homeodomain peptide, PY136, inhibited LIX1L-induced cell proliferation in vitro, and PY136 inhibited MKN45 cell proliferation in vivo. We also determined the miRNA-targeted genes and showed that was apoptosis induced through the reduction of pTyr136. Moreover, ROS1, HCK, ABL1, ABL2, JAK3, LCK and TYR03 were identified as candidate kinases responsible for the phosphorylation of Tyr136 of LIX1L. These data provide novel insights into the biological significance of LIX1L, suggesting that this protein might be an RBP, with implications for therapeutic approaches for targeting LIX1L in LIX1L-expressing cancer cells. PMID:26310847

  12. Novel roles for LIX1L in promoting cancer cell proliferation through ROS1-mediated LIX1L phosphorylation.

    PubMed

    Nakamura, Satoki; Kahyo, Tomoaki; Tao, Hong; Shibata, Kiyoshi; Kurabe, Nobuya; Yamada, Hidetaka; Shinmura, Kazuya; Ohnishi, Kazunori; Sugimura, Haruhiko

    2015-01-01

    Herein, we report the characterization of Limb expression 1-like, (LIX1L), a putative RNA-binding protein (RBP) containing a double-stranded RNA binding motif, which is highly expressed in various cancer tissues. Analysis of MALDI-TOF/TOF mass spectrometry and RNA immunoprecipitation-sequencing of interacting proteins and the microRNAs (miRNAs) bound to LIX1L revealed that LIX1L interacts with proteins (RIOK1, nucleolin and PABPC4) and miRNAs (has-miRNA-520a-5p, -300, -216b, -326, -190a, -548b-3p, -7-5p and -1296) in HEK-293 cells. Moreover, the reduction of phosphorylated Tyr(136) (pTyr(136)) in LIX1L through the homeodomain peptide, PY136, inhibited LIX1L-induced cell proliferation in vitro, and PY136 inhibited MKN45 cell proliferation in vivo. We also determined the miRNA-targeted genes and showed that was apoptosis induced through the reduction of pTyr(136). Moreover, ROS1, HCK, ABL1, ABL2, JAK3, LCK and TYR03 were identified as candidate kinases responsible for the phosphorylation of Tyr(136) of LIX1L. These data provide novel insights into the biological significance of LIX1L, suggesting that this protein might be an RBP, with implications for therapeutic approaches for targeting LIX1L in LIX1L-expressing cancer cells. PMID:26310847

  13. Shipper/receiver difference verification of spent fuel by use of PDET

    SciTech Connect

    Ham, Y. S.; Sitaraman, S.

    2011-07-01

    Spent fuel storage pools in most countries are rapidly approaching their design limits with the discharge of over 10,000 metric tons of heavy metal from global reactors. Countries like UK, France or Japan have adopted a closed fuel cycle by reprocessing spent fuel and recycling MOX fuel while many other countries opted for above ground interim dry storage for their spent fuel management strategy. Some countries like Finland and Sweden are already well on the way to setting up a conditioning plant and a deep geological repository for spent fuel. For all these situations, shipments of spent fuel are needed and the number of these shipments is expected to increase significantly. Although shipper/receiver difference (SRD) verification measurements are needed by IAEA when the recipient facility receives spent fuel, these are not being practiced to the level that IAEA has desired due to lack of a credible measurement methodology and instrument that can reliably perform these measurements to verify non-diversion of spent fuel during shipment and confirm facility operator declarations on the spent fuel. In this paper, we describe a new safeguards method and an associated instrument, Partial Defect Tester (PDET), which can detect pin diversion from Pressurized Water Reactor (PWR) Spent Fuel Assemblies in an in-situ condition. The PDET uses multiple tiny neutron and gamma detectors in the form of a cluster and a simple, yet highly precise, gravity-driven system to obtain underwater radiation measurements inside a Pressurized Water Reactor (PWR) spent fuel assembly. The method takes advantage of the PWR fuel design which contains multiple guide tubes which can be accessed from the top. The data obtained in such a manner can provide spatial distribution of neutron and gamma flux within a spent fuel assembly. Our simulation study as well as validation measurements indicated that the ratio of the gamma signal to the thermal neutron signal at each detector location normalized to

  14. Type A radioactive liquid sample packaging family

    SciTech Connect

    Edwards, W.S.

    1995-11-01

    Westinghouse Hanford Company (WHC) has developed two packagings that can be used to ship Type A quantities of radioactive liquids. WHC designed these packagings to take advantage of commercially available items where feasible to reduce the overall packaging cost. The Hedgehog packaging can ship up to one liter of Type A radioactive liquid with no shielding and 15 cm of distance between the liquid and the package exterior, or 30 ml of liquid with 3.8 cm of stainless steel shielding and 19 cm of distance between the liquid and the package exterior. The One Liter Shipper can ship up to one liter of Type A radioactive liquid that does not require shielding.

  15. 49 CFR 375.201 - What is my normal liability for loss and damage when I accept goods from an individual shipper?

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... TRANSPORTATION FEDERAL MOTOR CARRIER SAFETY REGULATIONS TRANSPORTATION OF HOUSEHOLD GOODS IN INTERSTATE COMMERCE.... The rate may be increased annually by the motor carrier based on the U.S. Department of Commerce's... of insurance. (e) You must, in a clear and concise manner, disclose to the individual shipper...

  16. 49 CFR 375.201 - What is my normal liability for loss and damage when I accept goods from an individual shipper?

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... TRANSPORTATION FEDERAL MOTOR CARRIER SAFETY REGULATIONS TRANSPORTATION OF HOUSEHOLD GOODS IN INTERSTATE COMMERCE.... The rate may be increased annually by the motor carrier based on the U.S. Department of Commerce's... of insurance. (e) You must, in a clear and concise manner, disclose to the individual shipper...

  17. 49 CFR 375.521 - What must I do if an individual shipper wants to know the actual weight or charges for a shipment...

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 49 Transportation 5 2012-10-01 2012-10-01 false What must I do if an individual shipper wants to know the actual weight or charges for a shipment before I tender delivery? 375.521 Section 375.521 Transportation Other Regulations Relating to Transportation (Continued) FEDERAL MOTOR CARRIER SAFETY ADMINISTRATION, DEPARTMENT OF...

  18. 49 CFR 375.521 - What must I do if an individual shipper wants to know the actual weight or charges for a shipment...

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 49 Transportation 5 2010-10-01 2010-10-01 false What must I do if an individual shipper wants to know the actual weight or charges for a shipment before I tender delivery? 375.521 Section 375.521 Transportation Other Regulations Relating to Transportation (Continued) FEDERAL MOTOR CARRIER SAFETY ADMINISTRATION, DEPARTMENT OF...

  19. 49 CFR 375.521 - What must I do if an individual shipper wants to know the actual weight or charges for a shipment...

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 49 Transportation 5 2011-10-01 2011-10-01 false What must I do if an individual shipper wants to know the actual weight or charges for a shipment before I tender delivery? 375.521 Section 375.521 Transportation Other Regulations Relating to Transportation (Continued) FEDERAL MOTOR CARRIER SAFETY ADMINISTRATION, DEPARTMENT OF...

  20. 49 CFR 375.521 - What must I do if an individual shipper wants to know the actual weight or charges for a shipment...

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 49 Transportation 5 2014-10-01 2014-10-01 false What must I do if an individual shipper wants to know the actual weight or charges for a shipment before I tender delivery? 375.521 Section 375.521 Transportation Other Regulations Relating to Transportation (Continued) FEDERAL MOTOR CARRIER SAFETY ADMINISTRATION, DEPARTMENT OF...

  1. The L1=L2 Hypotheses: A Reconsideration

    ERIC Educational Resources Information Center

    Ellis, Rod

    1985-01-01

    Discusses the L1=L2 hypothesis which states that, all other things except knowledge of language being equal, first language acquisition is the same as second language acquisition. Reviews the evidence for and against the hypothesis, looks at current research and considers the general distinction between formal and informal learning. (SED)

  2. L1-L2 Sentence Translation in Classroom Grammar Tests

    ERIC Educational Resources Information Center

    Salem, Ilana

    2012-01-01

    L1-L2 translation of separate sentences is one kind of task format used by mainstream EFL teachers to assess their learners' grammatical accuracy. Aimed at improving teacher-written translation items, this study analyses linguistic features potentially causing such decontextualized cues (and their target responses) to sound odd or untypical of…

  3. Evidence for Involvement of GNB1L in Autism

    PubMed Central

    Chen, Ying-Zhang; Matsushita, Mark; Girirajan, Santhosh; Lisowski, Mark; Sun, Elizabeth; Sul, Youngmee; Bernier, Raphael; Estes, Annette; Dawson, Geraldine; Minshew, Nancy; Shellenberg, Gerard D; Eichler, Evan E; Rieder, Mark J; Nickerson, Deborah A; Tsuang, Debby W; Tsuang, Ming T; Wijsman, Ellen M; Raskind, Wendy H; Brkanac, Zoran

    2012-01-01

    Structural variations in the chromosome 22q11.2 region mediated by nonallelic homologous recombination result in 22q11.2 deletion (del22q11.2) and 22q11.2 duplication (dup22q11.2) syndromes. The majority of del22q11.2 cases have facial and cardiac malformations, immunologic impairments, specific cognitive profile and increased risk for schizophrenia and autism spectrum disorders (ASDs). The phenotype of dup22q11.2 is frequently without physical features but includes the spectrum of neurocognitive abnormalities. Although there is substantial evidence that haploinsufficiency for TBX1 plays a role in the physical features of del22q11.2, it is not known which gene(s) in the critical 1.5 Mb region are responsible for the observed spectrum of behavioral phenotypes. We identified an individual with a balanced translocation 46,XY,t(1;22)(p36.1;q11.2) and a behavioral phenotype characterized by cognitive impairment, autism, and schizophrenia in the absence of congenital malformations. Using somatic cell hybrids and comparative genomic hybridization (CGH) we mapped the chromosome-22 breakpoint within intron 7 of the GNB1L gene. Copy number evaluations and direct DNA sequencing of GNB1L in 271 schizophrenia and 513 autism cases revealed dup22q11.2 in two families with autism and private GNB1L missense variants in conserved residues in three families (P = 0.036). The identified missense variants affect residues in the WD40 repeat domains and are predicted to have deleterious effects on the protein. Prior studies provided evidence that GNB1L may have a role in schizophrenia. Our findings support involvement of GNB1L in ASDs as well. © 2011 Wiley Periodicals, Inc. PMID:22095694

  4. Fitness cost of LINE-1 (L1) activity in humans

    PubMed Central

    Boissinot, Stephane; Davis, Jerel; Entezam, Ali; Petrov, Dimitri; Furano, Anthony V.

    2006-01-01

    The self-replicating LINE-1 (L1) retrotransposon family is the dominant retrotransposon family in mammals and has generated 30–40% of their genomes. Active L1 families are present in modern mammals but the important question of whether these currently active families affect the genetic fitness of their hosts has not been addressed. This issue is of particular relevance to humans as Homo sapiens contains the active L1 Ta1 subfamily of the human specific Ta (L1Pa1) L1 family. Although DNA insertions generated by the Ta1 subfamily can cause genetic defects in current humans, these are relatively rare, and it is not known whether Ta1-generated inserts or any other property of Ta1 elements have been sufficiently deleterious to reduce the fitness of humans. Here we show that full-length (FL) Ta1 elements, but not the truncated Ta1 elements or SINE (Alu) insertions generated by Ta1 activity, were subject to negative selection. Thus, one or more properties unique to FL L1 elements constitute a genetic burden for modern humans. We also found that the FL Ta1 elements became more deleterious as the expansion of Ta1 has proceeded. Because this expansion is ongoing, the Ta1 subfamily almost certainly continues to decrease the fitness of modern humans. PMID:16766655

  5. Vaccinia Virus N1l Protein Resembles a B Cell Lymphoma-2 (Bcl-2) Family Protein

    SciTech Connect

    Aoyagi, M.; Zhai, D.; Jin, C.; Aleshin, A.E.; Stec, B.; Reed, J.C.; Liddington, R.C.; /Burnham Inst.

    2007-07-03

    Poxviruses encode immuno-modulatory proteins capable of subverting host defenses. The poxvirus vaccinia expresses a small 14-kDa protein, N1L, that is critical for virulence. We report the crystal structure of N1L, which reveals an unexpected but striking resemblance to host apoptotic regulators of the B cell lymphoma-2 (Bcl-2) family. Although N1L lacks detectable Bcl-2 homology (BH) motifs at the sequence level, we show that N1L binds with high affinity to the BH3 peptides of pro-apoptotic Bcl-2 family proteins in vitro, consistent with a role for N1L in modulating host antiviral defenses.

  6. Implementation of the beryllium reflector filter concept in the LANSCE 1L target Mark-III upgrade.

    SciTech Connect

    Muhrer, G.; Pitcher, E. J.; Russell, G. J.

    2005-01-01

    We have investigated the possibility of implementing the idea of a beryllium reflector filter in the LANSCE 1L target Mark-III upgrade. We will present different concepts of beryllium reflector filters (megaphone, chevron and swiss-cheese concept) and their effect on the integrated cold flux and the time distribution of the lower tier hydrogen flux-trap moderator as a function of the three instruments, (SPEAR, LQD and Asterix) which are served by this moderator. As part of the LANSCE 1L target upgrade study it is the declared goal to increase the cold flux (E < 5meV) of the lower tier partially coupled liquid hydrogen moderator by a factor of two. This goal is proposed to be achieved by adding a pre-moderator system to the moderator and by implementing the cold beryllium reflector concept. The cold beryllium reflector filter concept was tested in an experiment at the weapons neutron research (WNR) facility at LANSCE in January 2003 by Pitcher et al. Based on the success of this experiment it was then decided to implement this concept into the 1L target Mark-III upgrade. In this context a series of Monte Carlo transport calculations was performed to optimize the cold neutron flux.

  7. Ezetimibe-sensitive cholesterol uptake by NPC1L1 protein does not require endocytosis

    PubMed Central

    Johnson, Tory A.; Pfeffer, Suzanne R.

    2016-01-01

    Human NPC1L1 protein mediates cholesterol absorption in the intestine and liver and is the target of the drug ezetimibe, which is used to treat hypercholesterolemia. Previous studies concluded that NPC1L1-GFP protein trafficking is regulated by cholesterol binding and that ezetimibe blocks NPC1L1-GFP function by inhibiting its endocytosis. We used cell surface biotinylation to monitor NPC1L1-GFP endocytosis and show that ezetimibe does not alter the rate of NPC1L1-GFP endocytosis in cultured rat hepatocytes grown under normal growth conditions. As expected, NPC1L1-GFP endocytosis depends in part on C-terminal, cytoplasmically oriented sequences, but endocytosis does not require cholesterol binding to NPC1L1’s N-terminal domain. In addition, two small- molecule inhibitors of general (and NPC1L1-GFP) endocytosis failed to inhibit the ezetimibe-sensitive uptake of [3H]cholesterol from taurocholate micelles. These experiments demonstrate that cholesterol uptake by NPC1L1 does not require endocytosis; moreover, ezetimibe interferes with NPC1L1’s cholesterol adsorption activity without blocking NPC1L1 internalization in RH7777 cells. PMID:27075173

  8. Ezetimibe-sensitive cholesterol uptake by NPC1L1 protein does not require endocytosis.

    PubMed

    Johnson, Tory A; Pfeffer, Suzanne R

    2016-06-01

    Human NPC1L1 protein mediates cholesterol absorption in the intestine and liver and is the target of the drug ezetimibe, which is used to treat hypercholesterolemia. Previous studies concluded that NPC1L1-GFP protein trafficking is regulated by cholesterol binding and that ezetimibe blocks NPC1L1-GFP function by inhibiting its endocytosis. We used cell surface biotinylation to monitor NPC1L1-GFP endocytosis and show that ezetimibe does not alter the rate of NPC1L1-GFP endocytosis in cultured rat hepatocytes grown under normal growth conditions. As expected, NPC1L1-GFP endocytosis depends in part on C-terminal, cytoplasmically oriented sequences, but endocytosis does not require cholesterol binding to NPC1L1's N-terminal domain. In addition, two small- molecule inhibitors of general (and NPC1L1-GFP) endocytosis failed to inhibit the ezetimibe-sensitive uptake of [(3)H]cholesterol from taurocholate micelles. These experiments demonstrate that cholesterol uptake by NPC1L1 does not require endocytosis; moreover, ezetimibe interferes with NPC1L1's cholesterol adsorption activity without blocking NPC1L1 internalization in RH7777 cells. PMID:27075173

  9. The transition zone protein Rpgrip1l regulates proteasomal activity at the primary cilium

    PubMed Central

    Lier, Johanna Maria; Burmühl, Stephan; Struchtrup, Andreas; Deutschmann, Kathleen; Vetter, Maik; Leu, Tristan; Reeg, Sandra; Grune, Tilman; Rüther, Ulrich

    2015-01-01

    Mutations in RPGRIP1L result in severe human diseases called ciliopathies. To unravel the molecular function of RPGRIP1L, we analyzed Rpgrip1l−/− mouse embryos, which display a ciliopathy phenotype and die, at the latest, around birth. In these embryos, cilia-mediated signaling was severely disturbed. Defects in Shh signaling suggested that the Rpgrip1l deficiency causes an impairment of protein degradation and protein processing. Indeed, we detected a cilia-dependent decreased proteasomal activity in the absence of Rpgrip1l. We found different proteasomal components localized to cilia and identified Psmd2, a component of the regulatory proteasomal 19S subunit, as an interaction partner for Rpgrip1l. Quantifications of proteasomal substrates demonstrated that Rpgrip1l regulates proteasomal activity specifically at the basal body. Our study suggests that Rpgrip1l controls ciliary signaling by regulating the activity of the ciliary proteasome via Psmd2. PMID:26150391

  10. Structure-Guided DOT1L Probe Optimization by Label-Free Ligand Displacement

    PubMed Central

    2014-01-01

    The DOT1L lysine methyltransferase has emerged as a validated therapeutic target in MLL-rearranged (MLLr) acute leukemias. Although S-adenosylmethionine competitive inhibitors have demonstrated pharmacological proof-of-principle in MLLr-leukemia, these compounds require further optimization to improve cellular potency and pharmacokinetic stability. Limiting DOT1L inhibitor discovery and ligand optimization have been complex biochemical methods often using radionucleotides and cellular methods requiring prolonged culture. We therefore developed a new suite of assay technologies that allows comparative assessment of chemical tools for DOT1L in a miniaturized format. Coupling these assays with structural information, we developed new insights into DOT1L ligand binding and identified several functionalized probes with increased cellular potency (IC50 values ∼10 nM) and excellent selectivity for DOT1L. Together these assay technologies define a platform capability for discovery and optimization of small-molecule DOT1L inhibitors. PMID:25397901

  11. Structure-guided DOT1L probe optimization by label-free ligand displacement.

    PubMed

    Yi, Joanna S; Federation, Alexander J; Qi, Jun; Dhe-Paganon, Sirano; Hadler, Michael; Xu, Xiang; St Pierre, Roodolph; Varca, Anthony C; Wu, Lei; Marineau, Jason J; Smith, William B; Souza, Amanda; Chory, Emma J; Armstrong, Scott A; Bradner, James E

    2015-03-20

    The DOT1L lysine methyltransferase has emerged as a validated therapeutic target in MLL-rearranged (MLLr) acute leukemias. Although S-adenosylmethionine competitive inhibitors have demonstrated pharmacological proof-of-principle in MLLr-leukemia, these compounds require further optimization to improve cellular potency and pharmacokinetic stability. Limiting DOT1L inhibitor discovery and ligand optimization have been complex biochemical methods often using radionucleotides and cellular methods requiring prolonged culture. We therefore developed a new suite of assay technologies that allows comparative assessment of chemical tools for DOT1L in a miniaturized format. Coupling these assays with structural information, we developed new insights into DOT1L ligand binding and identified several functionalized probes with increased cellular potency (IC50 values ∼10 nM) and excellent selectivity for DOT1L. Together these assay technologies define a platform capability for discovery and optimization of small-molecule DOT1L inhibitors. PMID:25397901

  12. NME1L Negatively Regulates IGF1-Dependent Proliferation of Breast Cancer Cells.

    PubMed

    You, Dong-Joo; Mander, Sunam; Park, Cho Rong; Koo, Okjae; Lee, Cheolju; Oh, Seong-Hyun; Ahn, Curie; Seong, Jae Young; Hwang, Jong-Ik

    2016-06-01

    Non-metastatic cells 1 (NME1) or nm23 is the first metastasis suppressor gene discovered. It functions through various enzymatic activities and interacts with many intracellular proteins. The NME1 gene encodes two splicing variants, NME1 and NME1L. Most studies have focused on NME1 because of its abundance in cells. We previously reported NME1L-mediated suppression of NF-κB signaling by interacting with and inhibiting IKKβ. In this study, we demonstrated that NME1L, but not NME1, mediated inhibition of cell proliferation, although both NME1 and NME1L were involved in suppressing metastasis. A reporter gene assay was performed to determine the growth signaling pathway regulated by NME1L but none of the growth factors tested could induce an NF-κB-dependent luciferase expression except TNFα. Interestingly, SRE-reporter gene activation by IGF1 was significantly downregulated, along with reduction of ERK phosphorylation in NME1L expressing cells, compared to vector or NME1 expressing cells. NME1L directly interacted with KSR1, which is a scaffold for Raf-1, MEK, and ERK, that regulates ERK activation. Hence, NME1L plays a crucial role in regulation of cell proliferation by inhibiting IGF1-stimulated ERK phosphorylation through N-terminal 25 amino acid-mediated interaction with KSR1. J. Cell. Biochem. 117: 1454-1463, 2016. © 2015 Wiley Periodicals, Inc. PMID:26565392

  13. Concatenative and Nonconcatenative Plural Formation in L1, L2, and Heritage Speakers of Arabic

    ERIC Educational Resources Information Center

    Albirini, Abdulkafi; Benmamoun, Elabbas

    2014-01-01

    This study compares Arabic L1, L2, and heritage speakers' (HS) knowledge of plural formation, which involves concatenative and nonconcatenative modes of derivation. Ninety participants (divided equally among L1, L2, and heritage speakers) completed two oral tasks: a picture naming task (to measure proficiency) and a plural formation task. The…

  14. Ash1l controls quiescence and self-renewal potential in hematopoietic stem cells

    PubMed Central

    Jones, Morgan; Chase, Jennifer; Brinkmeier, Michelle; Xu, Jing; Weinberg, Daniel N.; Schira, Julien; Friedman, Ann; Malek, Sami; Grembecka, Jolanta; Cierpicki, Tomasz; Dou, Yali; Camper, Sally A.; Maillard, Ivan

    2015-01-01

    Rapidly cycling fetal and neonatal hematopoietic stem cells (HSCs) generate a pool of quiescent adult HSCs after establishing hematopoiesis in the bone marrow. We report an essential role for the trithorax group gene absent, small, or homeotic 1-like (Ash1l) at this developmental transition. Emergence and expansion of Ash1l-deficient fetal/neonatal HSCs were preserved; however, in young adult animals, HSCs were profoundly depleted. Ash1l-deficient adult HSCs had markedly decreased quiescence and reduced cyclin-dependent kinase inhibitor 1b/c (Cdkn1b/1c) expression and failed to establish long-term trilineage bone marrow hematopoiesis after transplantation to irradiated recipients. Wild-type HSCs could efficiently engraft when transferred to unirradiated, Ash1l-deficient recipients, indicating increased availability of functional HSC niches in these mice. Ash1l deficiency also decreased expression of multiple Hox genes in hematopoietic progenitors. Ash1l cooperated functionally with mixed-lineage leukemia 1 (Mll1), as combined loss of Ash1l and Mll1, but not isolated Ash1l or Mll1 deficiency, induced overt hematopoietic failure. Our results uncover a trithorax group gene network that controls quiescence, niche occupancy, and self-renewal potential in adult HSCs. PMID:25866973

  15. CHD1L Regulates Cell Cycle, Apoptosis, and Migration in Glioma.

    PubMed

    Sun, Jie; Zhang, Li; Zhao, Hongyu; Qiu, Xiaojun; Chen, Wenjuan; Wang, Donglin; Ban, Na; Fan, Shaochen; Shen, Chaoyan; Xia, Xiaojie; Ji, Bin; Wang, Yuchan

    2016-05-01

    Chromodomain helicase/ATPase DNA binding protein 1-like (CHD1L) gene is a newly identified oncogene located at Chr1q21 and it is amplified in many solid tumors. In this study, we intended to investigate the clinical significance of CHD1L expression in human glioma and its biological function in glioma cells. Western blot and immunohistochemistry analysis showed that CHD1L was overexpressed in glioma tissues and glioma cell lines. In addition, the expression level of CHD1L was positively correlated with glioma pathological grade and Ki-67 expression. Kaplan-Meier curve indicated that high expression of CHD1L may result in poor prognosis of glioma patients. Accordingly, suppression of CHD1L in glioma cells was shown to induce cell cycle arrest and increase apoptosis. In addition, knockdown of CHD1L significantly accelerated migration and invasion ability of glioma cells. Together our findings suggest that CHD1L is involved in the progression of glioma and may be a novel target for further therapy. PMID:26162969

  16. The histone methyltransferase DOT1L: regulatory functions and a cancer therapy target

    PubMed Central

    Wong, Matthew; Polly, Patsie; Liu, Tao

    2015-01-01

    DOT1L is a unique histone methyltransferase that targets the histone H3 lysine 79 (H3K79) residue for mono-, di- and tri- methylation. Histone H3K79 mono- and di-methylation results in active gene transcription, while H3K79 tri-methylation is associated with gene repression. DOT1L has a critical role in regulating gene transcription, development, cell cycle progression, somatic reprogramming and DNA damage repair. DOT1L interacts with Mixed Lineage Leukemia (MLL) fusion proteins, leading to enhanced H3K79 methylation, maintenance of open chromatin, overexpression of downstream oncogenes and leukemogenesis. Importantly, small molecule DOT1L inhibitors have been recently developed, and one of the DOT1L inhibitors is already under investigation in a Phase I clinical trial in patients with MLL fusion gene-driven leukemia. PMID:26609488

  17. YME1L degradation reduces mitochondrial proteolytic capacity during oxidative stress

    PubMed Central

    Rainbolt, T Kelly; Saunders, Jaclyn M; Wiseman, R Luke

    2015-01-01

    Mitochondrial proteostasis is maintained by a network of ATP-dependent quality control proteases including the inner membrane protease YME1L. Here, we show that YME1L is a stress-sensitive mitochondrial protease that is rapidly degraded in response to acute oxidative stress. This degradation requires reductions in cellular ATP and involves the activity of the ATP-independent protease OMA1. Oxidative stress-dependent reductions in YME1L inhibit protective YME1L-dependent functions and increase cellular sensitivity to oxidative insult. Collectively, our results identify stress-induced YME1L degradation as a biologic process that attenuates protective regulation of mitochondrial proteostasis and promotes cellular death in response to oxidative stress. PMID:25433032

  18. Discovery of Novel Dot1L Inhibitors through a Structure-Based Fragmentation Approach.

    PubMed

    Chen, Chao; Zhu, Hugh; Stauffer, Frédéric; Caravatti, Giorgio; Vollmer, Susanne; Machauer, Rainer; Holzer, Philipp; Möbitz, Henrik; Scheufler, Clemens; Klumpp, Martin; Tiedt, Ralph; Beyer, Kim S; Calkins, Keith; Guthy, Daniel; Kiffe, Michael; Zhang, Jeff; Gaul, Christoph

    2016-08-11

    Oncogenic MLL fusion proteins aberrantly recruit Dot1L, a histone methyltransferase, to ectopic loci, leading to local hypermethylation of H3K79 and misexpression of HoxA genes driving MLL-rearranged leukemias. Inhibition of the methyltransferase activity of Dot1L in this setting is predicted to reverse aberrant H3K79 methylation, leading to repression of leukemogenic genes and tumor growth inhibition. In the context of our Dot1L drug discovery program, high-throughput screening led to the identification of 2, a weak Dot1L inhibitor with an unprecedented, induced pocket binding mode. A medicinal chemistry campaign, strongly guided by structure-based consideration and ligand-based morphing, enabled the discovery of 12 and 13, potent, selective, and structurally completely novel Dot1L inhibitors. PMID:27563395

  19. CLPTM1L promotes growth and enhances aneuploidy in pancreatic cancer cells

    PubMed Central

    Jia, Jinping; Bosley, Allen D.; Thompson, Abbey; Hoskins, Jason W.; Cheuk, Adam; Collins, Irene; Parikh, Hemang; Xiao, Zhen; Ylaya, Kris; Dzyadyk, Marta; Cozen, Wendy; Hernandez, Brenda Y.; Lynch, Charles F.; Loncarek, Jadranka; Altekruse, Sean F.; Zhang, Lizhi; Westlake, Christopher J.; Factor, Valentina M.; Thorgeirsson, Snorri; Bamlet, William R.; Hewitt, Stephen M.; Petersen, Gloria M.; Andresson, Thorkell; Amundadottir, Laufey T.

    2014-01-01

    Genome wide association studies (GWAS) of ten different cancers have identified pleiotropic cancer predisposition loci across a region of chromosome 5p15.33 that includes the TERT and CLPTM1L genes. Of these, susceptibility alleles for pancreatic cancer have mapped to the CLPTM1L gene, thus prompting an investigation of the function of CLPTM1L in the pancreas. Immunofluorescence analysis indicated that CLPTM1L localized to the endoplasmic reticulum (ER) where it is likely embedded in the membrane, in accord with multiple predicted trans-membrane domains. Overexpression of CLPTM1L enhanced growth of pancreatic cancer cells in vitro (1.3–1.5 fold, PDAY7<0.003) and in vivo (3.46 fold, PDAY68=0.039), suggesting a role in tumor growth; this effect was abrogated by deletion of two hydrophilic domains. Affinity purification followed by mass-spectrometry identified an interaction between CLPTM1L and non-muscle myosin II (NMM-II), a protein involved in maintaining cell shape, migration, and cytokinesis. The two proteins co-localized in the cytoplasm and, after treatment with a DNA damaging agent, at the centrosomes. Overexpression of CLPTM1L and depletion of NMM-II induced aneuploidy, indicating that CLPTM1L may interfere with normal NMM-II function in regulating cytokinesis. Immunohistochemical analysis revealed enhanced staining of CLPTM1L in human pancreatic ductal adenocarcinoma (n=378) as compared to normal pancreatic tissue samples (n=17) (P=1.7×10−4). Our results suggest that CLPTM1L functions as a growth promoting gene in the pancreas and that overexpression may lead to an abrogation of normal cytokinesis, indicating that it should be considered as a plausible candidate gene that could explain the effect of pancreatic cancer susceptibility alleles on chr5p15.33. PMID:24648346

  20. Upregulation of PKD1L2 provokes a complex neuromuscular disease in the mouse

    PubMed Central

    Mackenzie, Francesca E.; Romero, Rosario; Williams, Debbie; Gillingwater, Thomas; Hilton, Helen; Dick, Jim; Riddoch-Contreras, Joanna; Wong, Frances; Ireson, Lisa; Powles-Glover, Nicola; Riley, Genna; Underhill, Peter; Hough, Tertius; Arkell, Ruth; Greensmith, Linda; Ribchester, Richard R.; Blanco, Gonzalo

    2009-01-01

    Following a screen for neuromuscular mouse mutants, we identified ostes, a novel N-ethyl N-nitrosourea-induced mouse mutant with muscle atrophy. Genetic and biochemical evidence shows that upregulation of the novel, uncharacterized transient receptor potential polycystic (TRPP) channel PKD1L2 (polycystic kidney disease gene 1-like 2) underlies this disease. Ostes mice suffer from chronic neuromuscular impairments including neuromuscular junction degeneration, polyneuronal innervation and myopathy. Ectopic expression of PKD1L2 in transgenic mice reproduced the ostes myopathic changes and, indeed, caused severe muscle atrophy in Tg(Pkd1l2)/Tg(Pkd1l2) mice. Moreover, double-heterozygous mice (ostes/+, Tg(Pkd1l2)/0) suffer from myopathic changes more profound than each heterozygote, indicating positive correlation between PKD1L2 levels and disease severity. We show that, in vivo, PKD1L2 primarily associates with endogenous fatty acid synthase in normal skeletal muscle, and these proteins co-localize to costameric regions of the muscle fibre. In diseased ostes/ostes muscle, both proteins are upregulated, and ostes/ostes mice show signs of abnormal lipid metabolism. This work shows the first role for a TRPP channel in neuromuscular integrity and disease. PMID:19578180

  1. The potential role of CAMSAP1L1 in symptomatic epilepsy.

    PubMed

    Zhang, Shuai; Kwan, Patrick; Baum, Larry

    2013-11-27

    In a recent genome-wide association study (GWAS) of symptomatic epilepsy in the Chinese population, the most significant single nucleotide polymorphism (SNP) allele was rs2292096 [G] (P=1.0×10(-8), odds ratio [OR]=0.63), in the CAMSAP1L1 gene (also known as CAMSAP2). Here, we report that rs2292096 genotypes tended to associate with expression of CAMSAP1L1 RNA in the temporal lobe (p=0.054) and hippocampus (p=0.20) of epilepsy surgery patients, with expression tending to increase with the G allele. CAMSAP1L1 and β-tubulin double immunofluorescence exhibited partial overlap. CAMSAP1L1 siRNA transfection of human SH-SY5Y neuroblastoma cells treated with or without retinoic acid reduced the CAMSAP1L1 protein level nearly 60% and stimulated neurite outgrowth, as measured by total length, number of processes and number of branches. Therefore, the rs2292096 G allele of CAMSAP1L1, which was associated with reduced risk of symptomatic epilepsy, tended to associate with increased expression of CAMSAP1L1, which represses neurite outgrowth. Greater neurite growth in response to brain insults might increase formation of ectopic neural circuits and thus the risk of epileptogenesis. PMID:24148305

  2. Characterization of Functional Domains in NME1L Regulation of NF-κB Signaling.

    PubMed

    You, Dong-Joo; Park, Cho Rong; Mander, Sunam; Ahn, Curie; Seong, Jae Young; Hwang, Jong-Ik

    2016-05-31

    NME1 is a well-known metastasis suppressor which has been reported to be downregulated in some highly aggressive cancer cells. Although most studies have focused on NME1, the NME1 gene also encodes the protein (NME1L) containing N-terminal 25 extra amino acids by alternative splicing. According to previous studies, NME1L has potent anti-metastatic activity, in comparison with NME1, by interacting with IKKβ and regulating its activity. In the present study, we tried to define the role of the N-terminal 25 amino acids of NME1L in NF-κB activation signaling. Unfortunately, the sequence itself did not interact with IKKβ, suggesting that it may be not enough to constitute the functional structure. Further construction of NME1L fragments and biochemical analysis revealed that N-terminal 84 residues constitute minimal structure for homodimerization, IKKβ interaction and regulation of NF-κB signaling. The inhibitory effect of the fragment on cancer cell migration and NF-κB-stimulated gene expression was equivalent to that of whole NME1L. The data suggest that the N-terminal 84 residues may be a core region for the anti-metastatic activity of NME1L. Based on this result, further structural analysis of the binding between NME1L and IKKβ may help in understanding the anti-metastatic activity of NME1L and provide direction to NME1L and IKKβ-related anti-cancer drug design. PMID:27094059

  3. Characterization of Functional Domains in NME1L Regulation of NF-κB Signaling

    PubMed Central

    You, Dong-Joo; Park, Cho Rong; Mander, Sunam; Ahn, Curie; Seong, Jae Young; Hwang, Jong-Ik

    2016-01-01

    NME1 is a well-known metastasis suppressor which has been reported to be downregulated in some highly aggressive cancer cells. Although most studies have focused on NME1, the NME1 gene also encodes the protein (NME1L) containing N-terminal 25 extra amino acids by alternative splicing. According to previous studies, NME1L has potent anti-metastatic activity, in comparison with NME1, by interacting with IKKβ and regulating its activity. In the present study, we tried to define the role of the N-terminal 25 amino acids of NME1L in NF-κB activation signaling. Unfortunately, the sequence itself did not interact with IKKβ, suggesting that it may be not enough to constitute the functional structure. Further construction of NME1L fragments and biochemical analysis revealed that N-terminal 84 residues constitute minimal structure for homodimerization, IKKβ interaction and regulation of NF-κB signaling. The inhibitory effect of the fragment on cancer cell migration and NF-κB-stimulated gene expression was equivalent to that of whole NME1L. The data suggest that the N-terminal 84 residues may be a core region for the anti-metastatic activity of NME1L. Based on this result, further structural analysis of the binding between NME1L and IKKβ may help in understanding the anti-metastatic activity of NME1L and provide direction to NME1L and IKKβ-related anti-cancer drug design. PMID:27094059

  4. Genome-wide analysis of CrRLK1L gene family in Gossypium and identification of candidate CrRLK1L genes related to fiber development.

    PubMed

    Niu, Erli; Cai, Caiping; Zheng, Yongjie; Shang, Xiaoguang; Fang, Lei; Guo, Wangzhen

    2016-06-01

    Members of the CrRLK1L family, a subgroup of the receptor-like kinase (RLK) gene family, are thought to act as sensors for the integrity of the cell wall and regulators of polar elongation. To better understand the various functions in fiber development, we conducted genome-wide identification and characterization analyses of CrRLK1L family in cotton. Here 44, 40, and 79 CrRLK1L genes were identified from three cotton species: diploid G. raimondii (D5), diploid G. arboreum (A2), and tetraploid G. hirsutum TM-1 (AD1), respectively. The 44 CrRLK1Ls in G. raimondii were anchored to the 12 chromosomes unevenly and were classified into six groups (I-VI), with group II and group IV being further divided into two subgroups (groups IIa and IIb, and IVa and IVb, respectively). These CrRLK1Ls displayed a highly regular pattern of developmental and spatial regulation in cotton. Using the transcriptome data of five chromosomal segment introgression lines (CSILs) and the physical integration of CrRLK1Ls with the quantitative trait loci (QTLs) related to fiber quality traits, we revealed that six CrRLK1L genes were highly associated with fiber development. This study brings new insights into the integrated genome-wide identification of CrRLK1Ls in cotton and provides references for the genetic improvement of cotton fiber. PMID:26833484

  5. AFAP-1L1-mediated actin filaments crosslinks hinder Trypanosoma cruzi cell invasion and intracellular multiplication.

    PubMed

    de Araújo, Karine Canuto Loureiro; Teixeira, Thaise Lara; Machado, Fabrício Castro; da Silva, Aline Alves; Quintal, Amanda Pifano Neto; da Silva, Claudio Vieira

    2016-10-01

    Host actin cytoskeleton polymerization has been shown to play an important role during Trypanosoma cruzi internalization into mammalian cell. The structure and dynamics of the actin cytoskeleton in cells are regulated by a vast number of actin-binding proteins. Here we aimed to verify the impact of AFAP-1L1, during invasion and multiplication of T. cruzi. Knocking-down AFAP-1L1 increased parasite cell invasion and intracellular multiplication. Thus, we have shown that the integrity of the machinery formed by AFAP-1L1 in actin cytoskeleton polymerization is important to hinder parasite infection. PMID:27349187

  6. Structural Insight into BH3 Domain Binding of Vaccinia Virus Antiapoptotic F1L

    PubMed Central

    Campbell, Stephanie; Thibault, John; Mehta, Ninad; Colman, Peter M.

    2014-01-01

    ABSTRACT Apoptosis is a tightly regulated process that plays a crucial role in the removal of virus-infected cells, a process controlled by both pro- and antiapoptotic members of the Bcl-2 family. The proapoptotic proteins Bak and Bax are regulated by antiapoptotic Bcl-2 proteins and are also activated by a subset of proteins known as BH3-only proteins that perform dual functions by directly activating Bak and Bax or by sequestering and neutralizing antiapoptotic family members. Numerous viruses express proteins that prevent premature host cell apoptosis. Vaccinia virus encodes F1L, an antiapoptotic protein essential for survival of infected cells that bears no discernible sequence homology to mammalian cell death inhibitors. Despite the limited sequence similarities, F1L has been shown to adopt a novel dimeric Bcl-2-like fold that enables hetero-oligomeric binding to both Bak and the proapoptotic BH3-only protein Bim that ultimately prevents Bak and Bax homo-oligomerization. However, no structural data on the mode of engagement of F1L and its Bcl-2 counterparts are available. Here we solved the crystal structures of F1L in complex with two ligands, Bim and Bak. Our structures indicate that F1L can engage two BH3 ligands simultaneously via the canonical Bcl-2 ligand binding grooves. Furthermore, by structure-guided mutagenesis, we generated point mutations within the binding pocket of F1L in order to elucidate the residues responsible for both Bim and Bak binding and prevention of apoptosis. We propose that the sequestration of Bim by F1L is primarily responsible for preventing apoptosis during vaccinia virus infection. IMPORTANCE Numerous viruses have adapted strategies to counteract apoptosis by encoding proteins responsible for sequestering proapoptotic components. Vaccinia virus, the prototypical member of the family Orthopoxviridae, encodes a protein known as F1L that functions to prevent apoptosis by interacting with Bak and the BH3-only protein Bim. Despite

  7. 10. DETAILED INTERIOR VIEW OF WEB AT U1L1, SHOWING VERTICAL, ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    10. DETAILED INTERIOR VIEW OF WEB AT U1L1, SHOWING VERTICAL, ARCH RIB WELDED/BOLTED GUSSET PLATE AND GUARDRAIL, LOOKING SOUTH - Cottonville Bridge, County Road D-61 at Farmer's Creek, Maquoketa, Jackson County, IA

  8. Catalytic site remodelling of the DOT1L methyltransferase by selective inhibitors

    SciTech Connect

    Yu, Wenyu; Chory, Emma J.; Wernimont, Amy K.; Tempel, Wolfram; Scopton, Alex; Federation, Alexander; Marineau, Jason J.; Qi, Jun; Barsyte-Lovejoy, Dalia; Yi, Joanna; Marcellus, Richard; Iacob, Roxana E.; Engen, John R.; Griffin, Carly; Aman, Ahmed; Wienholds, Erno; Li, Fengling; Pineda, Javier; Estiu, Guillermina; Shatseva, Tatiana; Hajian, Taraneh; Al-awar, Rima; Dick, John E.; Vedadi, Masoud; Brown, Peter J.; Arrowsmith, Cheryl H.; Bradner, James E.; Schapira, Matthieu

    2012-12-18

    Selective inhibition of protein methyltransferases is a promising new approach to drug discovery. An attractive strategy towards this goal is the development of compounds that selectively inhibit binding of the cofactor, S-adenosylmethionine, within specific protein methyltransferases. Here we report the three-dimensional structure of the protein methyltransferase DOT1L bound toEPZ004777, the first S-adenosylmethionine-competitive inhibitor of a protein methyltransferase with in vivo efficacy. This structure and those of four new analogues reveal remodelling of the catalytic site. EPZ004777 and a brominated analogue, SGC0946, inhibit DOT1L in vitro and selectively kill mixed lineage leukaemia cells, in which DOT1L is aberrantly localized via interaction with an oncogenic MLL fusion protein. These data provide important new insight into mechanisms of cell-active S-adenosylmethionine-competitive protein methyltransferase inhibitors, and establish a foundation for the further development of drug-like inhibitors of DOT1L for cancer therapy.

  9. Essential role of spi-1–like (spi-1l) in zebrafish myeloid cell differentiation

    PubMed Central

    Bukrinsky, Alex; Griffin, Kevin J. P.; Zhao, Yan; Lin, Shuo

    2009-01-01

    The ETS protein Spi-1/Pu.1 plays a pivotal and widespread role throughout hematopoiesis in many species. This study describes the identification, characterization, and functional analysis of a new zebrafish spi transcription factor spi-1–like (spi-1l) that is expressed in primitive myeloid cells, erythro-myelo progenitor cells, and in the adult kidney. Spi-1l functions genetically downstream of etsrp, scl, and spi-1/pu.1 in myeloid differentiation. Spi-1l is coexpressed in a subset of spi-1/pu.1 cells and its function is necessary and sufficient for macrophage and granulocyte differentiation. These results establish a critical role for spi-1l in zebrafish myeloid cell differentiation. PMID:19131555

  10. Biallelic Mutations in DNM1L are Associated with a Slowly Progressive Infantile Encephalopathy.

    PubMed

    Nasca, Alessia; Legati, Andrea; Baruffini, Enrico; Nolli, Cecilia; Moroni, Isabella; Ardissone, Anna; Goffrini, Paola; Ghezzi, Daniele

    2016-09-01

    Mitochondria are highly dynamic organelles, undergoing continuous fission and fusion, and mitochondrial dynamics is important for several cellular functions. DNM1L is the most important mediator of mitochondrial fission, with a role also in peroxisome division. Few reports of patients with genetic defects in DNM1L have been published, most of them describing de novo dominant mutations. We identified compound heterozygous DNM1L variants in two brothers presenting with an infantile slowly progressive neurological impairment. One variant was a frame-shift mutation, the other was a missense change, the pathogenicity of which was validated in a yeast model. Fluorescence microscopy revealed abnormally elongated mitochondria and aberrant peroxisomes in mutant fibroblasts, indicating impaired fission of these organelles. In conclusion, we described a recessive disease caused by DNM1L mutations, with a clinical phenotype resembling mitochondrial disorders but without any biochemical features typical of these syndromes (lactic acidosis, respiratory chain complex deficiency) or indicating a peroxisomal disorder. PMID:27328748

  11. FSCB phosphorylation regulates mouse spermatozoa capacitation through suppressing SUMOylation of ROPN1/ROPN1L

    PubMed Central

    Zhang, Xinqi; Chen, Mingrui; Yu, Renyi; Liu, Benli; Tian, Zhiqiang; Liu, Shunli

    2016-01-01

    Fibrous sheath CABYR binding protein (FSCB) is regulated by protein kinase A (PKA)-mediated tyrosine phosphorylation in the spermatozoa capacitation. Recently, we showed that FSCB phosphorylation activated spermatozoa motility. Nevertheless, the underlying mechanisms have not been completely elucidated. Here, we showed that FSCB phosphorylation inhibited SUMOylation of two crucial proteins ROPN1/ROPN1L that are associated with PKA/A kinase activity and spermatozoa motility. Suppression of SUMOylation of ROPN1/ROPN1L mimicked the effects of FSCB phosphorylation on spermatozoa motility. Immunoprecipitation assay showed that phosphorylated FSCB had a significantly higher affinity to ROPN1/ROPN1L than non-phosphorylated FSCB. Together, our data suggest that FSCB phosphorylation may regulate mouse spermatozoa capacitation through suppressing SUMOylation of ROPN1/ROPN1L, which sheds new light on creating a therapeutic strategy targeting FSCB phosphorylation in the study of infertility.

  12. CpG Island Hypermethylation Frequently Silences FILIP1L Isoform 2 Expression in Prostate Cancer

    PubMed Central

    Desotelle, Joshua; Truong, Matthew; Ewald, Jonathan; Weeratunga, Pushpa; Yang, Bing; Huang, Wei; Jarrard, David

    2013-01-01

    Purpose Senescence related regulatory pathways serve as barriers to cancer immortalization and progression but they are currently not well defined. FILIP1L is a growth inhibitory gene with multiple isoforms whose expression is increased in senescent prostate and prostate cancer cells, and decreased in many cancers. We investigated whether DNA methylation regulates FILIP1L in senescence and in prostate cancer development. Materials and Methods FILIP1L mRNA expression was assessed in prostate cancer and associated normal prostate tissues using quantitative polymerase chain reaction. A tissue microarray was constructed using 95 prostate cancer specimens and 45 benign prostate specimens. Vectra™ imaging was used to quantitate nuclear and cytoplasmic FILIP1L protein expression. Bisulfite sequencing and Pyrosequencing® were used to assess methylation. Prostate cancer cell lines were treated with 2′-deoxy-5-azacytidine and mRNA expression was assessed. Results FILIP1L isoform 2 mRNA was increased in replicatively senescent human prostate epithelial cells and decreased in prostate cancer specimens. We verified a reduction in nuclear FILIP1L protein in prostate cancer using tissue microarrays (p = 0.006). A CpG island 5′ of the isoform 2 translational start site was identified that showed hypermethylation in prostate cancer cell lines and tumors compared to normal prostate cells and tissues. Pyrosequencing confirmed FILIP1L hypermethylation in all 14 tumors compared to paired normal tissues (p <0.0001). Isoform 2 expression was induced in prostate cancer cell lines using 2′-deoxy-5-azacytidine. Conclusions FILIP1L isoform 2 is one of the most commonly hypermethylated genes in prostate cancer. It may serve as an important marker of prostate cancer. Isoform 2 expression is associated with senescence and its down-regulation may represent an early important biological event in prostate cancer development. PMID:23174249

  13. Homozygous YME1L1 mutation causes mitochondriopathy with optic atrophy and mitochondrial network fragmentation

    PubMed Central

    Hartmann, Bianca; Wai, Timothy; Hu, Hao; MacVicar, Thomas; Musante, Luciana; Fischer-Zirnsak, Björn; Stenzel, Werner; Gräf, Ralph; van den Heuvel, Lambert; Ropers, Hans-Hilger; Wienker, Thomas F; Hübner, Christoph; Langer, Thomas; Kaindl, Angela M

    2016-01-01

    Mitochondriopathies often present clinically as multisystemic disorders of primarily high-energy consuming organs. Assembly, turnover, and surveillance of mitochondrial proteins are essential for mitochondrial function and a key task of AAA family members of metalloproteases. We identified a homozygous mutation in the nuclear encoded mitochondrial escape 1-like 1 gene YME1L1, member of the AAA protease family, as a cause of a novel mitochondriopathy in a consanguineous pedigree of Saudi Arabian descent. The homozygous missense mutation, located in a highly conserved region in the mitochondrial pre-sequence, inhibits cleavage of YME1L1 by the mitochondrial processing peptidase, which culminates in the rapid degradation of YME1L1 precursor protein. Impaired YME1L1 function causes a proliferation defect and mitochondrial network fragmentation due to abnormal processing of OPA1. Our results identify mutations in YME1L1 as a cause of a mitochondriopathy with optic nerve atrophy highlighting the importance of YME1L1 for mitochondrial functionality in humans. DOI: http://dx.doi.org/10.7554/eLife.16078.001 PMID:27495975

  14. Reciprocal Degradation of YME1L and OMA1 Adapts Mitochondrial Proteolytic Activity During Stress

    PubMed Central

    Rainbolt, T. Kelly; Lebeau, Justine; Puchades, Cristina; Wiseman, R. Luke

    2016-01-01

    SUMMARY The mitochondrial inner membrane proteases YME1L and OMA1 are critical regulators of essential mitochondrial functions including inner membrane proteostasis maintenance and mitochondrial dynamics. Here, we show that YME1L and OMA1 are reciprocally degraded in response to distinct types of cellular stress. OMA1 is degraded through a YME1L-dependent mechanism in response to toxic insults that depolarize the mitochondrial membrane. Alternatively, insults that depolarize mitochondria and deplete cellular ATP stabilize active OMA1 and promote YME1L degradation. We show that the differential degradation of YME1L and OMA1 alters their proteolytic processing of the dynamin-like GTPase OPA1, a critical regulator of mitochondrial inner membrane morphology, which influences the recovery of tubular mitochondria following membrane depolarization-induced fragmentation. Our results reveal the differential stress-induced degradation of YME1L and OMA1 as a mechanism to sensitively adapt mitochondrial inner membrane protease activity and function in response to distinct types of cellular insults. PMID:26923599

  15. The clathrin adaptor Numb regulates intestinal cholesterol absorption through dynamic interaction with NPC1L1.

    PubMed

    Li, Pei-Shan; Fu, Zhen-Yan; Zhang, Ying-Yu; Zhang, Jin-Hui; Xu, Chen-Qi; Ma, Yi-Tong; Li, Bo-Liang; Song, Bao-Liang

    2014-01-01

    Hypercholesterolemia, typically due to excessive cholesterol uptake, is a major risk factor for cardiovascular disease, which is responsible for ∼50% of all deaths in developed societies. Although it has been shown that intestinal cholesterol absorption is mediated by vesicular endocytosis of the Niemann-Pick C1-like 1 (NPC1L1) protein, the mechanism of sterol-stimulated NPC1L1 internalization is still mysterious. Here, we identified an endocytic peptide signal, YVNXXF (where X stands for any amino acid), in the cytoplasmic C-terminal tail of NPC1L1. Cholesterol binding on the N-terminal domain of NPC1L1 released the YVNXXF-containing region of NPC1L1 from association with the plasma membrane and enabled Numb binding. We also found that Numb, a clathrin adaptor, specifically recognized this motif and recruited clathrin for internalization. Disrupting the NPC1L1-Numb interaction decreased cholesterol uptake. Ablation of Numb in mouse intestine significantly reduced dietary cholesterol absorption and plasma cholesterol level. Together, these data show that Numb is a pivotal protein for intestinal cholesterol absorption and may provide a therapeutic target for hypercholesterolemia. PMID:24336247

  16. Homozygous YME1L1 mutation causes mitochondriopathy with optic atrophy and mitochondrial network fragmentation.

    PubMed

    Hartmann, Bianca; Wai, Timothy; Hu, Hao; MacVicar, Thomas; Musante, Luciana; Fischer-Zirnsak, Björn; Stenzel, Werner; Gräf, Ralph; van den Heuvel, Lambert; Ropers, Hans-Hilger; Wienker, Thomas F; Hübner, Christoph; Langer, Thomas; Kaindl, Angela M

    2016-01-01

    Mitochondriopathies often present clinically as multisystemic disorders of primarily high-energy consuming organs. Assembly, turnover, and surveillance of mitochondrial proteins are essential for mitochondrial function and a key task of AAA family members of metalloproteases. We identified a homozygous mutation in the nuclear encoded mitochondrial escape 1-like 1 gene YME1L1, member of the AAA protease family, as a cause of a novel mitochondriopathy in a consanguineous pedigree of Saudi Arabian descent. The homozygous missense mutation, located in a highly conserved region in the mitochondrial pre-sequence, inhibits cleavage of YME1L1 by the mitochondrial processing peptidase, which culminates in the rapid degradation of YME1L1 precursor protein. Impaired YME1L1 function causes a proliferation defect and mitochondrial network fragmentation due to abnormal processing of OPA1. Our results identify mutations in YME1L1 as a cause of a mitochondriopathy with optic nerve atrophy highlighting the importance of YME1L1 for mitochondrial functionality in humans. PMID:27495975

  17. Deletion of Mthfd1l causes embryonic lethality and neural tube and craniofacial defects in mice.

    PubMed

    Momb, Jessica; Lewandowski, Jordan P; Bryant, Joshua D; Fitch, Rebecca; Surman, Deborah R; Vokes, Steven A; Appling, Dean R

    2013-01-01

    Maternal supplementation with folic acid is known to reduce the incidence of neural tube defects (NTDs) by as much as 70%. Despite the strong clinical link between folate and NTDs, the biochemical mechanisms through which folic acid acts during neural tube development remain undefined. The Mthfd1l gene encodes a mitochondrial monofunctional 10-formyl-tetrahydrofolate synthetase, termed MTHFD1L. This gene is expressed in adults and at all stages of mammalian embryogenesis with localized regions of higher expression along the neural tube, developing brain, craniofacial structures, limb buds, and tail bud. In both embryos and adults, MTHFD1L catalyzes the last step in the flow of one-carbon units from mitochondria to cytoplasm, producing formate from 10-formyl-THF. To investigate the role of mitochondrial formate production during embryonic development, we have analyzed Mthfd1l knockout mice. All embryos lacking Mthfd1l exhibit aberrant neural tube closure including craniorachischisis and exencephaly and/or a wavy neural tube. This fully penetrant folate-pathway mouse model does not require feeding a folate-deficient diet to cause this phenotype. Maternal supplementation with sodium formate decreases the incidence of NTDs and partially rescues the growth defect in embryos lacking Mthfd1l. These results reveal the critical role of mitochondrially derived formate in mammalian development, providing a mechanistic link between folic acid and NTDs. In light of previous studies linking a common splice variant in the human MTHFD1L gene with increased risk for NTDs, this mouse model provides a powerful system to help elucidate the specific metabolic mechanisms that underlie folate-associated birth defects, including NTDs. PMID:23267094

  18. Identification of the α1L-adrenoceptor in rat cerebral cortex and possible relationship between α1L- and α1A-adrenoceptors

    PubMed Central

    Morishima, S; Suzuki, F; Yoshiki, H; Md Anisuzzaman, A S; Sathi, Z S; Tanaka, T; Muramatsu, I

    2008-01-01

    Background and purpose: In addition to α1A, α1B and α1D-adrenoceptors (ARs), putative α1L-ARs with a low affinity for prazosin have been proposed. The purpose of the present study was to identify the α1A-AR and clarify its pharmacological profile using a radioligand binding assay. Experimental approach: Binding experiments with [3H]-silodosin and [3H]-prazosin were performed in intact tissue segments and crude membrane preparations of rat cerebral cortex. Intact tissue binding assays were also conducted in rat tail artery. Key results: [3H]-silodosin at subnanomolar concentrations specifically bound to intact tissue segments and membrane preparations of rat cerebral cortex at the same density (approximately 150 fmol mg−1 total tissue protein). The binding sites in intact segments consisted of α1A and α1L-ARs that had different affinities for prazosin, while the binding sites in membranes showed an α1A-AR-like profile having single high affinity for prazosin. [3H]-prazosin also bound at subnanomolar concentrations to α1A and α1B-ARs but not α1L-ARs in cerebral cortex; the binding densities being approximately 200 and 290 fmol mg−1 protein in the segments and the membranes, respectively. In the segments of tail artery, [3H]-silodosin only recognized α1A-ARs, whereas [3H]-prazosin bound to α1A and α1B-ARs. Conclusions and implications: The present study clearly reveals the presence of α1L-ARs as a pharmacologically distinct entity from α1A and α1B-ARs in intact tissue segments of rat cerebral cortex but not tail artery. However, the α1L-ARs disappeared after tissue homogenization, suggesting their decomposition and/or their pharmacological profile changes to that of α1A-ARs. PMID:18223667

  19. Conservation of imprinting of MKRN3 and NAP1L5 in rabbits.

    PubMed

    Yuan, L; Lai, L; Duan, F; Chen, M; Deng, J; Li, Z

    2016-08-01

    Maternally imprinted genes of makorin ring finger protein 3 (MKRN3) and nucleosome assembly protein 1-like 5 (NAP1L5) have been identified in many species but have not yet been investigated in rabbits. In this study, a polymorphism-based approach and bisulfite-sequencing PCR (BSP) were used to determine the imprinting status of MKRN3 and NAP1L5 in rabbits. The single nucleotide polymorphism (SNP)-based sequencing results demonstrated that MKRN3 and NAP1L5 were expressed preferentially from the paternal allele. Furthermore, the BSP results showed the gamete-specific methylation patterns and hemimethylation in brain and full methylation in liver were observed in MKRN3 and NAP1L5 respectively. Thus, we provide the first evidence that MKRN3 and NAP1L5 are paternally expressed genes and that the CpG islands located in the promoter region may be the putative differentially methylated region of these two genes in rabbits. PMID:27091003

  20. Histone Methyltransferase DOT1L Drives Recovery of Gene Expression after a Genotoxic Attack

    PubMed Central

    Ziani, Salim; Mari, Pierre-Olivier; Eberova, Jitka; Nardo, Tiziana; Stefanini, Miria; Giglia-Mari, Giuseppina; Egly, Jean-Marc; Coin, Frédéric

    2013-01-01

    UV-induced DNA damage causes repression of RNA synthesis. Following the removal of DNA lesions, transcription recovery operates through a process that is not understood yet. Here we show that knocking-out of the histone methyltransferase DOT1L in mouse embryonic fibroblasts (MEFDOT1L) leads to a UV hypersensitivity coupled to a deficient recovery of transcription initiation after UV irradiation. However, DOT1L is not implicated in the removal of the UV-induced DNA damage by the nucleotide excision repair pathway. Using FRAP and ChIP experiments we established that DOT1L promotes the formation of the pre-initiation complex on the promoters of UV-repressed genes and the appearance of transcriptionally active chromatin marks. Treatment with Trichostatin A, relaxing chromatin, recovers both transcription initiation and UV-survival. Our data suggest that DOT1L secures an open chromatin structure in order to reactivate RNA Pol II transcription initiation after a genotoxic attack. PMID:23861670

  1. Histone methyltransferase DOT1L drives recovery of gene expression after a genotoxic attack.

    PubMed

    Oksenych, Valentyn; Zhovmer, Alexander; Ziani, Salim; Mari, Pierre-Olivier; Eberova, Jitka; Nardo, Tiziana; Stefanini, Miria; Giglia-Mari, Giuseppina; Egly, Jean-Marc; Coin, Frédéric

    2013-01-01

    UV-induced DNA damage causes repression of RNA synthesis. Following the removal of DNA lesions, transcription recovery operates through a process that is not understood yet. Here we show that knocking-out of the histone methyltransferase DOT1L in mouse embryonic fibroblasts (MEF(DOT1L)) leads to a UV hypersensitivity coupled to a deficient recovery of transcription initiation after UV irradiation. However, DOT1L is not implicated in the removal of the UV-induced DNA damage by the nucleotide excision repair pathway. Using FRAP and ChIP experiments we established that DOT1L promotes the formation of the pre-initiation complex on the promoters of UV-repressed genes and the appearance of transcriptionally active chromatin marks. Treatment with Trichostatin A, relaxing chromatin, recovers both transcription initiation and UV-survival. Our data suggest that DOT1L secures an open chromatin structure in order to reactivate RNA Pol II transcription initiation after a genotoxic attack. PMID:23861670

  2. Crystal structure of SEL1L: Insight into the roles of SLR motifs in ERAD pathway

    PubMed Central

    Jeong, Hanbin; Sim, Hyo Jung; Song, Eun Kyung; Lee, Hakbong; Ha, Sung Chul; Jun, Youngsoo; Park, Tae Joo; Lee, Changwook

    2016-01-01

    Terminally misfolded proteins are selectively recognized and cleared by the endoplasmic reticulum-associated degradation (ERAD) pathway. SEL1L, a component of the ERAD machinery, plays an important role in selecting and transporting ERAD substrates for degradation. We have determined the crystal structure of the mouse SEL1L central domain comprising five Sel1-Like Repeats (SLR motifs 5 to 9; hereafter called SEL1Lcent). Strikingly, SEL1Lcent forms a homodimer with two-fold symmetry in a head-to-tail manner. Particularly, the SLR motif 9 plays an important role in dimer formation by adopting a domain-swapped structure and providing an extensive dimeric interface. We identified that the full-length SEL1L forms a self-oligomer through the SEL1Lcent domain in mammalian cells. Furthermore, we discovered that the SLR-C, comprising SLR motifs 10 and 11, of SEL1L directly interacts with the N-terminus luminal loops of HRD1. Therefore, we propose that certain SLR motifs of SEL1L play a unique role in membrane bound ERAD machinery. PMID:27064360

  3. MLL1 and DOT1L cooperate with meningioma-1 to induce acute myeloid leukemia.

    PubMed

    Riedel, Simone S; Haladyna, Jessica N; Bezzant, Matthew; Stevens, Brett; Pollyea, Daniel A; Sinha, Amit U; Armstrong, Scott A; Wei, Qi; Pollock, Roy M; Daigle, Scott R; Jordan, Craig T; Ernst, Patricia; Neff, Tobias; Bernt, Kathrin M

    2016-04-01

    Meningioma-1 (MN1) overexpression is frequently observed in patients with acute myeloid leukemia (AML) and is predictive of poor prognosis. In murine models, forced expression of MN1 in hematopoietic progenitors induces an aggressive myeloid leukemia that is strictly dependent on a defined gene expression program in the cell of origin, which includes the homeobox genes Hoxa9 and Meis1 as key components. Here, we have shown that this program is controlled by two histone methyltransferases, MLL1 and DOT1L, as deletion of either Mll1 or Dot1l in MN1-expressing cells abrogated the cell of origin-derived gene expression program, including the expression of Hoxa cluster genes. In murine models, genetic inactivation of either Mll1 or Dot1l impaired MN1-mediated leukemogenesis. We determined that HOXA9 and MEIS1 are coexpressed with MN1 in a subset of clinical MN1hi leukemia, and human MN1hi/HOXA9hi leukemias were sensitive to pharmacologic inhibition of DOT1L. Together, these data point to DOT1L as a potential therapeutic target in MN1hi AML. In addition, our findings suggest that epigenetic modulation of the interplay between an oncogenic lesion and its cooperating developmental program has therapeutic potential in AML. PMID:26927674

  4. STIM1L traps and gates Orai1 channels without remodeling the cortical ER

    PubMed Central

    Saüc, Sophie; Bulla, Monica; Nunes, Paula; Orci, Lelio; Marchetti, Anna; Antigny, Fabrice; Bernheim, Laurent; Cosson, Pierre; Frieden, Maud; Demaurex, Nicolas

    2015-01-01

    STIM proteins populate and expand cortical endoplasmic reticulum (ER) sheets to mediate store-operated Ca2+ entry (SOCE) by trapping and gating Orai channels in ER-plasma membrane clusters. A longer splice variant, STIM1L, forms permanent ER-plasma membrane clusters and mediates rapid Ca2+ influx in muscle. Here, we used electron microscopy, total internal reflection fluorescence (TIRF) microscopy and Ca2+ imaging to establish the trafficking and signaling properties of the two STIM1 isoforms in Stim1−/−/Stim2−/− fibroblasts. Unlike STIM1, STIM1L was poorly recruited into ER-plasma membrane clusters and did not mediate store-dependent expansion of cortical ER cisternae. Removal of the STIM1 lysine-rich tail prevented store-dependent cluster enlargement, whereas inhibition of cytosolic Ca2+ elevations or removal of the STIM1L actin-binding domain had no impact on cluster expansion. Finally, STIM1L restored robust but not accelerated SOCE and clustered with Orai1 channels more slowly than STIM1 following store depletion. These results indicate that STIM1L does not mediate rapid SOCE but can trap and gate Orai1 channels efficiently without remodeling cortical ER cisternae. The ability of STIM proteins to induce cortical ER formation is dispensable for SOCE and requires the lysine-rich tail of STIM1 involved in binding to phosphoinositides. PMID:25736291

  5. MLL1 and DOT1L cooperate with meningioma-1 to induce acute myeloid leukemia

    PubMed Central

    Riedel, Simone S.; Haladyna, Jessica N.; Bezzant, Matthew; Stevens, Brett; Pollyea, Daniel A.; Sinha, Amit U.; Armstrong, Scott A.; Wei, Qi; Pollock, Roy M.; Daigle, Scott R.; Jordan, Craig T.; Ernst, Patricia; Bernt, Kathrin M.

    2016-01-01

    Meningioma-1 (MN1) overexpression is frequently observed in patients with acute myeloid leukemia (AML) and is predictive of poor prognosis. In murine models, forced expression of MN1 in hematopoietic progenitors induces an aggressive myeloid leukemia that is strictly dependent on a defined gene expression program in the cell of origin, which includes the homeobox genes Hoxa9 and Meis1 as key components. Here, we have shown that this program is controlled by two histone methyltransferases, MLL1 and DOT1L, as deletion of either Mll1 or Dot1l in MN1-expressing cells abrogated the cell of origin–derived gene expression program, including the expression of Hoxa cluster genes. In murine models, genetic inactivation of either Mll1 or Dot1l impaired MN1-mediated leukemogenesis. We determined that HOXA9 and MEIS1 are coexpressed with MN1 in a subset of clinical MN1hi leukemia, and human MN1hi/HOXA9hi leukemias were sensitive to pharmacologic inhibition of DOT1L. Together, these data point to DOT1L as a potential therapeutic target in MN1hi AML. In addition, our findings suggest that epigenetic modulation of the interplay between an oncogenic lesion and its cooperating developmental program has therapeutic potential in AML. PMID:26927674

  6. Vaccinia Virus Virulence Factor N1L is a Novel Promising Target for Antiviral Therapeutic Intervention

    PubMed Central

    Cheltsov, Anton V.; Aoyagi, Mika; Aleshin, Alexander; Chi-Wang, Yu Eric; Gilliland, Taylor; Zhai, Dayong; Bobkov, Andrey A.; Reed, John C.; Liddington, Robert C.; Abagyan, Ruben

    2010-01-01

    The 14 kDa homodimeric N1L protein is a potent vaccinia and variola (smallpox) virulence factor. It is not essential for viral replication, but it causes a strong attenuation of viral production in culture when deleted. The N1L protein is predicted to contain the BH3-like binding domain characteristic of Bcl-2 family proteins, and it is able to bind the BH3 peptides. Its overexpression has been reported to prevent infected cells from committing apoptosis. Therefore, interfering with the N1L apoptotic blockade may be a legitimate therapeutic strategy affecting the viral growth. By using in silico ligand docking and an array of in vitro assays, we have identified sub-micromolar (600 nM) N1L antagonists, belonging to the family of polyphenols. Their affinity is comparable to that of the BH3 peptides (70 nM ÷ 1000 nM). We have also identified the natural polyphenol resveratrol as a moderate N1L inhibitor. Finally, we show that our ligands efficiently inhibit growth of vaccinia virus. PMID:20441222

  7. Structure-function analysis of human protein Ero1-L{alpha}

    SciTech Connect

    Chu, Yanyan; Yang, Charles; Chen, Xianjun; Zheng, Wenyun; Yang, Yi; Tang, Yun

    2009-11-27

    Human Ero1-L{alpha} catalyzes the formation of disulfide bond and hence plays an essential role in protein folding. Understanding the mechanism of disulfide bond formation in mammals is important because of the involvement of protein misfolding in conditions such as diabetes, arthritis, cancer, and aging. However, the crystal structure of the enzyme is not available yet, which seriously hinders the understanding of biological function of Ero1-L{alpha}. Based on the crystal structure of yeast Ero1p, a rational three-dimensional structural model of Ero1-L{alpha} was built and the characteristics of the enzyme were hence investigated. The characteristic similarities and differences between Ero1-L{alpha} and Ero1p were compared on the basis of computational and experimental results, providing the first insight into the structure-function relationships of the enzymes. Both calculation and experiment got the concordant conclusion that FAD binds more tightly with Ero1-L{alpha} than Ero1p. In addition, the probable electron transfer pathway was proposed on the basis of the structural models.

  8. The ERdj5-Sel1L complex facilitates cholera toxin retrotranslocation.

    PubMed

    Williams, Jeffrey M; Inoue, Takamasa; Banks, Lindsey; Tsai, Billy

    2013-03-01

    Cholera toxin (CT) traffics from the host cell surface to the endoplasmic reticulum (ER), where the toxin's catalytic CTA1 subunit retrotranslocates to the cytosol to induce toxicity. In the ER, CT is captured by the E3 ubiquitin ligase Hrd1 via an undefined mechanism to prepare for retrotranslocation. Using loss-of-function and gain-of-function approaches, we demonstrate that the ER-resident factor ERdj5 promotes CTA1 retrotranslocation, in part, via its J domain. This Hsp70 cochaperone regulates binding between CTA and the ER Hsp70 BiP, a chaperone previously implicated in toxin retrotranslocation. Importantly, ERdj5 interacts with the Hrd1 adaptor Sel1L directly through Sel1L's N-terminal lumenal domain, thereby linking ERdj5 to the Hrd1 complex. Sel1L itself also binds CTA and facilitates toxin retrotranslocation. By contrast, EDEM1 and OS-9, two established Sel1L binding partners, do not play significant roles in CTA1 retrotranslocation. Our results thus identify two ER factors that promote ER-to-cytosol transport of CTA1. They also indicate that ERdj5, by binding to Sel1L, triggers BiP-toxin interaction proximal to the Hrd1 complex. We postulate this scenario enables the Hrd1-associated retrotranslocation machinery to capture the toxin efficiently once the toxin is released from BiP. PMID:23363602

  9. Impaired complex III assembly associated with BCS1L gene mutations in isolated mitochondrial encephalopathy.

    PubMed

    Fernandez-Vizarra, Erika; Bugiani, Marianna; Goffrini, Paola; Carrara, Franco; Farina, Laura; Procopio, Elena; Donati, Alice; Uziel, Graziella; Ferrero, Iliana; Zeviani, Massimo

    2007-05-15

    We investigated two unrelated children with an isolated defect of mitochondrial complex III activity. The clinical picture was characterized by a progressive encephalopathy featuring early-onset developmental delay, spasticity, seizures, lactic acidosis, brain atrophy and MRI signal changes in the basal ganglia. Both children were compound heterozygotes for novel mutations in the human bc1 synthesis like (BCS1L) gene, which encodes an AAA mitochondrial protein putatively involved in both iron homeostasis and complex III assembly. The pathogenic role of the mutations was confirmed by complementation assays, using a DeltaBcs1 strain of Saccharomyces cerevisiae. By investigating complex III assembly and the structural features of the BCS1L gene product in skeletal muscle, cultured fibroblasts and lymphoblastoid cell lines from our patients, we have demonstrated, for the first time in a mammalian system, that a major function of BCS1L is to promote the maturation of complex III and, more specifically, the incorporation of the Rieske iron-sulfur protein into the nascent complex. Defective BCS1L leads to the formation of a catalytically inactive, structurally unstable complex III. We have also shown that BCS1L is contained within a high-molecular-weight supramolecular complex which is clearly distinct from complex III intermediates. PMID:17403714

  10. Understanding CrRLK1L Function: Cell Walls and Growth Control.

    PubMed

    Nissen, Karen S; Willats, William G T; Malinovsky, Frederikke G

    2016-06-01

    To develop successfully in an ever-changing environment, it is essential for plants to monitor and control their growth. Therefore, cell expansion is carefully regulated to establish correct cell shape and size. In this review, we explore the role of the Catharanthus roseus receptor-like kinase (CrRLK1L) subfamily as regulators of cell expansion. Recently, the downstream signalling events of individual CrRLK1L pathways were discovered, implicating known modulators of cell expansion, such as reactive oxygen species (ROS) production, Ca(2+) dynamics, and exocytosis of cell wall material. Based on these intriguing new insights, we propose a model for a common pathway of CrRLK1L signalling that enables spatial and temporal control of cell wall extensibility throughout the plant. PMID:26778775

  11. Hypomorphism of Fto and Rpgrip1l causes obesity in mice.

    PubMed

    Stratigopoulos, George; Burnett, Lisa Cole; Rausch, Richard; Gill, Richard; Penn, David Barth; Skowronski, Alicja A; LeDuc, Charles A; Lanzano, Anthony J; Zhang, Pumin; Storm, Daniel R; Egli, Dieter; Leibel, Rudolph L

    2016-05-01

    Noncoding polymorphisms in the fat mass and obesity-associated (FTO) gene represent common alleles that are strongly associated with effects on food intake and adiposity in humans. Previous studies have suggested that the obesity-risk allele rs8050136 in the first intron of FTO alters a regulatory element recognized by the transcription factor CUX1, thereby leading to decreased expression of FTO and retinitis pigmentosa GTPase regulator-interacting protein-1 like (RPGRIP1L). Here, we evaluated the effects of rs8050136 and another potential CUX1 element in rs1421085 on expression of nearby genes in human induced pluripotent stem cell-derived (iPSC-derived) neurons. There were allele-dosage effects on FTO, RPGRIP1L, and AKT-interacting protein (AKTIP) expression, but expression of other vicinal genes, including IRX3, IRX5, and RBL2, which have been implicated in mediating functional effects, was not altered. In vivo manipulation of CUX1, Fto, and/or Rpgrip1l expression in mice affected adiposity in a manner that was consistent with CUX1 influence on adiposity via remote effects on Fto and Rpgrip1l expression. In support of a mechanism, mice hypomorphic for Rpgrip1l exhibited hyperphagic obesity, as the result of diminished leptin sensitivity in Leprb-expressing neurons. Together, the results of this study indicate that the effects of FTO-associated SNPs on energy homeostasis are due in part to the effects of these genetic variations on hypothalamic FTO, RPGRIP1L, and possibly other genes. PMID:27064284

  12. Auxiliary Subunit GSG1L Acts to Suppress Calcium-Permeable AMPA Receptor Function

    PubMed Central

    McGee, Thomas P.; Bats, Cécile

    2015-01-01

    AMPA-type glutamate receptors are ligand-gated cation channels responsible for a majority of the fast excitatory synaptic transmission in the brain. Their behavior and calcium permeability depends critically on their subunit composition and the identity of associated auxiliary proteins. Calcium-permeable AMPA receptors (CP-AMPARs) contribute to various forms of synaptic plasticity, and their dysfunction underlies a number of serious neurological conditions. For CP-AMPARs, the prototypical transmembrane AMPAR regulatory protein stargazin, which acts as an auxiliary subunit, enhances receptor function by increasing single-channel conductance, slowing channel gating, increasing calcium permeability, and relieving the voltage-dependent block by endogenous intracellular polyamines. We find that, in contrast, GSG1L, a transmembrane auxiliary protein identified recently as being part of the AMPAR proteome, acts to reduce the weighted mean single-channel conductance and calcium permeability of recombinant CP-AMPARs, while increasing polyamine-dependent rectification. To examine the effects of GSG1L on native AMPARs, we manipulated its expression in cerebellar and hippocampal neurons. Transfection of GSG1L into mouse cultured cerebellar stellate cells that lack this protein increased the inward rectification of mEPSCs. Conversely, shRNA-mediated knockdown of endogenous GSG1L in rat cultured hippocampal pyramidal neurons led to an increase in mEPSC amplitude and in the underlying weighted mean single-channel conductance, revealing that GSG1L acts to suppress current flow through native CP-AMPARs. Thus, our data suggest that GSG1L extends the functional repertoire of AMPAR auxiliary subunits, which can act not only to enhance but also diminish current flow through their associated AMPARs. SIGNIFICANCE STATEMENT Calcium-permeable AMPA receptors (CP-AMPARs) are an important group of receptors for the neurotransmitter glutamate. These receptors contribute to various forms of

  13. NDH-1L interacts with ferredoxin via the subunit NdhS in Thermosynechococcus elongatus.

    PubMed

    He, Zhihui; Zheng, Fangfang; Wu, Yaozong; Li, Qinghua; Lv, Jing; Fu, Pengcheng; Mi, Hualing

    2015-12-01

    The large size complex of cyanobacterial NAD(P)H dehydrogenase (NDH-1) complex (NDH-1L) plays crucial role in a variety of bioenergetic reactions such as respiration and cyclic electron flow around photosystem I. Although the complex has been isolated and identified, its biochemical function still remains to be clarified. Here, we highly purified the NDH-1L complex from the cells of Thermosynechococcus elongatus by Ni(2+) affinity chromatography and size-exclusion chromatography. The purified NDH-1L complex has an apparent total molecular mass of approximately 500 kDa. 14 known subunits were identified by mass spectrometry and immunoblotting, including the NdhS subunit containing ferredoxin (Fd)-docking site domain. Surface plasmon resonance measurement demonstrates that the NDH-1L complex could bind to Fd with the binding constant (K D) of 59 µM. Yeast two-hybrid system assay further confirmed the interaction of Fd with NdhS and indicated that NdhH is involved in the interaction. Our results provide direct biochemical evidence that the cyanobacterial NDH-1 complex catalyzes the electron transport from reduced Fd to plastoquinone via NdhS and NdhH. PMID:25630976

  14. Niemann-Pick C1-Like 1 (NPC1L1) Inhibition and Cardiovascular Diseases.

    PubMed

    Pirillo, A; Catapano, A L; Norata, G D

    2016-01-01

    Circulating levels of cholesterol are derived from either endogenous production or intestinal absorption of dietary and biliary cholesterol. Niemann-Pick C1-Like 1 (NPC1L1) is a transmembrane protein that plays a key role in the intestinal absorption of cholesterol by facilitating its uptake through vesicular endocytosis. NPC1L1 is the molecular target of ezetimibe which binds its extracellular loop and inhibits sterol absorption without affecting the absorption of other molecules. Ezetimibe significantly reduces plasma levels of total and low density lipoprotein cholesterol (LDL-C) as monotherapy or when added to statins, the association with a low dose of statin is of particular interest for patients experiencing statin-related side effects. The recent results of the IMPROVE-IT study, which evaluated the cardiovascular effect of ezetimibe added to simvastatin therapy in subjects who had had an acute coronary syndrome and with LDL-C levels within the recommended range, showed that a further LDL-C lowering reduced the incidence of cardiovascular events. To date, ezetimibe represents the only inhibitor of NPC1L1 available for clinical use, however, novel aminoß- lactam ezetimibe derivatives have been synthesized and their efficacy to inhibit NPC1L1 protein and decrease plasma cholesterol levels is under evaluation. PMID:26923679

  15. Mutation of kri1l causes definitive hematopoiesis failure via PERK-dependent excessive autophagy induction.

    PubMed

    Jia, Xiao-E; Ma, Ke; Xu, Tao; Gao, Lei; Wu, Shuang; Fu, Cong; Zhang, Wenjuan; Wang, Zhizhang; Liu, Kaiyu; Dong, Mei; Jing, Changbin; Ren, Chunguang; Dong, Zhiwei; Chen, Yi; Jin, Yi; Huang, Qiuhua; Chang, Xing; Deng, Min; Li, Li; Luo, Lingfei; Zhu, Jun; Dang, Yongjun; Chang, Hung-Chun; Zon, Leonard I; Zhou, Yi; Chen, Saijuan; Pan, Weijun

    2015-08-01

    Dysregulation of ribosome biogenesis causes human diseases, such as Diamond-Blackfan anemia, del (5q-) syndrome and bone marrow failure. However, the mechanisms of blood disorders in these diseases remain elusive. Through genetic mapping, molecular cloning and mechanism characterization of the zebrafish mutant cas002, we reveal a novel connection between ribosomal dysfunction and excessive autophagy in the regulation of hematopoietic stem/progenitor cells (HSPCs). cas002 carries a recessive lethal mutation in kri1l gene that encodes an essential component of rRNA small subunit processome. We show that Kri1l is required for normal ribosome biogenesis, expansion of definitive HSPCs and subsequent lineage differentiation. Through live imaging and biochemical studies, we find that loss of Kri1l causes the accumulation of misfolded proteins and excessive PERK activation-dependent autophagy in HSPCs. Blocking autophagy but not inhibiting apoptosis by Bcl2 overexpression can fully rescue hematopoietic defects, but not the lethality of kri1l(cas002) embryos. Treatment with autophagy inhibitors (3-MA and Baf A1) or PERK inhibitor (GSK2656157), or knockdown of beclin1 or perk can markedly restore HSPC proliferation and definitive hematopoietic cell differentiation. These results may provide leads for effective therapeutics that benefit patients with anemia or bone marrow failure caused by ribosome disorders. PMID:26138676

  16. L1/L2/L3 Writing Development: Longitudinal Case Study of a Japanese Multicompetent Writer

    ERIC Educational Resources Information Center

    Kobayashi, Hiroe; Rinnert, Carol

    2013-01-01

    This longitudinal case study, supplemented by cross-sectional comparisons among five groups of writers with differing backgrounds, investigates how Natsu, a Japanese multilingual writer, developed her L1, L2 (English), and L3 (Chinese) writing competence over two and a half years. To create a comprehensive picture of this multilingual writer, the…

  17. Cone Dystrophy in Patient with Homozygous RP1L1 Mutation

    PubMed Central

    Kikuchi, Sachiko; El Shamieh, Said; Akeo, Keiichiro; Sugawara, Yuko; Yamaki, Kunihiko; Takahashi, Hiroshi

    2015-01-01

    The purpose of this study was to determine whether an autosomal recessive cone dystrophy was caused by a homozygous RP1L1 mutation. A family including one subject affected with cone dystrophy and four unaffected members without evidence of consanguinity underwent detailed ophthalmic evaluations. The ellipsoid and interdigitation zones on the spectral-domain optical coherence tomography images were disorganized in the proband. The proband had a reduced amplitude of cone and flicker full-field electroretinograms (ERGs). Focal macular ERGs and multifocal ERGs were severely reduced in the proband. A homozygous RP1L1 mutation (c.3628T>C, p.S1210P) was identified in the proband. Family members who were heterozygous for the p.S1210P mutation had normal visual acuity and normal results of clinical evaluations. To investigate other putative pathogenic variant(s), a next-generation sequencing (NGS) approach was applied to the proband. NGS identified missense changes in the heterozygous state of the PCDH15, RPGRIP1, and GPR98 genes. None of these variants cosegregated with the phenotype and were predicted to be benign reinforcing the putative pathogenicity of the RP1L1 homozygous mutation. The AO images showed a severe reduction of the cone density in the proband. Our findings indicate that a homozygous p.S1210P exchange in the RP1L1 gene can cause cone dystrophy. PMID:25692141

  18. L1-L2 Convergence in Clausal Packaging in Japanese and English

    ERIC Educational Resources Information Center

    Brown, Amanda; Gullberg, Marianne

    2013-01-01

    This study investigates L1-L2 convergence among bilinguals at an intermediate (CEFR-B2) level of L2 proficiency, focusing on the clausal packaging of Manner and Path of motion. Previous research has shown cross-linguistic differences between English and Japanese in this domain (Allen et al., 2003; Kita & Ozyurek, 2003, though note Brown &…

  19. Histone methyltransferase Ash1L mediates activity-dependent repression of neurexin-1α

    PubMed Central

    Zhu, Τao; Liang, Chen; Li, Dongdong; Tian, Miaomiao; Liu, Sanxiong; Gao, Guanjun; Guan, Ji-Song

    2016-01-01

    Activity-dependent transcription is critical for the regulation of long-term synaptic plasticity and plastic rewiring in the brain. Here, we report that the transcription of neurexin1α (nrxn1α), a presynaptic adhesion molecule for synaptic formation, is regulated by transient neuronal activation. We showed that 10 minutes of firing at 50 Hz in neurons repressed the expression of nrxn1α for 24 hours in a primary cortical neuron culture through a transcriptional repression mechanism. By performing a screening assay using a synthetic zinc finger protein (ZFP) to pull down the proteins enriched near the nrxn1α promoter region in vivo, we identified that Ash1L, a histone methyltransferase, is enriched in the nrxn1α promoter. Neuronal activity triggered binding of Ash1L to the promoter and enriched the histone marker H3K36me2 at the nrxn1α promoter region. Knockout of Ash1L in mice completely abolished the activity-dependent repression of nrxn1α. Taken together, our results reveal that a novel process of activity-dependent transcriptional repression exists in neurons and that Ash1L mediates the long-term repression of nrxn1α, thus implicating an important role for epigenetic modification in brain functioning. PMID:27229316

  20. Integrating Meaning and Structure in L1-L2 and L2-L1 Translations

    ERIC Educational Resources Information Center

    Lim, Jung Hyun; Christianson, Kiel

    2013-01-01

    This article examined the integration of semantic and morphosyntactic information by Korean learners of English as a second language (L2). In Experiment 1, L2 learners listened to English active or passive sentences that were either plausible or implausible and translated them into Korean. A significant number of Korean translations maintained the…

  1. DNM1L-related mitochondrial fission defect presenting as refractory epilepsy.

    PubMed

    Vanstone, Jason R; Smith, Amanda M; McBride, Skye; Naas, Turaya; Holcik, Martin; Antoun, Ghadi; Harper, Mary-Ellen; Michaud, Jean; Sell, Erick; Chakraborty, Pranesh; Tetreault, Martine; Majewski, Jacek; Baird, Stephen; Boycott, Kym M; Dyment, David A; MacKenzie, Alex; Lines, Matthew A

    2016-07-01

    Mitochondrial fission and fusion are dynamic processes vital to mitochondrial quality control and the maintenance of cellular respiration. In dividing mitochondria, membrane scission is accomplished by a dynamin-related GTPase, DNM1L, that oligomerizes at the site of fission and constricts in a GTP-dependent manner. There is only a single previous report of DNM1L-related clinical disease: a female neonate with encephalopathy due to defective mitochondrial and peroxisomal fission (EMPF; OMIM #614388), a lethal disorder characterized by cerebral dysgenesis, seizures, lactic acidosis, elevated very long chain fatty acids, and abnormally elongated mitochondria and peroxisomes. Here, we describe a second individual, diagnosed via whole-exome sequencing, who presented with developmental delay, refractory epilepsy, prolonged survival, and no evidence of mitochondrial or peroxisomal dysfunction on standard screening investigations in blood and urine. EEG was nonspecific, showing background slowing with frequent epileptiform activity at the frontal and central head regions. Electron microscopy of skeletal muscle showed subtle, nonspecific abnormalities of cristal organization, and confocal microscopy of patient fibroblasts showed striking hyperfusion of the mitochondrial network. A panel of further bioenergetic studies in patient fibroblasts showed no significant differences versus controls. The proband's de novo DNM1L variant, NM_012062.4:c.1085G>A; NP_036192.2:p.(Gly362Asp), falls within the middle (oligomerization) domain of DNM1L, implying a likely dominant-negative mechanism. This disorder, which presents nonspecifically and affords few diagnostic clues, can be diagnosed by means of DNM1L sequencing and/or confocal microscopy. PMID:26604000

  2. Shipper's guide to energy conservation

    SciTech Connect

    Marien, E.J.

    1980-01-01

    Recommendations are presented for money-saving tips for the shipping industry. Tips are included for the warehousing and distribution center management; transportation management; materials management for product packaging and shipping; and deployment and management of inventories sections in a firm.

  3. Captive shippers issue: an update

    SciTech Connect

    Not Available

    1984-10-11

    For quite some while now, electric utilities have been complaining about the cost of coal transported by rail. Over half of the electricity produced in the United States comes from coal; about 60 per cent of that is shipped on railroads. And according to the Edison Electric Institute, the majority of power plants currently using rail transportation of coal have no alternative available and are thus captive to the railroads. Railroads seem to have taken advantage of their situation, charging utilities nearly $5 billion last year. In some cases, the cost of rail transportation is almost 80 per cent of the delivered price of coal.

  4. Constraints on the U(1)L gauge boson in a wide mass range

    NASA Astrophysics Data System (ADS)

    Jeong, Yu Seon; Kim, C. S.; Lee, Hye-Sung

    2016-04-01

    There is a growing interest for the search of new light gauge bosons. The small mass of a new boson can turn various kinds of low-energy experiments to a new discovery machine, depending on their couplings to the Standard Model particles. It is important to understand the properties of each type of gauge boson and their current constraints for a given mass. While the dark photon (which couples to the electric charges) and the U(1)B‑L gauge boson have been well studied in an extensive mass range, the U(1)L gauge boson has not been fully investigated yet. We consider the gauge boson of the U(1)L in a wide mass range mZ‧≈ 0-1012eV and investigate the constraints on its coupling from various experiments, discussing the similarities and differences from the dark photon and the U(1)B‑L gauge boson.

  5. 1 L-myo-Inositol 1-Phosphate Synthase from Arabidopsis thaliana.

    PubMed Central

    Johnson, M. D.; Sussex, I. M.

    1995-01-01

    A recombinant phage containing an Arabidopsis thaliana cDNA sequence encoding a protein with 1L-myo-inositol 1-phosphate synthase (EC 5.5.1.4) activity has been isolated and used for transcriptional and translational studies. The identification of the recombinant phage relied on the observations that (a) the clone complements a mutation in the structural gene for 1L-myo-inositol 1-phosphate synthase in the yeast Saccharomyces cerevisiae, (b) the in vitro synthesized polypeptide enzymatically converts glucose 6-phosphate into inositol 1-phosphate, (c) in vitro transcription and translation of this cDNA sequence produces a polypeptide that is recognized by anti-yeast myo-inositol 1-phosphate synthase antiserum, and (d) inositol regulates the expression of the corresponding gene in Arabidopsis. PMID:12228386

  6. Human CD180 Transmits Signals via the PIM-1L Kinase

    PubMed Central

    Egli, Nicole; Zajonz, Alexandra; Burger, Matthew T.; Schweighoffer, Tamas

    2015-01-01

    Toll-like receptors (TLRs) are important sensors of the innate immune system that recognize conserved structural motifs and activate cells via a downstream signaling cascade. The CD180/MD1 molecular complex is an unusual member of the TLR family, since it lacks the components that are normally required for signal transduction by other TLRs. Therefore the CD180/MD 1 complex has been considered of being incapable of independently initiating cellular signals. Using chemogenetic approaches we identified specifically the membrane bound long form of PIM-1 kinase, PIM-1L as the mediator of CD180-dependent signaling. A dominant negative isoform of PIM-1L, but not of other PIM kinases, inhibited signaling elicited by cross-linking of CD180, and this effect was phenocopied by PIM inhibitors. PIM-1L was directed to the cell membrane by its N-terminal extension, where it colocalized and physically associated with CD180. Triggering CD180 also induced increased phosphorylation of the anti-apoptotic protein BAD in a PIM kinase-dependent fashion. Also in primary human B cells, which are the main cells expressing CD180 in man, cross-linking of CD180 by monoclonal antibodies stimulated cell survival and proliferation that was abrogated by specific inhibitors. By associating with PIM-1L, CD180 can thus obtain autonomous signaling capabilities, and this complex is then channeling inflammatory signals into B cell survival programs. Pharmacological inhibition of PIM-1 should therefore provide novel therapeutic options in diseases that respond to innate immune stimulation with subsequently increased B cell activity, such as lupus erythematosus or myasthenia gravis. PMID:26555723

  7. Sipa1l1 is an early biomarker of liver fibrosis in CCl4-treated rats

    PubMed Central

    Marfà, Santiago; Morales-Ruiz, Manuel; Oró, Denise; Ribera, Jordi; Fernández-Varo, Guillermo; Jiménez, Wladimiro

    2016-01-01

    ABSTRACT At present, several procedures are used for staging liver fibrosis. However, these methods may involve clinical complications and/or present diagnostic uncertainty mainly in the early stages of the disease. Thus, this study was designed to unveil new non-invasive biomarkers of liver fibrosis in an in vivo model of fibrosis/cirrhosis induction by CCl4 inhalation by using a label-free quantitative LC-MS/MS approach. We analyzed 94 serum samples from adult Wistar rats with different degrees of liver fibrosis and 36 control rats. Firstly, serum samples from 18 CCl4-treated rats were clustered into three different groups according to the severity of hepatic and the serum proteome was characterized by label-free LC-MS/MS. Furthermore, three different pooled serum samples obtained from 16 control Wistar rats were also analyzed. Based on the proteomic data obtained, we performed a multivariate analysis which displayed three main cell signaling pathways altered in fibrosis. In cirrhosis, more biological imbalances were detected as well as multi-organ alterations. In addition, hemopexin and signal-induced proliferation-associated 1 like 1 (SIPA1L1) were selected as potential serum markers of liver fibrogenesis among all the analyzed proteins. The results were validated by ELISA in an independent group of 76 fibrotic/cirrhotic rats and 20 controls which confirmed SIPA1L1 as a potential non-invasive biomarker of liver fibrosis. In particular, SIPA1L1 showed a clear diminution in serum samples from fibrotic/cirrhotic rats and a great accuracy at identifying early fibrotic stages. In conclusion, the proteomic analysis of serum samples from CCl4-treated rats has enabled the identification of SIPA1L1 as a non-invasive marker of early liver fibrosis. PMID:27230648

  8. Human CD180 Transmits Signals via the PIM-1L Kinase.

    PubMed

    Egli, Nicole; Zajonz, Alexandra; Burger, Matthew T; Schweighoffer, Tamas

    2015-01-01

    Toll-like receptors (TLRs) are important sensors of the innate immune system that recognize conserved structural motifs and activate cells via a downstream signaling cascade. The CD180/MD1 molecular complex is an unusual member of the TLR family, since it lacks the components that are normally required for signal transduction by other TLRs. Therefore the CD180/MD 1 complex has been considered of being incapable of independently initiating cellular signals. Using chemogenetic approaches we identified specifically the membrane bound long form of PIM-1 kinase, PIM-1L as the mediator of CD180-dependent signaling. A dominant negative isoform of PIM-1L, but not of other PIM kinases, inhibited signaling elicited by cross-linking of CD180, and this effect was phenocopied by PIM inhibitors. PIM-1L was directed to the cell membrane by its N-terminal extension, where it colocalized and physically associated with CD180. Triggering CD180 also induced increased phosphorylation of the anti-apoptotic protein BAD in a PIM kinase-dependent fashion. Also in primary human B cells, which are the main cells expressing CD180 in man, cross-linking of CD180 by monoclonal antibodies stimulated cell survival and proliferation that was abrogated by specific inhibitors. By associating with PIM-1L, CD180 can thus obtain autonomous signaling capabilities, and this complex is then channeling inflammatory signals into B cell survival programs. Pharmacological inhibition of PIM-1 should therefore provide novel therapeutic options in diseases that respond to innate immune stimulation with subsequently increased B cell activity, such as lupus erythematosus or myasthenia gravis. PMID:26555723

  9. Rcan1-1L overexpression induces mitochondrial autophagy and improves cell survival in angiotensin II-exposed cardiomyocytes

    SciTech Connect

    Duan, Hongyan; Li, Yongqiang; Yan, Lijie; Yang, Haitao; Wu, Jintao; Qian, Peng; Li, Bing; Wang, Shanling

    2015-07-01

    Mitochondrial autophagy is an important adaptive stress response and can be modulated by various key molecules. A previous study found that the regulator of calcineurin 1-1L (Rcan1-1L) may regulate mitochondrial autophagy and cause mitochondria degradation in neurocytes. However, the effect of Rcan1-1L on cardiomyocytes has not been determined. In the present study, we aimed to investigate the role of Rcan1-1L in angiotensin II (Ang II)-exposed human cardiomyocytes. Above all, Human adult cardiac myocytes (HACMs) were exposed to 200 nmol/L Ang II for 4 days. Enhanced H{sub 2}O{sub 2} production, cytochrome C release and mitochondrial permeability were observed in these cells, which were blocked by valsartan. Consistently, Ang II exposure significantly reduced cardiomyocyte viability. However, transfection of Rcan1-1L vector promoted cell viability and ameliorated the apoptosis caused by Ang II. Rcan1-1L clearly promoted mitochondrial autophagy in HACMs, with elevated autophagy protein (ATG) 5 and light chain 3 (LC3) expression. Transient mitochondrial biogenesis and reduced cytochrome C release was also induced by Rcan1-1L. Additionally, Rcan1-1L significantly inhibited calcineurin/nuclear factor of activated T cells (NFAT) signaling. We thus conclude that Rcan1-1L may play a protective role in Ang II-treated cardiomyocytes through the induction of mitochondrial autophagy, and may be an alternative method of cardiac protection. - Highlights: • Transfection of Rcan1-1L into HACMs promoted cell viability and reduced apoptosis. • Transfection of Rcan1-1L promoted mitochondrial autophagy in HACMs. • Rcan1-1L inhibited the calcineurin/nuclear factor of activated T cells signaling.

  10. Epithelial Sel1L is required for the maintenance of intestinal homeostasis

    PubMed Central

    Sun, Shengyi; Lourie, Rohan; Cohen, Sara B.; Ji, Yewei; Goodrich, Julia K.; Poole, Angela C.; Ley, Ruth E.; Denkers, Eric Y.; McGuckin, Michael A.; Long, Qiaoming; Duhamel, Gerald E.; Simpson, Kenneth W.; Qi, Ling

    2016-01-01

    Inflammatory bowel disease (IBD) is an incurable chronic idiopathic disease that drastically decreases quality of life. Endoplasmic reticulum (ER)–associated degradation (ERAD) is responsible for the clearance of misfolded proteins; however, its role in disease pathogenesis remains largely unexplored. Here we show that the expression of SEL1L and HRD1, the most conserved branch of mammalian ERAD, is significantly reduced in ileal Crohn’s disease (CD). Consistent with this observation, laboratory mice with enterocyte-specific Sel1L deficiency (Sel1LΔIEC) develop spontaneous enteritis and have increased susceptibility to Toxoplasma gondii–induced ileitis. This is associated with profound defects in Paneth cells and a disproportionate increase of Ruminococcus gnavus, a mucolytic bacterium with known association with CD. Surprisingly, whereas both ER stress sensor IRE1α and effector CHOP are activated in the small intestine of Sel1LΔIEC mice, they are not solely responsible for ERAD deficiency–associated lesions seen in the small intestine. Thus our study points to a constitutive role of Sel1L-Hrd1 ERAD in epithelial cell biology and the pathogenesis of intestinal inflammation in CD. PMID:26631554

  11. A Mutation in the Mitochondrial Fission Gene Dnm1l Leads to Cardiomyopathy

    PubMed Central

    Ashrafian, Houman; Docherty, Louise; Leo, Vincenzo; Towlson, Christopher; Neilan, Monica; Steeples, Violetta; Lygate, Craig A.; Hough, Tertius; Townsend, Stuart; Williams, Debbie; Wells, Sara; Norris, Dominic; Glyn-Jones, Sarah; Land, John; Barbaric, Ivana; Lalanne, Zuzanne; Denny, Paul; Szumska, Dorota; Bhattacharya, Shoumo; Griffin, Julian L.; Hargreaves, Iain; Fernandez-Fuentes, Narcis; Cheeseman, Michael; Watkins, Hugh; Dear, T. Neil

    2010-01-01

    Mutations in a number of genes have been linked to inherited dilated cardiomyopathy (DCM). However, such mutations account for only a small proportion of the clinical cases emphasising the need for alternative discovery approaches to uncovering novel pathogenic mutations in hitherto unidentified pathways. Accordingly, as part of a large-scale N-ethyl-N-nitrosourea mutagenesis screen, we identified a mouse mutant, Python, which develops DCM. We demonstrate that the Python phenotype is attributable to a dominant fully penetrant mutation in the dynamin-1-like (Dnm1l) gene, which has been shown to be critical for mitochondrial fission. The C452F mutation is in a highly conserved region of the M domain of Dnm1l that alters protein interactions in a yeast two-hybrid system, suggesting that the mutation might alter intramolecular interactions within the Dnm1l monomer. Heterozygous Python fibroblasts exhibit abnormal mitochondria and peroxisomes. Homozygosity for the mutation results in the death of embryos midway though gestation. Heterozygous Python hearts show reduced levels of mitochondria enzyme complexes and suffer from cardiac ATP depletion. The resulting energy deficiency may contribute to cardiomyopathy. This is the first demonstration that a defect in a gene involved in mitochondrial remodelling can result in cardiomyopathy, showing that the function of this gene is needed for the maintenance of normal cellular function in a relatively tissue-specific manner. This disease model attests to the importance of mitochondrial remodelling in the heart; similar defects might underlie human heart muscle disease. PMID:20585624

  12. Genetic variations in the TERT and CLPTM1L gene region and gastrointestinal stromal tumors risk

    PubMed Central

    Zhang, Rui; Zhao, Jian; Xu, Jian; Liu, Fang; Xu, Yongqing; Bu, Xianmin; Dai, Chaoliu; Song, Chun

    2015-01-01

    Recent studies have suggested polymorphisms in the TERT and CLPTM1L region are associated with carcinogenesis of many distinct cancer types, including gastrointestinal cancers. However, the contribution of polymorphisms in the TERT and CLPTM1L gene region to gastrointestinal stromal tumors (GISTs) risk is still unknown. We tested the six tagSNPs on TERT and CLPTM1L region with GIST risk, using a population-based, two-stage, case-control study in 2,000 subjects. Functional validation was conducted to validate our findings of TERT rs2736098 and explore its influence on relative telomere length (RTL) in GIST cells. It showed that variant rs2736098 was significantly associated with increased risk of GIST (per allele OR = 1.29, 95% CI: 1.14–1.47, P = 7.03 × 10−5). The difference remain significant after Bonferroni correction (P = 7.03 × 10−5 * 6 = 4.2 × 10−4). Real-time PCR showed carriers of genotype CC have the longest RTL, following by carriers of genotype CT, while carriers of genotype TT have the shortest RTL in GIST tissues (P < 0.001). Our data provide evidence to implicate TERT rs2736098 polymorphism as a novel susceptibility factor for GIST risk. PMID:26372813

  13. Mutation of kri1l causes definitive hematopoiesis failure via PERK-dependent excessive autophagy induction

    PubMed Central

    Jia, Xiao-E; Ma, Ke; Xu, Tao; Gao, Lei; Wu, Shuang; Fu, Cong; Zhang, Wenjuan; Wang, Zhizhang; Liu, Kaiyu; Dong, Mei; Jing, Changbin; Ren, Chunguang; Dong, Zhiwei; Chen, Yi; Jin, Yi; Huang, Qiuhua; Chang, Xing; Deng, Min; Li, Li; Luo, Lingfei; Zhu, Jun; Dang, Yongjun; Chang, Hung-Chun; Zon, Leonard I; Zhou, Yi; Chen, Saijuan; Pan, Weijun

    2015-01-01

    Dysregulation of ribosome biogenesis causes human diseases, such as Diamond-Blackfan anemia, del (5q-) syndrome and bone marrow failure. However, the mechanisms of blood disorders in these diseases remain elusive. Through genetic mapping, molecular cloning and mechanism characterization of the zebrafish mutant cas002, we reveal a novel connection between ribosomal dysfunction and excessive autophagy in the regulation of hematopoietic stem/progenitor cells (HSPCs). cas002 carries a recessive lethal mutation in kri1l gene that encodes an essential component of rRNA small subunit processome. We show that Kri1l is required for normal ribosome biogenesis, expansion of definitive HSPCs and subsequent lineage differentiation. Through live imaging and biochemical studies, we find that loss of Kri1l causes the accumulation of misfolded proteins and excessive PERK activation-dependent autophagy in HSPCs. Blocking autophagy but not inhibiting apoptosis by Bcl2 overexpression can fully rescue hematopoietic defects, but not the lethality of kri1lcas002 embryos. Treatment with autophagy inhibitors (3-MA and Baf A1) or PERK inhibitor (GSK2656157), or knockdown of beclin1 or perk can markedly restore HSPC proliferation and definitive hematopoietic cell differentiation. These results may provide leads for effective therapeutics that benefit patients with anemia or bone marrow failure caused by ribosome disorders. PMID:26138676

  14. The ciliopathy gene Rpgrip1l is essential for hair follicle development

    PubMed Central

    Chen, Jiang; Laclef, Christine; Moncayo, Alejandra; Snedecor, Elizabeth R.; Yang, Ning; Li, Li; Takemaru, Ken-Ichi; Paus, Ralf; Schneider-Maunoury, Sylvie; Clark, Richard A

    2014-01-01

    The primary cilium is essential for skin morphogenesis through regulating the Notch, Wnt, and hedgehog signaling pathways. Prior studies on the functions of primary cilia in the skin were based on the investigations of genes that are essential for cilium formation. However, none of these ciliogenic genes has been linked to ciliopathy, a group of disorders caused by abnormal formation or function of cilia. To determine whether there is a genetic and molecular link between ciliopathies and skin morphogenesis, we investigated the role of RPGRIP1L, a gene mutated in Joubert (JBTS) and Meckel (MKS) syndromes, two severe forms of ciliopathy, in the context of skin development. We found that RPGRIP1L is essential for hair follicle morphogenesis. Specifically, disrupting the Rpgril1 gene in mice resulted in reduced proliferation and differentiation of follicular keratinocytes, leading to hair follicle developmental defects. These defects were associated with significantly decreased primary cilium formation and attenuated hedgehog signaling. In contrast, we found that hair follicle induction and polarization and the development of interfollicular epidermis were unaffected. This study indicates that RPGRIP1L, a ciliopathy gene, is essential for hair follicle morphogenesis likely through regulating primary cilia formation and the hedgehog signaling pathway. PMID:25398052

  15. Overexpression of Rcan1-1L inhibits hypoxia-induced cell apoptosis through induction of mitophagy.

    PubMed

    Sun, Lijun; Hao, Yuewen; An, Rui; Li, Haixun; Xi, Cong; Shen, Guohong

    2014-11-01

    Mitophagy, a cellular process that selectively targets dysfunctional mitochondria for degradation, is currently a hot topic in research into the pathogenesis and treatment of many human diseases. Considering that hypoxia causes mitochondrial dysfunction, which results in cell death, we speculated that selective activation of mitophagy might promote cell survival under hypoxic conditions. In the present study, we introduced the Regulator of calcineurin 1-1L (Rcan1-1L) to initiate the mitophagy pathway and aimed to evaluate the effect of Rcan1-1L-induced mitophagy on cell survival under hypoxic conditions. Recombinant adenovirus vectors carrying Rcan1-1L were transfected into human umbilical vein endothelial cells and human adult cardiac myocytes. Using the 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide MTT assay and Trypan blue exclusion assay, Rcan1-1L overexpression was found to markedly reverse cell growth inhibition induced by hypoxia. Additionally, Rcan1-1L overexpression inhibited cell apoptosis under hypoxic conditions, as detected by annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) apoptosis assay. Meanwhile, the mitochondria-mediated cell apoptotic pathway was inhibited by Rcan1-1L. In contrast, knockdown of Rcan1-1L accelerated hypoxia-induced cell apoptosis. Moreover, Rcan1-1L overexpression significantly reduced mitochondrial mass, decreased depolarized mitochondria, and downregulated ATP and reactive oxygen species production. We further delineated that the loss of mitochondrial mass was due to the activation of mitophagy induced by Rcan1-1L. Rcan1-1L overexpression activated autophagy flux and promoted translocation of the specific mitophagy receptor Parkin into mitochondria from the cytosol, whereas inhibition of autophagy flux resulted in the accumulation of Parkin-loaded mitochondria. Finally, we demonstrated that mitochondrial permeability transition pore opening was significantly increased by Rcan1-1L overexpression

  16. KSHV encoded LANA recruits Nucleosome Assembly Protein NAP1L1 for regulating viral DNA replication and transcription

    PubMed Central

    Gupta, Namrata; Thakker, Suhani; Verma, Subhash C.

    2016-01-01

    The establishment of latency is an essential for lifelong persistence and pathogenesis of Kaposi’s sarcoma-associated herpesvirus (KSHV). Latency-associated nuclear antigen (LANA) is the most abundantly expressed protein during latency and is important for viral genome replication and transcription. Replication-coupled nucleosome assembly is a major step in packaging the newly synthesized DNA into chromatin, but the mechanism of KSHV genome chromatinization post-replication is not understood. Here, we show that nucleosome assembly protein 1-like protein 1 (NAP1L1) associates with LANA. Our binding assays revealed an association of LANA with NAP1L1 in KSHV-infected cells, which binds through its amino terminal domain. Association of these proteins confirmed their localization in specific nuclear compartments of the infected cells. Chromatin immunoprecipitation assays from NAP1L1-depleted cells showed LANA-mediated recruitment of NAP1L1 at the terminal repeat (TR) region of the viral genome. Presence of NAP1L1 stimulated LANA-mediated DNA replication and persistence of a TR-containing plasmid. Depletion of NAP1L1 led to a reduced nucleosome positioning on the viral genome. Furthermore, depletion of NAP1L1 increased the transcription of viral lytic genes and overexpression decreased the promoter activities of LANA-regulated genes. These results confirmed that LANA recruitment of NAP1L1 helps in assembling nucleosome for the chromatinization of newly synthesized viral DNA. PMID:27599637

  17. HNF1α and SREBP2 are important regulators of NPC1L1 in human liver

    PubMed Central

    Pramfalk, Camilla; Jiang, Zhao-Yan; Cai, Qu; Hu, Hai; Zhang, Sheng-Dao; Han, Tian-Quan; Eriksson, Mats; Parini, Paolo

    2010-01-01

    Niemann-Pick C1-like 1 (NPC1L1), a key regulator of intestinal cholesterol absorption, is highly expressed in human liver. Here, we aimed to gain more insight into mechanisms participating in its hepatic regulation in humans. Correlation analysis in livers from Chinese patients with and without gallstone disease revealed strong positive correlations between NPC1L1 and sterol regulatory element binding protein 2 (SREBP2) (r = 0.74, P < 0.05) and between NPC1L1 and hepatic nuclear factor α (HNF4α) (r = 0.53, P < 0.05) mRNA expression. HNF4α is an upstream regulator of HNF1α; thus, we also tested whether HNF1α participates in the regulation of NPC1L1. We showed a dose-dependent regulation by SREBP2 on the NPC1L1 promoter activity and mRNA expression in HuH7 cells. Chromatin immunoprecipitation assay confirmed the binding of SREBP2 to the promoter in vivo. Surprisingly, HNF4α slightly decreased the NPC1L1 promoter activity but had no effect on its gene expression. By contrast, HNF1α increased the promoter activity and the gene expression, and an important HNF1 binding site was identified within the human NPC1L1 promoter. ChIP assays confirmed that HNF1α can bind to the NPC1L1 promoter in vivo. PMID:20460578

  18. The histone methyltransferase Wolf-Hirschhorn syndrome candidate 1-like 1 (WHSC1L1) is involved in human carcinogenesis.

    PubMed

    Kang, Daechun; Cho, Hyun-Soo; Toyokawa, Gouji; Kogure, Masaharu; Yamane, Yuka; Iwai, Yukiko; Hayami, Shinya; Tsunoda, Tatsuhiko; Field, Helen I; Matsuda, Koichi; Neal, David E; Ponder, Bruce A J; Maehara, Yoshihiko; Nakamura, Yusuke; Hamamoto, Ryuji

    2013-02-01

    Histone lysine methylation plays a fundamental role in chromatin organization. Although a set of histone methyltransferases have been identified and biochemically characterized, the pathological roles of their dysfunction in human cancers are still not well understood. In this study, we demonstrate important roles of WHSC1L1 in human carcinogenesis. Expression levels of WHSC1L1 transcript were significantly elevated in various human cancers including bladder carcinoma. Immunohistochemical analysis of bladder, lung, and liver cancers confirmed overexpression of WHSC1L1. WHSC1L1-specific small interfering RNAs significantly knocked down its expression and resulted in suppression of proliferation of bladder and lung cancer cell lines. WHSC1L1 knockdown induced cell cycle arrest at the G(2)/M phase followed by multinucleation of cancer cells. Expression profile analysis using Affymetrix GeneChip(®) showed that WHSC1L1 affected the expression of a number of genes including CCNG1 and NEK7, which are known to play crucial roles in the cell cycle progression at mitosis. As WHSC1L1 expression is significantly low in various normal tissues including vital organs, WHSC1L1 could be a good candidate molecule for development of novel treatment for various types of cancer. PMID:23011637

  19. KSHV encoded LANA recruits Nucleosome Assembly Protein NAP1L1 for regulating viral DNA replication and transcription.

    PubMed

    Gupta, Namrata; Thakker, Suhani; Verma, Subhash C

    2016-01-01

    The establishment of latency is an essential for lifelong persistence and pathogenesis of Kaposi's sarcoma-associated herpesvirus (KSHV). Latency-associated nuclear antigen (LANA) is the most abundantly expressed protein during latency and is important for viral genome replication and transcription. Replication-coupled nucleosome assembly is a major step in packaging the newly synthesized DNA into chromatin, but the mechanism of KSHV genome chromatinization post-replication is not understood. Here, we show that nucleosome assembly protein 1-like protein 1 (NAP1L1) associates with LANA. Our binding assays revealed an association of LANA with NAP1L1 in KSHV-infected cells, which binds through its amino terminal domain. Association of these proteins confirmed their localization in specific nuclear compartments of the infected cells. Chromatin immunoprecipitation assays from NAP1L1-depleted cells showed LANA-mediated recruitment of NAP1L1 at the terminal repeat (TR) region of the viral genome. Presence of NAP1L1 stimulated LANA-mediated DNA replication and persistence of a TR-containing plasmid. Depletion of NAP1L1 led to a reduced nucleosome positioning on the viral genome. Furthermore, depletion of NAP1L1 increased the transcription of viral lytic genes and overexpression decreased the promoter activities of LANA-regulated genes. These results confirmed that LANA recruitment of NAP1L1 helps in assembling nucleosome for the chromatinization of newly synthesized viral DNA. PMID:27599637

  20. GSG1L suppresses AMPA receptor-mediated synaptic transmission and uniquely modulates AMPA receptor kinetics in hippocampal neurons

    PubMed Central

    Gu, Xinglong; Mao, Xia; Lussier, Marc P.; Hutchison, Mary Anne; Zhou, Liang; Hamra, F. Kent; Roche, Katherine W.; Lu, Wei

    2016-01-01

    Regulation of AMPA receptor (AMPAR)-mediated synaptic transmission is a key mechanism for synaptic plasticity. In the brain, AMPARs assemble with a number of auxiliary subunits, including TARPs, CNIHs and CKAMP44, which are important for AMPAR forward trafficking to synapses. Here we report that the membrane protein GSG1L negatively regulates AMPAR-mediated synaptic transmission. Overexpression of GSG1L strongly suppresses, and GSG1L knockout (KO) enhances, AMPAR-mediated synaptic transmission. GSG1L-dependent regulation of AMPAR synaptic transmission relies on the first extracellular loop domain and its carboxyl-terminus. GSG1L also speeds up AMPAR deactivation and desensitization in hippocampal CA1 neurons, in contrast to the effects of TARPs and CNIHs. Furthermore, GSG1L association with AMPARs inhibits CNIH2-induced slowing of the receptors in heterologous cells. Finally, GSG1L KO rats have deficits in LTP and show behavioural abnormalities in object recognition tests. These data demonstrate that GSG1L represents a new class of auxiliary subunit with distinct functional properties for AMPARs. PMID:26932439

  1. Functional Analysis of SPECC1L in Craniofacial Development and Oblique Facial Cleft Pathogenesis

    PubMed Central

    Gfrerer, Lisa; Shubinets, Valeriy; Hoyos, Tatiana; Kong, Yawei; Nguyen, Christina; Pietschmann, Peter; Morton, Cynthia C.; Maas, Richard L.; Liao, Eric C.

    2015-01-01

    Background Oblique facial clefts, also known as Tessier clefts, are severe orofacial clefts, the genetic basis of which is poorly understood. Human genetics studies revealed that disruption in SPECC1L resulted in oblique facial clefts, demonstrating that oblique facial cleft malformation has a genetic basis. An important step toward innovation in treatment of oblique facial clefts would be improved understanding of its genetic pathogenesis. The authors exploit the zebrafish model to elucidate the function of SPECC1L by studying its homolog, specc1lb. Methods Gene and protein expression analysis was carried out by reverse-transcriptase polymerase chain reaction and immunohistochemistry staining. Morpholino knockdown, mRNA rescue, lineage tracing and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling assays were performed for functional analysis. Results Expression of specc1lb was detected in epithelia juxtaposed to chondrocytes. Knockdown of specc1lb resulted in bilateral clefts between median and lateral elements of the ethmoid plate, structures analogous to the frontonasal process and the paired maxillary processes. Lineage tracing analysis revealed that cranial neural crest cells contributing to the frontonasal prominence failed to integrate with the maxillary prominence populations. Cells contributing to lower jaw structures were able to migrate to their destined pharyngeal segment but failed to converge to form mandibular elements. Conclusions These results demonstrate that specc1lb is required for integration of frontonasal and maxillary elements and convergence of mandibular prominences. The authors confirm the role of SPECC1L in orofacial cleft pathogenesis in the first animal model of Tessier cleft, providing morphogenetic insight into the mechanisms of normal craniofacial development and oblique facial cleft pathogenesis. PMID:25357034

  2. Preliminary Design Considerations for Access and Operations in Earth-Moon L1/L2 Orbits

    NASA Technical Reports Server (NTRS)

    Folta, David C.; Pavlak, Thomas A.; Haapala, Amanda F.; Howell, Kathleen C.

    2013-01-01

    Within the context of manned spaceflight activities, Earth-Moon libration point orbits could support lunar surface operations and serve as staging areas for future missions to near-Earth asteroids and Mars. This investigation examines preliminary design considerations including Earth-Moon L1/L2 libration point orbit selection, transfers, and stationkeeping costs associated with maintaining a spacecraft in the vicinity of L1 or L2 for a specified duration. Existing tools in multi-body trajectory design, dynamical systems theory, and orbit maintenance are leveraged in this analysis to explore end-to-end concepts for manned missions to Earth-Moon libration points.

  3. Exome sequencing reveals novel BCS1L mutations in siblings with hearing loss and hypotrichosis.

    PubMed

    Zhang, Jie; Duo, Lina; Lin, Zhimiao; Wang, Huijun; Yin, Jinghua; Cao, Xu; Zhao, Jiahui; Dai, Lanlan; Liu, Xuanzhu; Zhang, Jianguo; Yang, Yong; Tang, Zhanli

    2015-07-15

    As a powerful tool to identify the molecular pathogenesis of Mendelian disorders, exome sequencing was used to identify the genetic basis of two siblings with hearing loss and hypotrichosis and clarify the diagnosis. No pathogenic mutations in GJB2, GJB3 and GJB6 genes were found in the siblings. By analysis of exome of the proband, we identified a novel missense (p.R306C) mutation and a nonsense (p.R186*) mutation in the BCS1L gene. Mutations were confirmed by Sanger sequencing. The siblings were compound heterozygotes, and the inheritance mode of autosomal recessive was postulated. BCS1L is the causative gene of Björnstad syndrome, which is characterized by sensorineural hearing loss and pili torti. The longitudinal gutters along the hair shaft were found by scanning electron microscopy in our patient. Therefore the diagnosis of Björnstad syndrome was eventually made for the patients. Our study extends the phenotypic spectrum of Björnstad syndrome and highlights the clinical applicability of exome sequencing as a diagnostic tool for atypical Mendelian disorders. PMID:25895478

  4. Ionospheric TEC variations at mid-latitudes obsereved using SBAS L1/L5 signals.

    NASA Astrophysics Data System (ADS)

    Padokhin, A. M.; Kunitsyn, V.; Kurbatov, G.; Yasyukevich, Y.

    2014-12-01

    With the development of Satellite based augmentation systems (SBAS) the dual frequency L1/L5 observations from a number of geostationary satellites are now available. It provides the possibility to retrieve ionospheric total electron content (TEC) from these observations using the same approach as for dual frequency GPS/GLONASS observations. In this work we study L1/L5 signals of Indian GAGAN geostationary satellites observed with geodetic GNSS receivers at several stations at mid-latitudes and estimate corresponding geostationary TEC and errors of such estimations. TEC RMS was found to reach up to 1.5 TECU with typical values of 0.25-0.5 TECU which is several times greater than for common GPS/GLONASS observations. TEC RMS also manifests UT-dynamics which is specific for the chosen geostationary satellite and not relevant to the receiver site and signal paths. SBAS TEC was found to be in good agreement with the data of nearest ionosondes taking into account low elevation angles of SBAS satellites already at mid-latitudes and spatial gradients of electron density along the ray paths. We also present the wavelet analysis of geostationary TEC, providing typical periods of observed variations at different time scales (from tens of minutes to tens of days) and discuss the capabilities of SBAS TEC observations in connection with ionospheric effects of solar flares.

  5. A sample introduction system for an inductively coupled plasma operating on an argon carrier gas flow of 0.1 l/min

    NASA Astrophysics Data System (ADS)

    Ripson, P. A. M.; de Galan, L.

    A sample introduction system is described for use with a water-cooled ICP torch described previously [ Anal. Chem.51, 2378 (1979)]. It consists of a narrow bore (100 μm) stainless steel Babington nebulizer operating on 0.05 to 0.2 l/min argon inserted into a small (10 ml) nebulizer chamber. The solvent is force-fed continuously by gas pressure or with a peristaltic pump. Liquid samples can be supplied continuously or in discrete quantities using a sample loop between the pump and the nebulizer. In the latter case only 25 s are required for sample change. The nebulization efficiency for water and organic solvents is comparable to that of conventional pneumatic nebulizers operating on 1 l/min argon.

  6. Association of the SEL1L protein transmembrane domain with HRD1 ubiquitin ligase regulates ERAD-L.

    PubMed

    Hosokawa, Nobuko; Wada, Ikuo

    2016-01-01

    Misfolded proteins in the endoplasmic reticulum (ER) are transported to the cytoplasm for degradation by the ubiquitin-proteasome system, a process otherwise known as ER-associated degradation (ERAD). Mammalian HRD1, an integral membrane ubiquitin ligase that ubiquitinates ERAD substrates, forms a large assembly in the ER membrane including SEL1L, a single-pass membrane protein, and additional components. The mechanism by which these molecules export misfolded proteins through the ER membrane remains unclear. Unlike Hrd3p, the homologue in Saccharomyces cerevisiae, human SEL1L is an unstable protein, which is restored by the association with HRD1. Here we report that the inherently unstable nature of the human SEL1L protein lies in its transmembrane domain, and that association of HRD1 with the SEL1L transmembrane domain restored its stability. On the other hand, we found that the SEL1L luminal domain escaped degradation, and inhibited the degradation of misfolded α1 -antitrypsin variant null Hong Kong by retaining the misfolded cargo in the ER. Overexpression of HRD1 inhibited the degradation of unfolded secretory cargo, which was restored by the interaction of HRD1 with the SEL1L transmembrane domain. Hence, we propose that SEL1L critically regulates HRD1-mediated disposal of misfolded cargo through its short membrane spanning stretch. PMID:26471130

  7. TOM1L1 drives membrane delivery of MT1-MMP to promote ERBB2-induced breast cancer cell invasion

    PubMed Central

    Chevalier, Clément; Collin, Guillaume; Descamps, Simon; Touaitahuata, Heiani; Simon, Valérie; Reymond, Nicolas; Fernandez, Laurent; Milhiet, Pierre-Emmanuel; Georget, Virginie; Urbach, Serge; Lasorsa, Laurence; Orsetti, Béatrice; Boissière-Michot, Florence; Lopez-Crapez, Evelyne; Theillet, Charles; Roche, Serge; Benistant, Christine

    2016-01-01

    ERBB2 overexpression in human breast cancer leads to invasive carcinoma but the mechanism is not clearly understood. Here we report that TOM1L1 is co-amplified with ERBB2 and defines a subgroup of HER2+/ER+ tumours with early metastatic relapse. TOM1L1 encodes a GAT domain-containing trafficking protein and is a SRC substrate that negatively regulates tyrosine kinase signalling. We demonstrate that TOM1L1 upregulation enhances the invasiveness of ERBB2-transformed cells. This pro-tumoural function does not involve SRC, but implicates membrane-bound membrane-type 1 MMP (MT1-MMP)-dependent activation of invadopodia, membrane protrusions specialized in extracellular matrix degradation. Mechanistically, ERBB2 elicits the indirect phosphorylation of TOM1L1 on Ser321. The phosphorylation event promotes GAT-dependent association of TOM1L1 with the sorting protein TOLLIP and trafficking of the metalloprotease MT1-MMP from endocytic compartments to invadopodia for tumour cell invasion. Collectively, these results show that TOM1L1 is an important element of an ERBB2-driven proteolytic invasive programme and that TOM1L1 amplification potentially enhances the metastatic progression of ERBB2-positive breast cancers. PMID:26899482

  8. SPECC1L deficiency results in increased adherens junction stability and reduced cranial neural crest cell delamination

    PubMed Central

    Wilson, Nathan R.; Olm-Shipman, Adam J.; Acevedo, Diana S.; Palaniyandi, Kanagaraj; Hall, Everett G.; Kosa, Edina; Stumpff, Kelly M.; Smith, Guerin J.; Pitstick, Lenore; Liao, Eric C.; Bjork, Bryan C.; Czirok, Andras; Saadi, Irfan

    2016-01-01

    Cranial neural crest cells (CNCCs) delaminate from embryonic neural folds and migrate to pharyngeal arches, which give rise to most mid-facial structures. CNCC dysfunction plays a prominent role in the etiology of orofacial clefts, a frequent birth malformation. Heterozygous mutations in SPECC1L have been identified in patients with atypical and syndromic clefts. Here, we report that in SPECC1L-knockdown cultured cells, staining of canonical adherens junction (AJ) components, β-catenin and E-cadherin, was increased, and electron micrographs revealed an apico-basal diffusion of AJs. To understand the role of SPECC1L in craniofacial morphogenesis, we generated a mouse model of Specc1l deficiency. Homozygous mutants were embryonic lethal and showed impaired neural tube closure and CNCC delamination. Staining of AJ proteins was increased in the mutant neural folds. This AJ defect is consistent with impaired CNCC delamination, which requires AJ dissolution. Further, PI3K-AKT signaling was reduced and apoptosis was increased in Specc1l mutants. In vitro, moderate inhibition of PI3K-AKT signaling in wildtype cells was sufficient to cause AJ alterations. Importantly, AJ changes induced by SPECC1L-knockdown were rescued by activating the PI3K-AKT pathway. Together, these data indicate SPECC1L as a novel modulator of PI3K-AKT signaling and AJ biology, required for neural tube closure and CNCC delamination. PMID:26787558

  9. Genetic demonstration of intestinal NPC1L1 as a major determinant of hepatic cholesterol and blood atherogenic lipoprotein levels

    PubMed Central

    Xie, Ping; Zhu, Hongling; Jia, Lin; Ma, Yinyan; Tang, Weiqing; Wang, Youlin; Xue, Bingzhong; Shi, Hang; Yu, Liqing

    2014-01-01

    Objective The correlation between intestinal cholesterol absorption values and plasma low-density lipoprotein-cholesterol (LDL-C) levels remains controversial. Niemann-Pick-C1-Like 1 (NPC1L1) is essential for intestinal cholesterol absorption, and is the target of ezetimibe, a cholesterol absorption inhibitor. However, studies with NPC1L1 knockout mice or ezetimibe cannot definitively clarify this correlation because NPC1L1 expression is not restricted to intestine in humans and mice. In this study we sought to genetically address this issue. Methods and results We developed a mouse model that lacks endogenous (NPC1L1) and LDL receptor (LDLR) (DKO), but transgenically expresses human NPC1L1 in gastrointestinal tract only (DKO/L1IntOnly mice). Our novel model eliminated potential effects of non-intestinal NPC1L1 on cholesterol homeostasis. We found that human NPC1L1 was localized at the intestinal brush border membrane of DKO/L1IntOnly mice. Cholesterol feeding induced formation of NPC1L1-positive vesicles beneath this membrane in an ezetimibe-sensitive manner. Compared to DKO mice, DKO/L1IntOnly mice showed significant increases in cholesterol absorption and blood/hepatic/biliary cholesterol. Increased blood cholesterol was restricted to very low-density lipoprotein (VLDL) and LDL fractions, which was associated with increased secretion and plasma levels of apolipoproteins B100 and B48. Additionally, DKO/L1IntOnly mice displayed decreased fecal cholesterol excretion and hepatic/intestinal expression of cholesterologenic genes. Ezetimibe treatment virtually reversed all of the transgene-related phenotypes in DKO/L1IntOnly mice. Conclusion Our findings from DKO/L1IntOnly mice clearly demonstrate that NPC1L1-mediated cholesterol absorption is a major determinant of blood levels of apolipoprotein B-containing atherogenic lipoproteins, at least in mice. PMID:25463095

  10. Mutations in SIPA1L3 cause eye defects through disruption of cell polarity and cytoskeleton organization.

    PubMed

    Greenlees, Rebecca; Mihelec, Marija; Yousoof, Saira; Speidel, Daniel; Wu, Selwin K; Rinkwitz, Silke; Prokudin, Ivan; Perveen, Rahat; Cheng, Anson; Ma, Alan; Nash, Benjamin; Gillespie, Rachel; Loebel, David A F; Clayton-Smith, Jill; Lloyd, I Christopher; Grigg, John R; Tam, Patrick P L; Yap, Alpha S; Becker, Thomas S; Black, Graeme C M; Semina, Elena; Jamieson, Robyn V

    2015-10-15

    Correct morphogenesis and differentiation are critical in development and maintenance of the lens, which is a classic model system for epithelial development and disease. Through germline genomic analyses in patients with lens and eye abnormalities, we discovered functional mutations in the Signal Induced Proliferation Associated 1 Like 3 (SIPA1L3) gene, which encodes a previously uncharacterized member of the Signal Induced Proliferation Associated 1 (SIPA1 or SPA1) family, with a role in Rap1 signalling. Patient 1, with a de novo balanced translocation, 46,XY,t(2;19)(q37.3;q13.1), had lens and ocular anterior segment abnormalities. Breakpoint mapping revealed transection of SIPA1L3 at 19q13.1 and reduced SIPA1L3 expression in patient lymphoblasts. SIPA1L3 downregulation in 3D cell culture revealed morphogenetic and cell polarity abnormalities. Decreased expression of Sipa1l3 in zebrafish and mouse caused severe lens and eye abnormalities. Sipa1l3(-/-) mice showed disrupted epithelial cell organization and polarity and, notably, abnormal epithelial to mesenchymal transition in the lens. Patient 2 with cataracts was heterozygous for a missense variant in SIPA1L3, c.442G>T, p.Asp148Tyr. Examination of the p.Asp148Tyr mutation in an epithelial cell line showed abnormal clustering of actin stress fibres and decreased formation of adherens junctions. Our findings show that abnormalities of SIPA1L3 in human, zebrafish and mouse contribute to lens and eye defects, and we identify a critical role for SIPA1L3 in epithelial cell morphogenesis, polarity, adhesion and cytoskeletal organization. PMID:26231217

  11. Niemann-Pick C1-Like 1 (NPC1L1) Protein in Intestinal and Hepatic Cholesterol Transport

    PubMed Central

    Jia, Lin; Betters, Jenna L.; Yu, Liqing

    2014-01-01

    Increased blood cholesterol is an independent risk factor for atherosclerotic cardiovascular disease. Cholesterol homeostasis in the body is controlled mainly by endogenous synthesis, intestinal absorption, and hepatic excretion. Niemann-Pick C1-Like 1 (NPC1L1) is a polytopic transmembrane protein localized at the apical membrane of enterocytes and the canalicular membrane of hepatocytes. It functions as a sterol transporter to mediate intestinal cholesterol absorption and counterbalances hepatobiliary cholesterol excretion. NPC1L1 is the molecular target of ezetimibe, a potent cholesterol absorption inhibitor that is widely used in treating hypercholesterolemia. Recent findings suggest that NPC1L1 deficiency or ezetimibe treatment also prevents diet-induced hepatic steatosis and obesity in addition to reducing blood cholesterol. Future studies should focus on molecular mechanisms underlying NPC1L1-dependent cholesterol transport and elucidation of how a cholesterol transporter modulates the pathogenesis of metabolic diseases. PMID:20809793

  12. Gtsf1l and Gtsf2 Are Specifically Expressed in Gonocytes and Spermatids but Are Not Essential for Spermatogenesis

    PubMed Central

    Takemoto, Noriaki; Yoshimura, Takuji; Miyazaki, Satsuki; Tashiro, Fumi; Miyazaki, Jun-ichi

    2016-01-01

    The unknown protein family 0224 (UPF0224) includes three members that are expressed in germ-line cells in mice: Gtsf1, Gtsf1l, and BC048502 (Gtsf2). These genes produce proteins with two repeats of the CHHC Zn-finger domain, a predicted RNA-binding motif, in the N terminus. We previously reported that Gtsf1 is essential for spermatogenesis and retrotransposon suppression. In this study, we investigated the expression patterns and functions of Gtsf1l and Gtsf2. Interestingly, Gtsf1l and Gtsf2 were found to be sequentially but not simultaneously expressed in gonocytes and spermatids. Pull-down experiments showed that both GTSF1L and GTSF2 can interact with PIWI-protein complexes. Nevertheless, knocking out Gtsf1, Gtsf2, or both did not cause defects in spermatogenesis or retrotransposon suppression in mice. PMID:26930067

  13. Essential and synergistic roles of RP1 and RP1L1 in rod photoreceptor axoneme and retinitis pigmentosa

    PubMed Central

    Yamashita, Tetsuji; Liu, Jiewu; Gao, Jiangang; LeNoue, Sean; Wang, Changguan; Kaminoh, Jack; Bowne, Sara J.; Sullivan, Lori S.; Daiger, Stephen P.; Zhang, Kang; Fitzgerald, Malinda E.C.; Kefalov, Vladimir J.; Zuo, Jian

    2009-01-01

    Retinitis pigmentosa 1 (RP1) is a common inherited retinopathy with variable onset and severity. The RP1 gene encodes a photoreceptor-specific, microtubule-associated ciliary protein containing the doublecortin (DCX) domain. Here we show that another photoreceptor-specific Rp1-like protein (Rp1L1) in mice is also localized to the axoneme of outer segments (OS) and connecting cilia in rod photoreceptors, overlapping with Rp1. Rp1L1−/− mice display scattered OS disorganization, reduced electroretinogram amplitudes, and progressive photoreceptor degeneration, less severe and slower than in Rp1−/− mice. In single rods of Rp1L1−/−, photosensitivity is reduced, similar to that of Rp1−/−. While individual heterozygotes are normal, double heterozygotes of Rp1 and Rp1L1 exhibit abnormal OS morphology and reduced single rod photosensitivity and dark currents. The electroretinogram amplitudes of double heterozygotes are more reduced than those of individual heterozygotes combined. In support, Rp1L1 interacts with Rp1 in transfected cells and in retina pull-down experiments. Interestingly, phototransduction kinetics are normal in single rods and whole retinas of individual or double Rp1 and Rp1L1 mutant mice. Together, Rp1 and Rp1L1 play essential and synergistic roles in affecting photosensitivity and OS morphogenesis of rod photoreceptors. Our findings suggest that mutations in RP1L1 could underlie retinopathy or modify RP1 disease expression in humans. PMID:19657028

  14. Synthesis, Activity and Metabolic Stability of Non-Ribose Containing Inhibitors of Histone Methyltransferase DOT1L

    PubMed Central

    Deng, Lisheng; Zhang, Li; Yao, Yuan; Wang, Cong; Redell, Michele S.; Dong, Shuo

    2013-01-01

    Histone methyltransferase DOT1L is a drug target for MLL leukemia. We report an efficient synthesis of a cyclopentane-containing compound that potently and selectively inhibits DOT1L (Ki = 1.1 nM) as well as H3K79 methylation (IC50 ~ 200 nM). Importantly, this compound exhibits a high stability in plasma and liver microsomes, suggesting it is a better drug candidate. PMID:23795283

  15. Association between ALDH1L1 gene polymorphism and neural tube defects in the Chinese Han population.

    PubMed

    Wu, Lihua; Lu, Xiaolin; Guo, Jin; Zhang, Ting; Wang, Fang; Bao, Yihua

    2016-07-01

    We investigated single-nucleotide polymorphisms (SNPs) in the aldehyde dehydrogenase family1 L1 gene (ALDH1L1) and their association with neural tube defects (NTDs) in the Chinese population. A total of 271 NTDs cases and 192 healthy controls were used in this study. A total of 112 selected SNPs in the ALDH1L1 gene were analyzed using the next-generation sequencing method. Statistical analysis was carried out to investigate the correlation between SNPs and patient susceptibility to NTDs. Statistical analysis revealed a significant correlation between the SNP sites rs4646733, rs2305225, and rs2276731 in the ALDH1L1 gene and NTDs. The TT genotype and T allele of rs4646733 in ALDH1L1 were associated with a significantly increased incidence of NTDs [odds ratio (OR) = 2.16, 95 % confidence interval (CI) 1.199-3.896 for genotype; and OR = 1.46, 95 % CI 1.092-1.971 for allele]. The AA genotype and A allele of rs2305225 in ALDH1L1 were associated with a significantly increased incidence of NTDs (OR = 2.03, 95 % CI 1.202-3.646 for genotype, and OR = 1.44, 95 % CI 1.096-1.905 for allele). The CT genotype and C allele of rs2276731 in ALDH1L1 significantly were associated with an increased incidence of NTDs (OR = 1.67, 95 % CI 1.129-2.491 with genotype, and OR = 1.32, 95 % CI 0.956-1.816 with allele).The polymorphic loci rs4646733, rs2305225, and rs2276731 in the ALDH1L1 gene maybe potential risk factors for NTDs in the Chinese population. PMID:26993122

  16. Regulation of sarcoma cell migration, invasion and invadopodia formation by AFAP1L1 through a phosphotyrosine-dependent pathway.

    PubMed

    Tie, S R; McCarthy, D J; Kendrick, T S; Louw, A; Le, C; Satiaputra, J; Kucera, N; Phillips, M; Ingley, E

    2016-04-21

    Invasion and metastasis are controlled by the invadopodia, which delivers matrix-degrading enzymes to the invasion interface permitting cancer cell penetration and spread into healthy tissue. We have identified a novel pathway that directs Lyn/Src family tyrosine kinase signals to the invadopodia to regulate sarcoma cell invasion via the molecule AFAP-1-like-1 (AFAP1L1), a new member of the AFAP (actin filament-associated protein) family. We show that AFAP1L1 can transform cells, promote migration and co-expression with active Lyn profoundly influences cell morphology and movement. AFAP1L1 intersects several invadopodia pathway components through its multiple domains and motifs, including the following (i) pleckstrin homology domains that bind phospholipids generated at the plasma membrane by phosphoinositide 3-kinase, (ii) a direct filamentous-actin binding domain and (iii) phospho-tyrosine motifs (pY136 and pY566) that specifically bind Vav2 and Nck2 SH2 domains, respectively. These phosphotyrosine motifs are essential for AFAP1L1-mediated cytoskeleton regulation. Through its interaction with Vav2, AFAP1L1 regulates Rac activity and downstream control of PAK1/2/3 (p21-activated kinases) phosphorylation of myosin light chain (MLC) kinase and MLC2. AFAP1L1 interaction with Nck2 recruits actin-nucleating complexes. Significantly, in osteosarcoma cell lines, knockdown of AFAP1L1 inhibits phosphorylated MLC2 recruitment to filamentous-actin structures, disrupts invadopodia formation, cell attachment, migration and invasion. These data define a novel pathway that directs Lyn/Src family tyrosine kinase signals to sarcoma cell invadopodia through specific recruitment of Vav2 and Nck2 to phosphorylated AFAP1L1, to control cell migration and invasion. PMID:26212012

  17. Dot1l deficiency leads to increased intercalated cells and upregulation of V-ATPase B1 in mice.

    PubMed

    Xiao, Zhou; Chen, Lihe; Zhou, Qiaoling; Zhang, Wenzheng

    2016-06-10

    The collecting duct in the mammalian kidney consists of principal cells (PCs) and intercalated cells (ICs), which regulate electrolyte/fluid and acid/base balance, respectively. The epigenetic regulators of PC and IC differentiation remain obscure. We previously used Aqp2 and V-ATPase B1B2 to label PCs and ICs, respectively. We found that mice with histone H3 K79 methyltransferase Dot1l disrupted in Aqp2-expressing cells (Dot1l(AC)) vs. Dot1l(f/f) possessed ~20% more ICs coupled with a similar decrease in PCs. Here, we performed multiple double immunofluorescence staining using various PC and IC markers and confirmed that this finding. Both α-IC and β-IC populations were significantly expanded in Dot1l(AC) vs. Dot1l(f/f). These changes are associated with significantly upregulated V-ATPase B1 and B2, but not Aqp2, AE1, and Pendrin. Chromatin immunoprecipitation assay unveiled a significant reduction of Dot1l and H3K79 di-methylation bound at the Atp6v1b1 5' flanking region. Overexpression of Dot1a significantly downregulated a stably-transfected luciferase reporter driven by the Atp6v1b1 promoter in IMCD3 cells. This downregulation was impaired, but not completely abolished when a methyltransferase-dead mutant was overexpressed. Taken together, our data suggest that Dot1l is a new epigenetic regulator of PC and IC differentiation and Atp6v1b1 is a new transcriptional target of Dot1l. PMID:26404731

  18. DOT1L as a therapeutic target for the treatment of DNMT3A-mutant acute myeloid leukemia.

    PubMed

    Rau, Rachel E; Rodriguez, Benjamin A; Luo, Min; Jeong, Mira; Rosen, Allison; Rogers, Jason H; Campbell, Carly T; Daigle, Scott R; Deng, Lishing; Song, Yongcheng; Sweet, Steve; Chevassut, Timothy; Andreeff, Michael; Kornblau, Steven M; Li, Wei; Goodell, Margaret A

    2016-08-18

    Mutations in DNA methyltransferase 3A (DNMT3A) are common in acute myeloid leukemia and portend a poor prognosis; thus, new therapeutic strategies are needed. The likely mechanism by which DNMT3A loss contributes to leukemogenesis is altered DNA methylation and the attendant gene expression changes; however, our current understanding is incomplete. We observed that murine hematopoietic stem cells (HSCs) in which Dnmt3a had been conditionally deleted markedly overexpress the histone 3 lysine 79 (H3K79) methyltransferase, Dot1l. We demonstrate that Dnmt3a(-/-) HSCs have increased H3K79 methylation relative to wild-type (WT) HSCs, with the greatest increases noted at DNA methylation canyons, which are regions highly enriched for genes dysregulated in leukemia and prone to DNA methylation loss with Dnmt3a deletion. These findings led us to explore DOT1L as a therapeutic target for the treatment of DNMT3A-mutant AML. We show that pharmacologic inhibition of DOT1L resulted in decreased expression of oncogenic canyon-associated genes and led to dose- and time-dependent inhibition of proliferation, induction of apoptosis, cell-cycle arrest, and terminal differentiation in DNMT3A-mutant cell lines in vitro. We show in vivo efficacy of the DOT1L inhibitor EPZ5676 in a nude rat xenograft model of DNMT3A-mutant AML. DOT1L inhibition was also effective against primary patient DNMT3A-mutant AML samples, reducing colony-forming capacity (CFC) and inducing terminal differentiation in vitro. These studies suggest that DOT1L may play a critical role in DNMT3A-mutant leukemia. With pharmacologic inhibitors of DOT1L already in clinical trials, DOT1L could be an immediately actionable therapeutic target for the treatment of this poor prognosis disease. PMID:27335278

  19. Genetic variation at the NPC1L1 gene locus, plasma lipoproteins, and heart disease risk in the elderly

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Niemann-Pick C1-like 1 protein (NPC1L1) plays a critical role in intestinal cholesterol absorption. Our objective was to examine whether five variants (-133A>G, -18A>C, L272L, V1296V, and U3_28650A>G) at the NPC1L1 gene have effects on lipid levels, prevalence, and incidence of coronary heart diseas...

  20. EHBP1L1 coordinates Rab8 and Bin1 to regulate apical-directed transport in polarized epithelial cells

    PubMed Central

    Nakajo, Atsuhiro; Togawa, Hiroko; Kunii, Masataka; Iwano, Tomohiko; Izumi, Ayaka; Noguchi, Yuria; Watanabe, Ayako; Goto, Ayako; Sato, Toshiro

    2016-01-01

    The highly conserved Rab guanosine triphosphatase (GTPase) Rab8 plays a role in exocytosis toward the polarized plasma membrane in eukaryotic cells. In murine Rab8-deficient small intestine cells, apical proteins are missorted into lysosomes. In this study, we identified a novel Rab8-interacting protein complex containing an EH domain–binding protein 1–like 1 (EHBP1L1), Bin1/amphiphysin II, and dynamin. Biochemical analyses showed that EHBP1L1 directly bound to GTP-loaded Rab8 and Bin1. The spatial dependency of these complexes at the endocytic recycling compartment (ERC) was demonstrated through overexpression and knockdown experiments. EHBP1L1- or Bin1-depleted or dynamin-inhibited small intestine organoids significantly accumulated apical membrane proteins but not basolateral membrane proteins in lysosomes. Furthermore, in EHBP1L1-deficient mice, small intestine cells displayed truncated and sparse microvilli, suggesting that EHBP1L1 maintains the apical plasma membrane by regulating apical transport. In summary, our data demonstrate that EHBP1L1 links Rab8 and the Bin1–dynamin complex, which generates membrane curvature and excises the vesicle at the ERC for apical transport. PMID:26833786

  1. The identification of two regulatory ESCC susceptibility genetic variants in the TERT-CLPTM1L loci

    PubMed Central

    Wei, Jinyu; Shi, Juan; Pan, Wenting; Ren, Yanli; Xiong, Xiangyu; Xia, Jianhong; Shen, Yue; Li, Hongliang; Yang, Ming

    2016-01-01

    The chromosome 5p15.33 TERT-CLPTM1L region has been identified by genome-wide association studies as a susceptibility locus of multiple malignancies. However, the involvement of this locus in esophageal squamous cell carcinoma (ESCC) development is still largely unclear. We fine-mapped the TERT-CLPTM1L region through genotyping 15 haplotype-tagging single nucleotide polymorphisms (htSNPs) using a two stage case-control strategy. After analyzing 2098 ESCC patients and frequency-matched 2150 unaffected controls, we found that rs2853691, rs2736100 and rs451360 genetic polymorphisms are significantly associated with ESCC risk in Chinese (all P<0.05). Reporter gene assays indicated that the ESCC susceptibility SNP rs2736100 locating in a potential TERT intronic promoter has a genotype-specific effect on TERT expression. Similarly, the CLPTM1L rs451360 SNP also showed allelic impacts on gene expression. After measuring TERT and CLPTM1L expression in sixty-six pairs of esophageal cancer and normal tissues, we observed that the rs2736100 G risk allele carriers showed elevated oncogene TERT expression. Also, subjects with the rs451360 protective T allele had much lower oncogene CLPTM1L expression than those with G allele in tissue specimens. Results of these analyses underline the complexity of genetic regulation of telomere biology and further support the important role of telomerase in carcinogenesis. Our data also support the involvement of CLPTM1L in ESCC susceptibility. PMID:26716642

  2. The NPC1L1 Polymorphism 1679C>G Is Associated with Gallstone Disease in Chinese Patients

    PubMed Central

    Fei, Jian; Zhang, Sheng-Dao; Han, Tian-Quan; Hu, Hai; Jiang, Zhao-Yan

    2016-01-01

    Niemann Pick Type C1 Like 1 (NPC1L1) protein plays a key role in intestinal and hepatic cholesterol metabolism in humans. Genetic variation in NPC1L1 has been widely studied in recent years. We analyzed NPC1L1 single nucleotide polymorphisms in Chinese gallstone disease patients to investigate their association with gallstone disease. NPC1L1 mRNA expression was also measured in liver biopsies from patients with cholesterol gallstone disease and compared between genotypes. The G allele of the g1679C>G (rs2072183) polymorphism was significantly more prevalent in patients with gallstones compared with gallstone-free subjects. Moreover, patients carrying the G allele had lower hepatic NPC1L1 mRNA expression and higher biliary cholesterol (molar percentages) and cholesterol saturation index. Our study suggests that the G allele of the NPC1L1 polymorphism g1679C>G may be a positive marker of gallstone formation risk. PMID:26800364

  3. A novel de novo dominant negative mutation in DNM1L impairs mitochondrial fission and presents as childhood epileptic encephalopathy.

    PubMed

    Fahrner, Jill A; Liu, Raymond; Perry, Michael Scott; Klein, Jessica; Chan, David C

    2016-08-01

    DNM1L encodes dynamin-related protein 1 (DRP1/DLP1), a key component of the mitochondrial fission machinery that is essential for proper functioning of the mammalian brain. Previously reported probands with de novo missense mutations in DNM1L presented in the first year of life with severe encephalopathy and refractory epilepsy, with several dying within the first several weeks after birth. In contrast, we report identical novel missense mutations in DNM1L in two unrelated probands who experienced normal development for several years before presenting with refractory focal status epilepticus and subsequent rapid neurological decline. We expand the phenotype of DNM1L-related mitochondrial fission defects, reveal common unique clinical characteristics and imaging findings, and compare the cellular impact of this novel mutation to the previously reported A395D lethal variant. We demonstrate that our R403C mutation, which resides in the assembly region of DRP1, acts by a dominant-negative mechanism and reduces oligomerization, mitochondrial fission activity, and mitochondrial recruitment of DRP1, but to a lesser extent compared to the A395D mutation. In contrast to the initial report of neonatal lethality resulting from DNM1L mutation and DRP1 dysfunction, our results show that milder DRP1 impairment is compatible with normal early development and subsequently results in a distinct set of neurological findings. In addition, we identify a common pathogenic mechanism whereby DNM1L mutations impair mitochondrial fission. © 2016 Wiley Periodicals, Inc. PMID:27145208

  4. Chromosome Condensation 1-Like (Chc1L) Is a Novel Tumor Suppressor Involved in Development of Histiocyte-Rich Neoplasms

    PubMed Central

    Newbigging, Susan; Wang, Youdong; Shi, Chang-Xin; Cho, Hae-Ra; Shimizu, Hiroki; Gramolini, Anthony; Liu, Mingyao; Wen, Xiao-Yan

    2015-01-01

    Human chromosomal region 13q14 is a deletion hotspot in prostate cancer, multiple myeloma, and chronic lymphocytic leukemia. This region is believed to host multiple tumor suppressors. Chromosome Condensation 1-like (CHC1L) is located at 13q14, and found within the smallest common region of loss of heterozygosity in prostate cancer. Decreased expression of CHC1L is linked to pathogenesis and progression of both prostate cancer and multiple myeloma. However, there is no direct evidence for CHC1L’s putative tumor suppressing role in current literature. Presently, we describe the generation and characterization of Chc1L knockout mice. Chc1L-/- mice do not develop cancer at a young age, but bone marrow and spleen cells from 8–12 week-old mice display an exaggerated proliferative response. By approximately two years of age, knockout and heterozygote mice have a markedly increased incidence of tumorigenesis compared to wild-type controls, with tumors occurring mainly in the spleen, mesenteric lymph nodes, liver and intestinal tract. Histopathological analysis found that most heterozygote and knockout mice succumb to either Histiocytic Sarcoma or Histiocyte-Associated Lymphoma. Our study suggests that Chc1L is involved in suppression of these two histiocyte-rich neoplasms in mice and supports clinical data suggesting that CHC1L loss of function is an important step in the pathogenesis of cancers containing 13q14 deletion. PMID:26291700

  5. EHBP1L1 coordinates Rab8 and Bin1 to regulate apical-directed transport in polarized epithelial cells.

    PubMed

    Nakajo, Atsuhiro; Yoshimura, Shin-ichiro; Togawa, Hiroko; Kunii, Masataka; Iwano, Tomohiko; Izumi, Ayaka; Noguchi, Yuria; Watanabe, Ayako; Goto, Ayako; Sato, Toshiro; Harada, Akihiro

    2016-02-01

    The highly conserved Rab guanosine triphosphatase (GTPase) Rab8 plays a role in exocytosis toward the polarized plasma membrane in eukaryotic cells. In murine Rab8-deficient small intestine cells, apical proteins are missorted into lysosomes. In this study, we identified a novel Rab8-interacting protein complex containing an EH domain-binding protein 1-like 1 (EHBP1L1), Bin1/amphiphysin II, and dynamin. Biochemical analyses showed that EHBP1L1 directly bound to GTP-loaded Rab8 and Bin1. The spatial dependency of these complexes at the endocytic recycling compartment (ERC) was demonstrated through overexpression and knockdown experiments. EHBP1L1- or Bin1-depleted or dynamin-inhibited small intestine organoids significantly accumulated apical membrane proteins but not basolateral membrane proteins in lysosomes. Furthermore, in EHBP1L1-deficient mice, small intestine cells displayed truncated and sparse microvilli, suggesting that EHBP1L1 maintains the apical plasma membrane by regulating apical transport. In summary, our data demonstrate that EHBP1L1 links Rab8 and the Bin1-dynamin complex, which generates membrane curvature and excises the vesicle at the ERC for apical transport. PMID:26833786

  6. The NPC1L1 Polymorphism 1679C>G Is Associated with Gallstone Disease in Chinese Patients.

    PubMed

    Wu, Jian; Cui, Wei; Cai, Qu; Fei, Jian; Zhang, Sheng-Dao; Han, Tian-Quan; Hu, Hai; Jiang, Zhao-Yan

    2016-01-01

    Niemann Pick Type C1 Like 1 (NPC1L1) protein plays a key role in intestinal and hepatic cholesterol metabolism in humans. Genetic variation in NPC1L1 has been widely studied in recent years. We analyzed NPC1L1 single nucleotide polymorphisms in Chinese gallstone disease patients to investigate their association with gallstone disease. NPC1L1 mRNA expression was also measured in liver biopsies from patients with cholesterol gallstone disease and compared between genotypes. The G allele of the g1679C>G (rs2072183) polymorphism was significantly more prevalent in patients with gallstones compared with gallstone-free subjects. Moreover, patients carrying the G allele had lower hepatic NPC1L1 mRNA expression and higher biliary cholesterol (molar percentages) and cholesterol saturation index. Our study suggests that the G allele of the NPC1L1 polymorphism g1679C>G may be a positive marker of gallstone formation risk. PMID:26800364

  7. The identification of two regulatory ESCC susceptibility genetic variants in the TERT-CLPTM1L loci.

    PubMed

    Zhou, Liqing; Fu, Guobin; Wei, Jinyu; Shi, Juan; Pan, Wenting; Ren, Yanli; Xiong, Xiangyu; Xia, Jianhong; Shen, Yue; Li, Hongliang; Yang, Ming

    2016-02-01

    The chromosome 5p15.33 TERT-CLPTM1L region has been identified by genome-wide association studies as a susceptibility locus of multiple malignancies. However, the involvement of this locus in esophageal squamous cell carcinoma (ESCC) development is still largely unclear. We fine-mapped the TERT-CLPTM1L region through genotyping 15 haplotype-tagging single nucleotide polymorphisms (htSNPs) using a two stage case-control strategy. After analyzing 2098 ESCC patients and frequency-matched 2150 unaffected controls, we found that rs2853691, rs2736100 and rs451360 genetic polymorphisms are significantly associated with ESCC risk in Chinese (all P<0.05). Reporter gene assays indicated that the ESCC susceptibility SNP rs2736100 locating in a potential TERT intronic promoter has a genotype-specific effect on TERT expression. Similarly, the CLPTM1L rs451360 SNP also showed allelic impacts on gene expression. After measuring TERT and CLPTM1L expression in sixty-six pairs of esophageal cancer and normal tissues, we observed that the rs2736100 G risk allele carriers showed elevated oncogene TERT expression. Also, subjects with the rs451360 protective T allele had much lower oncogene CLPTM1L expression than those with G allele in tissue specimens. Results of these analyses underline the complexity of genetic regulation of telomere biology and further support the important role of telomerase in carcinogenesis. Our data also support the involvement of CLPTM1L in ESCC susceptibility. PMID:26716642

  8. LIQUID-LIQUID EXTRACTION COLUMNS

    DOEpatents

    Thornton, J.D.

    1957-12-31

    This patent relates to liquid-liquid extraction columns having a means for pulsing the liquid in the column to give it an oscillatory up and down movement, and consists of a packed column, an inlet pipe for the dispersed liquid phase and an outlet pipe for the continuous liquid phase located in the direct communication with the liquid in the lower part of said column, an inlet pipe for the continuous liquid phase and an outlet pipe for the dispersed liquid phase located in direct communication with the liquid in the upper part of said column, a tube having one end communicating with liquid in the lower part of said column and having its upper end located above the level of said outlet pipe for the dispersed phase, and a piston and cylinder connected to the upper end of said tube for applying a pulsating pneumatic pressure to the surface of the liquid in said tube so that said surface rises and falls in said tube.

  9. Alpha1L-adrenoceptors mediate contractions of the isolated mouse prostate.

    PubMed

    Gray, Katherine T; Ventura, Sabatino

    2006-07-01

    The subtype of alpha1-adrenoceptor mediating noradrenaline-induced contractile responses in isolated mouse prostate glands was investigated. Adrenoceptor agonists were able to produce concentration-dependent contractions with the following rank order of potency: adrenaline > or = noradrenaline > or = clonidine = phenylephrine > dopamine > or = isoprenaline. Concentration-response curves to noradrenaline of the prostatic smooth muscle were antagonised by prazosin, N-[2-(2-cyclopropylmethoxyphenoxy)ethyl]-5-chloro-alpha, alpha-dimethyl-1H-indole-3-ethanamine (RS-17053), 2-(2,6-dimethoxyphenoxyethyl)aminomethyl-1,4-benzodioxane (WB 4101), tamsulosin and yohimbine with mean antagonist affinity estimates (pA2 or apparent pKB) of 8.12+/-0.10, 6.56+/-0.11, 8.38+/-0.06, 10.14+/-0.19 and 7.38+/-1.36 respectively. Propranolol (1 microM) had no antagonist activity (P = 0.994, n = 6). Yohimbine (0.01, 0.1, 1 microM) had no antagonist activity in the presence of prazosin (0.1 microM) (P > or = 0.059). The results obtained indicate that alpha1-adrenoceptors mediate the contractile response in isolated preparations of the mouse prostate. Furthermore, the particular subtype of alpha1-adrenoceptor mediating the response to exogenously administered noradrenaline corresponds to the alpha1L-subtype, the same subtype as that which has been shown to mediate noradrenaline-induced contractile activity in the human prostate. PMID:16716294

  10. Quantification of the Host Response Proteome after Mammalian Reovirus T1L Infection

    PubMed Central

    Berard, Alicia R.; Cortens, John P.; Krokhin, Oleg; Wilkins, John A.; Severini, Alberto; Coombs, Kevin M.

    2012-01-01

    All viruses are dependent upon host cells for replication. Infection can induce profound changes within cells, including apoptosis, morphological changes, and activation of signaling pathways. Many of these alterations have been analyzed by gene arrays to measure the cellular “transcriptome.” We used SILAC (stable isotope labeling by amino acids in cell culture), combined with high-throughput 2-D HPLC/mass spectrometry, to determine relative quantitative differences in host proteins at 6 and 24 hours after infecting HEK293 cells with reovirus serotype 1 Lang (T1L). 3,076 host proteins were detected at 6hpi, of which 132 and 68 proteins were significantly up or down regulated, respectively. 2,992 cellular proteins, of which 104 and 49 were up or down regulated, respectively, were identified at 24hpi. IPA and DAVID analyses indicated proteins involved in cell death, cell growth factors, oxygen transport, cell structure organization and inflammatory defense response to virus were up-regulated, whereas proteins involved in apoptosis, isomerase activity, and metabolism were down-regulated. These proteins and pathways may be suitable targets for intervention to either attenuate virus infection or enhance oncolytic potential. PMID:23240068

  11. Expression of 1L-Myoinositol-1-Phosphate Synthase in Organelles1

    PubMed Central

    Lackey, Kimberly Helms; Pope, Patricia Marie; Johnson, Margaret Dean

    2003-01-01

    We have studied the expression of 1l-myoinositol-1-phosphate synthase (MIPS; EC 5.5.1.4) in developing organs of Phaseolus vulgaris to define genetic controls that spatially regulate inositol phosphate biosynthesis. MIPS, the pivotal biosynthetic enzyme in inositol metabolism, is the only enzyme known to catalyze the conversion of glucose 6-phosphate to inositol phosphate. It is found in unicellular and multicellular eukaryotes and has been isolated as a soluble enzyme from both. Thus, it is widely accepted that inositol phosphate biosynthesis is largely restricted to the cytosol. Here, we report findings that suggest the enzyme is also expressed in membrane-bound organelles. Microscopic and biochemical analyses detected MIPS expression in plasma membranes, plastids, mitochondria, endoplasmic reticula, nuclei, and cell walls of bean. To address mechanisms by which the enzyme could be targeted to or through membranes, MIPS genes were analyzed for sorting signals within primary structures and upstream open reading frames that we discovered through our sequence analyses. Comprehensive computer analyses revealed putative transit peptides that are predicted to target the enzyme to different cellular compartments. Reverse transcriptase PCR experiments suggest that these putative targeting peptides are expressed in bean roots and leaves. PMID:12913178

  12. Adenovirus RIDα uncovers a novel pathway requiring ORP1L for lipid droplet formation independent of NPC1

    PubMed Central

    Cianciola, Nicholas L.; Greene, Diane J.; Morton, Richard E.; Carlin, Cathleen R.

    2013-01-01

    Niemann–Pick disease type C (NPC) is caused by mutations in NPC1 or NPC2, which coordinate egress of low-density-lipoprotein (LDL)-cholesterol from late endosomes. We previously reported that the adenovirus-encoded protein RIDα rescues the cholesterol storage phenotype in NPC1-mutant fibroblasts. We show here that RIDα reconstitutes deficient endosome-to-endoplasmic reticulum (ER) transport, allowing excess LDL-cholesterol to be esterified by acyl-CoA:cholesterol acyltransferase and stored in lipid droplets (LDs) in NPC1-deficient cells. Furthermore, the RIDα pathway is regulated by the oxysterol-binding protein ORP1L. Studies have classified ORP1L as a sterol sensor involved in LE positioning downstream of GTP-Rab7. Our data, however, suggest that ORP1L may play a role in transport of LDL-cholesterol to a specific ER pool designated for LD formation. In contrast to NPC1, which is dispensable, the RIDα/ORP1L-dependent route requires functional NPC2. Although NPC1/NPC2 constitutes the major pathway, therapies that amplify minor egress routes for LDL-cholesterol could significantly improve clinical management of patients with loss-of-function NPC1 mutations. The molecular identity of putative alternative pathways, however, is poorly characterized. We propose RIDα as a model system for understanding physiological egress routes that use ORP1L to activate ER feedback responses involved in LD formation. PMID:24025716

  13. Incidence and clinical features of X-linked Cornelia de Lange syndrome due to SMC1L1 mutations.

    PubMed

    Borck, Guntram; Zarhrate, Mohamed; Bonnefont, Jean-Paul; Munnich, Arnold; Cormier-Daire, Valérie; Colleaux, Laurence

    2007-02-01

    Cornelia de Lange syndrome (CdLS) is a multisystem developmental disorder characterized by facial dysmorphism, growth and mental retardation, microcephaly, and various malformations. Heterozygous mutations in the NIPBL gene have been detected in approximately 45% of affected individuals. Recently, a second CdLS gene, mapping to the X chromosome, has been identified: SMC1L1 (structural maintenance of chromosomes 1-like 1; or SMC1A). In order to estimate the incidence and refine the clinical presentation of X-linked CdLS, we have screened a series of 11 CdLS boys carrying no NIPBL anomaly. We have identified two novel de novo SMC1L1 missense mutations (c.587G>A [p.Arg196His] and c.3254A>G [p.Tyr1085Cys]). Our results confirm that SMC1L1 mutations cause CdLS and support the view that SMC1L1 accounts for a significant fraction of boys with unexplained CdLS. Furthermore, we suggest that SMC1L1 mutations have milder effects than NIPBL mutations with respect to pre- and postnatal growth retardation and associated malformations. If confirmed, these data may have important implications for directing mutation screening in CdLS. PMID:17221863

  14. DOT1L inhibits SIRT1-mediated epigenetic silencing to maintain leukemic gene expression in MLL-rearranged leukemia

    PubMed Central

    Chen, C.W.; Koche, R.P.; Sinha, A.U.; Deshpande, A.J.; Zhu, N.; Eng, R.; Doench, J.G.; Xu, H.; Chu, S.H.; Qi, J.; Wang, X.; Delaney, C.; Bernt, K.M.; Root, D.E.; Hahn, W.C.; Bradner, J.E.; Armstrong, S.A.

    2015-01-01

    MLL -rearrangements generate MLL-fusion proteins that bind DNA and drive leukemogenic gene expression. This gene expression program is dependent on the histone 3 lysine 79 (H3K79) methyltransferase DOT1L, and small molecule DOT1L inhibitors show promise as therapeutics for these leukemias. However, the mechanisms underlying this dependency are unclear. We conducted a genome-scale RNAi screen and found that the histone deacetylase SIRT1 is required for the establishment of a heterochromatin-like state around MLL-fusion target genes after DOT1L inhibition. DOT1L inhibits chromatin localization of a repressive complex composed of SIRT1 and SUV39H1, thereby maintaining an open chromatin state with elevated H3K9 acetylation and minimal H3K9 methylation at MLL-fusion target genes. Furthermore, the combination of SIRT1 activators and DOT1L inhibitors shows enhanced activity against MLL-rearranged leukemia cells. These results indicate that the dynamic interplay between chromatin regulators controlling activation and repression of gene expression could provide novel opportunities for combination therapy. PMID:25822366

  15. Orlistat limits cholesterol intestinal absorption by Niemann-pick C1-like 1 (NPC1L1) inhibition.

    PubMed

    Alqahtani, Saeed; Qosa, Hisham; Primeaux, Brian; Kaddoumi, Amal

    2015-09-01

    The known mechanism by which orlistat decreases the absorption of dietary cholesterol is by inhibition of intestinal lipases. The aim of this study was to investigate the ability of orlistat to limit cholesterol absorption by inhibition of the cholesterol transport protein Niemann-Pick C1-like 1 (NPC1L1) as another mechanism of action. In situ rat intestinal perfusion studies were conducted to study the effect of orlistat on jejunal cholesterol absorption. Inhibition kinetic parameters were calculated from in vitro inhibition studies using Caco2 and NPC1L1 transfected cell lines. The in situ studies demonstrated that intestinal perfusion of orlistat (100µM) was able to reduce cholesterol absorption by three-fold when compared to control (i.e. in the absence of orlistat, P<0.01). In vitro studies using Caco2 cells demonstrated orlistat to reduce the cellular uptake of cholesterol by 30%. Additionally, orlistat reduced the cellular uptake of cholesterol in dose dependent manner in NPC1L1 transfected cell line with an IC50=1.2µM. Lineweaver-Burk plot indicated a noncompetitive inhibition of NPC1L1 by orlistat. Beside the already established mechanism by which orlistat reduces the absorption of cholesterol, we demonstrated for the first time that orlistat limits cholesterol absorption by the inhibition of NPC1L1 transport protein. PMID:26048312

  16. LIQUID TARGET

    DOEpatents

    Martin, M.D.; Salsig, W.W. Jr.

    1959-01-13

    A liquid handling apparatus is presented for a liquid material which is to be irradiated. The apparatus consists essentially of a reservoir for the liquid, a target element, a drain tank and a drain lock chamber. The target is in the form of a looped tube, the upper end of which is adapted to be disposed in a beam of atomic particles. The lower end of the target tube is in communication with the liquid in the reservoir and a means is provided to continuously circulate the liquid material to be irradiated through the target tube. Means to heat the reservoir tank is provided in the event that a metal is to be used as the target material. The apparatus is provided with suitable valves and shielding to provide maximum safety in operation.

  17. Intestinal and hepatic Niemann-Pick C1L1 proteins: future therapeutic targets for cholesterol gallstones disease?

    PubMed

    Castro-Torres, Ibrahim Guillermo; De la O-Arciniega, Minarda; Bravo-Duarte, Gustavo Adolfo; Gallegos-Estudillo, Janeth; Domínguez-Ortíz, Miguel Ángel; Martínez-Vázquez, Mariano

    2014-04-01

    The formation of cholesterol gallstones is a very complex and polygenic disorder that involves an alteration of the secretion of bile lipids, cholesterol crystallization, important immunological reactions in the gallbladder tissue, formation of biliary sludge composed of mucin, and inadequate gallbladder motility. The search for a therapeutic target is oriented towards decreasing bile secretion and intestinal absorption of cholesterol, in which Niemann-Pick C1L1 (NPC1L1) proteins play an important role. In basic and clinical studies, regulating the expression of these proteins can reduce intestinal, liver, plasma and bile cholesterol levels, a therapeutic effect that would be useful not only for treating the disease, but to prevent it, given the large quantity of risk factors. We discuss these effects in this review and propose NPC1L1 proteins as future therapeutic targets of cholesterol gallstones disease. PMID:24525336

  18. Identification of α1L-adrenoceptor in mice and its abolition by α1A-adrenoceptor gene knockout

    PubMed Central

    Muramatsu, I; Morishima, S; Suzuki, F; Yoshiki, H; Anisuzzaman, A S M; Tanaka, T; Rodrigo, M C; Myagmar, B E; Simpson, P C

    2008-01-01

    Background and purpose: The α1L-adrenoceptor has pharmacological properties that distinguish it from three classical α1-adrenoceptors (α1A, α1B and α1D). The purpose of this was to identify α1L-adrenoceptors in mice and to examine their relationship to classical α1-adrenoceptors. Experimental approach: Radioligand binding and functional bioassay experiments were performed on the cerebral cortex, vas deferens and prostate of wild-type (WT) and α1A-, α1B- and α1D-adrenoceptor gene knockout (AKO, BKO and DKO) mice. Key results: The radioligand [3H]-silodosin bound to intact segments of the cerebral cortex, vas deferens and prostate of WT, BKO and DKO but not of AKO mice. The binding sites were composed of two components with high and low affinities for prazosin or RS-17053, indicating the pharmacological profiles of α1A-adrenoceptors and α1L-adrenoceptors. In membrane preparations of WT mouse cortex, however, [3H]-silodosin bound to a single population of prazosin high-affinity sites, suggesting the presence of α1A-adrenoceptors alone. In contrast, [3H]-prazosin bound to two components having α1A-adrenoceptor and α1B-adrenoceptor profiles in intact segments of WT and DKO mouse cortices, but AKO mice lacked α1A-adrenoceptor profiles and BKO mice lacked α1B-adrenoceptor profiles. Noradrenaline produced contractions through α1L-adrenoceptors with low affinity for prazosin in the vas deferens and prostate of WT, BKO and DKO mice. However, the contractions were abolished or markedly attenuated in AKO mice. Conclusions and implications: α1L-Adrenoceptors were identified as binding and functional entities in WT, BKO and DKO mice but not in AKO mice, suggesting that the α1L-adrenoceptor is one phenotype derived from the α1A-adrenoceptor gene. PMID:18806813

  19. AKT activation drives the nuclear localization of CSE1L and a pro-oncogenic transcriptional activation in ovarian cancer cells

    SciTech Connect

    Lorenzato, Annalisa; Biolatti, Marta; Delogu, Giuseppe; Capobianco, Giampiero; Farace, Cristiano; Dessole, Salvatore; Cossu, Antonio; Tanda, Francesco; Madeddu, Roberto; Olivero, Martina; Di Renzo, Maria Flavia

    2013-10-15

    The human homolog of the yeast cse1 gene (CSE1L) is over-expressed in ovarian cancer. CSE1L forms complex with Ran and importin-α and has roles in nucleocytoplasmic traffic and gene expression. CSE1L accumulated in the nucleus of ovarian cancer cell lines, while it was localized also in the cytoplasm of other cancer cell lines. Nuclear localization depended on AKT, which was constitutively active in ovarian cancer cells, as the CSE1L protein translocated to the cytoplasm when AKT was inactivated. Moreover, the expression of a constitutively active AKT forced the translocation of CSE1L from the cytoplasm to the nucleus in other cancer cells. Nuclear accrual of CSE1L was associated to the nuclear accumulation of the phosphorylated Ran Binding protein 3 (RanBP3), which depended on AKT as well. Also in samples of human ovarian cancer, AKT activation was associated to nuclear accumulation of CSE1L and phosphorylation of RanBP3. Expression profiling of ovarian cancer cells after CSE1L silencing showed that CSE1L was required for the expression of genes promoting invasion and metastasis. In agreement, CSE1L silencing impaired motility and invasiveness of ovarian cancer cells. Altogether these data show that in ovarian cancer cells activated AKT by affecting RanBP3 phosphorylation determines the nuclear accumulation of CSE1L and likely the nuclear concentration of transcription factors conveying pro-oncogenic signals. - highlights: • CSE1L is a key player in nucleocytoplasmic traffic by forming complex with Ran. • AKT phosphorylates RanBP3 that regulates the nucleocytoplasmic gradient of Ran. • The activated oncogenic AKT drives the nuclear accumulation of CSE1L. • CSE1L in the nucleus up-regulates genes conveying pro-oncogenic signals. • CSE1L might contribute to tumor progression driven by the activated oncogenic AKT.

  20. AKT activation drives the nuclear localization of CSE1L and a pro-oncogenic transcriptional activation in ovarian cancer cells.

    PubMed

    Lorenzato, Annalisa; Biolatti, Marta; Delogu, Giuseppe; Capobianco, Giampiero; Farace, Cristiano; Dessole, Salvatore; Cossu, Antonio; Tanda, Francesco; Madeddu, Roberto; Olivero, Martina; Di Renzo, Maria Flavia

    2013-10-15

    The human homolog of the yeast cse1 gene (CSE1L) is over-expressed in ovarian cancer. CSE1L forms complex with Ran and importin-α and has roles in nucleocytoplasmic traffic and gene expression. CSE1L accumulated in the nucleus of ovarian cancer cell lines, while it was localized also in the cytoplasm of other cancer cell lines. Nuclear localization depended on AKT, which was constitutively active in ovarian cancer cells, as the CSE1L protein translocated to the cytoplasm when AKT was inactivated. Moreover, the expression of a constitutively active AKT forced the translocation of CSE1L from the cytoplasm to the nucleus in other cancer cells. Nuclear accrual of CSE1L was associated to the nuclear accumulation of the phosphorylated Ran Binding protein 3 (RanBP3), which depended on AKT as well. Also in samples of human ovarian cancer, AKT activation was associated to nuclear accumulation of CSE1L and phosphorylation of RanBP3. Expression profiling of ovarian cancer cells after CSE1L silencing showed that CSE1L was required for the expression of genes promoting invasion and metastasis. In agreement, CSE1L silencing impaired motility and invasiveness of ovarian cancer cells. Altogether these data show that in ovarian cancer cells activated AKT by affecting RanBP3 phosphorylation determines the nuclear accumulation of CSE1L and likely the nuclear concentration of transcription factors conveying pro-oncogenic signals. PMID:23948303

  1. Cholesterol and ORP1L-mediated ER contact sites control autophagosome transport and fusion with the endocytic pathway

    PubMed Central

    Wijdeven, Ruud H.; Janssen, Hans; Nahidiazar, Leila; Janssen, Lennert; Jalink, Kees; Berlin, Ilana; Neefjes, Jacques

    2016-01-01

    Autophagy is the main homeostatic pathway guiding cytosolic materials for degradation by the lysosome. Maturation of autophagosomes requires their transport towards the perinuclear region of the cell, with key factors underlying both processes still poorly understood. Here we show that transport and positioning of late autophagosomes depends on cholesterol by way of the cholesterol-sensing Rab7 effector ORP1L. ORP1L localizes to late autophagosomes and—under low-cholesterol conditions—contacts the ER protein VAP-A, forming ER-autophagosome contact sites, which prevent minus-end transport by the Rab7–RILP–dynein complex. ORP1L-mediated contact sites also inhibit localization of PLEKHM1 to Rab7. PLEKHM1, together with RILP, then recruits the homotypic fusion and vacuole protein-sorting (HOPS) complex for fusion of autophagosomes with late endosomes and lysosomes. Thus, ORP1L, via its liganding by lipids and the formation of contacts between autophagic vacuoles and the ER, governs the last steps in autophagy that lead to the lysosomal degradation of cytosolic material. PMID:27283760

  2. Loss of Drosophila i-AAA protease, dYME1L, causes abnormal mitochondria and apoptotic degeneration.

    PubMed

    Qi, Y; Liu, H; Daniels, M P; Zhang, G; Xu, H

    2016-02-01

    Mitochondrial AAA (ATPases Associated with diverse cellular Activities) proteases i-AAA (intermembrane space-AAA) and m-AAA (matrix-AAA) are closely related and have major roles in inner membrane protein homeostasis. Mutations of m-AAA proteases are associated with neuromuscular disorders in humans. However, the role of i-AAA in metazoans is poorly understood. We generated a deletion affecting Drosophila i-AAA, dYME1L (dYME1L(del)). Mutant flies exhibited premature aging, progressive locomotor deficiency and neurodegeneration that resemble some key features of m-AAA diseases. dYME1L(del) flies displayed elevated mitochondrial unfolded protein stress and irregular cristae. Aged dYME1L(del) flies had reduced complex I (NADH/ubiquinone oxidoreductase) activity, increased level of reactive oxygen species (ROS), severely disorganized mitochondrial membranes and increased apoptosis. Furthermore, inhibiting apoptosis by targeting dOmi (Drosophila Htra2/Omi) or DIAP1, or reducing ROS accumulation suppressed retinal degeneration. Our results suggest that i-AAA is essential for removing unfolded proteins and maintaining mitochondrial membrane architecture. Loss of i-AAA leads to the accumulation of oxidative damage and progressive deterioration of membrane integrity, which might contribute to apoptosis upon the release of proapoptotic molecules such as dOmi. Containing ROS level could be a potential strategy to manage mitochondrial AAA protease deficiency. PMID:26160069

  3. Disruption of the Hbs1l-Myb locus causes hereditary persistence of fetal hemoglobin in a mouse model.

    PubMed

    Suzuki, Mikiko; Yamazaki, Hiromi; Mukai, Harumi Y; Motohashi, Hozumi; Shi, Lihong; Tanabe, Osamu; Engel, James Douglas; Yamamoto, Masayuki

    2013-04-01

    The human β-globin locus is comprised of embryonic, fetal, and adult globin genes, each of which is expressed at distinct stages of pre- and postnatal development. Functional defects in globin proteins or expression results in mild to severe anemia, such as in sickle-cell disease or β-thalassemia, but the clinical symptoms of both disorders are ameliorated by persistent expression of the fetal globin genes. Recent genome-wide association studies (GWAS) identified the intergenic region between the HBS1L and MYB loci as a candidate modifier of fetal hemoglobin expression in adults. However, it remains to be clarified whether the enhancer activity within the HBS1L-MYB regulatory domain contributes to the production of fetal hemoglobin in adults. Here we report a new mouse model of hereditary persistence of fetal hemoglobin (HPFH) in which a transgene was randomly inserted into the orthologous murine Hbs1l-Myb locus. This mutant mouse exhibited typically elevated expression of embryonic globins and hematopoietic parameters similar to those observed in human HPFH. These results support the contention that mutation of the HBS1L-MYB genomic domain is responsible for elevated expression of the fetal globin genes, and this model serves as an important means for the analysis of networks that regulate fetal globin gene expression. PMID:23428869

  4. Late endosomal transport and tethering are coupled processes controlled by RILP and the cholesterol sensor ORP1L.

    PubMed

    van der Kant, Rik; Fish, Alexander; Janssen, Lennert; Janssen, Hans; Krom, Sabine; Ho, Nataschja; Brummelkamp, Thijn; Carette, Jan; Rocha, Nuno; Neefjes, Jacques

    2013-08-01

    Late endosomes and lysosomes are dynamic organelles that constantly move and fuse to acquire cargo from early endosomes, phagosomes and autophagosome. Defects in lysosomal dynamics cause severe neurodegenerative and developmental diseases, such as Niemann-Pick type C disease and ARC syndrome, yet little is known about the regulation of late endosomal fusion in a mammalian system. Mammalian endosomes destined for fusion need to be transported over very long distances before they tether to initiate contact. Here, we describe that lysosomal tethering and transport are combined processes co-regulated by one multi-protein complex: RAB7-RILP-ORP1L. We show that RILP directly and concomitantly binds the tethering HOPS complex and the p150(Glued) subunit of the dynein motor. ORP1L then functions as a cholesterol-sensing switch controlling RILP-HOPS-p150(Glued) interactions. We show that RILP and ORP1L control Ebola virus infection, a process dependent on late endosomal fusion. By combining recruitment and regulation of both the dynein motor and HOPS complex into a single multiprotein complex, the RAB7-RILP-ORP1L complex efficiently couples and regulates the timing of microtubule minus-end transport and fusion, two major events in endosomal biology. PMID:23729732

  5. Ataxin1L is a regulator of HSC function highlighting the utility of cross-tissue comparisons for gene discovery.

    PubMed

    Kahle, Juliette J; Souroullas, George P; Yu, Peng; Zohren, Fabian; Lee, Yoontae; Shaw, Chad A; Zoghbi, Huda Y; Goodell, Margaret A

    2013-03-01

    Hematopoietic stem cells (HSCs) are rare quiescent cells that continuously replenish the cellular components of the peripheral blood. Observing that the ataxia-associated gene Ataxin-1-like (Atxn1L) was highly expressed in HSCs, we examined its role in HSC function through in vitro and in vivo assays. Mice lacking Atxn1L had greater numbers of HSCs that regenerated the blood more quickly than their wild-type counterparts. Molecular analyses indicated Atxn1L null HSCs had gene expression changes that regulate a program consistent with their higher level of proliferation, suggesting that Atxn1L is a novel regulator of HSC quiescence. To determine if additional brain-associated genes were candidates for hematologic regulation, we examined genes encoding proteins from autism- and ataxia-associated protein-protein interaction networks for their representation in hematopoietic cell populations. The interactomes were found to be highly enriched for proteins encoded by genes specifically expressed in HSCs relative to their differentiated progeny. Our data suggest a heretofore unappreciated similarity between regulatory modules in the brain and HSCs, offering a new strategy for novel gene discovery in both systems. PMID:23555280

  6. Cholesterol and ORP1L-mediated ER contact sites control autophagosome transport and fusion with the endocytic pathway.

    PubMed

    Wijdeven, Ruud H; Janssen, Hans; Nahidiazar, Leila; Janssen, Lennert; Jalink, Kees; Berlin, Ilana; Neefjes, Jacques

    2016-01-01

    Autophagy is the main homeostatic pathway guiding cytosolic materials for degradation by the lysosome. Maturation of autophagosomes requires their transport towards the perinuclear region of the cell, with key factors underlying both processes still poorly understood. Here we show that transport and positioning of late autophagosomes depends on cholesterol by way of the cholesterol-sensing Rab7 effector ORP1L. ORP1L localizes to late autophagosomes and-under low-cholesterol conditions-contacts the ER protein VAP-A, forming ER-autophagosome contact sites, which prevent minus-end transport by the Rab7-RILP-dynein complex. ORP1L-mediated contact sites also inhibit localization of PLEKHM1 to Rab7. PLEKHM1, together with RILP, then recruits the homotypic fusion and vacuole protein-sorting (HOPS) complex for fusion of autophagosomes with late endosomes and lysosomes. Thus, ORP1L, via its liganding by lipids and the formation of contacts between autophagic vacuoles and the ER, governs the last steps in autophagy that lead to the lysosomal degradation of cytosolic material. PMID:27283760

  7. Proposed Ames M2-F1, M1-L half-cone, and Langley lenticular bodies.

    NASA Technical Reports Server (NTRS)

    1962-01-01

    Dale Reed, who inaugurated the lifting-body flight research at NASA's Flight Research Center (later, Dryden Flight Research Center, Edwards, CA), originally proposed that three wooden outer shells be built. These would then be attached to the single internal steel structure. The three shapes were (viewer's left to right) the M2-F1, the M1-L, and a lenticular shape. Milt Thompson, who supported Reed's advocacy for a lifting-body research project, recommended that only the M2-F1 shell be built, believing that the M1-L shape was 'too radical,' while the lenticular one was 'too exotic.' Although the lenticular shape was often likened to that of a flying saucer, Reed's wife Donna called it the 'powder puff.' The wingless, lifting body aircraft design was initially conceived as a means of landing an aircraft horizontally after atmospheric reentry. The absence of wings would make the extreme heat of re-entry less damaging to the vehicle. In 1962, Dryden management approved a program to build a lightweight, unpowered lifting body as a prototype to flight test the wingless concept. It would look like a 'flying bathtub,' and was designated the M2-F1, the 'M' referring to 'manned' and 'F' referring to 'flight' version. It featured a plywood shell placed over a tubular steel frame crafted at Dryden. Construction was completed in 1963. The first flight tests of the M2-F1 were over Rogers Dry Lake at the end of a tow rope attached to a hopped-up Pontiac convertible driven at speeds up to about 120 mph. This vehicle needed to be able to tow the M2-F1 on the Rogers Dry Lakebed adjacent to NASA's Flight Research Center (FRC) at a minimum speed of 100 miles per hour. To do that, it had to handle the 400-pound pull of the M2-F1. Walter 'Whitey' Whiteside, who was a retired Air Force maintenance officer working in the FRC's Flight Operations Division, was a dirt-bike rider and hot-rodder. Together with Boyden 'Bud' Bearce in the Procurement and Supply Branch of the FRC, Whitey

  8. Liquid atomization

    NASA Astrophysics Data System (ADS)

    Bayvel, L.; Orzechowski, Z.

    The present text defines the physical processes of liquid atomization, the primary types of atomizers and their design, and ways of measuring spray characteristics; it also presents experimental investigation results on atomizers and illustrative applications for them. Attention is given to the macrostructural and microstructural parameters of atomized liquids; swirl, pneumatic, and rotary atomizers; and optical drop sizing methods, with emphasis on nonintrusive optical methods.

  9. NPC1L1 is a key regulator of intestinal vitamin K absorption and a modulator of warfarin therapy.

    PubMed

    Takada, Tappei; Yamanashi, Yoshihide; Konishi, Kentaro; Yamamoto, Takehito; Toyoda, Yu; Masuo, Yusuke; Yamamoto, Hideaki; Suzuki, Hiroshi

    2015-02-18

    Vitamin K (VK) is a micronutrient that facilitates blood coagulation. VK antagonists, such as warfarin, are used in the clinic to prevent thromboembolism. Because VK is not synthesized in the body, its intestinal absorption is crucial for maintaining whole-body VK levels. However, the molecular mechanism of this absorption is unclear. We demonstrate that Niemann-Pick C1-like 1 (NPC1L1) protein, a cholesterol transporter, plays a central role in intestinal VK uptake and modulates the anticoagulant effect of warfarin. In vitro studies using NPC1L1-overexpressing intestinal cells and in vivo studies with Npc1l1-knockout mice revealed that intestinal VK absorption is NPC1L1-dependent and inhibited by ezetimibe, an NPC1L1-selective inhibitor clinically used for dyslipidemia. In addition, in vivo pharmacological studies demonstrated that the coadministration of ezetimibe and warfarin caused a reduction in hepatic VK levels and enhanced the pharmacological effect of warfarin. Adverse events caused by the coadministration of ezetimibe and warfarin were rescued by oral VK supplementation, suggesting that the drug-drug interaction effects observed were the consequence of ezetimibe-mediated VK malabsorption. This mechanism was supported by a retrospective evaluation of clinical data showing that, in more than 85% of warfarin-treated patients, the anticoagulant activity was enhanced by cotreatment with ezetimibe. Our findings provide insight into the molecular mechanism of VK absorption. This new drug-drug interaction mechanism between ezetimibe (a cholesterol transport inhibitor) and warfarin (a VK antagonist and anticoagulant) could inform clinical care of patients on these medications, such as by altering the kinetics of essential, fat-soluble vitamins. PMID:25696002

  10. Occult macular dystrophy in an Italian family carrying a mutation in the RP1L1 gene.

    PubMed

    Piermarocchi, Stefano; Segato, Tatiana; Leon, Alberta; Colavito, Davide; Miotto, Stefania

    2016-03-01

    Occult macular dystrophy (OMD) is an inherited macular disease characterized by progressive visual decline with the absence of visible retinal abnormalities. Typical alterations of the retinal structure are detectable by spectral domain optical coherence tomography (SD‑OCT). Mutations in the RP1L1 gene have been identified to be responsible for the disease in Asian subjects. The present study assessed the role of mutations in the RP1L1 gene in an Italian family with OMD. One patient with OMD and five related subjects (two male offspring affected by progressive visual decline and three asymptomatic siblings of the patient) were subjected to complete ophthalmological examination. SD‑OCT was also performed. All subjects were screened for OMD‑associated genetic mutations in the RP1L1 gene. The OMD patient and the two symptomatic offspring presented with a reduced best‑corrected visual acuity. Although no fundus abnormalities were observed, SD‑OCT examination showed that the external limiting membrane and the inner segment/outer segment band were not clearly identifiable and a focal disruption of the photoreceptor layer was present. The degree of photoreceptor alterations was correlated with the severity of visual impairment. Clinical and tomographic results in the three asymptomatic relatives were normal. A p.Arg45Trp mutation in the RP1L1 gene was identified in the OMD patient, in the two symptomatic offspring and also in two of the asymptomatic siblings of the patient. The identification of RP1L1 mutations in subjects with OMD may improve the accuracy of diagnosis of this rare condition and may aid in enhancing the efficacy of genetic counseling. PMID:26782618

  11. YME1L controls the accumulation of respiratory chain subunits and is required for apoptotic resistance, cristae morphogenesis, and cell proliferation.

    PubMed

    Stiburek, Lukas; Cesnekova, Jana; Kostkova, Olga; Fornuskova, Daniela; Vinsova, Kamila; Wenchich, Laszlo; Houstek, Josef; Zeman, Jiri

    2012-03-01

    Mitochondrial ATPases associated with diverse cellular activities (AAA) proteases are involved in the quality control and processing of inner-membrane proteins. Here we investigate the cellular activities of YME1L, the human orthologue of the Yme1 subunit of the yeast i-AAA complex, using stable short hairpin RNA knockdown and expression experiments. Human YME1L is shown to be an integral membrane protein that exposes its carboxy-terminus to the intermembrane space and exists in several complexes of 600-1100 kDa. The stable knockdown of YME1L in human embryonic kidney 293 cells led to impaired cell proliferation and apoptotic resistance, altered cristae morphology, diminished rotenone-sensitive respiration, and increased susceptibility to mitochondrial membrane protein carbonylation. Depletion of YME1L led to excessive accumulation of nonassembled respiratory chain subunits (Ndufb6, ND1, and Cox4) in the inner membrane. This was due to a lack of YME1L proteolytic activity, since the excessive accumulation of subunits was reversed by overexpression of wild-type YME1L but not a proteolytically inactive YME1L variant. Similarly, the expression of wild-type YME1L restored the lamellar cristae morphology of YME1L-deficient mitochondria. Our results demonstrate the importance of mitochondrial inner-membrane proteostasis to both mitochondrial and cellular function and integrity and reveal a novel role for YME1L in the proteolytic regulation of respiratory chain biogenesis. PMID:22262461

  12. YME1L controls the accumulation of respiratory chain subunits and is required for apoptotic resistance, cristae morphogenesis, and cell proliferation

    PubMed Central

    Stiburek, Lukas; Cesnekova, Jana; Kostkova, Olga; Fornuskova, Daniela; Vinsova, Kamila; Wenchich, Laszlo; Houstek, Josef; Zeman, Jiri

    2012-01-01

    Mitochondrial ATPases associated with diverse cellular activities (AAA) proteases are involved in the quality control and processing of inner-membrane proteins. Here we investigate the cellular activities of YME1L, the human orthologue of the Yme1 subunit of the yeast i‑AAA complex, using stable short hairpin RNA knockdown and expression experiments. Human YME1L is shown to be an integral membrane protein that exposes its carboxy-terminus to the intermembrane space and exists in several complexes of 600–1100 kDa. The stable knockdown of YME1L in human embryonic kidney 293 cells led to impaired cell proliferation and apoptotic resistance, altered cristae morphology, diminished rotenone-sensitive respiration, and increased susceptibility to mitochondrial membrane protein carbonylation. Depletion of YME1L led to excessive accumulation of nonassembled respiratory chain subunits (Ndufb6, ND1, and Cox4) in the inner membrane. This was due to a lack of YME1L proteolytic activity, since the excessive accumulation of subunits was reversed by overexpression of wild-type YME1L but not a proteolytically inactive YME1L variant. Similarly, the expression of wild-type YME1L restored the lamellar cristae morphology of YME1L-deficient mitochondria. Our results demonstrate the importance of mitochondrial inner-membrane proteostasis to both mitochondrial and cellular function and integrity and reveal a novel role for YME1L in the proteolytic regulation of respiratory chain biogenesis. PMID:22262461

  13. Liquid marbles

    NASA Astrophysics Data System (ADS)

    Aussillous, Pascale; Quéré, David

    2001-06-01

    The transport of a small amount of liquid on a solid is not a simple process, owing to the nature of the contact between the two phases. Setting a liquid droplet in motion requires non-negligible forces (because the contact-angle hysteresis generates a force opposing the motion), and often results in the deposition of liquid behind the drop. Different methods of levitation-electrostatic, electromagnetic, acoustic, or even simpler aerodynamic techniques-have been proposed to avoid this wetting problem, but all have proved to be rather cumbersome. Here we propose a simple alternative, which consists of encapsulating an aqueous liquid droplet with a hydrophobic powder. The resulting `liquid marbles' are found to behave like a soft solid, and show dramatically reduced adhesion to a solid surface. As a result, motion can be generated using gravitational, electrical and magnetic fields. Moreover, because the viscous friction associated with motion is very small, we can achieve quick displacements of the droplets without any leaks. All of these features are of potential benefit in microfluidic applications, and also permit the study of a drop in a non-wetting situation-an issue of renewed interest following the recent achievement of super-hydrophobic substrates.

  14. Liquid Crystals

    NASA Astrophysics Data System (ADS)

    1990-01-01

    Thermochromic liquid crystals, or TLCs, are a type of liquid crystals that react to changes in temperature by changing color. The Hallcrest/NASA collaboration involved development of a new way to visualize boundary layer transition in flight and in wind tunnel testing of aircraft wing and body surfaces. TLCs offered a new and potentially better method of visualizing the boundary layer transition in flight. Hallcrest provided a liquid crystal formulation technique that afforded great control over the sensitivity of the liquid crystals to varying conditions. Method is of great use to industry, government and universities for aerodynamic and hydrodynamic testing. Company's principal line is temperature indicating devices for industrial use, such as non-destructive testing and flaw detection in electric/electronic systems, medical application, such as diagnostic systems, for retail sale, such as room, refrigerator, baby bath and aquarium thermometers, and for advertising and promotion specials. Additionally, Hallcrest manufactures TLC mixtures for cosmetic applications, and liquid crystal battery tester for Duracell batteries.

  15. Liquid marbles.

    PubMed

    Aussillous, P; Quéré, D

    2001-06-21

    The transport of a small amount of liquid on a solid is not a simple process, owing to the nature of the contact between the two phases. Setting a liquid droplet in motion requires non-negligible forces (because the contact-angle hysteresis generates a force opposing the motion), and often results in the deposition of liquid behind the drop. Different methods of levitation-electrostatic, electromagnetic, acoustic, or even simpler aerodynamic techniques-have been proposed to avoid this wetting problem, but all have proved to be rather cumbersome. Here we propose a simple alternative, which consists of encapsulating an aqueous liquid droplet with a hydrophobic powder. The resulting 'liquid marbles' are found to behave like a soft solid, and show dramatically reduced adhesion to a solid surface. As a result, motion can be generated using gravitational, electrical and magnetic fields. Moreover, because the viscous friction associated with motion is very small, we can achieve quick displacements of the droplets without any leaks. All of these features are of potential benefit in microfluidic applications, and also permit the study of a drop in a non-wetting situation-an issue of renewed interest following the recent achievement of super-hydrophobic substrates. PMID:11418851

  16. β-Cryptoxanthin modulates the response to phytosterols in post-menopausal women carrying NPC1L1 L272L and ABCG8 A632 V polymorphisms: an exploratory study.

    PubMed

    Granado-Lorencio, F; de Las Heras, L; Millán, C San; Garcia-López, F J; Blanco-Navarro, I; Pérez-Sacristán, B; Domínguez, G

    2014-09-01

    Phytosterol (PS) intake may be used for hypercholesterolaemia in some groups although the presence of non-responders is well known. Carotenoids and PS/cholesterol may compete for the same transporters during absorption. As part of a randomized, double-blind, crossover, multiple-dose supplementation study with β-cryptoxanthin (β-Cx) and PS, single and combined, polymorphisms of ABCG8 (A632V) and NCPL1 (L272L) were determined in 19 post-menopausal women. Subjects carrying CC polymorphism for NCP1L1 (L272L) showed a net increase in total cholesterol and LDL after PS intake but, interestingly, displayed a decrease in both lipid fractions after consuming PS plus β-Cx. For the ABCG8 (A632V) gene, CT/TT carriers consuming PS also displayed an increase in total cholesterol and LDL, but this increment was much lower after the intake of PS plus β-Cx. Additionally, in CC carriers for ABCG8 (A632V), a greater decrease in total cholesterol and LDL was found after the intake of PS plus β-Cx compared to that observed after PS alone. Overall, our results suggest that β-Cx improves the response to PS in individuals carrying specific genetic polymorphisms (i.e. non-responders), opening the possibility to modulate the response to PS by food technology. (ClinicalTrials.gov NCT01074723). PMID:25163590

  17. Macromolecular liquids

    SciTech Connect

    Safinya, C.R.; Safran, S.A. ); Pincus, P.A. )

    1990-01-01

    Liquids include a broad range of material systems which are of high scientific and technological interest. Generally speaking, these are partially ordered or disordered phases where the individual molecular species have organized themselves on length scales which are larger than simple fluids, typically between 10 Angstroms and several microns. The specific systems reported on in this book include membranes, microemulsions, micelles, liquid crystals, colloidal suspensions, and polymers. They have a major impact on a broad spectrum of technological industries such as displays, plastics, soap and detergents, chemicals and petroleum, and pharmaceuticals.

  18. Human PSENEN and U2AF1L4 genes are concertedly regulated by a genuine bidirectional promoter.

    PubMed

    Didych, D A; Shamsutdinov, M F; Smirnov, N A; Akopov, S B; Monastyrskaya, G S; Uspenskaya, N Y; Nikolaev, L G; Sverdlov, E D

    2013-02-15

    Head-to-head genes with a short distance between their transcription start sites may constitute up to 10% of all genes in the genomes of various species. It was hypothesized that this intergenic space may represent bidirectional promoters which are able to initiate transcription of both genes, but the true bidirectionality was proved only for a few of them. We present experimental evidence that, according to several criteria, a 269 bp region located between the PSENEN and U2AF1L4 human genes is a genuine bidirectional promoter regulating a concerted divergent transcription of these genes. Concerted transcription of PSENEN and U2AF1L4 can be necessary for regulation of T-cell activity. PMID:23246698

  19. Crystallization and preliminary X-ray diffraction analysis of the Sel1-like repeats of SEL1L.

    PubMed

    Jeong, Hanbin; Lee, Hakbong; Lee, Changwook

    2014-12-01

    Terminally misfolded or unassembled proteins are selectively recognized and cleared by the ER-associated degradation (ERAD) pathway. Suppressor/enhancer of lin-12-like (SEL1L), a component of the dislocation machinery containing the E3 ubiquitin ligase Hrd1, plays an important role in selecting and transporting ERAD substrates for degradation in the endoplasmic reticulum. In this study, the purification, crystallization and preliminary X-ray diffraction analysis of recombinant mouse SEL1L (residues 348-533) are reported. The crystals were obtained by the hanging-drop vapour-diffusion method at pH 8.5 and 277 K using 30% 2-propanol as a precipitant. Optimized crystals diffracted to 3.3 Å resolution at a synchrotron-radiation source. Preliminary X-ray diffraction analysis revealed that the crystals belonged to space group P21 and contained four molecules per asymmetric unit, with a solvent content of 44%. PMID:25484212

  20. Optimization of a Fragment-Based Screening Hit toward Potent DOT1L Inhibitors Interacting in an Induced Binding Pocket.

    PubMed

    Scheufler, Clemens; Möbitz, Henrik; Gaul, Christoph; Ragot, Christian; Be, Céline; Fernández, César; Beyer, Kim S; Tiedt, Ralph; Stauffer, Frédéric

    2016-08-11

    Mixed lineage leukemia (MLL) gene rearrangement induces leukemic transformation by ectopic recruitment of disruptor of telomeric silencing 1-like protein (DOT1L), a lysine histone methyltransferase, leading to local hypermethylation of H3K79 and misexpression of genes (including HoxA), which drive the leukemic phenotype. A weak fragment-based screening hit identified by SPR was cocrystallized with DOT1L and optimized using structure-based ligand optimization to yield compound 8 (IC50 = 14 nM). This series of inhibitors is structurally not related to cofactor SAM and is not interacting within the SAM binding pocket but induces a pocket adjacent to the SAM binding site. PMID:27563394

  1. aura (mid1ip1l) regulates the cytoskeleton at the zebrafish egg-to-embryo transition.

    PubMed

    Eno, Celeste; Solanki, Bharti; Pelegri, Francisco

    2016-05-01

    Embryos from females homozygous for a recessive maternal-effect mutation in the gene aura exhibit defects including reduced cortical integrity, defective cortical granule (CG) release upon egg activation, failure to complete cytokinesis, and abnormal cell wound healing. We show that the cytokinesis defects are associated with aberrant cytoskeletal reorganization during furrow maturation, including abnormal F-actin enrichment and microtubule reorganization. Cortical F-actin prior to furrow formation fails to exhibit a normal transition into F-actin-rich arcs, and drug inhibition is consistent with aura function promoting F-actin polymerization and/or stabilization. In mutants, components of exocytic and endocytic vesicles, such as Vamp2, Clathrin and Dynamin, are sequestered in unreleased CGs, indicating a need for CG recycling in the normal redistribution of these factors. However, the exocytic targeting factor Rab11 is recruited to the furrow plane normally at the tip of bundling microtubules, suggesting an alternative anchoring mechanism independent of membrane recycling. A positional cloning approach indicates that the mutation in aura is associated with a truncation of Mid1 interacting protein 1 like (Mid1ip1l), previously identified as an interactor of the X-linked Opitz G/BBB syndrome gene product Mid1. A Cas9/CRISPR-induced mutant allele in mid1ip1l fails to complement the originally isolated aura maternal-effect mutation, confirming gene assignment. Mid1ip1l protein localizes to cortical F-actin aggregates, consistent with a direct role in cytoskeletal regulation. Our studies indicate that maternally provided aura (mid1ip1l) acts during the reorganization of the cytoskeleton at the egg-to-embryo transition and highlight the importance of cytoskeletal dynamics and membrane recycling during this developmental period. PMID:26965374

  2. Potential effects of NPC1L1 polymorphisms in protecting against clinical disease in a chinese family with sitosterolaemia.

    PubMed

    Hu, Miao; Yuen, Yuet-Ping; Kwok, Jeffrey Ss; Griffith, James F; Tomlinson, Brian

    2014-01-01

    Sitosterolaemia is caused by mutations in either ABCG5 or ABCG8. Chinese and Japanese individuals usually have mutations in ABCG5. We herein report a known and a novel mutation in ABCG8 and their potential interaction with NPC1L1 polymorphisms in a Chinese family with sitosterolaemia. We sequenced ABCG5 and ABCG8 and measured the levels of plasma plant sterols in a 15-year-old Chinese girl with clinical sitosterolaemia (xanthomas with elevated low-density lipoprotein cholesterol (LDL-C) and plant sterols) and her apparently healthy family members. NPC1L1 was sequenced in the genetically affected sibling and other family members. A known mutation, c.490C>T (p. Arg164(*)), in exon 4 and a novel mutation, c.1949T>G (p.Leu650Arg), in exon 13 of ABCG8 were detected in the proband and her sister, who had elevated sterols but low LDL-C levels and no xanthomas. The genetically affected sister, but not the proband, carried two additional heterozygous changes in NPC1L1 (rs2072183 C>G, rs2301935 A>C), which were inherited from the mother, who also had a low LDL-C level. In this study, we detected a known and a novel mutation in ABCG8 in a Chinese patient with sitosterolaemia. The same mutations were found in her clinically normal sister, suggesting that the contrasting features with the proband may be related to different variants in NPC1L1 and/or some other undetermined lipid-related genetic factors. PMID:25056759

  3. Dynamics of DOT1L localization and H3K79 methylation during meiotic prophase I in mouse spermatocytes

    PubMed Central

    Ontoso, David; Kauppi, Liisa; Keeney, Scott; San-Segundo, Pedro A.

    2013-01-01

    During meiotic prophase I, interactions between maternal and paternal chromosomes, under checkpoint surveillance, establish connections between homologs that promote their accurate distribution to meiotic progeny. In human, faulty meiosis causes aneuploidy resulting in miscarriages and genetic diseases. Meiotic processes occur in the context of chromatin, therefore histone post-translational modifications are expected to play important roles. Here, we report the cytological distribution of the evolutionarily conserved DOT1L methyltransferase and the different H3K79 methylation states resulting from its activity (mono-, di- and tri-methylation; H3K79me1, me2 and me3, respectively) during meiotic prophase I in mouse spermatocytes. In the wild type, whereas low amounts of H3K79me1 are rather uniformly present throughout prophase I, levels of DOT1L, H3K79me2 and H3K79me3 exhibit a notable increase from pachynema onwards, but with differential subnuclear distribution patterns. The heterochromatic centromeric regions and the sex body are enriched for H3K79me3. In contrast, H3K79me2 is present all over the chromatin, but is largely excluded from the sex body despite the accumulation of DOT1L. In meiosis-defective mouse mutants, the increase of DOT1L and H3K79me is blocked at the same stage where meiosis is arrested. H3K79me patterns, combined with the cytological analysis of the H3.3, γH2AX, macroH2A and H2A.Z histone variants, are consistent with a differential role for these epigenetic marks in male mouse meiotic prophase I. We propose that H3K79me2 is related to transcriptional reactivation on autosomes during pachynema, whereas H3K79me3 may contribute to the maintenance of repressive chromatin at centromeric regions and the sex body. PMID:24105599

  4. L1/L2 Differences in the Acquisition of Form-Meaning Pairings in a Second Language

    ERIC Educational Resources Information Center

    McManus, Kevin

    2015-01-01

    This paper examines the impact of L1/L2 form-meaning differences in the domain of aspect to investigate whether L2 learners are able to acquire properties of the L2 that are different from the L1. Oral data were collected from English- and German-speaking university learners of French L2 (n = 75) at two different levels of proficiency. The results…

  5. A natural product inspired tetrahydropyran collection yields mitosis modulators that synergistically target CSE1L and tubulin.

    PubMed

    Voigt, Tobias; Gerding-Reimers, Claas; Ngoc Tran, Tuyen Thi; Bergmann, Sabrina; Lachance, Hugo; Schölermann, Beate; Brockmeyer, Andreas; Janning, Petra; Ziegler, Slava; Waldmann, Herbert

    2013-01-01

    A Prins cyclization between a polymer-bound aldehyde and a homoallylic alcohol served as the key step in the synthesis of tetrahydropyran derivatives. A phenotypic screen led to the identification of compounds that inhibit mitosis (as seen by the accumulation of round cells with condensed DNA and membrane blebs). These compounds were termed tubulexins as they target the CSE1L protein and the vinca alkaloid binding site of tubulin. PMID:23080551

  6. SEL1L SNP rs12435998, a predictor of glioblastoma survival and response to radio-chemotherapy

    PubMed Central

    Storaci, Alessandra Maria; Annovazzi, Laura; Cassoni, Paola; Melcarne, Antonio; De Blasio, Pasquale; Schiffer, Davide; Biunno, Ida

    2015-01-01

    The suppressor of Lin-12-like (C. elegans) (SEL1L) is involved in the endoplasmic reticulum (ER)-associated degradation pathway, malignant transformation and stem cells. In 412 formalin-fixed and paraffin-embedded brain tumors and 39 Glioblastoma multiforme (GBM) cell lines, we determined the frequency of five SEL1L single nucleotide genetic variants with regulatory and coding functions by a SNaPShot™ assay. We tested their possible association with brain tumor risk, prognosis and therapy. We studied the in vitro cytotoxicity of valproic acid (VPA), temozolomide (TMZ), doxorubicin (DOX) and paclitaxel (PTX), alone or in combination, on 11 GBM cell lines, with respect to the SNP rs12435998 genotype. The SNP rs12435998 was prevalent in anaplastic and malignant gliomas, and in meningiomas of all histologic grades, but unrelated to brain tumor risks. In GBM patients, the SNP rs12435998 was associated with prolonged overall survival (OS) and better response to TMZ-based radio-chemotherapy. GBM stem cells with this SNP showed lower levels of SEL1L expression and enhanced sensitivity to VPA. PMID:25948789

  7. Increased cellular apoptosis susceptibility (CSE1L/CAS) protein expression promotes protrusion extension and enhances migration of MCF-7 breast cancer cells

    SciTech Connect

    Tai, Cheng-Jeng; Shen, Shing-Chuan; Lee, Woan-Ruoh; Liao, Ching-Fong; Deng, Win-Ping; Chiou, Hung-Yi; Hsieh, Cheng-I; Tung, Jai-Nien; Chen, Ching-Shyang; Chiou, Jeng-Fong; Li, Li-Tzu; Lin, Chuang-Yu; Hsu, Chung-Huei; Jiang, Ming-Chung

    2010-10-15

    Microtubules are part of cell structures that play a role in regulating the migration of cancer cells. The cellular apoptosis susceptibility (CSE1L/CAS) protein is a microtubule-associated protein that is highly expressed in cancer. We report here that CSE1L regulates the association of {alpha}-tubulin with {beta}-tubulin and promotes the migration of MCF-7 breast cancer cells. CSE1L was associated with {alpha}-tubulin and {beta}-tubulin in GST (glutathione S-transferase) pull-down and immunoprecipitation assays. CSE1L-GFP (green fluorescence protein) fusion protein experiments showed that the N-terminal of CSE1L interacted with microtubules. Increased CSE1L expression resulted in decreased tyrosine phosphorylation of {alpha}-tubulin and {beta}-tubulin, increased {alpha}-tubulin and {beta}-tubulin association, and enhanced assembly of microtubules. Cell protrusions or pseudopodia are temporary extensions of the plasma membrane and are implicated in cancer cell migration and invasion. Increased CSE1L expression increased the extension of MCF-7 cell protrusions. In vitro migration assay showed that enhanced CSE1L expression increased the migration of MCF-7 cells. Our results indicate that CSE1L plays a role in regulating the extension of cell protrusions and promotes the migration of cancer cells.

  8. Liquid atomization

    SciTech Connect

    Walzel, P. )

    1993-01-01

    A systematic review of different liquid atomizers is presented, accompanied by a discussion of various mechanisms of droplet formation in a gas atmosphere as a function of the liquid flow-regime and the geometry of the atomizer. Equations are presented for the calculation of the mean droplet-diameter. In many applications, details of the droplet size distribution are, also, important, e.g., approximate values of the breadth of the droplet formation are given. The efficiency of utilization of mechanical energy in droplet formation is indicated for the different types of atomizers. Atomization is used, in particular, for the following purposes: (1) atomization of fuels; (2) making granular products; (3) carrying out mass-transfer operations; and (4) coating of surfaces.

  9. Liquid electrode

    DOEpatents

    Ekechukwu, Amy A.

    1994-01-01

    A dropping electrolyte electrode for use in electrochemical analysis of non-polar sample solutions, such as benzene or cyclohexane. The liquid electrode, preferably an aqueous salt solution immiscible in the sample solution, is introduced into the solution in dropwise fashion from a capillary. The electrolyte is introduced at a known rate, thus, the droplets each have the same volume and surface area. The electrode is used in making standard electrochemical measurements in order to determine properties of non-polar sample solutions.

  10. Epigenetic modulation of intestinal cholesterol transporter Niemann-Pick C1-like 1 (NPC1L1) gene expression by DNA methylation.

    PubMed

    Malhotra, Pooja; Soni, Vinay; Kumar, Anoop; Anbazhagan, Arivarasu N; Dudeja, Amish; Saksena, Seema; Gill, Ravinder K; Dudeja, Pradeep K; Alrefai, Waddah A

    2014-08-15

    Intestinal NPC1L1 transporter is essential for cholesterol absorption and the maintenance of cholesterol homeostasis in the body. NPC1L1 is differentially expressed along the gastrointestinal tract with very low levels in the colon as compared with the small intestine. This study was undertaken to examine whether DNA methylation was responsible for segment-specific expression of NPC1L1. Treatment of mice with 5-azacytidine (i.p.) resulted in a significant dose-dependent increase in NPC1L1 mRNA expression in the colon. The lack of expression of NPC1L1 in the normal colon was associated with high levels of methylation in the area flanking the 3-kb fragment upstream of the initiation site of the mouse NPC1L1 gene in mouse colon as analyzed by EpiTYPER® MassARRAY®. The high level of methylation in the colon was observed in specific CpG dinucleotides and was significantly decreased in response to 5-azacytidine. Similar to mouse NPC1L1, 5-azacytidine treatment also increased the level of human NPC1L1 mRNA expression in the intestinal HuTu-80 cell line in a dose- and time-dependent manner. Silencing the expression of DNA methyltransferase DNMT1, -2, -3A, and -3B alone by siRNA did not affect NPC1L1 expression in HuTu-80 cells. However, the simultaneous attenuation of DNMT1 and -3B expression caused a significant increase in NPC1L1 mRNA expression as compared with control. Also, in vitro methylation of the human NPC1L1 promoter significantly decreased NPC1L1 promoter activity in human intestinal Caco2 cells. In conclusion, our data demonstrated for the first time that DNA methylation in the promoter region of the NPC1L1 gene appears to be a major mechanism underlying differential expression of NPC1L1 along the length of the gastrointestinal tract. PMID:24904062

  11. Receptor-like cytoplasmic kinase MARIS functions downstream of CrRLK1L-dependent signaling during tip growth.

    PubMed

    Boisson-Dernier, Aurélien; Franck, Christina Maria; Lituiev, Dmytro S; Grossniklaus, Ueli

    2015-09-29

    Growing plant cells need to rigorously coordinate external signals with internal processes. For instance, the maintenance of cell wall (CW) integrity requires the coordination of CW sensing with CW remodeling and biosynthesis to avoid growth arrest or integrity loss. Despite the involvement of receptor-like kinases (RLKs) of the Catharanthus roseus RLK1-like (CrRLK1L) subfamily and the reactive oxygen species-producing NADPH oxidases, it remains largely unknown how this coordination is achieved. ANXUR1 (ANX1) and ANX2, two redundant members of the CrRLK1L subfamily, are required for tip growth of the pollen tube (PT), and their closest homolog, FERONIA, controls root-hair tip growth. Previously, we showed that ANX1 overexpression mildly inhibits PT growth by oversecretion of CW material, whereas pollen tubes of anx1 anx2 double mutants burst spontaneously after germination. Here, we report the identification of suppressor mutants with improved fertility caused by the rescue of anx1 anx2 pollen tube bursting. Mapping of one these mutants revealed an R240C nonsynonymous substitution in the activation loop of a receptor-like cytoplasmic kinase (RLCK), which we named MARIS (MRI). We show that MRI is a plasma membrane-localized member of the RLCK-VIII subfamily and is preferentially expressed in both PTs and root hairs. Interestingly, mri-knockout mutants display spontaneous PT and root-hair bursting. Moreover, expression of the MRI(R240C) mutant, but not its wild-type form, partially rescues the bursting phenotypes of anx1 anx2 PTs and fer root hairs but strongly inhibits wild-type tip growth. Thus, our findings identify a novel positive component of the CrRLK1L-dependent signaling cascade that coordinates CW integrity and tip growth. PMID:26378127

  12. Receptor-like cytoplasmic kinase MARIS functions downstream of CrRLK1L-dependent signaling during tip growth

    PubMed Central

    Boisson-Dernier, Aurélien; Franck, Christina Maria; Lituiev, Dmytro S.; Grossniklaus, Ueli

    2015-01-01

    Growing plant cells need to rigorously coordinate external signals with internal processes. For instance, the maintenance of cell wall (CW) integrity requires the coordination of CW sensing with CW remodeling and biosynthesis to avoid growth arrest or integrity loss. Despite the involvement of receptor-like kinases (RLKs) of the Catharanthus roseus RLK1-like (CrRLK1L) subfamily and the reactive oxygen species-producing NADPH oxidases, it remains largely unknown how this coordination is achieved. ANXUR1 (ANX1) and ANX2, two redundant members of the CrRLK1L subfamily, are required for tip growth of the pollen tube (PT), and their closest homolog, FERONIA, controls root-hair tip growth. Previously, we showed that ANX1 overexpression mildly inhibits PT growth by oversecretion of CW material, whereas pollen tubes of anx1 anx2 double mutants burst spontaneously after germination. Here, we report the identification of suppressor mutants with improved fertility caused by the rescue of anx1 anx2 pollen tube bursting. Mapping of one these mutants revealed an R240C nonsynonymous substitution in the activation loop of a receptor-like cytoplasmic kinase (RLCK), which we named MARIS (MRI). We show that MRI is a plasma membrane-localized member of the RLCK-VIII subfamily and is preferentially expressed in both PTs and root hairs. Interestingly, mri-knockout mutants display spontaneous PT and root-hair bursting. Moreover, expression of the MRIR240C mutant, but not its wild-type form, partially rescues the bursting phenotypes of anx1 anx2 PTs and fer root hairs but strongly inhibits wild-type tip growth. Thus, our findings identify a novel positive component of the CrRLK1L-dependent signaling cascade that coordinates CW integrity and tip growth. PMID:26378127

  13. Inhibiting intestinal NPC1L1 activity prevents diet-induced increase in biliary cholesterol in Golden Syrian hamsters.

    PubMed

    Valasek, Mark A; Repa, Joyce J; Quan, Gang; Dietschy, John M; Turley, Stephen D

    2008-10-01

    Niemann-Pick C1-like 1 (NPC1L1) facilitates the uptake of sterols into the enterocyte and is the target of the novel cholesterol absorption inhibitor, ezetimibe. These studies used the Golden Syrian hamster as a model to delineate the changes in the relative mRNA expression of NPC1L1 and other proteins that regulate sterol homeostasis in the enterocyte during and following cessation of ezetimibe treatment and also to address the clinically important question of whether the marked inhibition of cholesterol absorption alters biliary lipid composition. In hamsters fed a low-cholesterol, low-fat basal diet, the abundance of mRNA for NPC1L1 in the small intestine far exceeded that in other regions of the gastrointestinal tract, liver, and gallbladder. In the first study, female hamsters were fed the basal diet containing ezetimibe at doses up to 2.0 mg.day(-1).kg body wt(-1). At this dose, cholesterol absorption fell by 82%, fecal neutral sterol excretion increased by 5.3-fold, and hepatic and intestinal cholesterol synthesis increased more than twofold, but there were no significant changes in either fecal bile acid excretion or biliary lipid composition. The ezetimibe-induced changes in intestinal cholesterol handling were reversed when treatment was withdrawn. In a second study, male hamsters were given a diet enriched in cholesterol and safflower oil without or with ezetimibe. The lipid-rich diet raised the absolute and relative cholesterol levels in bile more than fourfold. This increase was largely prevented by ezetimibe. These data are consistent with the recent finding that ezetimibe treatment significantly reduced biliary cholesterol saturation in patients with gallstones. PMID:18718997

  14. Structure-Activity Analysis of the Dermcidin-derived Peptide DCD-1L, an Anionic Antimicrobial Peptide Present in Human Sweat*

    PubMed Central

    Paulmann, Maren; Arnold, Thomas; Linke, Dirk; Özdirekcan, Suat; Kopp, Annika; Gutsmann, Thomas; Kalbacher, Hubert; Wanke, Ines; Schuenemann, Verena J.; Habeck, Michael; Bürck, Jochen; Ulrich, Anne S.; Schittek, Birgit

    2012-01-01

    Dermcidin encodes the anionic amphiphilic peptide DCD-1L, which displays a broad spectrum of antimicrobial activity under conditions resembling those in human sweat. Here, we have investigated its mode of antimicrobial activity. We found that DCD-1L interacts preferentially with negatively charged bacterial phospholipids with a helix axis that is aligned flat on a lipid bilayer surface. Upon interaction with lipid bilayers DCD-1L forms oligomeric complexes that are stabilized by Zn2+. DCD-1L is able to form ion channels in the bacterial membrane, and we propose that Zn2+-induced self-assembly of DCD-1L upon interaction with bacterial lipid bilayers is a prerequisite for ion channel formation. These data allow us for the first time to propose a molecular model for the antimicrobial mechanism of a naturally processed human anionic peptide that is active under the harsh conditions present in human sweat. PMID:22262861

  15. Pump for Saturated Liquids

    NASA Technical Reports Server (NTRS)

    Elliott, D. G.

    1986-01-01

    Boiling liquids pumped by device based on proven components. Expanding saturated liquid in nozzle and diverting its phases along separate paths in liquid/vapor separator raises pressure of liquid. Liquid cooled in process. Pump makes it unnecessary to pressurize cryogenic liquids in order to pump them. Problems of introducing noncondensable pressurizing gas avoided.

  16. JNK1/2 regulate Bid by direct phosphorylation at Thr59 in response to ALDH1L1

    PubMed Central

    Prakasam, A; Ghose, S; Oleinik, N V; Bethard, J R; Peterson, Y K; Krupenko, N I; Krupenko, S A

    2014-01-01

    BH3 interacting-domain death agonist (Bid) is a BH3-only pro-apoptotic member of the Bcl-2 family of proteins. Its function in apoptosis is associated with the proteolytic cleavage to the truncated form tBid, mainly by caspase-8. tBid translocates to mitochondria and assists Bax and Bak in induction of apoptosis. c-Jun N-terminal kinase (JNK)-dependent alternative processing of Bid to jBid was also reported. We have previously shown that the folate stress enzyme 10-formyltetrahydrofolate dehydrogenase (ALDH1L1) activates JNK1 and JNK2 in cancer cells as a pro-apoptotic response. Here we report that in PC-3 prostate cancer cells, JNK1/2 phosphorylate Bid at Thr59 within the caspase cleavage site in response to ALDH1L1. In vitro, all three JNK isoforms, JNK 1–3, phosphorylated Thr59 of Bid with JNK1 being the least active. Thr59 phosphorylation protected Bid from cleavage by caspase-8, resulting in strong accumulation of the full-length protein and its translocation to mitochondria. Interestingly, although we did not observe jBid in response to ALDH1L1 in PC-3 cells, transient expression of Bid mutants lacking the caspase-8 cleavage site resulted in strong accumulation of jBid. Of note, a T59D mutant mimicking constitutive phosphorylation revealed more profound cleavage of Bid to jBid. JNK-driven Bid accumulation had a pro-apoptotic effect in our study: small interfering RNA silencing of either JNK1/2 or Bid prevented Bid phosphorylation and accumulation, and rescued ALDH1L1-expressing cells. As full-length Bid is a weaker apoptogen than tBid, we propose that the phosphorylation of Bid by JNKs, followed by the accumulation of the full-length protein, delays attainment of apoptosis, and allows the cell to evaluate the stress and make a decision regarding the response strategy. This mechanism perhaps can be modified by the alternative cleavage of phospho-T59 Bid to jBid at some conditions. PMID:25077544

  17. Liquid electrode

    DOEpatents

    Ekechukwu, A.A.

    1994-07-05

    A dropping electrolyte electrode is described for use in electrochemical analysis of non-polar sample solutions, such as benzene or cyclohexane. The liquid electrode, preferably an aqueous salt solution immiscible in the sample solution, is introduced into the solution in dropwise fashion from a capillary. The electrolyte is introduced at a known rate, thus, the droplets each have the same volume and surface area. The electrode is used in making standard electrochemical measurements in order to determine properties of non-polar sample solutions. 2 figures.

  18. Functional evaluation of TERT-CLPTM1L genetic variants associated with susceptibility of papillary thyroid carcinoma

    PubMed Central

    Ge, Minghua; Shi, Meng; An, Changming; Yang, Wenjun; Nie, Xilin; Zhang, Jian; Lv, Zheng; Li, Jinliang; Zhou, Liqing; Du, Zhongli; Yang, Ming

    2016-01-01

    TERT is the catalytic subunit of telomerase which plays an essential part in cellular immortality by maintaining telomere integrity. TERT is commonly over-expressed in human malignancies, indicating its key role in cell transformation. The chromosome 5p15.33 TERT-CLPTM1L region has been associated with susceptibility of multiple cancers via a genome-wide association approach. However, the involvement of this locus in papillary thyroid carcinoma (PTC) etiology is still largely unknown. We analyzed 15 haplotype-tagging single nucleotide polymorphisms (htSNPs) of the TERT-CLPTM1L region in a two stage case-control design. After genotyping 2300 PTC patients and frequency-matched 2300 unaffected controls, we found that TERT rs2736100 genetic variant is significantly associated with elevated PTC risk. Ex vivo reporter gene assays indicated that the PTC susceptibility rs2736100 polymorphism locating in a potential TERT intronic enhancer has a genotype-specific effect on TERT expression. Correlations between rs2736100 genotypes and tissue-specific TERT expression supported the regulatory function of this genetic variant in vivo. Our data demonstrated that the functional TERT rs2736100 SNP as a novel genetic component of PTC etiology. This study, together with recent studies in other cancers, unequivocally establishes an essential role of TERT in cancers. PMID:27185198

  19. Biallelic expression of Tssc4, Nap1l4, Phlda2 and Osbpl5 in adult cattle.

    PubMed

    Wang, Mengnan; Li, Dongjie; Zhang, Mingyue; Yang, Wenzhi; Wu, Guojiang; Cui, Yali; Li, Shijie

    2015-09-01

    Genomic imprinting of the Cdkn1c/Kcnq1ot1 region shows lack of conservation between human and mouse. This region has been reported to be associated with Beckwith-Wiedemann syndrome (BWS) and cancer. To increase our understanding of imprinted genes in bovine Cdkn1c/Kcnq1ot1 imprinting cluster, we assessed the imprinting status of four cattle genes (Tssc4,Nap1l4, Phlda2 and Osbpl5) in seven types of tissues: heart, liver, spleen, lung, kidney, skeletal muscle and subcutaneous fat using polymorphism-based sequencing approach. It was found that all the four genes showed biallelic expression in tissues in which transcripts were detected. Nap1l4 and Tssc4 were detected in all examined tissues, while the expression of Phlda2 and Osbpl5 was tissue-specific. Phlda2 was not detected in heart and subcutaneous fat, and Osbpl5 was not detected in spleen and skeletal muscle. In addition, identification of species-specific imprinted genes is necessary to understand the evolution of genomic imprinting and to elucidate mechanisms leading to allele-specific expression. PMID:26440077

  20. Caenorhabditis elegans ciliary protein NPHP-8, the homologue of human RPGRIP1L, is required for ciliogenesis and chemosensation.

    PubMed

    Liu, Lin; Zhang, Mingshu; Xia, Zhiping; Xu, Pingyong; Chen, Liangyi; Xu, Tao

    2011-07-01

    Nephronophthisis (NPHP) is the most frequent genetic cause of end-stage renal failure in children and young adults. NPHP8/RPGRIP1L is a novel ciliary gene that, when mutated, in addition to causing NPHP, also causes Joubert syndrome (JBTS) and Meckel syndrome (MKS). The exact function of NPHP8 and how defects in NPHP8 lead to human diseases are poorly understood. Here, we studied the Caenorhabditis elegans homolog nphp-8 (C09G5.8) and explored the possible function of NPHP-8 in ciliated sensory neurons. We determined the gene structure of nphp-8 through rapid amplification of cDNA ends (RACE) analysis and discovered an X-box motif that had been previously overlooked. Moreover, NPHP-8 co-localized with NPHP-4 at the transition zone at the base of cilia. Mutation of nphp-8 led to abnormal dye filling (Dyf) and shorter cilia lengths in a subset of ciliary neurons. In addition, chemotaxis to several volatile attractants was significantly impaired in nphp-8 mutants. Our data suggest that NPHP-8/RPGRIP1L plays an important role in cilia formation and cilia-mediated chemosensation in a cell type-specific manner. PMID:21689635

  1. Crystal Structure of the CTP1L Endolysin Reveals How Its Activity Is Regulated by a Secondary Translation Product*

    PubMed Central

    Dunne, Matthew; Leicht, Stefan; Krichel, Boris; Thompson, Andrew; Gómez-Torres, Natalia; Garde, Sonia; Narbad, Arjan; Mayer, Melinda J.

    2016-01-01

    Bacteriophages produce endolysins, which lyse the bacterial host cell to release newly produced virions. The timing of lysis is regulated and is thought to involve the activation of a molecular switch. We present a crystal structure of the activated endolysin CTP1L that targets Clostridium tyrobutyricum, consisting of a complex between the full-length protein and an N-terminally truncated C-terminal cell wall binding domain (CBD). The truncated CBD is produced through an internal translation start site within the endolysin gene. Mutants affecting the internal translation site change the oligomeric state of the endolysin and reduce lytic activity. The activity can be modulated by reconstitution of the full-length endolysin-CBD complex with free CBD. The same oligomerization mechanism applies to the CD27L endolysin that targets Clostridium difficile and the CS74L endolysin that targets Clostridium sporogenes. When the CTP1L endolysin gene is introduced into the commensal bacterium Lactococcus lactis, the truncated CBD is also produced, showing that the alternative start codon can be used in other bacterial species. The identification of a translational switch affecting oligomerization presented here has implications for the design of effective endolysins for the treatment of bacterial infections. PMID:26683375

  2. Evidence for a pathogenic role of BRCA1 L1705P and W1837X germ-line mutations.

    PubMed

    Sokolenko, Anna P; Volkov, Nikita M; Preobrazhenskaya, Elena V; Suspitsin, Evgeny N; Garifullina, Aigul R; Ivantsov, Alexandr V; Togo, Alexandr V; Imyanitov, Evgeny N

    2016-05-01

    BRCA1 L1705P (c.5114T>C) has been classified in the NCBI SNP database as the variant with uncertain significance and is absent in major BRCA1 databases. BRCA1 W1837X (c.5511G>A) results in a loss of only last 27 residues of BRCA1 protein, thus its pathogenic role still requires a confirmation. This report describes two breast cancer (BC) patients carrying BRCA1 L1705P and W1837X germ-line mutations, respectively. Significant evidence for BC-predisposing impact of the mentioned mutations have been obtained: (1) both index cases presented with the triple-negative receptor status of BC disease; (2) complete segregation with BRCA1-related cancers was observed in the families of these patients; (3) somatic loss of the remaining (wild-type) BRCA1 allele was detected in tumor tissues of the affected women. The results of this study have to be taken into account while providing genetic counseling to cancer patients and while considering the use of BRCA1-specific therapeutic compounds for BC treatment. PMID:26951538

  3. Functional evaluation of TERT-CLPTM1L genetic variants associated with susceptibility of papillary thyroid carcinoma.

    PubMed

    Ge, Minghua; Shi, Meng; An, Changming; Yang, Wenjun; Nie, Xilin; Zhang, Jian; Lv, Zheng; Li, Jinliang; Zhou, Liqing; Du, Zhongli; Yang, Ming

    2016-01-01

    TERT is the catalytic subunit of telomerase which plays an essential part in cellular immortality by maintaining telomere integrity. TERT is commonly over-expressed in human malignancies, indicating its key role in cell transformation. The chromosome 5p15.33 TERT-CLPTM1L region has been associated with susceptibility of multiple cancers via a genome-wide association approach. However, the involvement of this locus in papillary thyroid carcinoma (PTC) etiology is still largely unknown. We analyzed 15 haplotype-tagging single nucleotide polymorphisms (htSNPs) of the TERT-CLPTM1L region in a two stage case-control design. After genotyping 2300 PTC patients and frequency-matched 2300 unaffected controls, we found that TERT rs2736100 genetic variant is significantly associated with elevated PTC risk. Ex vivo reporter gene assays indicated that the PTC susceptibility rs2736100 polymorphism locating in a potential TERT intronic enhancer has a genotype-specific effect on TERT expression. Correlations between rs2736100 genotypes and tissue-specific TERT expression supported the regulatory function of this genetic variant in vivo. Our data demonstrated that the functional TERT rs2736100 SNP as a novel genetic component of PTC etiology. This study, together with recent studies in other cancers, unequivocally establishes an essential role of TERT in cancers. PMID:27185198

  4. Vertical guidance performance analysis of the L1-L5 dual-frequency GPS/WAAS user avionics sensor.

    PubMed

    Jan, Shau-Shiun

    2010-01-01

    This paper investigates the potential vertical guidance performance of global positioning system (GPS)/wide area augmentation system (WAAS) user avionics sensor when the modernized GPS and Galileo are available. This paper will first investigate the airborne receiver code noise and multipath (CNMP) confidence (σair). The σair will be the dominant factor in the availability analysis of an L1-L5 dual-frequency GPS/WAAS user avionics sensor. This paper uses the MATLAB Algorithm Availability Simulation Tool (MAAST) to determine the required values for the σair, so that an L1-L5 dual-frequency GPS/WAAS user avionics sensor can meet the vertical guidance requirements of APproach with Vertical guidance (APV) II and CATegory (CAT) I over conterminous United States (CONUS). A modified MAAST that includes the Galileo satellite constellation is used to determine under what user configurations WAAS could be an APV II system or a CAT I system over CONUS. Furthermore, this paper examines the combinations of possible improvements in signal models and the addition of Galileo to determine if GPS/WAAS user avionics sensor could achieve 10 m Vertical Alert Limit (VAL) within the service volume. Finally, this paper presents the future vertical guidance performance of GPS user avionics sensor for the United States' WAAS, Japanese MTSAT-based satellite augmentation system (MSAS) and European geostationary navigation overlay service (EGNOS). PMID:22319263

  5. Tetrahymena macronuclear genome mapping: colinearity Of macronuclear coassortment groups and the micronuclear map on chromosome 1l.

    PubMed Central

    Wickert, S; Nangle, L; Shevel, S; Orias, E

    2000-01-01

    The genetics of the ciliate Tetrahymena thermophila are richer than for most other eukaryotic cells, because Tetrahymena possesses two genomes: a germline (micronuclear) genome that follows a Mendelian model of genetic transmission and a somatic (macronuclear) genome, derived from the micronuclear genome by fragmentation, which follows a different genetic transmission model called phenotypic assortment. While genetic markers in the micronucleus fall into classical linkage groups under meiotic recombination and segregation, the same markers in the macronucleus fall into coassortment groups (CAGs) under phenotypic assortment by the random distribution of MAC chromosome pieces. We set out to determine whether genomic mapping in the macronucleus by genetic means is feasible. To investigate the relationship between the micronuclear map and coassortment groups, we systematically placed into CAGs all of the markers lying on chromosome 1L that are also found in the macronucleus. Sixteen CAGs were identified, 7 of which contain at least two loci. We have concluded that CAGs represent a fundamental genetic feature of the MAC. The MIC and MAC maps on 1L are colinear; that is, CAGs consist exclusively of markers that map to a continuous segment in a given region of the micronuclear map, with no intervening markers from other CAGs. These findings provide a solid foundation for exploiting the MAC chromosome pieces to build a physical map of the Tetrahymena genome. PMID:10757760

  6. 49 CFR 173.22 - Shipper's responsibility.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... in 49 CFR 385.403 unless that motor carrier holds a safety permit issued by the Federal Motor Carrier Safety Administration. (c) Prior to each shipment of fissile radioactive materials, and Type B or highway... Transportation Other Regulations Relating to Transportation PIPELINE AND HAZARDOUS MATERIALS...

  7. Shippers face new, rigid hazmat rules

    SciTech Connect

    Alvarez, L.

    1994-10-01

    New regulations on packaging being adopted in the United States and internationally should significantly reduce the incidence of hazardous materials accidents and improve shipping safety. Packaging manufacturers, companies that dispatch hazardous materials, transporters, forwarders, and every person that handles packages along the transportation chain will be affected by the new regulations.

  8. 49 CFR 173.22 - Shipper's responsibility.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... motor carrier any hazardous material specifiedin 49 CFR 385.403 unless that motor carrier holds a safety... Federal Register citations affecting § 173.22, see the List of CFR Sections Affected, which appears in...

  9. 49 CFR 172.204 - Shipper's certification.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... affecting § 172.204, see the List of CFR Sections Affected, which appears in the Finding Aids section of the... medical diagnosis or treatment. (d) Signature. The certifications required by paragraph (a) or (c) of...

  10. 49 CFR 172.204 - Shipper's certification.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... affecting § 172.204, see the List of CFR Sections Affected, which appears in the Finding Aids section of the... medical diagnosis or treatment. (d) Signature. The certifications required by paragraph (a) or (c) of...

  11. 49 CFR 173.22 - Shipper's responsibility.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... in 49 CFR 385.403 unless that motor carrier holds a safety permit issued by the Federal Motor Carrier... § 173.22, see the List of CFR Sections Affected which appears in the Finding Aids section of the printed... international regulations based on the UN Recommendations (IBR, see § 171.7 of this subchapter), as...

  12. 49 CFR 172.204 - Shipper's certification.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... affecting § 172.204, see the List of CFR Sections Affected, which appears in the Finding Aids section of the... (manually or mechanically) on the shipping paper containing the required shipping description the...) Rail only certifications. For transportation by rail, the shipping paper certification may also...

  13. Railroads and shippers clash over coal dust

    SciTech Connect

    Buchsbaum, L.

    2007-11-15

    In an effort to reduce coal spillage from railcars, mines in the Powder River Basin (PRB) now load coal with a loaf profile but, reportedly, beginning in 2008, Burlington Northern Santa Fe (BNSF) will announce guidelines requiring all PRB coal loads to be sprayed with a chemical surfactant. If this does not fix the problem, greater measures will be taken. At the time of going to press, the details of how this would be implemented and regulated were unresolved. 1 photo.

  14. 49 CFR 172.204 - Shipper's certification.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... citations affecting § 172.204, see the List of CFR Sections Affected which appears in the Finding Aids... certification requirements. Effective October 1, 2006, each person who offers a hazardous material for... transport requirements, including appropriate types of packaging that conform to the packing...

  15. DOT1L cooperates with the c-Myc-p300 complex to epigenetically derepress CDH1 transcription factors in breast cancer progression

    PubMed Central

    Cho, Min-Hyung; Park, Ji-Hye; Choi, Hee-Joo; Park, Mi-Kyung; Won, Hee-Young; Park, Yeon-Ji; Lee, Chang Hoon; Oh, Seung-Hyun; Song, Young-Soo; Kim, Hyun Sung; Oh, Young-Ha; Lee, Jeong-Yeon; Kong, Gu

    2015-01-01

    DOT1L has emerged as an anticancer target for MLL-associated leukaemias; however, its functional role in solid tumours is largely unknown. Here we identify that DOT1L cooperates with c-Myc and p300 acetyltransferase to epigenetically activate epithelial–mesenchymal transition (EMT) regulators in breast cancer progression. DOT1L recognizes SNAIL, ZEB1 and ZEB2 promoters via interacting with the c-Myc-p300 complex and facilitates lysine-79 methylation and acetylation towards histone H3, leading to the dissociation of HDAC1 and DNMT1 in the regions. The upregulation of these EMT regulators by the DOT1L-c-Myc-p300 complex enhances EMT-induced breast cancer stem cell (CSC)-like properties. Furthermore, in vivo orthotopic xenograft models show that DOT1L is required for malignant transformation of breast epithelial cells and breast tumour initiation and metastasis. Clinically, DOT1L expression is associated with poorer survival and aggressiveness of breast cancers. Collectively, we suggest that cooperative effect of DOT1L and c-Myc-p300 is critical for acquisition of aggressive phenotype of breast cancer by promoting EMT/CSC. PMID:26199140

  16. Variola virus F1L is a Bcl-2-like protein that unlike its vaccinia virus counterpart inhibits apoptosis independent of Bim

    PubMed Central

    Marshall, B; Puthalakath, H; Caria, S; Chugh, S; Doerflinger, M; Colman, P M; Kvansakul, M

    2015-01-01

    Subversion of host cell apoptosis is an important survival strategy for viruses to ensure their own proliferation and survival. Certain viruses express proteins homologous in sequence, structure and function to mammalian pro-survival B-cell lymphoma 2 (Bcl-2) proteins, which prevent rapid clearance of infected host cells. In vaccinia virus (VV), the virulence factor F1L was shown to be a potent inhibitor of apoptosis that functions primarily be engaging pro-apoptotic Bim. Variola virus (VAR), the causative agent of smallpox, harbors a homolog of F1L of unknown function. We show that VAR F1L is a potent inhibitor of apoptosis, and unlike all other characterized anti-apoptotic Bcl-2 family members lacks affinity for the Bim Bcl-2 homology 3 (BH3) domain. Instead, VAR F1L engages Bid BH3 as well as Bak and Bax BH3 domains. Unlike its VV homolog, variola F1L only protects against Bax-mediated apoptosis in cellular assays. Crystal structures of variola F1L bound to Bid and Bak BH3 domains reveal that variola F1L forms a domain-swapped Bcl-2 fold, which accommodates Bid and Bak BH3 in the canonical Bcl-2-binding groove, in a manner similar to VV F1L. Despite the observed conservation of structure and sequence, variola F1L inhibits apoptosis using a startlingly different mechanism compared with its VV counterpart. Our results suggest that unlike during VV infection, Bim neutralization may not be required during VAR infection. As molecular determinants for the human-specific tropism of VAR remain essentially unknown, identification of a different mechanism of action and utilization of host factors used by a VAR virulence factor compared with its VV homolog suggest that studying VAR directly may be essential to understand its unique tropism. PMID:25766319

  17. The N Terminus of the Vaccinia Virus Protein F1L Is an Intrinsically Unstructured Region That Is Not Involved in Apoptosis Regulation.

    PubMed

    Caria, Sofia; Marshall, Bevan; Burton, Robyn-Lee; Campbell, Stephanie; Pantaki-Eimany, Delara; Hawkins, Christine J; Barry, Michele; Kvansakul, Marc

    2016-07-01

    Subversion of host cell apoptotic responses is a prominent feature of viral immune evasion strategies to prevent premature clearance of infected cells. Numerous poxviruses encode structural and functional homologs of the Bcl-2 family of proteins, and vaccinia virus harbors antiapoptotic F1L that potently inhibits the mitochondrial apoptotic checkpoint. Recently F1L has been assigned a caspase-9 inhibitory function attributed to an N-terminal α helical region of F1L spanning residues 1-15 (1) preceding the domain-swapped Bcl-2-like domains. Using a reconstituted caspase inhibition assay in yeast we found that unlike AcP35, a well characterized caspase-9 inhibitor from the insect virus Autographa californica multiple nucleopolyhedrovirus, F1L does not prevent caspase-9-mediated yeast cell death. Furthermore, we found that deletion of the F1L N-terminal region does not impede F1L antiapoptotic activity in the context of a viral infection. Solution analysis of the F1L N-terminal regions using small angle x-ray scattering indicates that the region of F1L spanning residues 1-50 located N-terminally from the Bcl-2 fold is an intrinsically unstructured region. We conclude that the N terminus of F1L is not involved in apoptosis inhibition and may act as a regulatory element in other signaling pathways in a manner reminiscent of other unstructured regulatory elements commonly found in mammalian prosurvival Bcl-2 members including Bcl-xL and Mcl-1. PMID:27151220

  18. Mutation in Glycerol-3-Phosphate Dehydrogenase 1–Like Gene (GPD1-L) Decreases Cardiac Na+ Current and Causes Inherited Arrhythmias

    PubMed Central

    London, Barry; Michalec, Michael; Mehdi, Haider; Zhu, Xiaodong; Kerchner, Laurie; Sanyal, Shamarendra; Viswanathan, Prakash C.; Pfahnl, Arnold E.; Shang, Lijuan L.; Madhusudanan, Mohan; Baty, Catherine J.; Lagana, Stephen; Aleong, Ryan; Gutmann, Rebecca; Ackerman, Michael J.; McNamara, Dennis M.; Weiss, Raul; Dudley, Samuel C.

    2011-01-01

    Background Brugada syndrome is a rare, autosomal-dominant, male-predominant form of idiopathic ventricular fibrillation characterized by a right bundle-branch block and ST elevation in the right precordial leads of the surface ECG. Mutations in the cardiac Na+ channel SCN5A on chromosome 3p21 cause ≈20% of the cases of Brugada syndrome; most mutations decrease inward Na+ current, some by preventing trafficking of the channels to the surface membrane. We previously used positional cloning to identify a new locus on chromosome 3p24 in a large family with Brugada syndrome and excluded SCN5A as a candidate gene. Methods and Results We used direct sequencing to identify a mutation (A280V) in a conserved amino acid of the glycerol-3-phosphate dehydrogenase 1–like (GPD1-L) gene. The mutation was present in all affected individuals and absent in >500 control subjects. GPD1-L RNA and protein are abundant in the heart. Compared with wild-type GPD1-L, coexpression of A280V GPD1-L with SCN5A in HEK cells reduced inward Na+ currents by ≈50% (P<0.005). Wild-type GPD1-L localized near the cell surface to a greater extent than A280V GPD1-L. Coexpression of A280V GPD1-L with SCN5A reduced SCN5A cell surface expression by 31±5% (P=0.01). Conclusions GPD1-L is a novel gene that may affect trafficking of the cardiac Na+ channel to the cell surface. A GPD1-L mutation decreases SCN5A surface membrane expression, reduces inward Na+ current, and causes Brugada syndrome. PMID:17967977

  19. Effects of the order of reading text or viewing a film and L1/L2 captions on reading comprehension.

    PubMed

    Chen, Mei-Ling

    2012-08-01

    The purpose of the study was to investigate the effects of order of reading text or viewing a film and L1/L2 captions on participants' comprehension. The participants were college freshmen (N = 236) enrolled in English as a Foreign Language (EFL) courses. In the experiment, there were four conditions: the text was read first, and then the film version viewed with either Chinese or English subtitles, or the film version was viewed first with Chinese or English subtitles. Pre-test and post-test scores on the General English Proficiency Test (GEPT) indicated that the read-first group viewing the film with Chinese subtitles outperformed the read-first and view-first groups viewing the film version with English subtitles. PMID:23033742

  20. Functional analysis of related CrRLK1L receptor-like kinases in pollen tube reception.

    PubMed

    Kessler, Sharon A; Lindner, Heike; Jones, Daniel S; Grossniklaus, Ueli

    2015-01-01

    The Catharanthus roseus Receptor-Like Kinase 1-like (CrRLK1L) family of 17 receptor-like kinases (RLKs) has been implicated in a variety of signaling pathways in Arabidopsis, ranging from pollen tube (PT) reception and tip growth to hormonal responses. The extracellular domains of these RLKs have malectin-like domains predicted to bind carbohydrate moieties. Domain swap analysis showed that the extracellular domains of the three members analyzed (FER, ANX1, HERK1) are not interchangeable, suggesting distinct upstream components, such as ligands and/or co-factors. In contrast, their intercellular domains are functionally equivalent for PT reception, indicating that they have common downstream targets in their signaling pathways. The kinase domain is necessary for FER function, but kinase activity itself is not, indicating that other kinases may be involved in signal transduction during PT reception. PMID:25490905

  1. Functional analysis of related CrRLK1L receptor-like kinases in pollen tube reception

    PubMed Central

    Kessler, Sharon A; Lindner, Heike; Jones, Daniel S; Grossniklaus, Ueli

    2015-01-01

    The Catharanthus roseus Receptor-Like Kinase 1-like (CrRLK1L) family of 17 receptor-like kinases (RLKs) has been implicated in a variety of signaling pathways in Arabidopsis, ranging from pollen tube (PT) reception and tip growth to hormonal responses. The extracellular domains of these RLKs have malectin-like domains predicted to bind carbohydrate moieties. Domain swap analysis showed that the extracellular domains of the three members analyzed (FER, ANX1, HERK1) are not interchangeable, suggesting distinct upstream components, such as ligands and/or co-factors. In contrast, their intercellular domains are functionally equivalent for PT reception, indicating that they have common downstream targets in their signaling pathways. The kinase domain is necessary for FER function, but kinase activity itself is not, indicating that other kinases may be involved in signal transduction during PT reception. PMID:25490905

  2. Exchange spring in A1/L10 FePt composite and its application in magnetic force microscope

    NASA Astrophysics Data System (ADS)

    Li, Guoqing; Zhu, Yanyan; Zhang, Yong; Zhao, Hujun; Zeng, Daofu; Li, Yuhui; Lu, Wei

    2015-02-01

    This paper reported fabrication of FexPt100-x films with (001) epitaxy on MgO(100) substrates. The atomic percentage of Fe was changed within the range of x = 10-85 in order to search the optimal atomic ratio for achieving both high and isotropic-like coercivity. It was found that the Fe60Pt40 film exhibited large coercivities exceeding 5 kOe along both in-plane and out-of-plane directions due to the formation of A1/L10 FePt composite. A penta-domain model for hard/soft/hard exchange spring system was proposed to interpret the anomalous magnetization behaviors observed in Fe60Pt40 sample. By using Fe60Pt40 as the magnetic coating layer on a probe of magnetic force microscope, the flux changes at a linear density of 1000 kfci could be readily observed at a resolution of ˜13 nm.

  3. The CD27L and CTP1L Endolysins Targeting Clostridia Contain a Built-in Trigger and Release Factor

    PubMed Central

    Dunne, Matthew; Mertens, Haydyn D. T.; Garefalaki, Vasiliki; Jeffries, Cy M.; Thompson, Andrew; Lemke, Edward A.; Svergun, Dmitri I.; Mayer, Melinda J.; Narbad, Arjan; Meijers, Rob

    2014-01-01

    The bacteriophage ΦCD27 is capable of lysing Clostridium difficile, a pathogenic bacterium that is a major cause for nosocomial infection. A recombinant CD27L endolysin lyses C. difficile in vitro, and represents a promising alternative as a bactericide. To better understand the lysis mechanism, we have determined the crystal structure of an autoproteolytic fragment of the CD27L endolysin. The structure covers the C-terminal domain of the endolysin, and represents a novel fold that is identified in a number of lysins that target Clostridia bacteria. The structure indicates endolysin cleavage occurs at the stem of the linker connecting the catalytic domain with the C-terminal domain. We also solved the crystal structure of the C-terminal domain of a slow cleaving mutant of the CTP1L endolysin that targets C. tyrobutyricum. Two distinct dimerization modes are observed in the crystal structures for both endolysins, despite a sequence identity of only 22% between the domains. The dimers are validated to be present for the full length protein in solution by right angle light scattering, small angle X-ray scattering and cross-linking experiments using the cross-linking amino acid p-benzoyl-L-phenylalanine (pBpa). Mutagenesis on residues contributing to the dimer interfaces indicates that there is a link between the dimerization modes and the autocleavage mechanism. We show that for the CTP1L endolysin, there is a reduction in lysis efficiency that is proportional to the cleavage efficiency. We propose a model for endolysin triggering, where the extended dimer presents the inactive state, and a switch to the side-by-side dimer triggers the cleavage of the C-terminal domain. This leads to the release of the catalytic portion of the endolysin, enabling the efficient digestion of the bacterial cell wall. PMID:25058163

  4. Functional and radioligand binding characterization of the α1L-adrenoceptor subtype of the human vas deferens.

    PubMed

    Davis, B J; Wiener, M; Chapple, C R; Sellers, D J; Chess-Williams, R

    2015-04-01

    Alpha1 -adrenoceptor antagonists can cause ejaculatory dysfunction as an adverse effect. Contractions of the human vas deferens are mediated via α1A -adrenoceptors, and this study investigated whether the low affinity state of this receptor (α1L -adrenoceptor) is involved in mediating contractions of this tissue. The potency of agonists and the affinity of receptor subtype selective antagonists were determined in functional experiments and in [(3) H]tamsulosin binding experiments to identify the α1 -adrenoceptor subtype population present in the human vas deferens. The α1A -adrenoceptor selective agonist A61603 was a full agonist and was 250-fold more potent than noradrenaline. Prazosin antagonized contractile responses to phenylephrine with a low affinity (pKd = 8.6). Only high concentrations of RS17053 antagonized responses to phenylephrine and yielded a relatively low affinity estimate of 7.0. BMY7378 (α1D -adrenoceptor selective) gave a low affinity estimate (pKd = 6.7), whilst tamsulosin (α1A - and α1D -adrenoceptor selective) had a high affinity (pKd = 9.9). [(3) H]Tamsulosin bound to human vas deferens membranes with a high affinity (pKd = 10.0). Prazosin, RS17053 and BMY7378 competed with [(3) H]tamsulosin with low affinities for a single population of binding sites (pKd values of 8.5, 7.2 and 6.3, respectively). These functional and radioligand binding data indicate that the human vas deferens possesses a homogeneous population of α1 -adrenoceptors which have the pharmacological properties of the putative α1L -adrenoceptor, the same functional receptor previously identified in the human prostate. PMID:25790239

  5. Impact of physicochemical and structural properties on the pharmacokinetics of a series of alpha1L-adrenoceptor antagonists.

    PubMed

    Betts, Alison; Atkinson, Fidelma; Gardner, Iain; Fox, David; Webster, Rob; Beaumont, Kevin; Morgan, Paul

    2007-08-01

    A rational drug discovery process was initiated to design a potent and prostate-selective alpha1(L)-adrenoceptor antagonist with pharmacokinetic properties suitable for once a day administration after oral dosing, for the treatment of benign prostatic hyperplasia. Two series of compounds based on a quinoline or quinazoline template were identified with appropriate pharmacology. A series of high molecular weight cations with high hydrogen-bonding potential had extensive in vivo clearance, despite demonstrating metabolic stability. Studies in the isolated perfused rat liver and fresh rat hepatocytes indicated that active transport protein-mediated hepatobiliary elimination is an efficient clearance process for these compounds. A reduction in molecular weight and hydrogen-bonding potential resulted in a second series of compounds with in vivo hepatic clearance predictable from in vitro metabolic clearance. Initially, lipophilicity was reduced within this second series to reduce metabolic clearance and increase elimination half-life. However, this strategy also resulted in a concomitant reduction in volume of distribution and a negligible effect on prolonging half-life. An alternative strategy was to increase the intrinsic metabolic stability of the molecule by careful structural modifications while maintaining lipophilicity. Replacement of the metabolically vulnerable morpholine side chain resulted in identification of UK-338,003, (N-[2-(4-amino-6,7-dimethoxy-5-pyridin-2-yl-quinazolin-2-yl)-1,2,3,4-tetrahydro-isoquinolin-5-yl]-methanesulfonamide), which fulfilled the objectives of the discovery program with suitable pharmacology (human prostate alpha1(L) pA(2) of 9.2 with 25-fold selectivity over rat aorta alpha1(D)) and sufficiently long elimination half-life in human volunteers (11-17 h) for once a day administration. PMID:17502340

  6. Rs6922269 marker at the MTHFD1L gene predict cardiovascular mortality in males after acute coronary syndrome.

    PubMed

    Hubacek, J A; Staněk, V; Gebauerová, M; Poledne, R; Aschermann, M; Skalická, H; Matoušková, J; Kruger, A; Pěnička, M; Hrabáková, H; Veselka, J; Hájek, P; Lánská, V; Adámková, V; Pitˇha, J

    2015-08-01

    Myocardial infarction (MI) is the leading cause of death in industrialized countries. All the traditional risk factors for MI are responsible for approximately 50% of cases of MI cases. Attention therefore has recently focused on genetic variants that are not associated with conventional risk factors. One of them is the marker rs6922269, which has been suggested as a risk factor for development of MI in Western populations. We analyzed the relationship between rs6922269 variant on MTHFD1L gene and (i) risk of the acute coronary syndrome (ACS) in the Czech population and (ii) mortality in 7 years follow up. Rs6922269 (G>A) variant was analyzed (CR 99.3% for patients and 98.0% for controls) by PCR-RFLP in consecutively examined 1614 men and 503 women with ACS (age below 65 years) and in population-based controls--1191 men and 1368 women (aged up to 65 years). ANOVA and Chi square were used for statistical analysis. The genotype frequencies were almost identical (P=0.87) in the ACS patients and in controls and no differences were observed, if males (P=0.73) and females (P=0.93) were analysed separately. In addition, rs6922269 polymorphism was not associated with the classical risk factors (dyslipidemia, hypertension, obesity, smoking, diabetes) in control population. Cardiovascular mortality was significantly higher in males, carriers of the AA genotype (P<0.001, OR 2.52, 95% CI 1.40-4.55, for AA vs. +G). We conclude, that rs6922269 variant at MTHFD1L gene could be an important prognostic factor for cardiovascular mortality in patients after ACS. PMID:25809277

  7. New small molecule inhibitors of histone methyl transferase DOT1L with a nitrile as a non-traditional replacement for heavy halogen atoms.

    PubMed

    Spurr, Sophie S; Bayle, Elliott D; Yu, Wenyu; Li, Fengling; Tempel, Wolfram; Vedadi, Masoud; Schapira, Matthieu; Fish, Paul V

    2016-09-15

    A number of new nucleoside derivatives are disclosed as inhibitors of DOT1L activity. SARs established that DOT1L inhibition could be achieved through incorporation of polar groups and small heterocycles at the 5-position (5, 6, 12) or by the application of alternative nitrogenous bases (18). Based on these results, CN-SAH (19) was identified as a potent and selective inhibitor of DOT1L activity where the polar 5-nitrile group was shown by crystallography to bind in the hydrophobic pocket of DOT1L. In addition, we show that a polar nitrile group can be used as a non-traditional replacement for heavy halogen atoms. PMID:27485386

  8. Discovery of imatinib-responsive FIP1L1-PDGFRA mutation during refractory acute myeloid leukemia transformation of chronic myelomonocytic leukemia

    PubMed Central

    2014-01-01

    The FIP1L1-PDGFRA rearrangement results in constitutive activation of the tyrosine kinase PDGFRA. Neoplasms harboring this rearrangement are responsive to imatinib mesylate at doses much lower than those recommended for the treatment of chronic myelogenous leukemia. Only a single report has described the identification of FIP1L1-PDGFRA in chronic myelomonocytic leukemia (CMML). Herein, we present a case report of a patient in whom the FIP1L1-PDGFRA was discovered as he evolved from CMML to acute myeloid leukemia (AML). The presence of a dominant neoplastic clone with FIP1L1-PDGFRA rearrangement was suspected on the basis of sudden onset of peripheral and bone marrow eosinophilia and confirmed by fluorescence in situ hybridization and molecular diagnostic tests. Whereas the patient was initially refractory to chemotherapy before the rearrangement was detected, subsequent therapy with imatinib led to complete remission. PMID:24669761

  9. MAD1L1 Arg558His and MAD2L1 Leu84Met interaction with smoking increase the risk of colorectal cancer

    PubMed Central

    Zhong, Rong; Chen, Xiaohua; Chen, Xueqin; Zhu, Beibei; Lou, Jiao; Li, Jiaoyuan; Shen, Na; Yang, Yang; Gong, Yajie; Zhu, Ying; Yuan, Jing; Xia, Xiaoping; Miao, Xiaoping

    2015-01-01

    The spindle assembly checkpoint (SAC) has been established as an important mechanism of driving aneuploidy, which occurs at a high frequency in the colorectal tumorigenesis. Two important components of SAC are MAD1L1 and MAD2L1, which function together in an interactive manner to initiate the checkpoint signal. We hypothesize that genetic variants in the binding domains of MAD1L1 and MAD2L1 may modulate protein structures and eventually contribute to CRC susceptibility. A case-control study including 710 CRC cases and 735 controls was performed to examine MAD1L1 Arg558His and MAD2L1 Leu84Met’s conferring susceptibility to CRC. Cytokinesis-block micronucleus cytome assays were applied to assess the effect of two functional variants on chromosomal instability (CIN). Significant associations with CRC risk were observed for MAD1L1 Arg558His (OR = 1.38,95% CI: 1.09–1.75) and MAD2L1 Leu84Met in a dominant model (OR = 1.48,95% CI: 1.09–2.01). Moreover, significant multiplicative gene-smoking interactions were found in MAD1L1 Arg558His (P = 0.019) and MAD2L184 Leu/Met (P = 0.016) to enhance CRC risk. Additionally, the frequencies of lymphocytic micro-nucleated binucleated cells for MAD1L1 Arg558His polymorphism were significantly different in the exposed group (P = 0.013), but not in the control group. The study emphasized that MAD1L1 Arg558His and MAD2L1 Leu84Met can significantly interact with smoking to enhance CRC risk, and the genetic effects of MAD1L1Arg558His on CIN need to be further clarified in follow-up studies. PMID:26183163

  10. Change of function of the wheat stress-responsive transcriptional repressor TaRAP2.1L by repressor motif modification.

    PubMed

    Amalraj, Amritha; Luang, Sukanya; Kumar, Manoj Yadav; Sornaraj, Pradeep; Eini, Omid; Kovalchuk, Nataliya; Bazanova, Natalia; Li, Yuan; Yang, Nannan; Eliby, Serik; Langridge, Peter; Hrmova, Maria; Lopato, Sergiy

    2016-02-01

    Plants respond to abiotic stresses by changes in gene regulation, including stress-inducible expression of transcriptional activators and repressors. One of the best characterized families of drought-related transcription factors are dehydration-responsive element binding (DREB) proteins, known as C-repeat binding factors (CBF). The wheat DREB/CBF gene TaRAP2.1L was isolated from drought-affected tissues using a dehydration-responsive element (DRE) as bait in a yeast one-hybrid screen. TaRAP2.1L is induced by elevated abscisic acid, drought and cold. A C-terminal ethylene responsive factor-associated amphiphilic repression (EAR) motif, known to be responsible for active repression of target genes, was identified in the TaRAP2.1L protein. It was found that TaRAP2.1L has a unique selectivity of DNA-binding, which differs from that of DREB activators. This binding selectivity remains unchanged in a TaRAP2.1L variant with an inactivated EAR motif (TaRAP2.1Lmut). To study the role of the TaRAP2.1L repressor activity associated with the EAR motif in planta, transgenic wheat overexpressing native or mutated TaRAP2.1L was generated. Overexpression of TaRAP2.1L under constitutive and stress-inducible promoters in transgenic wheat and barley led to dwarfism and decreased frost tolerance. By contrast, constitutive overexpression of the TaRAP2.1Lmut gene had little or no negative influence on wheat development or grain yield. Transgenic lines with the TaRAP2.1Lmut transgene had an enhanced ability to survive frost and drought. The improved stress tolerance is attributed to up-regulation of several stress-related genes known to be downstream genes of DREB/CBF activators. PMID:26150199

  11. Functional Characterization of Nupr1L, A Novel p53-Regulated Isoform of the High-Mobility Group (HMG)-Related Protumoral Protein Nupr1.

    PubMed

    Lopez, Maria Belen; Garcia, Maria Noé; Grasso, Daniel; Bintz, Jennifer; Molejon, Maria Inés; Velez, Gabriel; Lomberk, Gwen; Neira, Jose Luis; Urrutia, Raul; Iovanna, Juan

    2015-12-01

    We have previously demonstrated a crucial role of nuclear protein 1 (NUPR1) in tumor development and progression. In this work, we report the functional characterization of a novel Nupr1-like isoform (NUPR1L) and its functional interaction with the protumoral factor NUPR1. Through the use of primary sequence analysis, threading, and homology-based molecular modeling, as well as expression and immunolocalization, studies reveal that NUPR1L displays properties, which are similar to member of the HMG-like family of chromatin regulators, including its ability to translocate to the cell nucleus and bind to DNA. Analysis of the NUPR1L promoter showed the presence of two p53-response elements at positions -37 and -7, respectively. Experiments using reporter assays combined with site-directed mutagenesis and using cells with controllable p53 expression demonstrate that both of these sequences are responsible for the regulation of NUPR1L expression by p53. Congruently, NUPR1L gene expression is activated in response to DNA damage induced by oxaliplatin treatment or cell cycle arrest induced by serum starvation, two well-validated methods to achieve p53 activation. Interestingly, expression of NUPR1L downregulates the expression of NUPR1, its closely related protumoral isoform, by a mechanism that involves the inhibition of its promoter activity. At the cellular level, overexpression of NUPR1L induces G1 cell cycle arrest and a decrease in their cell viability, an effect that is mediated, at least in part, by downregulating NUPR1 expression. Combined, these experiments constitute the first functional characterization of NUPR1L as a new p53-induced gene, which negatively regulates the protumoral factor NUPR1. PMID:25899918

  12. Embryonic expression of Xenopus SGLT-1L, a novel member of the solute carrier family 5 (SLC5), is confined to tubules of the pronephric kidney.

    PubMed

    Eid, Samer R; Terrettaz, Anne; Nagata, Katsumi; Brändli, André W

    2002-01-01

    Plasma membrane proteins of the solute carrier family 5 (SLC5) are responsible for sodium-coupled uptake of ions, sugars and nutrients in the vertebrate body. Mutations in SLC5 genes are the cause of several inherited human disorders. We have recently reported the cloning and transport properties of SGLT-1L, a Xenopus homologue of the sodium-dependent glucose cotransporter 1 (SGLT-1) [Nagata et al. (1999) Am. J. Physiol. 276: G1251 -G 1259]. Here, we describe the phylogenetic relationship of SGLT-1L with other members of the SLC5 family and characterize its expression during Xenopus embryogenesis and in organ cultures. Sequence comparisons and phylogenetic analyses of all known vertebrate SLC5 sequences indicated that Xenopus SGLT-1L encodes a novel SLC5 member, which shares highest amino acid identity with mammalian ST-1 proteins. Temporal and spatial expression of SGLT-1L during Xenopus embryogenesis was examined by whole mount in situ hybridization. Initiation of SGLT-1L expression occurred in the late tailbud embryo. Remarkably, expression was restricted to the developing pronephric kidney. SGLT-1L was highly expressed in tubular epithelia, but completely absent from the epithelia of the duct. Analysis of growth factor-treated animal caps indicated that expression of SGLT-1L could also be induced in organ cultures. Taken together, our findings indicate that the expression of sodium-dependent solute cotransporter genes in early segments of the excretory system appears to be conserved between pronephric and metanephric kidneys. Furthermore, we establish SGLT-1L as a novel, highly specific molecular marker for pronephric tubule epithelia undergoing maturation and terminal differentiation in Xenopus. PMID:11902681

  13. Transcription Enhancer Factor 3 (TEF3) Mediates the Expression of Down Syndrome Candidate Region 1 Isoform 1 (DSCR1-1L) in Endothelial Cells*

    PubMed Central

    Liu, Xin; Zhao, Dezheng; Qin, Liuliang; Li, Jian; Zeng, Huiyan

    2008-01-01

    The Down syndrome candidate region 1 gene (DSCR1) can be expressed as four isoforms, one of which is the well-studied isoform 4 (DSCR1-4) that is induced by VEGF-A165 to provide a negative feedback loop in the VEGF-A165-induced angiogenesis. We reported previously that another DSCR1 isoform, DSCR1-1L, was also up-regulated by VEGF-A165 in cultured endothelial cells and in several in vivo models of pathological angiogenesis and that different from DSCR1-4, DSCR1-1L overexpression alone induced cultured endothelial cell proliferation and promoted angiogenesis in Matrigel assays. It was reported recently that tumor growth was greatly repressed in DSCR1 knock-out mice. Although DSCR1-4 transcription was primarily regulated by NFAT, the mechanism regulating DSCR1-1L expression was still unknown. We developed human DSCR1-1L promoter-driven luciferase system and found that deletion of a putative conserved M-CAT site located 1426-bp upstream of the translation start site blunted promoter activity. We further showed that knockdown of TEF3, not other members of TEF family inhibited VEGF-A165-induced DSCR1-1L expression. We also demonstrated that TEF3 directly interacted with the putative M-CAT site in the DSCR1-1L promoter in vitro and in vivo. Finally, overexpression of TEF3 isoform 1, not isoform 3, in HUVEC was sufficient to induce DSCR1-1L expression even in the absence of VEGF-A165 stimulation. Taken together, we elucidated a novel function of transcriptional factor TEF3. TEF3 was required for DSCR1-1L expression through binding to the M-CAT site in its promoter and could be an attractive target for anti-angiogenesis therapy. PMID:18840614

  14. Liquid Crystal Devices.

    ERIC Educational Resources Information Center

    Bradshaw, Madeline J.

    1983-01-01

    The nature of liquid crystals and several important liquid crystal devices are described. Ideas for practical experiments to illustrate the properties of liquid crystals and their operation in devices are also described. (Author/JN)

  15. Liquid Crystal Inquiries.

    ERIC Educational Resources Information Center

    Marroum, Renata-Maria

    1996-01-01

    Discusses the properties and classification of liquid crystals. Presents a simple experiment that illustrates the structure of liquid crystals and the differences between the various phases liquid crystals can assume. (JRH)

  16. Structure-activity relationship study of non-steroidal NPC1L1 ligands identified through cell-based assay using pharmacological chaperone effect as a readout.

    PubMed

    Karaki, Fumika; Ohgane, Kenji; Fukuda, Hiromitsu; Nakamura, Masahiko; Dodo, Kosuke; Hashimoto, Yuichi

    2014-07-15

    Niemann-Pick type C1-like 1 (NPC1L1) is an intestinal cholesterol transporter that is known to be the target of the cholesterol absorption inhibitor ezetimibe. We previously discovered steroidal NPC1L1 ligands by using a novel cell-based assay that employs pharmacological chaperone effect as a readout. Those steroid derivatives bound to a site different from both the sterol-binding domain and the ezetimibe-binding site, implying that they may be a novel class of NPC1L1 inhibitors with a distinct mode of action. As an extension of that work, we aimed here to find non-steroidal NPC1L1 ligands, which may be better candidates for clinical application than steroidal ligands, by using the same assay to screen our focused library of ligands for liver X receptor (LXR), a nuclear receptor that recognizes oxysterols as endogenous ligands. Here we describe identification of a novel class of NPC1L1 ligands with a ring-fused quinolinone scaffold, and an analysis of the structure-activity relationships of their derivatives as NPC1L1 ligands. PMID:24906511

  17. Endoplasmic Reticulum Oxidoreductin-1-Like β (ERO1lβ) Regulates Susceptibility to Endoplasmic Reticulum Stress and Is Induced by Insulin Flux in β-Cells

    PubMed Central

    Khoo, Cynthia; Yang, Juxiang; Rajpal, Gautam; Wang, You; Liu, Jiangying; Arvan, Peter

    2011-01-01

    Hyperglycemia increases insulin flux through the endoplasmic reticulum (ER) of pancreatic β-cells, and the unfolded protein response pathway is required to enhance insulin processing. Pancreatic and duodenal homeobox 1 (PDX1), a key pancreatic transcription factor, regulates insulin along with targets involved in insulin processing and secretion. Here we find that PDX1 is a direct transcriptional regulator of ER oxidoreductin-1-like β (Ero1lβ), which maintains the oxidative environment of the ER to facilitate disulfide bond formation. PDX1 deficiency reduced Ero1lβ transcript levels in mouse islets and mouse insulinoma (MIN6) cells; moreover, PDX1 occupied the Ero1lβ promoter in β-cells. ERO1lβ levels were induced by high glucose concentrations and by the reducing agent dithiothreitol, indicating potential roles in adaptation to increased oxidative protein folding load in the β-cell ER. In MIN6 cells, small interfering RNA-mediated silencing of Ero1lβ decreased insulin content and increased susceptibility to ER stress-induced apoptosis. These findings demonstrate roles for the PDX1 target ERO1lβ in maintaining insulin content and regulating cell survival during ER stress. PMID:21540283

  18. Human Vitamin K 2,3-Epoxide Reductase Complex Subunit 1-like 1 (VKORC1L1) Mediates Vitamin K-dependent Intracellular Antioxidant Function*

    PubMed Central

    Westhofen, Philipp; Watzka, Matthias; Marinova, Milka; Hass, Moritz; Kirfel, Gregor; Müller, Jens; Bevans, Carville G.; Müller, Clemens R.; Oldenburg, Johannes

    2011-01-01

    Human vitamin K 2,3-epoxide reductase complex subunit 1-like 1 (VKORC1L1), expressed in HEK 293T cells and localized exclusively to membranes of the endoplasmic reticulum, was found to support both vitamin K 2,3-epoxide reductase (VKOR) and vitamin K reductase enzymatic activities. Michaelis-Menten kinetic parameters for dithiothreitol-driven VKOR activity were: Km (μm) = 4.15 (vitamin K1 epoxide) and 11.24 (vitamin K2 epoxide); Vmax (nmol·mg−1·hr−1) = 2.57 (vitamin K1 epoxide) and 13.46 (vitamin K2 epoxide). Oxidative stress induced by H2O2 applied to cultured cells up-regulated VKORC1L1 expression and VKOR activity. Cell viability under conditions of no induced oxidative stress was increased by the presence of vitamins K1 and K2 but not ubinquinone-10 and was specifically dependent on VKORC1L1 expression. Intracellular reactive oxygen species levels in cells treated with 2,3-dimethoxy-1,4-naphthoquinone were mitigated in a VKORC1L1 expression-dependent manner. Intracellular oxidative damage to membrane intrinsic proteins was inversely dependent on VKORC1L1 expression and the presence of vitamin K1. Taken together, our results suggest that VKORC1L1 is responsible for driving vitamin K-mediated intracellular antioxidation pathways critical to cell survival. PMID:21367861

  19. REDISTRIBUTOR FOR LIQUID-LIQUID EXTRACTION COLUMNS

    DOEpatents

    Bradley, J.G.

    1957-10-29

    An improved baffle plate construction to intimately mix immiscible liquid solvents for solvent extraction processes in a liquid-liquid pulse column is described. To prevent the light and heavy liquids from forming separate continuous homogeneous vertical channels through sections of the column, a baffle having radially placed rectangular louvers with deflection plates opening upon alternate sides of the baffle is placed in the column, normal to the axis. This improvement substantially completely reduces strippiig losses due to poor mixing.

  20. Liquid supercoiling

    NASA Astrophysics Data System (ADS)

    Ribe, Neil; Habibi, Mehdi; Hosseini, Hossein; Hassan Khatami, Mohammad

    2011-11-01

    Supercoiling is defined as the large-scale secondary coiling of a slender body that is already coiled at a smaller scale (e.g., telephone cords and DNA strands). We demonstrate experimentally a novel fluid-mechanical form of supercoiling that occurs in the context of the familiar ``liquid rope coiling'' instability of a thin thread of viscous fluid falling onto a rigid surface. Under appropriate conditions, the coiling instability generates a tall pile of coils in the form of a hollow cylindrical column, which in turn becomes unstable to a secondary coiling instability with a frequency ~ 10 % of the primary one. To place this phenomenon in a broader context, we determine experimentally the phase diagram for the different possible behaviors of the thread (stagnation flow, simple coiling, rotatory folding, periodic column collapse, supercoiling) in the space of the fluid viscosity, the flow rate, and the fall height. We formulate a mathematical model for supercoiling by combining a thin-shell description of the column wall with a slender-thread description of the column as a whole. This leads to a set of coupled ordinary differential equations in one space dimension (the arclength along the axis of the coiling column) that we solve numerically using a continuation method. A comparison of the predicted and observed frequencies of secondary coiling will be shown.

  1. Liquid annulus

    NASA Technical Reports Server (NTRS)

    Ludewig, Hans

    1991-01-01

    It is shown that the specific impulse varies with the square root of the temperature and inversely with the square root of the molecular weight of the propellant. Typical values for specific impulse corresponding to various rocket concepts are shown. The Liquid Annulus core concept consists of a fuel element which will be arranged in a moderator block. The advantages as seen for the system are: high specific impulse; structural material will all run at low temperature; and lower fission product inventory because of evaporation. It is felt that this concept is worth at least a first look because of the promise of very high specific impulse. Because of the low thrust, one would probably need a cluster of engines. This is not necessarily bad because there would be some redundancy, but because of the low thrust one might have to refuel while running. Depending on the fuel vaporization, material can be included in the uranium that is injected as one is running along.

  2. Minimizing liquid contaminants in natural gas liquids

    SciTech Connect

    Brown, R.L.; Wines, T.H.; Williamson, K.M.

    1996-12-31

    In processing natural gas liquids, significant contamination occurs with liquid dispersions and emulsions. Natural gas liquids (NGL) and liquid petroleum gas (LPG) streams are treated with caustic to remove residual organic sulfur compounds such as mercaptans and with amines to remove hydrogen sulfide. In both cases a liquid/liquid contactor is used. Significant amounts of the caustic or amine can be carried over into the product stream in process units that are running at rates above design capacity, are treating high sulfur feed stocks, or have other operational problems. The carried over liquid results in off-spec products, excessive loses of caustic or amine, and can cause operating problems in downstream processes. In addition, water is a significant contaminant which can cause LPG and natural gasoline to be off-specification. This paper discusses a new technique for separating very stable liquid dispersions of caustic, amine, or water from natural gas liquids using liquid/liquid cartridge coalescers constructed with specially formulated polymer and fluoropolymer medium with enhanced surface properties. In addition, factors influencing the coalescer mechanism will be discussed including interfacial tension, concentration of surface active compounds, steric repulsion, and electrostatic charge affects. Results from field tests, operating data from commercial installations, and economic benefits will also be presented.

  3. The c.-133A > G polymorphism in NPC1L1 gene influences the efficacy of ezetimibe monotherapy on apolipoprotein A1 in hyperlipidemic patients.

    PubMed

    Zsíros, N; Bodor, M; Varga, V; Berta, E; Balogh, I; Seres, I; Paragh, G; Harangi, M

    2014-06-01

    Niemann-Pick C1-like 1 protein (NPC1L1) plays a critical role in intestinal cholesterol absorption. Previous studies found that the NPC1L1 c.-133A > G SNP, but not other NPC1L1 SNPs, was associated with response to statin treatment and statin-ezetimibe combinations. To date effect of NPC1L1 c.-133A > G SNP on ezetimibe monotherapy has not been studied. Our objective was to examine whether SNP c.-133A > G at the NPC1L1 gene has effects on lipid levels and on the efficacy of 3, 6 and 12 months of 10 mg daily ezetimibe monotherapy in hyperlipidemic patients with statin induced adverse effects. One hundred and one type IIa and IIb hyperlipidemic patients (72 females, 29 males; age: 61.23 +/- 9.87 ys; BMI: 28.18 +/- 4.29 kg/m2) were enrolled. The genotype frequencies were conformed to Hardy-Weinberg equilibrium. We could not find significant differences in initial lipid levels between AA and AG + GG patients. While plasma levels of apolipoprotein A1 (ApoA1) did not significantly decrease after ezetimibe treatment (1.96; 3.39 and 2.74%) in AA patients, a significant elevation in ApoA1 levels has been found after treatment in AG + GG patients (9.15; 8.54 and 13.58%). The effect of NPC1L1 c.-133A > G on the ApoA1 levels was found significant (p < 0.05). Efficacy of treatment with ezetimibe on other plasma lipid parameters after 3, 6 or 12 months did not differ significantly. NPC1L1-133A > G SNP influences the ApoA1 response to ezetimibe monotherapy, therefore, may alter the effect of ezetimibe on the structure and function of the high-density lipoprotein particles. PMID:24974575

  4. Knockdown of human Oxa1l impairs the biogenesis of F1Fo-ATP synthase and NADH:ubiquinone oxidoreductase.

    PubMed

    Stiburek, Lukas; Fornuskova, Daniela; Wenchich, Laszlo; Pejznochova, Martina; Hansikova, Hana; Zeman, Jiri

    2007-11-23

    The Oxa1 protein is a founding member of the evolutionarily conserved Oxa1/Alb3/YidC protein family, which is involved in the biogenesis of membrane proteins in mitochondria, chloroplasts and bacteria. The predicted human homologue, Oxa1l, was originally identified by partial functional complementation of the respiratory growth defect of the yeast oxa1 mutant. Here we demonstrate that both the endogenous human Oxa1l, with an apparent molecular mass of 42 kDa, and the Oxa1l-FLAG chimeric protein localize exclusively to mitochondria in HEK293 cells. Furthermore, human Oxa1l was found to be an integral membrane protein, and, using two-dimensional blue native/denaturing PAGE, the majority of the protein was identified as part of a 600-700 kDa complex. The stable short hairpin (sh)RNA-mediated knockdown of Oxa1l in HEK293 cells resulted in markedly decreased steady-state levels and ATP hydrolytic activity of the F(1)F(o)-ATP synthase and moderately reduced levels and activity of NADH:ubiquinone oxidoreductase (complex I). However, no significant accumulation of corresponding sub-complexes could be detected on blue native immunoblots. Intriguingly, the achieved depletion of Oxa1l protein did not adversely affect the assembly or activity of cytochrome c oxidase or the cytochrome bc(1) complex. Taken together, our results indicate that human Oxa1l represents a mitochondrial integral membrane protein required for the correct biogenesis of F(1)F(o)-ATP synthase and NADH:ubiquinone oxidoreductase. PMID:17936786

  5. Loss of R2D2 proteins ROPN1 and ROPN1L causes defects in murine sperm motility, phosphorylation, and fibrous sheath integrity.

    PubMed

    Fiedler, Sarah E; Dudiki, Tejasvi; Vijayaraghavan, Srinivasan; Carr, Daniel W

    2013-02-01

    The fibrous sheath (FS) is a flagellar cytoskeletal structure unique to sperm that surrounds the outer dense fibers and axoneme. Its primary components are A-kinase anchoring proteins (AKAPs) 3 and 4, which suggests that the FS affects flagellar beating via the scaffolding of signaling pathways necessary for motility. Sperm proteins ROPN1 and ROPN1L bind AKAP3. To determine the role of ROPN1 and ROPN1L in sperm function, we created mice deficient in ROPN1 (RKO), mice deficient in ROPN1L (RLKO), and double knockout mice (DKO). All three strains of mice had normal testicular morphology and spermatogenesis. Only the DKOs had obvious defects in sperm morphology (thinning and shredding of the principal piece), which was accompanied by a reduction in AKAP3 levels. RLKO mice had slightly reduced sperm motility and increased levels of ROPN1. RKO mice had moderately impaired motility and increased levels of ROPN1L. DKO sperm were immotile. We have previously determined that RKO male mice are subfertile, and DKO males are infertile. Together these data indicate that ROPN1L and ROPN1 compensate for each other in the absence of the opposing protein, possibly to maintain AKAP3 incorporation in the FS. Sperm from mice lacking ROPN1L exhibited reductions in both cAMP-dependent protein kinase (PKA) phosphorylation of a 270-kDa protein (perhaps FSCB), and in capacitation-induced tyrosine phosphorylation. Sperm from mice lacking ROPN1 had reduced levels of FSCB and increased tyrosine phosphorylation of noncapacitated sperm. These data demonstrate that mutations in ROPN1 and ROPN1L can cause defects in FS integrity, sperm motility, and PKA-dependent signaling processes, leading to male infertility. PMID:23303679

  6. Age-related change of the role of alpha1L-adrenoceptor in canine urethral smooth muscle.

    PubMed

    Suzuki, Y; Moriyama, N; Okaya, Y; Nishimatsu, H; Kawabe, K; Aisaka, K

    1999-10-01

    To examine age-related alteration of the role of alpha1L-adrenoceptor in the urethra, young non-parous and aged parous female dogs were used. In a functional study, we evaluated phenylephrine-induced contraction and antagonistic effects of JTH-601, a newly synthesized alpha1-adrenoceptor antagonist, and prazosin; in a localization survey using autoradiographic technique, we investigated specific [3H]JTH-601 and [3H]tamsulosin binding. Concentration-response curves were obtained for phenylephrine (pD2 = 5.0-5.3). JTH-601 and prazosin antagonized this contraction with pA2 values of 8.2-8.3 and 8.0-8.1, respectively. Specific binding of both [3H]JTH-601 and [3H]tamsulosin were observed in the bladder neck and proximal section of urethra. There were no significant differences of the pD2, pA2, and radio ligand binding between young non-parous and aged parous dogs. PMID:10523074

  7. Control of steroid receptor dynamics and function by genomic actions of the cochaperones p23 and Bag-1L

    PubMed Central

    Cato, Laura; Neeb, Antje; Brown, Myles

    2014-01-01

    Molecular chaperones encompass a group of unrelated proteins that facilitate the correct assembly and disassembly of other macromolecular structures, which they themselves do not remain a part of. They associate with a large and diverse set of coregulators termed cochaperones that regulate their function and specificity. Amongst others, chaperones and cochaperones regulate the activity of several signaling molecules including steroid receptors, which upon ligand binding interact with discrete nucleotide sequences within the nucleus to control the expression of diverse physiological and developmental genes. Molecular chaperones and cochaperones are typically known to provide the correct conformation for ligand binding by the steroid receptors. While this contribution is widely accepted, recent studies have reported that they further modulate steroid receptor action outside ligand binding. They are thought to contribute to receptor turnover, transport of the receptor to different subcellular localizations, recycling of the receptor on chromatin and even stabilization of the DNA-binding properties of the receptor. In addition to these combined effects with molecular chaperones, cochaperones are reported to have additional functions that are independent of molecular chaperones. Some of these functions also impact on steroid receptor action. Two well-studied examples are the cochaperones p23 and Bag-1L, which have been identified as modulators of steroid receptor activity in nuclei. Understanding details of their regulatory action will provide new therapeutic opportunities of controlling steroid receptor action independent of the widespread effects of molecular chaperones. PMID:25422595

  8. Genetic variation at MECOM, TERT, JAK2 and HBS1L-MYB predisposes to myeloproliferative neoplasms.

    PubMed

    Tapper, William; Jones, Amy V; Kralovics, Robert; Harutyunyan, Ashot S; Zoi, Katerina; Leung, William; Godfrey, Anna L; Guglielmelli, Paola; Callaway, Alison; Ward, Daniel; Aranaz, Paula; White, Helen E; Waghorn, Katherine; Lin, Feng; Chase, Andrew; Baxter, E Joanna; Maclean, Cathy; Nangalia, Jyoti; Chen, Edwin; Evans, Paul; Short, Michael; Jack, Andrew; Wallis, Louise; Oscier, David; Duncombe, Andrew S; Schuh, Anna; Mead, Adam J; Griffiths, Michael; Ewing, Joanne; Gale, Rosemary E; Schnittger, Susanne; Haferlach, Torsten; Stegelmann, Frank; Döhner, Konstanze; Grallert, Harald; Strauch, Konstantin; Tanaka, Toshiko; Bandinelli, Stefania; Giannopoulos, Andreas; Pieri, Lisa; Mannarelli, Carmela; Gisslinger, Heinz; Barosi, Giovanni; Cazzola, Mario; Reiter, Andreas; Harrison, Claire; Campbell, Peter; Green, Anthony R; Vannucchi, Alessandro; Cross, Nicholas C P

    2015-01-01

    Clonal proliferation in myeloproliferative neoplasms (MPN) is driven by somatic mutations in JAK2, CALR or MPL, but the contribution of inherited factors is poorly characterized. Using a three-stage genome-wide association study of 3,437 MPN cases and 10,083 controls, we identify two SNPs with genome-wide significance in JAK2(V617F)-negative MPN: rs12339666 (JAK2; meta-analysis P=1.27 × 10(-10)) and rs2201862 (MECOM; meta-analysis P=1.96 × 10(-9)). Two additional SNPs, rs2736100 (TERT) and rs9376092 (HBS1L/MYB), achieve genome-wide significance when including JAK2(V617F)-positive cases. rs9376092 has a stronger effect in JAK2(V617F)-negative cases with CALR and/or MPL mutations (Breslow-Day P=4.5 × 10(-7)), whereas in JAK2(V617F)-positive cases rs9376092 associates with essential thrombocythemia (ET) rather than polycythemia vera (allelic χ(2) P=7.3 × 10(-7)). Reduced MYB expression, previously linked to development of an ET-like disease in model systems, associates with rs9376092 in normal myeloid cells. These findings demonstrate that multiple germline variants predispose to MPN and link constitutional differences in MYB expression to disease phenotype. PMID:25849990

  9. Exchange spring in A1/L1{sub 0} FePt composite and its application in magnetic force microscope

    SciTech Connect

    Li, Guoqing Zhu, Yanyan Zhang, Yong; Zhao, Hujun; Zeng, Daofu; Li, Yuhui; Lu, Wei

    2015-02-23

    This paper reported fabrication of Fe{sub x}Pt{sub 100-x} films with (001) epitaxy on MgO(100) substrates. The atomic percentage of Fe was changed within the range of x = 10–85 in order to search the optimal atomic ratio for achieving both high and isotropic-like coercivity. It was found that the Fe{sub 60}Pt{sub 40} film exhibited large coercivities exceeding 5 kOe along both in-plane and out-of-plane directions due to the formation of A1/L1{sub 0} FePt composite. A penta-domain model for hard/soft/hard exchange spring system was proposed to interpret the anomalous magnetization behaviors observed in Fe{sub 60}Pt{sub 40} sample. By using Fe{sub 60}Pt{sub 40} as the magnetic coating layer on a probe of magnetic force microscope, the flux changes at a linear density of 1000 kfci could be readily observed at a resolution of ∼13 nm.

  10. A common variant at the TERT-CLPTM1L locus is associated with estrogen receptor–negative breast cancer

    PubMed Central

    Haiman, Christopher A; Chen, Gary K; Vachon, Celine M; Canzian, Federico; Dunning, Alison; Millikan, Robert C; Wang, Xianshu; Ademuyiwa, Foluso; Ahmed, Shahana; Ambrosone, Christine B; Baglietto, Laura; Balleine, Rosemary; Bandera, Elisa V; Beckmann, Matthias W; Berg, Christine D; Bernstein, Leslie; Blomqvist, Carl; Blot, William J; Brauch, Hiltrud; Buring, Julie E; Carey, Lisa A; Carpenter, Jane E; Chang-Claude, Jenny; Chanock, Stephen J; Chasman, Daniel I; Clarke, Christine L; Cox, Angela; Cross, Simon S; Deming, Sandra L; Diasio, Robert B; Dimopoulos, Athanasios M; Driver, W Ryan; Dünnebier, Thomas; Durcan, Lorraine; Eccles, Diana; Edlund, Christopher K; Ekici, Arif B; Fasching, Peter A; Feigelson, Heather S; Flesch-Janys, Dieter; Fostira, Florentia; Försti, Asta; Fountzilas, George; Gerty, Susan M; Giles, Graham G; Godwin, Andrew K; Goodfellow, Paul; Graham, Nikki; Greco, Dario; Hamann, Ute; Hankinson, Susan E; Hartmann, Arndt; Hein, Rebecca; Heinz, Judith; Holbrook, Andrea; Hoover, Robert N; Hu, Jennifer J; Hunter, David J; Ingles, Sue A; Irwanto, Astrid; Ivanovich, Jennifer; John, Esther M; Johnson, Nicola; Jukkola-Vuorinen, Arja; Kaaks, Rudolf; Ko, Yon-Dschun; Kolonel, Laurence N; Konstantopoulou, Irene; Kosma, Veli-Matti; Kulkarni, Swati; Lambrechts, Diether; Lee, Adam M; Le Marchand, Loïc; Lesnick, Timothy; Liu, Jianjun; Lindstrom, Sara; Mannermaa, Arto; Margolin, Sara; Martin, Nicholas G; Miron, Penelope; Montgomery, Grant W; Nevanlinna, Heli; Nickels, Stephan; Nyante, Sarah; Olswold, Curtis; Palmer, Julie; Pathak, Harsh; Pectasides, Dimitrios; Perou, Charles M; Peto, Julian; Pharoah, Paul D P; Pooler, Loreall C; Press, Michael F; Pylkäs, Katri; Rebbeck, Timothy R; Rodriguez-Gil, Jorge L; Rosenberg, Lynn; Ross, Eric; Rüdiger, Thomas; Silva, Isabel dos Santos; Sawyer, Elinor; Schmidt, Marjanka K; Schulz-Wendtland, Rüdiger; Schumacher, Fredrick; Severi, Gianluca; Sheng, Xin; Signorello, Lisa B; Sinn, Hans-Peter; Stevens, Kristen N; Southey, Melissa C; Tapper, William J; Tomlinson, Ian; Hogervorst, Frans B L; Wauters, Els; Weaver, JoEllen; Wildiers, Hans; Winqvist, Robert; Van Den Berg, David; Wan, Peggy; Xia, Lucy Y; Yannoukakos, Drakoulis; Zheng, Wei; Ziegler, Regina G; Siddiq, Afshan; Slager, Susan L; Stram, Daniel O; Easton, Douglas; Kraft, Peter; Henderson, Brian E; Couch, Fergus J

    2012-01-01

    Estrogen receptor (ER)-negative breast cancer shows a higher incidence in women of African ancestry compared to women of European ancestry. In search of common risk alleles for ER-negative breast cancer, we combined genome-wide association study (GWAS) data from women of African ancestry (1,004 ER-negative cases and 2,745 controls) and European ancestry (1,718 ER-negative cases and 3,670 controls), with replication testing conducted in an additional 2,292 ER-negative cases and 16,901 controls of European ancestry. We identified a common risk variant for ER-negative breast cancer at the TERT-CLPTM1L locus on chromosome 5p15 (rs10069690: per-allele odds ratio (OR) = 1.18 per allele, P = 1.0 × 10−10). The variant was also significantly associated with triple-negative (ER-negative, progesterone receptor (PR)-negative and human epidermal growth factor-2 (HER2)-negative) breast cancer (OR = 1.25, P = 1.1 × 10−9), particularly in younger women (<50 years of age) (OR = 1.48, P = 1.9 × 10−9). Our results identify a genetic locus associated with estrogen receptor negative breast cancer subtypes in multiple populations. PMID:22037553

  11. Point-spread function reconstruction in ground-based astronomy by l(1)-l(p) model.

    PubMed

    Chan, Raymond H; Yuan, Xiaoming; Zhang, Wenxing

    2012-11-01

    In ground-based astronomy, images of objects in outer space are acquired via ground-based telescopes. However, the imaging system is generally interfered by atmospheric turbulence, and hence images so acquired are blurred with unknown point-spread function (PSF). To restore the observed images, the wavefront of light at the telescope's aperture is utilized to derive the PSF. A model with the Tikhonov regularization has been proposed to find the high-resolution phase gradients by solving a least-squares system. Here we propose the l(1)-l(p) (p=1, 2) model for reconstructing the phase gradients. This model can provide sharper edges in the gradients while removing noise. The minimization models can easily be solved by the Douglas-Rachford alternating direction method of a multiplier, and the convergence rate is readily established. Numerical results are given to illustrate that the model can give better phase gradients and hence a more accurate PSF. As a result, the restored images are much more accurate when compared to the traditional Tikhonov regularization model. PMID:23201786

  12. Genetic variation at MECOM, TERT, JAK2 and HBS1L-MYB predisposes to myeloproliferative neoplasms

    PubMed Central

    Tapper, William; Jones, Amy V.; Kralovics, Robert; Harutyunyan, Ashot S.; Zoi, Katerina; Leung, William; Godfrey, Anna L.; Guglielmelli, Paola; Callaway, Alison; Ward, Daniel; Aranaz, Paula; White, Helen E.; Waghorn, Katherine; Lin, Feng; Chase, Andrew; Joanna Baxter, E.; Maclean, Cathy; Nangalia, Jyoti; Chen, Edwin; Evans, Paul; Short, Michael; Jack, Andrew; Wallis, Louise; Oscier, David; Duncombe, Andrew S.; Schuh, Anna; Mead, Adam J.; Griffiths, Michael; Ewing, Joanne; Gale, Rosemary E.; Schnittger, Susanne; Haferlach, Torsten; Stegelmann, Frank; Döhner, Konstanze; Grallert, Harald; Strauch, Konstantin; Tanaka, Toshiko; Bandinelli, Stefania; Giannopoulos, Andreas; Pieri, Lisa; Mannarelli, Carmela; Gisslinger, Heinz; Barosi, Giovanni; Cazzola, Mario; Reiter, Andreas; Harrison, Claire; Campbell, Peter; Green, Anthony R.; Vannucchi, Alessandro; Cross, Nicholas C.P.

    2015-01-01

    Clonal proliferation in myeloproliferative neoplasms (MPN) is driven by somatic mutations in JAK2, CALR or MPL, but the contribution of inherited factors is poorly characterized. Using a three-stage genome-wide association study of 3,437 MPN cases and 10,083 controls, we identify two SNPs with genome-wide significance in JAK2V617F-negative MPN: rs12339666 (JAK2; meta-analysis P=1.27 × 10−10) and rs2201862 (MECOM; meta-analysis P=1.96 × 10−9). Two additional SNPs, rs2736100 (TERT) and rs9376092 (HBS1L/MYB), achieve genome-wide significance when including JAK2V617F-positive cases. rs9376092 has a stronger effect in JAK2V617F-negative cases with CALR and/or MPL mutations (Breslow–Day P=4.5 × 10−7), whereas in JAK2V617F-positive cases rs9376092 associates with essential thrombocythemia (ET) rather than polycythemia vera (allelic χ2 P=7.3 × 10−7). Reduced MYB expression, previously linked to development of an ET-like disease in model systems, associates with rs9376092 in normal myeloid cells. These findings demonstrate that multiple germline variants predispose to MPN and link constitutional differences in MYB expression to disease phenotype. PMID:25849990

  13. Ionic Liquids Database- (ILThermo)

    National Institute of Standards and Technology Data Gateway

    SRD 147 Ionic Liquids Database- (ILThermo) (Web, free access)   IUPAC Ionic Liquids Database, ILThermo, is a free web research tool that allows users worldwide to access an up-to-date data collection from the publications on experimental investigations of thermodynamic, and transport properties of ionic liquids as well as binary and ternary mixtures containing ionic liquids.

  14. Liquid crystal polyester thermosets

    SciTech Connect

    Benicewicz, B.C.; Hoyt, A.E.

    1990-01-01

    The present invention relates to the field of curable liquid crystal polyester monomers and to thermoset liquid crystalline polyester compositions prepared therefrom. It is an object of this invention to provide curable liquid crystalline polyester materials. Another object of this invention is to provide a process of preparing curable liquid crystal polyester monomers. Yet another object of this invention is to provide liquid crystalline blends of polyester materials. It is a further object of this invention to provide thermoset liquid crystalline polyester compositions. It is a still further object of this invention to provide thermoset liquid crystalline polyester compositions having a high heat resistance. 1 fig.

  15. Liquid crystal polyester thermosets

    SciTech Connect

    Benicewicz, B.C.; Hoyt, A.E.

    1990-12-31

    The present invention relates to the field of curable liquid crystal polyester monomers and to thermoset liquid crystalline polyester compositions prepared therefrom. It is an object of this invention to provide curable liquid crystalline polyester materials. Another object of this invention is to provide a process of preparing curable liquid crystal polyester monomers. Yet another object of this invention is to provide liquid crystalline blends of polyester materials. It is a further object of this invention to provide thermoset liquid crystalline polyester compositions. It is a still further object of this invention to provide thermoset liquid crystalline polyester compositions having a high heat resistance. 1 fig.

  16. The Sel1L-Hrd1 Endoplasmic Reticulum-Associated Degradation Complex Manages a Key Checkpoint in B Cell Development.

    PubMed

    Ji, Yewei; Kim, Hana; Yang, Liu; Sha, Haibo; Roman, Christopher A; Long, Qiaoming; Qi, Ling

    2016-09-01

    Endoplasmic reticulum (ER)-associated degradation (ERAD) is a principal mechanism that targets ER-associated proteins for cytosolic proteasomal degradation. Here, our data demonstrate a critical role for the Sel1L-Hrd1 complex, the most conserved branch of ERAD, in early B cell development. Loss of Sel1L-Hrd1 ERAD in B cell precursors leads to a severe developmental block at the transition from large to small pre-B cells. Mechanistically, we show that Sel1L-Hrd1 ERAD selectively recognizes and targets the pre-B cell receptor (pre-BCR) for proteasomal degradation in a BiP-dependent manner. The pre-BCR complex accumulates both intracellularly and at the cell surface in Sel1L-deficient pre-B cells, leading to persistent pre-BCR signaling and pre-B cell proliferation. This study thus implicates ERAD mediated by Sel1L-Hrd1 as a key regulator of B cell development and reveals the molecular mechanism underpinning the transient nature of pre-BCR signaling. PMID:27568564

  17. The oncogenic FIP1L1-PDGFRα fusion protein displays skewed signaling properties compared to its wild-type PDGFRα counterpart

    PubMed Central

    Haan, Serge; Bahlawane, Christelle; Wang, Jiali; Nazarov, Petr V; Muller, Arnaud; Eulenfeld, René; Haan, Claude; Rolvering, Catherine; Vallar, Laurent; Satagopam, Venkata P; Sauter, Thomas; Wiesinger, Monique Yvonne

    2015-01-01

    Aberrant activation of oncogenic kinases is frequently observed in human cancers, but the underlying mechanism and resulting effects on global signaling are incompletely understood. Here, we demonstrate that the oncogenic FIP1L1-PDGFRα kinase exhibits a significantly different signaling pattern compared to its PDGFRα wild type counterpart. Interestingly, the activation of primarily membrane-based signal transduction processes (such as PI3-kinase- and MAP-kinase- pathways) is remarkably shifted toward a prominent activation of STAT factors. This diverging signaling pattern compared to classical PDGF-receptor signaling is partially coupled to the aberrant cytoplasmic localization of the oncogene, since membrane targeting of FIP1L1-PDGFRα restores activation of MAPK- and PI3K-pathways. In stark contrast to the classical cytokine-induced STAT activation process, STAT activation by FIP1L1-PDGFRα does neither require Janus kinase activity nor Src kinase activity. Furthermore, we investigated the mechanism of STAT5 activation via FIP1L1-PDGFRα in more detail and found that STAT5 activation does not involve an SH2-domain-mediated binding mechanism. We thus demonstrate that STAT5 activation occurs via a non-canonical activation mechanism in which STAT5 may be subject to a direct phosphorylation by FIP1L1-PDGFRα. PMID:26413425

  18. Conserved Roles of CrRLK1L Receptor-Like Kinases in Cell Expansion and Reproduction from Algae to Angiosperms.

    PubMed

    Galindo-Trigo, Sergio; Gray, Julie E; Smith, Lisa M

    2016-01-01

    Receptor-like kinases (RLKs) are regulators of plant development through allowing cells to sense their extracellular environment. They facilitate detection of local endogenous signals, in addition to external biotic and abiotic stimuli. The Catharanthus roseus RLK1-like (CrRLK1L) protein kinase subfamily, which contains FERONIA, plays a central role in regulating fertilization and in cell expansion mechanisms such as cell elongation and tip growth, as well as having indirect links to plant-pathogen interactions. Several components of CrRLK1L signaling pathways have been identified, including an extracellular ligand, coreceptors, and downstream signaling elements. The presence and abundance of the CrRLK1L proteins in the plant kingdom suggest an origin within the Streptophyta lineage, with a notable increase in prevalence in the seeded land plants. Given the function of the sole CrRLK1L protein in a charophycean alga, the possibility of a conserved role in detection and/or regulation of cell wall integrity throughout the Strephtophytes is discussed. Orthologs of signaling pathway components are also present in extant representatives of non-vascular land plants and early vascular land plants including the liverwort Marchantia polymorpha, the moss Physcomitrella patens and the lycophyte Selaginella moellendorffii. Deciphering the roles in development of the CrRLK1L protein kinases in early diverging land plants will provide insights into their ancestral function, furthering our understanding of this diversified subfamily of receptors in higher plants. PMID:27621737

  19. Conserved Roles of CrRLK1L Receptor-Like Kinases in Cell Expansion and Reproduction from Algae to Angiosperms

    PubMed Central

    Galindo-Trigo, Sergio; Gray, Julie E.; Smith, Lisa M.

    2016-01-01

    Receptor-like kinases (RLKs) are regulators of plant development through allowing cells to sense their extracellular environment. They facilitate detection of local endogenous signals, in addition to external biotic and abiotic stimuli. The Catharanthus roseus RLK1-like (CrRLK1L) protein kinase subfamily, which contains FERONIA, plays a central role in regulating fertilization and in cell expansion mechanisms such as cell elongation and tip growth, as well as having indirect links to plant–pathogen interactions. Several components of CrRLK1L signaling pathways have been identified, including an extracellular ligand, coreceptors, and downstream signaling elements. The presence and abundance of the CrRLK1L proteins in the plant kingdom suggest an origin within the Streptophyta lineage, with a notable increase in prevalence in the seeded land plants. Given the function of the sole CrRLK1L protein in a charophycean alga, the possibility of a conserved role in detection and/or regulation of cell wall integrity throughout the Strephtophytes is discussed. Orthologs of signaling pathway components are also present in extant representatives of non-vascular land plants and early vascular land plants including the liverwort Marchantia polymorpha, the moss Physcomitrella patens and the lycophyte Selaginella moellendorffii. Deciphering the roles in development of the CrRLK1L protein kinases in early diverging land plants will provide insights into their ancestral function, furthering our understanding of this diversified subfamily of receptors in higher plants. PMID:27621737

  20. Identification of Novel Disruptor of Telomeric Silencing 1-like (DOT1L) Inhibitors through Structure-Based Virtual Screening and Biological Assays.

    PubMed

    Chen, Shijie; Li, Linjuan; Chen, Yantao; Hu, Junchi; Liu, Jingqiu; Liu, Yu-Chih; Liu, Rongfeng; Zhang, Yuanyuan; Meng, Fanwang; Zhu, Kongkai; Lu, Junyan; Zheng, Mingyue; Chen, Kaixian; Zhang, Jin; Jiang, Hualiang; Yao, Zhiyi; Luo, Cheng

    2016-03-28

    Histone methyltransferases are involved in many important biological processes, and abnormalities in these enzymes are associated with tumorigenesis and progression. Disruptor of telomeric silencing 1-like (DOT1L), a key hub in histone lysine methyltransferases, has been reported to play an important role in the processes of mixed-lineage leukemia (MLL)-rearranged leukemias and validated to be a potential therapeutic target. In this study, we identified a novel DOT1L inhibitor, DC_L115 (CAS no. 1163729-79-0), by combining structure-based virtual screening with biochemical analyses. This potent inhibitor DC_L115 shows high inhibitory activity toward DOT1L (IC50 = 1.5 μM). Through a process of surface plasmon resonance-based binding assays, DC_L115 was founded to bind to DOT1L with a binding affinity of 0.6 μM in vitro. Moreover, this compound selectively inhibits MLL-rearranged cell proliferation with an IC50 value of 37.1 μM. We further predicted the binding modes of DC_L115 through molecular docking analysis and found that the inhibitor competitively occupies the binding site of S-adenosylmethionine. Overall, this study demonstrates the development of potent DOT1L inhibitors with novel scaffolds. PMID:26914852

  1. Postnatal microcephaly and pain insensitivity due to a de novo heterozygous DNM1L mutation causing impaired mitochondrial fission and function.

    PubMed

    Sheffer, Ruth; Douiev, Liza; Edvardson, Simon; Shaag, Avraham; Tamimi, Khaled; Soiferman, Devorah; Meiner, Vardiella; Saada, Ann

    2016-06-01

    An emerging class of mitochondrial disorders is caused by mutations in nuclear genes affecting mitochondrial dynamics and function. One of these is the DNM1L gene encoding the dynamin-related protein 1 (DRP1), which is pivotal in the mitochondrial fission process. Here, we describe a patient with a novel dominant-negative, de novo DNM1L mutation, which expands the clinical spectrum. The patient reported here exhibits a chronic neurological disorder, characterized by postnatal microcephaly, developmental delay, and pain insensitivity. Muscle biopsy disclosed decreased respiratory chain complex IV activity. Exome sequencing showed a de novo heterozygous c.1084G>A (p.G362S) mutation. Subsequent studies of patient skin fibroblasts showed markedly impaired mitochondrial fission and a partial respiratory chain defect while peroxisomal morphology remained intact. Human foreskin fibroblasts over-expressing the mutant DNM1L gene displayed aberrant mitochondrial morphology. © 2016 Wiley Periodicals, Inc. PMID:26992161

  2. The HR97 (NR1L) group of nuclear receptors: a new group of nuclear receptors discovered in Daphnia species.

    PubMed

    Li, Yangchun; Ginjupalli, Gautam K; Baldwin, William S

    2014-09-15

    The recently sequenced Daphnia pulex genome revealed the NR1L nuclear receptor group consisting of three novel receptors. Phylogenetic studies show that this group is related to the NR1I group (CAR/PXR/VDR) and the NR1J group (HR96), and were subsequently named HR97a/b/g. Each of the HR97 paralogs from Daphnia magna, a commonly used crustacean in toxicity testing, was cloned, sequenced, and partially characterized. Phylogenetic analysis indicates that the HR97 receptors are present in primitive arthropods such as the chelicerates but lost in insects. qPCR and immunohistochemistry demonstrate that each of the receptors is expressed near or at reproductive maturity, and that HR97g, the most ancient of the HR97 receptors, is primarily expressed in the gastrointestinal tract, mandibular region, and ovaries, consistent with a role in reproduction. Transactivation assays using an HR97a/b/g-GAL4 chimera indicate that unlike Daphnia HR96 that is promiscuous with respect to ligand recognition, the HR97 receptors do not respond to many of the ligands that activate CAR/PXR/HR96 nuclear receptors. Only three putative ligands of HR97 receptors were identified in this study: pyriproxyfen, methyl farnesoate, and arachidonic acid. Only arachidonic acid, which acts as an inverse agonist, alters HR97g activity at concentrations that would be considered within physiologically relevant ranges. Overall, this study demonstrates that, although closely related to the promiscuous receptors in the NR1I and NR1J groups, the HR97 receptors are mostly likely not multi-xenobiotic sensors, but rather may perform physiological functions, potentially in reproduction, unique to crustaceans and other non-insect arthropod groups. PMID:25092536

  3. Antitumor activity of S116836, a novel tyrosine kinase inhibitor, against imatinib-resistant FIP1L1-PDGFRα-expressing cells

    PubMed Central

    Ding, Ke; Zhang, Zhang; Wang, Deping; Pan, Jingxuan

    2014-01-01

    The FIP1-like-1-platelet-derived growth factor receptor alpha (FIP1L1-PDGFRα) fusion oncogene is the driver factor in a subset of patients with hypereosinophilic syndrome (HES)/chronic eosinophilic leukemia (CEL). Most FIP1L1-PDGFRα-positive patients respond well to the tyrosine kinase inhibitor (TKI) imatinib. Resistance to imatinib in HES/CEL has been described mainly due to the T674I mutation in FIP1L1-PDGFRα, which is homologous to the imatinib-resistant T315I mutation in BCR-ABL. Development of novel TKIs is imperative to overcome resistance to imatinib. We synthesized S116836, a novel TKI. In this study, we evaluated the antitumor activity of S116836 in FIP1L1-PDGFRα-expressing cells. The results showed that S116836 potently inhibited PDGFRα and its downstream signaling molecules such as STAT3, AKT, and Erk1/2. S116836 effectively inhibited the growth of the WT and T674I FIP1L1-PDGFRα-expressing neoplastic cells in vitro and in nude mouse xenografts. Moreover, S116836 induced intrinsic pathway of apoptosis as well as the death receptor pathway, coincided with up-regulation of the proapoptotic BH3-only protein Bim-EL through the Erk1/2 pathway. In conclusion, S116836 is active against WT and T674I FIP1L1-PDGFRα-expressing cells, and may be a prospective agent for the treatment of HES/CEL. PMID:25431951

  4. VKORC1L1, an Enzyme Rescuing the Vitamin K 2,3-Epoxide Reductase Activity in Some Extrahepatic Tissues during Anticoagulation Therapy*

    PubMed Central

    Hammed, Abdessalem; Matagrin, Benjamin; Spohn, Gabriele; Prouillac, Caroline; Benoit, Etienne; Lattard, Virginie

    2013-01-01

    Vitamin K is involved in the γ-carboxylation of the vitamin K-dependent proteins, and vitamin K epoxide is a by-product of this reaction. Due to the limited intake of vitamin K, its regeneration is necessary and involves vitamin K 2,3-epoxide reductase (VKOR) activity. This activity is known to be supported by VKORC1 protein, but recently a second gene, VKORC1L1, appears to be able to support this activity when the encoded protein is expressed in HEK293T cells. Nevertheless, this protein was described as being responsible for driving the vitamin K-mediated antioxidation pathways. In this paper we precisely analyzed the catalytic properties of VKORC1L1 when expressed in Pichia pastoris and more particularly its susceptibility to vitamin K antagonists. Vitamin K antagonists are also inhibitors of VKORC1L1, but this enzyme appears to be 50-fold more resistant to vitamin K antagonists than VKORC1. The expression of Vkorc1l1 mRNA was observed in all tissues assayed, i.e. in C57BL/6 wild type and VKORC1-deficient mouse liver, lung, and testis and rat liver, lung, brain, kidney, testis, and osteoblastic cells. The characterization of VKOR activity in extrahepatic tissues demonstrated that a part of the VKOR activity, more or less important according to the tissue, may be supported by VKORC1L1 enzyme especially in testis, lung, and osteoblasts. Therefore, the involvement of VKORC1L1 in VKOR activity partly explains the low susceptibility of some extrahepatic tissues to vitamin K antagonists and the lack of effects of vitamin K antagonists on the functionality of the vitamin K-dependent protein produced by extrahepatic tissues such as matrix Gla protein or osteocalcin. PMID:23928358

  5. In Situ Proteome Profiling and Bioimaging Applications of Small-Molecule Affinity-Based Probes Derived From DOT1L Inhibitors.

    PubMed

    Zhu, Biwei; Zhang, Hailong; Pan, Sijun; Wang, Chenyu; Ge, Jingyan; Lee, Jun-Seok; Yao, Shao Q

    2016-06-01

    DOT1L is the sole protein methyltransferase that methylates histone H3 on lysine 79 (H3K79), and is a promising drug target against cancers. Small-molecule inhibitors of DOT1L such as FED1 are potential anti-cancer agents and useful tools to investigate the biological roles of DOT1L in human diseases. FED1 showed excellent in vitro inhibitory activity against DOT1L, but its cellular effect was relatively poor. In this study, we designed and synthesized photo-reactive and "clickable" affinity-based probes (AfBPs), P1 and P2, which were cell-permeable and structural mimics of FED1. The binding and inhibitory effects of these two probes against DOT1L protein were extensively investigated in vitro and in live mammalian cells (in situ). The cellular uptake and sub-cellular localization properties of the probes were subsequently studied in live-cell imaging experiments, and our results revealed that, whereas both P1 and P2 readily entered mammalian cells, most of them were not able to reach the cell nucleus where functional DOT1L resides. This offers a plausible explanation for the poor cellular activity of FED1. Finally with P1/P2, large-scale cell-based proteome profiling, followed by quantitative LC-MS/MS, was carried out to identify potential cellular off-targets of FED1. Amongst the more than 100 candidate off-targets identified, NOP2 (a putative ribosomal RNA methyltransferase) was further confirmed to be likely a genuine off-target of FED1 by preliminary validation experiments including pull-down/Western blotting (PD/WB) and cellular thermal shift assay (CETSA). PMID:27115831

  6. Analysis of Niemann-Pick C1-like 1 (NPC1L1) genetic polymorphisms and haplotypes in Chinese Han population.

    PubMed

    Lou, Xiao-Ya; Qin, Chong-Zhen; Bao, Mei-Hua; Hu, Dong-Li; Chen, Feng; Sun, Hong; Chen, Yao; Guo, Dong; Zhang, Wei; Cheng, Yu; Zhou, Hong-Hao

    2015-09-01

    Niemann-Pick C1-like 1 (NPC1L1i) protein is the key transporter responsible for dietary cholesterol absorption. Recent studies indicated that several functional polymorphisms of NPC1L1 were associated with coronary heart disease (CHD) and response to ezetimibe therapy. The aim of the present study was to analyze the allele frequency and haplotype distribution of NPC1L1 polymorphisms in Chinese Hans and to compare them with those of other ethnic populations reported before. Blood samples were collected from 424 unrelated Chinese Hans (246 males and 178 females). Ten NPC1L1 polymorphisms (-762T > C, -133A > G, -18C > A, 1721C > T, 1735C > G, 1764T > C, 1767G > A, 27677T > C, 25342A > C and 28650A > G) were genotyped by direct sequencing or polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay. Among the variants, the minor allele frequency of -762T > C and 1735C > G were 35.0% and 37.0%, respectively. Furthermore, these two polymorphisms were highly linked with a D' value of 0.80. The observed frequencies of two major haplotypes were 59.1% for T-762/C1735 and 30.1% for C-762/G1735, respectively. The frequencies of the rest variants were extremely low (1.8% for - 133G, 1.5% for -18A, 0.9% for 1721T and only 0.2% for 27677C allele, respectively) or even not detected (1764T > C, 1767G > A, 25342A > C and 28650A > G) in our study population. Comparison with other ethnic populations revealed a remarkable genetic variability in the incidences of NPC1L1 polymorphisms. The frequencies of NPC1L1 polymorphisms in Chinese Hans are comparable to Japanese population but totally different from Caucasians, African-Americans and Hispanic individuals. This is the first study to report the ethnic difference in the frequencies of NPC1L1 functional polymorphisms in detail. -762T > C and 1735C > G are two prevalent NPC1L1 variants which need further studies to explore their clinical impact on CHD prevalence and response to ezetimibe therapy in Chinese Hans

  7. Semiconductor nanorod liquid crystals

    SciTech Connect

    Li, Liang-shi; Walda, Joost; Manna, Liberato; Alivisatos, A. Paul

    2002-01-28

    Rodlike molecules form liquid crystalline phases with orientational order and positional disorder. The great majority of materials in which liquid crystalline phases have been observed are comprised of organic molecules or polymers, even though there has been continuing and growing interest in inorganic liquid crystals. Recent advances in the control of the sizes and shapes of inorganic nanocrystals allow for the formation of a broad class of new inorganic liquid crystals. Here we show the formation of liquid crystalline phases of CdSe semiconductor nanorods. These new liquid crystalline phases may have great importance for both application and fundamental study.

  8. Liquid level detector

    DOEpatents

    Grasso, A.P.

    1984-02-21

    A liquid level detector for low pressure boilers. A boiler tank, from which vapor, such as steam, normally exits via a main vent, is provided with a vertical side tube connected to the tank at the desired low liquid level. When the liquid level falls to the level of the side tube vapor escapes therethrough causing heating of a temperature sensitive device located in the side tube, which, for example, may activate a liquid supply means for adding liquid to the boiler tank. High liquid level in the boiler tank blocks entry of vapor into the side tube, allowing the temperature sensitive device to cool, for example, to ambient temperature.

  9. Liquid level detector

    DOEpatents

    Grasso, Albert P.

    1986-01-01

    A liquid level detector for low pressure boilers. A boiler tank, from which apor, such as steam, normally exits via a main vent, is provided with a vertical side tube connected to the tank at the desired low liquid level. When the liquid level falls to the level of the side tube vapor escapes therethrough causing heating of a temperature sensitive device located in the side tube, which, for example, may activate a liquid supply means for adding liquid to the boiler tank. High liquid level in the boiler tank blocks entry of vapor into the side tube, allowing the temperature sensitive device to cool, for example, to ambient temperature.

  10. OPTIMIZATION OF PURGING EFFICIENCY AND QUANTIFICATION OF ORGANIC CONTAMINANTS FROM WATER USING A 1-L CLOSED-LOOP-STRIPPING APPARATUS AND COMPUTERIZED CAPILLARY COLUMN GC/MS

    EPA Science Inventory

    Optimization of a HERL-designed 1-L closed-loop-stripping (CLS) system, using variations of sample water bath temperatures and purging times, was accomplished by spiking five 1-chloroalkanes (C6, C8, C12, C16, and C18) into low organic Milli-Q reagent water to determine the recov...

  11. SIPA1L3 identified by linkage analysis and whole-exome sequencing as a novel gene for autosomal recessive congenital cataract.

    PubMed

    Evers, Christina; Paramasivam, Nagarajan; Hinderhofer, Katrin; Fischer, Christine; Granzow, Martin; Schmidt-Bacher, Annette; Eils, Roland; Steinbeisser, Herbert; Schlesner, Matthias; Moog, Ute

    2015-12-01

    Congenital cataract (CC) is one of the most important causes for blindness or visual impairment in infancy. A substantial proportion of isolated CCs has monogenic causes. The disease is genetically heterogeneous, and all Mendelian modes of inheritance have been reported. We mapped a locus for isolated CC on 19p13.1-q13.2 in a distantly consanguineous German family with two sisters affected by dense white cataracts. Whole-exome sequencing identified a homozygous nonsense variant c.4489C>T (p.(R1497*)) in SIPA1L3 (signal-induced proliferation-associated 1 like 3) in both affected children. SIPA1L3 encodes a GTPase-activating protein (GAP), which interacts with small GTPases of the Rap family via its Rap-GAP-domain. The suggested role of Rap GTPases in cell growth, differentiation and organization of the cytoskeleton in the human lens, and lens-enriched expression of the murine ortholog gene Sipa1l3 in embryonic mice indicates that this gene is crucial for early lens development. Our results provide evidence that sequence variants in human SIPA1L3 cause autosomal recessive isolated CC and give new insight into the molecular pathogenesis underlying human cataracts. PMID:25804400

  12. Evidence that ubiquitylated H2B corrals hDot1L on the nucleosomal surface to induce H3K79 methylation

    PubMed Central

    Zhou, Linjiao; Holt, Matthew T.; Ohashi, Nami; Zhao, Aishan; Müller, Manuel M.; Wang, Boyuan; Muir, Tom W.

    2016-01-01

    Ubiquitylation of histone H2B at lysine 120 (H2B-Ub), a post-translational modification first discovered in 1980, plays a critical role in diverse nuclear processes including the regulation of transcription and DNA damage repair. Herein, we use a suite of protein chemistry methods to explore how H2B-Ub stimulates hDot1L-mediated methylation of histone H3 on lysine 79 (H3K79me). By using semisynthetic ‘designer' chromatin containing H2B-Ub bearing a site-specifically installed photocrosslinker, here we report an interaction between a functional hotspot on ubiquitin and the N-terminus of histone H2A. Our biochemical studies indicate that this interaction is required for stimulation of hDot1L activity and leads to a repositioning of hDot1L on the nucleosomal surface, which likely places the active site of the enzyme proximal to H3K79. Collectively, our data converge on a possible mechanism for hDot1L stimulation in which H2B-Ub physically ‘corrals' the enzyme into a productive binding orientation. PMID:26830124

  13. Exploring drug delivery for the DOT1L inhibitor pinometostat (EPZ-5676): Subcutaneous administration as an alternative to continuous IV infusion, in the pursuit of an epigenetic target.

    PubMed

    Waters, Nigel J; Daigle, Scott R; Rehlaender, Bruce N; Basavapathruni, Aravind; Campbell, Carly T; Jensen, Tyler B; Truitt, Brett F; Olhava, Edward J; Pollock, Roy M; Stickland, Kim A; Dovletoglou, Angelos

    2015-12-28

    Protein methyltransferases are emerging as promising drug targets for therapeutic intervention in human cancers. Pinometostat (EPZ-5676) is a small molecule inhibitor of the DOT1L enzyme, a histone methyltransferase that methylates lysine 79 of histone H3. DOT1L activity is dysregulated in the pathophysiology of rearranged mixed lineage leukemia (MLL-r). Pinometostat is currently in Phase 1 clinical trials in relapsed refractory acute leukemia patients and is administered as a continuous IV infusion (CIV). The studies herein investigated alternatives to CIV administration of pinometostat to improve patient convenience. Various sustained release technologies were considered, and based on the required dose size as well as practical considerations, subcutaneous (SC) bolus administration of a solution formulation was selected for further evaluation in preclinical studies. SC administration offered improved exposure and complete bioavailability of pinometostat relative to CIV and oral administration. These findings warranted further evaluation in rat xenograft models of MLL-r leukemia. SC dosing in xenograft models demonstrated inhibition of MLL-r tumor growth and inhibition of pharmacodynamic markers of DOT1L activity. However, a dosing frequency of thrice daily (t.i.d) was required in these studies to elicit optimal inhibition of DOT1L target genes and tumor growth inhibition. Development of an extended release formulation may prove useful in the further optimization of the SC delivery of pinometostat, moving towards a more convenient dosing paradigm for patients. PMID:26385168

  14. Evidence that ubiquitylated H2B corrals hDot1L on the nucleosomal surface to induce H3K79 methylation.

    PubMed

    Zhou, Linjiao; Holt, Matthew T; Ohashi, Nami; Zhao, Aishan; Müller, Manuel M; Wang, Boyuan; Muir, Tom W

    2016-01-01

    Ubiquitylation of histone H2B at lysine 120 (H2B-Ub), a post-translational modification first discovered in 1980, plays a critical role in diverse nuclear processes including the regulation of transcription and DNA damage repair. Herein, we use a suite of protein chemistry methods to explore how H2B-Ub stimulates hDot1L-mediated methylation of histone H3 on lysine 79 (H3K79me). By using semisynthetic 'designer' chromatin containing H2B-Ub bearing a site-specifically installed photocrosslinker, here we report an interaction between a functional hotspot on ubiquitin and the N-terminus of histone H2A. Our biochemical studies indicate that this interaction is required for stimulation of hDot1L activity and leads to a repositioning of hDot1L on the nucleosomal surface, which likely places the active site of the enzyme proximal to H3K79. Collectively, our data converge on a possible mechanism for hDot1L stimulation in which H2B-Ub physically 'corrals' the enzyme into a productive binding orientation. PMID:26830124

  15. The CEACAM1-L Glycoprotein Associates with the Actin Cytoskeleton and Localizes to Cell–Cell Contact through Activation of Rho-like GTPases

    PubMed Central

    Sadekova, Svetlana; Lamarche-Vane, Nathalie; Li, Xiaodong; Beauchemin, Nicole

    2000-01-01

    Associations between plasma membrane-linked proteins and the actin cytoskeleton play a crucial role in defining cell shape and determination, ensuring cell motility and facilitating cell–cell or cell–substratum adhesion. Here, we present evidence that CEACAM1-L, a cell adhesion molecule of the carcinoembryonic antigen family, is associated with the actin cytoskeleton. We have delineated the regions involved in actin cytoskeleton association to the distal end of the CEACAM1-L long cytoplasmic domain. We have demonstrated that CEACAM1-S, an isoform of CEACAM1 with a truncated cytoplasmic domain, does not interact with the actin cytoskeleton. In addition, a major difference in subcellular localization of the two CEACAM1 isoforms was observed. Furthermore, we have established that the localization of CEACAM1-L at cell–cell boundaries is regulated by the Rho family of GTPases. The retention of the protein at the sites of intercellular contacts critically depends on homophilic CEACAM1–CEACAM1 interactions and association with the actin cytoskeleton. Our results provide new evidence on how the Rho family of GTPases can control cell adhesion: by directing an adhesion molecule to its proper cellular destination. In addition, these results provide an insight into the mechanisms of why CEACAM1-L, but not CEACAM1-S, functions as a tumor cell growth inhibitor. PMID:10637291

  16. Precision liquid level sensor

    DOEpatents

    Field, M.E.; Sullivan, W.H.

    A precision liquid level sensor utilizes a balanced bridge, each arm including an air dielectric line. Changes in liquid level along one air dielectric line imbalance the bridge and create a voltage which is directly measurable across the bridge.

  17. Liquid medication administration

    MedlinePlus

    ... easily. Oral syringes have some advantages for giving liquid medicines. They are accurate. They are easy to ... cups are also a handy way to give liquid medicines. However, dosing errors have occurred with them. ...

  18. Zero gravity liquid mixer

    NASA Technical Reports Server (NTRS)

    Booth, F. W.; Bruce, R. A. (Inventor)

    1973-01-01

    An apparatus for mixing liquids under conditions of zero gravity is disclosed. The apparatus is comprised of a closed reservoir for the liquids, with a means for maintaining a positive pressure on the liquids in the reservoir. A valved liquid supply line is connected to the reservoir for supplying the reservoir with the liquids to be mixed in the reservoir. The portion of the reservoir containing the liquids to be mixed is in communication with a pump which alternately causes a portion of the liquids to flow out of the pump and into the reservoir to mix the liquids. The fluids in the reservoir are in communication through a conduit with the pump which alternately causes a portion of the fluids to flow out of the pump and into the sphere. The conduit connecting the pump and sphere may contain a nozzle or other jet-forming structure such as a venturi for further mixing the fluids.

  19. Liquid level sensing device

    DOEpatents

    Tokarz, Richard D.

    1983-01-01

    A liquid level sensing device comprising a load cell supporting a column or stack of segments freely resting on one another. The density of each element is substantially identical to that of the surrounding liquid. The elements are freely guided within a surrounding tube. As each element is exposed above the liquid level, its weight will be impressed through the column to the load cell, thereby providing a signal at the load cell directly proportional to the liquid level elevation.

  20. Distinct effects of folate pathway genes MTHFR and MTHFD1L on ruminative response style: a potential risk mechanism for depression

    PubMed Central

    Eszlari, N; Kovacs, D; Petschner, P; Pap, D; Gonda, X; Elliott, R; Anderson, I M; Deakin, J F W; Bagdy, G; Juhasz, G

    2016-01-01

    Alterations in the folate pathway have been related to both major depression and cognitive inflexibility; however, they have not been investigated in the genetic background of ruminative response style, which is a form of perseverative cognition and a risk factor for depression. In the present study, we explored the association of rumination (measured by the Ruminative Responses Scale) with polymorphisms of two distinct folate pathway genes, MTHFR rs1801133 (C677T) and MTHFD1L rs11754661, in a combined European white sample from Budapest, Hungary (n=895) and Manchester, United Kingdom (n=1309). Post hoc analysis investigated whether the association could be replicated in each of the two samples, and the relationship between folate pathway genes, rumination, lifetime depression and Brief Symptom Inventory depression score. Despite its functional effect on folate metabolism, the MTHFR rs1801133 showed no effect on rumination. However, the A allele of MTHFD1L rs11754661 was significantly associated with greater rumination, and this effect was replicated in both the Budapest and Manchester samples. In addition, rumination completely mediated the effects of MTHFD1L rs11754661 on depression phenotypes. These findings suggest that the MTHFD1L gene, and thus the C1-THF synthase enzyme of the folate pathway localized in mitochondria, has an important effect on the pathophysiology of depression through rumination, and maybe via this cognitive intermediate phenotype on other mental and physical disorders. Further research should unravel whether the reversible metabolic effect of MTHFD1L is responsible for increased rumination or other long-term effects on brain development. PMID:26926881

  1. Mitochondrial C1-Tetrahydrofolate Synthase (MTHFD1L) Supports the Flow of Mitochondrial One-carbon Units into the Methyl Cycle in Embryos*

    PubMed Central

    Pike, Schuyler T.; Rajendra, Rashmi; Artzt, Karen; Appling, Dean R.

    2010-01-01

    Mitochondrial folate-dependent one-carbon (1-C) metabolism converts 1-C donors such as serine and glycine to formate, which is exported and incorporated into the cytoplasmic tetrahydrofolate (THF) 1-C pool. Developing embryos depend on this mitochondrial pathway to provide 1-C units for cytoplasmic process such as de novo purine biosynthesis and the methyl cycle. This pathway is composed of sequential methylene-THF dehydrogenase, methenyl-THF cyclohydrolase, and 10-formyl-THF synthetase activities. In embryonic mitochondria, the bifunctional MTHFD2 enzyme catalyzes the dehydrogenase and cyclohydrolase reactions, but the enzyme responsible for the mitochondrial synthetase reaction has not been identified in embryos. A monofunctional 10-formyl-THF synthetase (MTHFD1L gene product) functions in adult mitochondria and is a likely candidate for the embryonic activity. Here we show that the MTHFD1L enzyme is present in mitochondria from normal embryonic tissues and embryonic fibroblast cell lines, and embryonic mitochondria possess the ability to synthesize formate from glycine. The MTHFD1L transcript was detected at all stages of mouse embryogenesis examined. In situ hybridizations showed that MTHFD1L was expressed ubiquitously throughout the embryo but with localized regions of higher expression. The spatial pattern of MTHFD1L expression was virtually indistinguishable from that of MTHFD2 and MTHFD1 (cytoplasmic C1-THF synthase) in embryonic day 9.5 mouse embryos, suggesting coordinated regulation. Finally, we show using stable isotope labeling that in an embryonic mouse cell line, greater than 75% of 1-C units entering the cytoplasmic methyl cycle are mitochondrially derived. Thus, a complete pathway of enzymes for supplying 1-C units from the mitochondria to the methyl cycle in embryonic tissues is established. PMID:19948730

  2. Common variations in TERT-CLPTM1L locus are reproducibly associated with the risk of nasopharyngeal carcinoma in Chinese populations

    PubMed Central

    Zhang, Hongxing; Zhai, Yun; Wang, Zhifu; Li, Peiyao; Yu, Lixia; Xia, Xia; Zhang, Ying; Zeng, Yixin; He, Fuchu; Zhou, Gangqiao

    2016-01-01

    Associations between single nucleotide polymorphisms (SNPs) at 5p15 (TERT-CLPTM1L) and multiple cancer types have been reported. We examined whether polymorphisms in the TERT-CLPTM1L locus were related to the risk of developing nasopharyngeal carcinoma (NPC) among Chinese populations. In the first stage, 26 tag SNPs were genotyped in a Guangxi population (855 patients and 1036 controls). In the second stage, the SNPs, which showed significant association, were further genotyped in a Guangdong population (997 patients and 972 controls). Functional analyses were conducted to verify the biological relevance of the associated polymorphism. In the 1st stage, four SNPs (rs2736098, rs2735845, rs402710, and rs401681) were significantly associated with the risk of developing NPC. After the 2nd stage validation, rs2735845 and rs401681 were independently associated with the risk of developing NPC in the additive model (rs2735845, OR = 1.19, 95% CI = 1.04–1.37, P = 0.011; rs401681, OR = 0.85, 95% CI = 0.74–0.99, P = 0.034). Furthermore, we observed higher CLPTM1L messenger RNA levels in fetal mesenchymal stem cells from the rs2735845 G allele carriers compared with that from non-carriers. In addition, using an immunohistochemistry assay, we observed higher TERT and CLPTM1L levels in NPC tissues compared with that in non-cancerous nasopharyngeal tissues. Our findings suggest that polymorphisms in the TERT-CLPTM1L locus may play a role in mediating the susceptibility to NPC in Chinese populations. PMID:26621837

  3. Distinct effects of folate pathway genes MTHFR and MTHFD1L on ruminative response style: a potential risk mechanism for depression.

    PubMed

    Eszlari, N; Kovacs, D; Petschner, P; Pap, D; Gonda, X; Elliott, R; Anderson, I M; Deakin, J F W; Bagdy, G; Juhasz, G

    2016-01-01

    Alterations in the folate pathway have been related to both major depression and cognitive inflexibility; however, they have not been investigated in the genetic background of ruminative response style, which is a form of perseverative cognition and a risk factor for depression. In the present study, we explored the association of rumination (measured by the Ruminative Responses Scale) with polymorphisms of two distinct folate pathway genes, MTHFR rs1801133 (C677T) and MTHFD1L rs11754661, in a combined European white sample from Budapest, Hungary (n=895) and Manchester, United Kingdom (n=1309). Post hoc analysis investigated whether the association could be replicated in each of the two samples, and the relationship between folate pathway genes, rumination, lifetime depression and Brief Symptom Inventory depression score. Despite its functional effect on folate metabolism, the MTHFR rs1801133 showed no effect on rumination. However, the A allele of MTHFD1L rs11754661 was significantly associated with greater rumination, and this effect was replicated in both the Budapest and Manchester samples. In addition, rumination completely mediated the effects of MTHFD1L rs11754661 on depression phenotypes. These findings suggest that the MTHFD1L gene, and thus the C1-THF synthase enzyme of the folate pathway localized in mitochondria, has an important effect on the pathophysiology of depression through rumination, and maybe via this cognitive intermediate phenotype on other mental and physical disorders. Further research should unravel whether the reversible metabolic effect of MTHFD1L is responsible for increased rumination or other long-term effects on brain development. PMID:26926881

  4. Missense variants in the middle domain of DNM1L in cases of infantile encephalopathy alter peroxisomes and mitochondria when assayed in Drosophila.

    PubMed

    Chao, Yu-Hsin; Robak, Laurie A; Xia, Fan; Koenig, Mary K; Adesina, Adekunle; Bacino, Carlos A; Scaglia, Fernando; Bellen, Hugo J; Wangler, Michael F

    2016-05-01

    Defects in organelle dynamics underlie a number of human degenerative disorders, and whole exome sequencing (WES) is a powerful tool for studying genetic changes that affect the cellular machinery. WES may uncover variants of unknown significance (VUS) that require functional validation. Previously, a pathogenic de novo variant in the middle domain of DNM1L (p.A395D) was identified in a single patient with a lethal defect of mitochondrial and peroxisomal fission. We identified two additional patients with infantile encephalopathy and partially overlapping clinical features, each with a novel VUS in the middle domain of DNM1L (p.G350R and p.E379K). To evaluate pathogenicity, we generated transgenic Drosophila expressing wild-type or variant DNM1L. We find that human wild-type DNM1L rescues the lethality as well as specific phenotypes associated with the loss of Drp1 in Drosophila. Neither the p.A395D variant nor the novel variant p.G350R rescue lethality or other phenotypes. Moreover, overexpression of p.A395D and p.G350R in Drosophila neurons, salivary gland and muscle strikingly altered peroxisomal and mitochondrial morphology. In contrast, the other novel variant (p.E379K) rescued lethality and did not affect organelle morphology, although it was associated with a subtle mitochondrial trafficking defect in an in vivo assay. Interestingly, the patient with the p.E379K variant also has a de novo VUS in pyruvate dehydrogenase 1 (PDHA1) affecting the same amino acid (G150) as another case of PDHA1 deficiency suggesting the PDHA1 variant may be pathogenic. In summary, detailed clinical evaluation and WES with functional studies in Drosophila can distinguish different functional consequences of newly-described DNM1L alleles. PMID:26931468

  5. Genetic tracing of the gustatory neural pathway originating from Pkd1l3-expressing type III taste cells in circumvallate and foliate papillae

    PubMed Central

    Yamamoto, Kurumi; Ishimaru, Yoshiro; Ohmoto, Makoto; Matsumoto, Ichiro; Asakura, Tomiko; Abe, Keiko

    2011-01-01

    Polycystic kidney disease 1-like 3 (Pkd1l3) is expressed specifically in sour-sensing type III taste cells that have synaptic contacts with afferent nerve fibers in circumvallate and foliate papillae located in the posterior region of the tongue, though not in fungiform papillae or the palate. To visualize the gustatory neural pathways that originate from type III taste cells in circumvallate and foliate papillae, we established transgenic mouse lines that express the transneuronal tracer wheat germ agglutinin (WGA) under the control of the mouse Pkd1l3 gene promoter/enhancer. The WGA transgene was accurately expressed in Pkd1l3-expressing type III taste cells in circumvallate and foliate papillae. Punctate WGA protein signals appeared to be detected specifically in type III taste cells but not in other types of taste cells. WGA protein was transferred primarily to a subset of neurons located in close proximity to the glossopharyngeal nerve bundles in the nodose/petrosal ganglion. WGA signals were also observed in a small population of neurons in the geniculate ganglion. This result demonstrates the anatomical connection between taste receptor cells in the foliate papillae and the chorda tympani nerves. WGA protein was further conveyed to neurons in a rostro-central subdivision of the nucleus of the solitary tract. These findings demonstrate that the approximately 10 kb 5’-flanking region of the mouse Pkd1l3 gene functions as a type III taste cell-specific promoter/enhancer. In addition, experiments using the pkd1l3-WGA transgenic mice reveal a sour gustatory pathway that originates from taste receptor cells in the posterior region of the tongue. PMID:21883212

  6. DOT1L Activity Promotes Proliferation and Protects Cortical Neural Stem Cells from Activation of ATF4-DDIT3-Mediated ER Stress In Vitro.

    PubMed

    Roidl, Deborah; Hellbach, Nicole; Bovio, Patrick P; Villarreal, Alejandro; Heidrich, Stefanie; Nestel, Sigrun; Grüning, Björn A; Boenisch, Ulrike; Vogel, Tanja

    2016-01-01

    Growing evidence suggests that the lysine methyltransferase DOT1L/KMT4 has important roles in proliferation, survival, and differentiation of stem cells in development and in disease. We investigated the function of DOT1L in neural stem cells (NSCs) of the cerebral cortex. The pharmacological inhibition and shRNA-mediated knockdown of DOT1L impaired proliferation and survival of NSCs. DOT1L inhibition specifically induced genes that are activated during the unfolded protein response (UPR) in the endoplasmic reticulum (ER). Chromatin-immunoprecipitation analyses revealed that two genes encoding for central molecules involved in the ER stress response, Atf4 and Ddit3 (Chop), are marked with H3K79 methylation. Interference with DOT1L activity resulted in transcriptional activation of both genes accompanied by decreased levels of H3K79 dimethylation. Although downstream effectors of the UPR, such as Ppp1r15a/Gadd34, Atf3, and Tnfrsf10b/Dr5 were also transcriptionally activated, this most likely occurred in response to increased ATF4 expression rather than as a direct consequence of altered H3K79 methylation. While stem cells are particularly vulnerable to stress, the UPR and ER stress have not been extensively studied in these cells yet. Since activation of the ER stress program is also implicated in directing stem cells into differentiation or to maintain a proliferative status, the UPR must be tightly regulated. Our and published data suggest that histone modifications, including H3K4me3, H3K14ac, and H3K79me2, are implicated in the control of transcriptional activation of ER stress genes. In this context, the loss of H3K79me2 at the Atf4- and Ddit3-promoters appears to mark a point-of-no-return that activates the death program in NSCs. PMID:26299268

  7. Renewable liquid reflection grating

    DOEpatents

    Ryutov, Dmitri D.; Toor, Arthur

    2003-10-07

    A renewable liquid reflection grating. Electrodes are operatively connected to a conducting liquid in an arrangement that produces a reflection grating and driven by a current with a resonance frequency. In another embodiment, the electrodes create the grating by a resonant electrostatic force acting on a dielectric liquid.

  8. Liquid detection circuit

    DOEpatents

    Regan, Thomas O.

    1987-01-01

    Herein is a circuit which is capable of detecting the presence of liquids, especially cryogenic liquids, and whose sensor will not overheat in a vacuum. The circuit parameters, however, can be adjusted to work with any liquid over a wide range of temperatures.

  9. Liquid metal cold trap

    DOEpatents

    Hundal, Rolv

    1976-01-01

    A cold trap assembly for removing impurities from a liquid metal being provided with a hole between the incoming impure liquid metal and purified outgoing liquid metal which acts as a continuous bleed means and thus prevents the accumulation of cover gases within the cold trap assembly.

  10. Variation in PTCHD2, CRISP3, NAP1L4, FSCB, and AP3B2 associated with spherical equivalent

    PubMed Central

    Chen, Fei; Duggal, Priya; Klein, Barbara E.K.; Lee, Kristine E.; Truitt, Barbara; Klein, Ronald; Iyengar, Sudha K.

    2016-01-01

    Purpose Ocular refraction is measured in spherical equivalent as the power of the external lens required to focus images on the retina. Myopia (nearsightedness) and hyperopia (farsightedness) are the most common refractive errors, and the leading causes of visual impairment and blindness in the world. The goal of this study is to identify rare and low-frequency variants that influence spherical equivalent. Methods We conducted variant-level and gene-level quantitative trait association analyses for mean spherical equivalent, using data from 1,560 individuals in the Beaver Dam Eye Study. Genotyping was conducted using the Illumina exome array. We analyzed 34,976 single nucleotide variants and 11,571 autosomal genes across the genome, using single-variant tests as well as gene-based tests. Results Spherical equivalent was significantly associated with five genes in gene-based analysis: PTCHD2 at 1p36.22 (p = 3.6 × 10−7), CRISP3 at 6p12.3 (p = 4.3 × 10−6), NAP1L4 at 11p15.5 (p = 3.6 × 10−6), FSCB at 14q21.2 (p = 1.5 × 10−7), and AP3B2 at 15q25.2 (p = 1.6 × 10−7). The variant-based tests identified evidence suggestive of association with two novel variants in linkage disequilibrium (pairwise r2 = 0.80) in the TCTE1 gene region at 6p21.1 (rs2297336, minor allele frequency (MAF) = 14.1%, β = –0.62 p = 3.7 × 10−6; rs324146, MAF = 16.9%, β = –0.55, p = 1.4 × 10−5). In addition to these novel findings, we successfully replicated a previously reported association with rs634990 near GJD2 at 15q14 (MAF = 47%, β = –0.29, p=1.8 × 10−3). We also found evidence of association with spherical equivalent on 2q37.1 in PRSS56 at rs1550094 (MAF = 31%, β = –0.33, p = 1.7 × 10−3), a region previously associated with myopia. Conclusions We identified several novel candidate genes that may play a role in the control of spherical equivalent. However, further studies are needed to replicate these findings. In addition, our results contribute to the

  11. Supported liquid membranes

    SciTech Connect

    Danesi, P.R.

    1984-01-01

    The possibility of utilizing thin layers of organic solutions of solvent extraction reagents, immobilized on microporous inert supports interposed between two aqueous solutions, for selectively removing metal ions from a mixture represents an attractive alternative to liquid-liquid extraction. A detailed knowledge of the liquid-liquid extraction equilibria and mass transfer kinetics is required to understand and to describe quantitatively the rate laws which control the permeation of metal species through Supported Liquid Membranes (SLM) and to exploit them for separation processes. This paper attempts to understand the mechanism of transport through SLM.

  12. Radiation monitor for liquids

    DOEpatents

    Koster, James E.; Bolton, Richard D.

    1999-01-01

    A radiation monitor for use with liquids that utilizes air ions created by alpha radiation emitted by the liquids as its detectable element. A signal plane, held at an electrical potential with respect to ground, collects these air ions. A guard plane or guard rings is used to limit leakage currents. In one embodiment, the monitor is used for monitoring liquids retained in a tank. Other embodiments monitor liquids flowing through a tank, and bodies of liquids, such as ponds, lakes, rivers and oceans.

  13. Radiation monitor for liquids

    DOEpatents

    Koster, J.E.; Bolton, R.D.

    1999-03-02

    A radiation monitor for use with liquids that utilizes air ions created by alpha radiation emitted by the liquids as its detectable element. A signal plane, held at an electrical potential with respect to ground, collects these air ions. A guard plane or guard rings is used to limit leakage currents. In one embodiment, the monitor is used for monitoring liquids retained in a tank. Other embodiments monitor liquids flowing through a tank, and bodies of liquids, such as ponds, lakes, rivers and oceans. 4 figs.

  14. Liquid Wall Chambers

    SciTech Connect

    Meier, W R

    2011-02-24

    The key feature of liquid wall chambers is the use of a renewable liquid layer to protect chamber structures from target emissions. Two primary options have been proposed and studied: wetted wall chambers and thick liquid wall (TLW) chambers. With wetted wall designs, a thin layer of liquid shields the structural first wall from short ranged target emissions (x-rays, ions and debris) but not neutrons. Various schemes have been proposed to establish and renew the liquid layer between shots including flow-guiding porous fabrics (e.g., Osiris, HIBALL), porous rigid structures (Prometheus) and thin film flows (KOYO). The thin liquid layer can be the tritium breeding material (e.g., flibe, PbLi, or Li) or another liquid metal such as Pb. TLWs use liquid jets injected by stationary or oscillating nozzles to form a neutronically thick layer (typically with an effective thickness of {approx}50 cm) of liquid between the target and first structural wall. In addition to absorbing short ranged emissions, the thick liquid layer degrades the neutron flux and energy reaching the first wall, typically by {approx}10 x x, so that steel walls can survive for the life of the plant ({approx}30-60 yrs). The thick liquid serves as the primary coolant and tritium breeding material (most recent designs use flibe, but the earliest concepts used Li). In essence, the TLW places the fusion blanket inside the first wall instead of behind the first wall.

  15. Acidic Ionic Liquids.

    PubMed

    Amarasekara, Ananda S

    2016-05-25

    Ionic liquid with acidic properties is an important branch in the wide ionic liquid field and the aim of this article is to cover all aspects of these acidic ionic liquids, especially focusing on the developments in the last four years. The structural diversity and synthesis of acidic ionic liquids are discussed in the introduction sections of this review. In addition, an unambiguous classification system for various types of acidic ionic liquids is presented in the introduction. The physical properties including acidity, thermo-physical properties, ionic conductivity, spectroscopy, and computational studies on acidic ionic liquids are covered in the next sections. The final section provides a comprehensive review on applications of acidic ionic liquids in a wide array of fields including catalysis, CO2 fixation, ionogel, electrolyte, fuel-cell, membrane, biomass processing, biodiesel synthesis, desulfurization of gasoline/diesel, metal processing, and metal electrodeposition. PMID:27175515

  16. Duality of liquids

    PubMed Central

    Trachenko, K.; Brazhkin, V. V.

    2013-01-01

    Liquids flow, and in this sense are close to gases. At the same time, interactions in liquids are strong as in solids. The combination of these two properties is believed to be the ultimate obstacle to constructing a general theory of liquids. Here, we adopt a new approach: instead of focusing on the problem of strong interactions, we zero in on the relative contributions of vibrational and diffusional motion. We show that liquid energy and specific heat are given, to a very good approximation, by their vibrational contributions as in solids over almost entire range of relaxation time in which liquids exist as such, and demonstrate that this result is consistent with liquid entropy exceeding solid entropy. Our analysis therefore reveals an interesting duality of liquids not hitherto known: they are close to solids from the thermodynamic perspective and to flowing gases. We discuss several implications of this result. PMID:23851971

  17. Nanowire liquid pumps

    NASA Astrophysics Data System (ADS)

    Huang, Jian Yu; Lo, Yu-Chieh; Niu, Jun Jie; Kushima, Akihiro; Qian, Xiaofeng; Zhong, Li; Mao, Scott X.; Li, Ju

    2013-04-01

    The ability to form tiny droplets of liquids and control their movements is important in printing or patterning, chemical reactions and biological assays. So far, such nanofluidic capabilities have principally used components such as channels, nozzles or tubes, where a solid encloses the transported liquid. Here, we show that liquids can flow along the outer surface of solid nanowires at a scale of attolitres per second and the process can be directly imaged with in situ transmission electron microscopy. Microscopy videos show that an ionic liquid can be pumped along tin dioxide, silicon or zinc oxide nanowires as a thin precursor film or as beads riding on the precursor film. Theoretical analysis suggests there is a critical film thickness of ~10 nm below which the liquid flows as a flat film and above which it flows as discrete beads. This critical thickness is the result of intermolecular forces between solid and liquid, which compete with liquid surface energy and Rayleigh-Plateau instability.

  18. Liquid/Gas Vortex Separator

    NASA Technical Reports Server (NTRS)

    Morris, B. G.

    1986-01-01

    Liquid/gas separator vents gas from tank of liquid that contains gas randomly distributed in bubbles. Centrifugal force separates liquid and gas, forcing liquid out of vortex tube through venturi tube. Gas vented through exhaust port. When liquid detected in vent tube, exhaust port closed, and liquid/gas mixture in vent tube drawn back into tank through venturi.

  19. Recent developments in nucleon spin structure with focus on h_{1L}^{\\perp} and pretzelosity h_{1T}^\\perp

    SciTech Connect

    Avagyan, Harutyun; Efremov, A.V.; Schweitzer, Peter; Metz, Andreas; Teckentrup, T.

    2008-07-20

    The leading twist transverse momentum dependent parton distribution functions h_{1L}^{\\perp} and h_{1T}^\\perp, which is sometimes called "pretzelosity," are studied. For h_{1L}^{\\perp} we consider a "Wandzura-Wilczek-type" approximation, which follows from QCD equations of motion upon the neglect of pure twist-3 terms and allows to express it in terms of transversity. On the basis of available data from HERMES we test the practical usefulness of this approximation and discuss how it can be further tested by future CLAS and COMPASS data. We review the theoretical properties of retzelosity and observe an interesting relation valid in a large class of relativistic models: The difference between helicity and transversity distributions, which is often said to be a 'measure of relativistic effects' in the nucleon, is nothing but the transverse moment of pretzelosity. We discuss preliminary deuteron target data from COMPASS on the single spin asymmetry (SSA) in semi-incl

  20. Low thrust spacecraft transfers optimization method with the stepwise control structure in the Earth-Moon system in terms of the L1-L2 transfer

    NASA Astrophysics Data System (ADS)

    Fain, M. K.; Starinova, O. L.

    2016-04-01

    The paper outlines the method for determination of the locally optimal stepwise control structure in the problem of the low thrust spacecraft transfer optimization in the Earth-Moon system, including the L1-L2 transfer. The total flight time as an optimization criterion is considered. The optimal control programs were obtained by using the Pontryagin's maximum principle. As a result of optimization, optimal control programs, corresponding trajectories, and minimal total flight times were determined.

  1. Expansion of a 12-kb VNTR containing the REXO1L1 gene cluster underlies the microscopically visible euchromatic variant of 8q21.2

    PubMed Central

    Tyson, Christine; Sharp, Andrew J; Hrynchak, Monica; Yong, Siu L; Hollox, Edward J; Warburton, Peter; Barber, John CK

    2014-01-01

    Copy number variants visible with the light microscope have been described as euchromatic variants (EVs) and EVs with extra G-light material at 8q21.2 have been reported only once before. We report four further patients with EVs of 8q21.2 ascertained for clinical (3) or reproductive reasons (1). Enhanced signal strength from two overlapping bacterial artificial chromosomes (BACs) and microarray analysis mapped the EV to a 284-kb interval in the reference genome. This interval consists of a sequence gap flanked by segmental duplications that contain the 12-kb components of one of the largest Variable Number Tandem Repeat arrays in the human genome. Using digital NanoString technology with a custom probe for the RNA exonuclease 1 homologue (S. cerevisiae)-like 1 (REXO1L1) gene within each 12-kb repeat, significantly enhanced diploid copy numbers of 270 and 265 were found in an EV family and a median diploid copy number of 166 copies in 216 controls. These 8q21.2 EVs are not thought to have clinical consequences as the phenotypes of the probands were inconsistent, those referred for reproductive reasons were otherwise phenotypically normal and the REXO1L1 gene has no known disease association. This EV was found in 4/3078 (1 in 770) consecutive referrals for chromosome analysis and needs to be distinguished from pathogenic imbalances of medial 8q. The REXO1L1 gene product is a marker of hepatitis C virus (HCV) infection and a possible association between REXO1L1 copy number and susceptibility to HCV infection, progression or response to treatment has not yet been excluded. PMID:24045839

  2. Investigating the Impact of a Genome-Wide Supported Bipolar Risk Variant of MAD1L1 on the Human Reward System.

    PubMed

    Trost, Sarah; Diekhof, Esther K; Mohr, Holger; Vieker, Henning; Krämer, Bernd; Wolf, Claudia; Keil, Maria; Dechent, Peter; Binder, Elisabeth B; Gruber, Oliver

    2016-10-01

    Recent genome-wide association studies have identified MAD1L1 (mitotic arrest deficient-like 1) as a susceptibility gene for bipolar disorder and schizophrenia. The minor allele of the single-nucleotide polymorphism (SNP) rs11764590 in MAD1L1 was associated with bipolar disorder. Both diseases, bipolar disorder and schizophrenia, are linked to functional alterations in the reward system. We aimed at investigating possible effects of the MAD1L1 rs11764590 risk allele on reward systems functioning in healthy adults. A large homogenous sample of 224 young (aged 18-31 years) participants was genotyped and underwent functional magnetic resonance imaging (fMRI). All participants performed the 'Desire-Reason Dilemma' paradigm investigating the neural correlates that underlie reward processing and active reward dismissal in favor of a long-term goal. We found significant hypoactivations of the ventral tegmental area (VTA), the bilateral striatum and bilateral frontal and parietal cortices in response to conditioned reward stimuli in the risk allele carriers compared with major allele carriers. In the dilemma situation, functional connectivity between prefrontal brain regions and the ventral striatum was significantly diminished in the risk allele carriers. Healthy risk allele carriers showed a significant deficit of their bottom-up response to conditioned reward stimuli in the bilateral VTA and striatum. Furthermore, functional connectivity between the ventral striatum and prefrontal areas exerting top-down control on the mesolimbic reward system was reduced in this group. Similar alterations in reward processing and disturbances of prefrontal control mechanisms on mesolimbic brain circuits have also been reported in bipolar disorder and schizophrenia. Together, these findings suggest the existence of an intermediate phenotype associated with MAD1L1. PMID:27184339

  3. Germline mosaic transmission of a novel duplication of PXDN and MYT1L to two male half-siblings with autism

    PubMed Central

    Meyer, Kacie J.; Axelsen, Michael S.; Sheffield, Val C.; Patil, Shivanand R.; Wassink, Thomas H.

    2011-01-01

    Objective Autism is a neurodevelopmental disorder with a strong genetic component to susceptibility. Here, we report the molecular characterization of an apparent de novo 281 kb duplication of Chromosome 2p25.3 in two male half-siblings with autism. Methods The 2p25.3 duplication was first identified through a low-density microarray, validated with FISH, and duplication breakpoints were delineated using an Affymetrix 6.0 SNP microarray. Results FISH results validated the novel copy number variant and revealed the mother to be mosaic, with ~33% of her lymphoblast cells carrying the duplication. Therefore, the duplication was transmitted through the mechanism of germline mosaicism. Additionally, duplication breakpoints were refined and show that PXDN is fully duplicated, while seven exons of the terminal portion of the 25 exon gene MYT1L are within the duplicated region. Conclusions MYT1L, a gene predominately expressed in the brain, has recently been linked to other neuropsychiatric illness such as schizophrenia and depression. Results from this study indicate that the 2p25.3 duplication disrupting PXDN and MYT1L is a potential autism-causing variant in the pedigree reported here and should receive further consideration as a candidate gene for autism. PMID:22157634

  4. A Case Report on Extruded Disc Acting as Buttonhole reventing Reduction in Traumatic Fracture and Lateral Dislocation of L1-l2 Vertebrae

    PubMed Central

    Sundaram, P.Shanmuga; Kanniyan, Kalaivanan; Bijarnia1, Isha

    2014-01-01

    Introduction: There is a scanty literature support describing the incidence, mechanism of lateral dislocations of thoracolumbar spine and its management describing the reduction techniques. Hereby we present an interesting case of extruded disc acting as buttonhole preventing the reduction of lateral dislocation of L1-L2 vertebrae and this would be the first of its variety to be described ever in literature. Case Report: A 30 year old female was referred to our hospital on post trauma day 7 with bilateral fascet fracture and lateral dislocation of L1-L2 vertebrae and fracture humerus on right arm following a road traffic accident. She presented with clinical signs consistent of cauda equina lesion. She underwent surgical reduction and TLIF L1-L2. On one year follow-up X-Rays showed maintenance of dorsolumbar saggital alignment without collapse. Conclusion: Dislocations of spine are three column injuries, are highly unstable requiring surgical stabilisation. Posterior instrumentation is the routinely followed technique to achieve reduction and for posterolateral fusion. In this case lateral dislocation was reduced only after removal of the laterally extruded disc. After complete discectomy TLIF was considered a good option for restoring disc space height and maintaining saggital balance. PMID:27298983

  5. A Homozygous Mutation in LYRM7/MZM1L Associated with Early Onset Encephalopathy, Lactic Acidosis, and Severe Reduction of Mitochondrial Complex III Activity

    PubMed Central

    Invernizzi, Federica; Tigano, Marco; Dallabona, Cristina; Donnini, Claudia; Ferrero, Ileana; Cremonte, Maurizio; Ghezzi, Daniele; Lamperti, Costanza; Zeviani, Massimo

    2013-01-01

    Mutations in nuclear genes associated with defective complex III (cIII) of the mitochondrial respiratory chain are rare, having been found in only two cIII assembly factors and, as private changes in single families, three cIII structural subunits. Recently, human LYRM7/MZM1L, the ortholog of yeast MZM1, has been identified as a new assembly factor for cIII. In a baby patient with early onset, severe encephalopathy, lactic acidosis and profound, isolated cIII deficiency in skeletal muscle, we identified a disease-segregating homozygous mutation (c.73G>A) in LYRM7/MZM1L, predicting a drastic change in a highly conserved amino-acid residue (p.Asp25Asn). In a mzm1Δ yeast strain, the expression of a mzm1D25N mutant allele caused temperature-sensitive respiratory growth defect, decreased oxygen consumption, impaired maturation/stabilization of the Rieske Fe–S protein, and reduced complex III activity and amount. LYRM7/MZM1L is a novel disease gene, causing cIII-defective, early onset, severe mitochondrial encephalopathy. PMID:24014394

  6. AG490 and PF431396 Sensitive Tyrosine Kinase Control the Population Heterogeneity of Basal STAT1 Activity in Ube1l Deficient Cells

    PubMed Central

    Now, Hesung; Yoo, Joo-Yeon

    2016-01-01

    A population often contains distinct sub-populations, thereby increasing the complexity of the overall heterogeneity. However, the cellular origin and biological relevance of sub-populations in cell population have not been clearly identified. Here we demonstrated the novel roles of ISGylation, which is an IFN-induced post-translational modification, controlling heterogeneity at the population level in cultured adherent cells. Without UBE1L, an E1 enzyme of ISGylation, mouse embryonic fibroblasts (MEF) exhibited low viral resistance despite high STAT1 and ISG expression compared with the wild-type MEF. We observe that Ube1l−/− MEF populations consist of two behaviorally distinguishable sub-populations with distinct basal STAT1 activity, while wild-type MEF populations are unimodal. This population heterogeneity in Ube1l knock-out cells was perturbed by tyrosine kinase inhibitors, AG490 and PF431396. In contrast, the neutralization of type I IFN did not affect population heterogeneity. Based on these results, we concluded that UBE1L functions to adjust basal immunological states with the regulation of population heterogeneity. PMID:27427993

  7. No association of TGFB1 L10P genotypes and breast cancer risk in BRCA1 and BRCA2 mutation carriers: a multi-center cohort study

    PubMed Central

    Antoniou, Antonis C.; Llopis, Trinidad Caldes; Nevanlinna, Heli; Aittomäki, Kristiina; Simard, Jacques; Spurdle, Amanda B.; Couch, Fergus J.; Pereira, Lutecia H. Mateus; Greene, Mark H.; Andrulis, Irene L.; Pasche, Boris; Kaklamani, Virginia; Hamann, Ute; Szabo, Csilla; Peock, Susan; Cook, Margaret; Harrington, Patricia A.; Donaldson, Alan; Male, Allison M.; Gardiner, Carol Anne; Gregory, Helen; Side, Lucy E.; Robinson, Anne C.; Emmerson, Louise; Ellis, Ian; Peyrat, Jean-Philippe; Fournier, Joëlle; Vennin, Philippe; Adenis, Claude; Muller, Danièle; Fricker, Jean-Pierre; Longy, Michel; Sinilnikova, Olga M.; Stoppa-Lyonnet, Dominique; Schmutzler, Rita K.; Versmold, Beatrix; Engel, Christoph; Meindl, Alfons; Kast, Karin; Schaefer, Dieter; Froster, Ursula G.; Chenevix-Trench, Georgia; Easton, Douglas F.

    2008-01-01

    Background The transforming growth factor β-1 gene (TGFB1) is a plausible candidate for breast cancer susceptibility. The L10P variant of TGFB1 is associated with higher circulating levels and secretion of TGF-β, and recent large-scale studies suggest strongly that this variant is associated with breast cancer risk in the general population. Methods To evaluate whether TGFB1 L10P also modifies the risk of breast cancer in BRCA1 or BRCA2 mutation carriers, we undertook a multi-center study of 3,442 BRCA1 and 2,095 BRCA2 mutation carriers. Results We found no evidence of association between TGFB1 L10P and breast cancer risk in either BRCA1 or BRCA2 mutation carriers. The per-allele HR for the L10P variant was 1.01 (95%CI: 0.92–1.11) in BRCA1 carriers and 0.92 (95%CI: 0.81–1.04) in BRCA2 mutation carriers. Conclusions These results do not support the hypothesis that TGFB1 L10P genotypes modify the risk of breast cancer in BRCA1 or BRCA2 mutation carriers. PMID:18523885

  8. Liquid Sloshing Dynamics

    NASA Astrophysics Data System (ADS)

    Ibrahim, Raouf A.

    2005-06-01

    The problem of liquid sloshing in moving or stationary containers remains of great concern to aerospace, civil, and nuclear engineers; physicists; designers of road tankers and ship tankers; and mathematicians. Beginning with the fundamentals of liquid sloshing theory, this book takes the reader systematically from basic theory to advanced analytical and experimental results in a self-contained and coherent format. The book is divided into four sections. Part I deals with the theory of linear liquid sloshing dynamics; Part II addresses the nonlinear theory of liquid sloshing dynamics, Faraday waves, and sloshing impacts; Part III presents the problem of linear and nonlinear interaction of liquid sloshing dynamics with elastic containers and supported structures; and Part IV considers the fluid dynamics in spinning containers and microgravity sloshing. This book will be invaluable to researchers and graduate students in mechanical and aeronautical engineering, designers of liquid containers, and applied mathematicians.

  9. Monogroove liquid heat exchanger

    NASA Technical Reports Server (NTRS)

    Brown, Richard F. (Inventor); Edelstein, Fred (Inventor)

    1990-01-01

    A liquid supply control is disclosed for a heat transfer system which transports heat by liquid-vapor phase change of a working fluid. An assembly (10) of monogroove heat pipe legs (15) can be operated automatically as either heat acquisition devices or heat discharge sources. The liquid channels (27) of the heat pipe legs (15) are connected to a reservoir (35) which is filled and drained by respective filling and draining valves (30, 32). Information from liquid level sensors (50, 51) on the reservoir (35) is combined (60) with temperature information (55) from the liquid heat exchanger (12) and temperature information (56) from the assembly vapor conduit (42) to regulate filling and draining of the reservoir (35), so that the reservoir (35) in turn serves the liquid supply/drain needs of the heat pipe legs (15), on demand, by passive capillary action (20, 28).

  10. Liquid level detector

    DOEpatents

    Tshishiku, Eugene M.

    2011-08-09

    A liquid level detector for conductive liquids for vertical installation in a tank, the detector having a probe positioned within a sheath and insulated therefrom by a seal so that the tip of the probe extends proximate to but not below the lower end of the sheath, the lower end terminating in a rim that is provided with notches, said lower end being tapered, the taper and notches preventing debris collection and bubble formation, said lower end when contacting liquid as it rises will form an airtight cavity defined by the liquid, the interior sheath wall, and the seal, the compression of air in the cavity preventing liquid from further entry into the sheath and contact with the seal. As a result, the liquid cannot deposit a film to form an electrical bridge across the seal.

  11. Harvesting contaminants from liquid

    DOEpatents

    Simpson, John T.; Hunter, Scott R.

    2016-05-31

    Disclosed are examples of apparatuses for evaporative purification of a contaminated liquid. In each example, there is a vessel for storing the contaminated fluid. The vessel includes a surface coated with a layer of superhydrophobic material and the surface is at least partially in contact with the contaminated liquid. The contaminants do not adhere to the surface as the purified liquid evaporates, thus allowing the contaminants to be harvested.

  12. LIQUID CYCLONE CONTACTOR

    DOEpatents

    Whatley, M.E.; Woods, W.M.

    1962-09-01

    This invention relates to liquid-liquid extraction systems. The invention, an improved hydroclone system, comprises a series of serially connected, axially aligned hydroclones, each of which is provided with an axially aligned overflow chamber. The chambers are so arranged that rotational motion of a fluid being passed through the system is not lost in passing from chamber to chamber; consequently, this system is highly efficient in contacting and separating two immiscible liquids. (AEC)

  13. The l1-l2 regularization framework unmasks the hypoxia signature hidden in the transcriptome of a set of heterogeneous neuroblastoma cell lines

    PubMed Central

    Fardin, Paolo; Barla, Annalisa; Mosci, Sofia; Rosasco, Lorenzo; Verri, Alessandro; Varesio, Luigi

    2009-01-01

    Background Gene expression signatures are clusters of genes discriminating different statuses of the cells and their definition is critical for understanding the molecular bases of diseases. The identification of a gene signature is complicated by the high dimensional nature of the data and by the genetic heterogeneity of the responding cells. The l1-l2 regularization is an embedded feature selection technique that fulfills all the desirable properties of a variable selection algorithm and has the potential to generate a specific signature even in biologically complex settings. We studied the application of this algorithm to detect the signature characterizing the transcriptional response of neuroblastoma tumor cell lines to hypoxia, a condition of low oxygen tension that occurs in the tumor microenvironment. Results We determined the gene expression profile of 9 neuroblastoma cell lines cultured under normoxic and hypoxic conditions. We studied a heterogeneous set of neuroblastoma cell lines to mimic the in vivo situation and to test the robustness and validity of the l1-l2 regularization with double optimization. Analysis by hierarchical, spectral, and k-means clustering or supervised approach based on t-test analysis divided the cell lines on the bases of genetic differences. However, the disturbance of this strong transcriptional response completely masked the detection of the more subtle response to hypoxia. Different results were obtained when we applied the l1-l2 regularization framework. The algorithm distinguished the normoxic and hypoxic statuses defining signatures comprising 3 to 38 probesets, with a leave-one-out error of 17%. A consensus hypoxia signature was established setting the frequency score at 50% and the correlation parameter ε equal to 100. This signature is composed by 11 probesets representing 8 well characterized genes known to be modulated by hypoxia. Conclusion We demonstrate that l1-l2 regularization outperforms more conventional

  14. PEP liquid level system

    SciTech Connect

    Lauritzen, T.; Sah, R.C.

    1981-03-01

    A liquid level system has been installed in the accelerator housing of the PEP storage ring. This instrument spans the entire 2.2 km circumference of the PEP project, and over one hundred readouts provide reference elevations which are used for the accurate alignment of accelerator components. The liquid level has proven to be extremely precise (+-0.10 mm) and quick to use, and it has contributed to the accurate alignment of PEP before beam turn-on. Since the liquid level readouts are rigidly attached to the accelerator housing, the liquid level has been a convenient means to monitor the settling of the accelerator housing.

  15. Liquid Crystal Optofluidics

    SciTech Connect

    Vasdekis, Andreas E.; Cuennet, J. G.; Psaltis, D.

    2012-10-11

    By employing anisotropic fluids and namely liquid crystals, fluid flow becomes an additional degree of freedom in designing optofluidic devices. In this paper, we demonstrate optofluidic liquid crystal devices based on the direct flow of nematic liquid crystals in microfluidic channels. Contrary to previous reports, in the present embodiment we employ the effective phase delay acquired by light travelling through flowing liquid crystal, without analysing the polarisation state of the transmitted light. With this method, we demonstrate the variation in the diffraction pattern of an array of microfluidic channels acting as a grating. We also discuss our recent activities in integrating mechanical oscillators for on-chip peristaltic pumping.

  16. Liquid explosives detection

    NASA Astrophysics Data System (ADS)

    Burnett, Lowell J.

    1994-03-01

    A Liquid Explosives Screening System capable of scanning unopened bottles for liquid explosives has been developed. The system can be operated to detect specific explosives directly, or to verify the labeled or bar-coded contents of the container. In this system nuclear magnetic resonance (NMR) is used to interrogate the liquid. NMR produces an extremely rich data set and many parameters of the NMR response can be determined simultaneously. As a result, multiple NMR signatures may be defined for any given set of liquids, and the signature complexity then selected according to the level of threat.

  17. Liquid level detector

    DOEpatents

    Tokarz, Richard D.

    1982-01-01

    A liquid level sensor having a pair of upright conductors spaced by an insulator defining a first high resistance path between the conductors. An electrically conductive path is interposed between the upright conductors at a discrete location at which liquid level is to be measured. It includes a liquid accessible gap of a dimension such that the electrical resistance across the conductor when the gap is filled with the liquid is detectably less than when the gap is emptied. The conductor might also be physically altered by temperature changes to serve also as an indicator of elevated temperature.

  18. RENEWABLE LIQUID GETTERING PUMP

    DOEpatents

    Batzer, T.H.

    1962-08-21

    A method and structure were developed for pumping gases by simple absorption into a liquid gettering material. The invention comprises means ror continuously pumping a liquid getterrng material from a reservoir to the top of a generally vertical surface disposed in a vacuum pumping chamber to receive gaseous and other particles in the liquid gettering material which continuously flows downward over the vertical suiface. Means are provided for continuous removal, degassing, and return of a portion of the liquid gettering material from the reservoir connected with collectrng means at the base of the generally vertical plate. (AEC)

  19. Ultrasonic liquid level detector

    DOEpatents

    Kotz, Dennis M.; Hinz, William R.

    2010-09-28

    An ultrasonic liquid level detector for use within a shielded container, the detector being tubular in shape with a chamber at its lower end into which liquid from in the container may enter and exit, the chamber having an ultrasonic transmitter and receiver in its top wall and a reflector plate or target as its bottom wall whereby when liquid fills the chamber a complete medium is then present through which an ultrasonic wave may be transmitted and reflected from the target thus signaling that the liquid is at chamber level.

  20. Liquid heat capacity lasers

    DOEpatents

    Comaskey, Brian J.; Scheibner, Karl F.; Ault, Earl R.

    2007-05-01

    The heat capacity laser concept is extended to systems in which the heat capacity lasing media is a liquid. The laser active liquid is circulated from a reservoir (where the bulk of the media and hence waste heat resides) through a channel so configured for both optical pumping of the media for gain and for light amplification from the resulting gain.

  1. INEEL Liquid Effluent Inventory

    SciTech Connect

    Major, C.A.

    1997-06-01

    The INEEL contractors and their associated facilities are required to identify all liquid effluent discharges that may impact the environment at the INEEL. This liquid effluent information is then placed in the Liquid Effluent Inventory (LEI) database, which is maintained by the INEEL prime contractor. The purpose of the LEI is to identify and maintain a current listing of all liquid effluent discharge points and to identify which discharges are subject to federal, state, or local permitting or reporting requirements and DOE order requirements. Initial characterization, which represents most of the INEEL liquid effluents, has been performed, and additional characterization may be required in the future to meet regulations. LEI information is made available to persons responsible for or concerned with INEEL compliance with liquid effluent permitting or reporting requirements, such as the National Pollutant Discharge Elimination System, Wastewater Land Application, Storm Water Pollution Prevention, Spill Prevention Control and Countermeasures, and Industrial Wastewater Pretreatment. The State of Idaho Environmental Oversight and Monitoring Program also needs the information for tracking liquid effluent discharges at the INEEL. The information provides a baseline from which future liquid discharges can be identified, characterized, and regulated, if appropriate. The review covered new and removed buildings/structures, buildings/structures which most likely had new, relocated, or removed LEI discharge points, and at least 10% of the remaining discharge points.

  2. Precision liquid level sensor

    DOEpatents

    Field, M.E.; Sullivan, W.H.

    1985-01-29

    A precision liquid level sensor utilizes a balanced R. F. bridge, each arm including an air dielectric line. Changes in liquid level along one air dielectric line imbalance the bridge and create a voltage which is directly measurable across the bridge. 2 figs.

  3. Precision liquid level sensor

    DOEpatents

    Field, Michael E.; Sullivan, William H.

    1985-01-01

    A precision liquid level sensor utilizes a balanced R. F. bridge, each arm including an air dielectric line. Changes in liquid level along one air dielectric line imbalance the bridge and create a voltage which is directly measurable across the bridge.

  4. LIGHT NONAQUEOUS PHASE LIQUIDS

    EPA Science Inventory

    Nonaqueous phase liquids (NAPLS) are hydrocarbons that exist as a separate, immiscible phase when in contact with water and/or air. ifferences in the physical and chemical properties of water and NAPL result in the formation of a physical interface between the liquids which preve...

  5. Column Liquid Chromatography.

    ERIC Educational Resources Information Center

    Majors, Ronald E.; And Others

    1984-01-01

    Reviews literature covering developments of column liquid chromatography during 1982-83. Areas considered include: books and reviews; general theory; columns; instrumentation; detectors; automation and data handling; multidimensional chromatographic and column switching techniques; liquid-solid chromatography; normal bonded-phase, reversed-phase,…

  6. Synthesis of ionic liquids

    DOEpatents

    Dai, Sheng; Luo, Huimin

    2011-11-01

    Ionic compounds which are liquids at room temperature are formed by the method of mixing a neutral organic ligand with the salt of a metal cation and its conjugate anion. The liquids are hydrophobic, conductive and stable and have uses as solvents and in electrochemical devices.

  7. Synthesis of ionic liquids

    DOEpatents

    Dai, Sheng [Knoxville, TN; Luo, Huimin [Knoxville, TN

    2008-09-09

    Ionic compounds which are liquids at room temperature are formed by the method of mixing a neutral organic liqand with the salt of a metal cation and its conjugate anion. The liquids are hydrophobic, conductive and stable and have uses as solvents and in electrochemical devices.

  8. Advanced proteomic liquid chromatography

    SciTech Connect

    Xie, Fang; Smith, Richard D.; Shen, Yufeng

    2012-10-26

    Liquid chromatography coupled with mass spectrometry is the predominant platform used to analyze proteomics samples consisting of large numbers of proteins and their proteolytic products (e.g., truncated polypeptides) and spanning a wide range of relative concentrations. This review provides an overview of advanced capillary liquid chromatography techniques and methodologies that greatly improve separation resolving power and proteomics analysis coverage, sensitivity, and throughput.

  9. Liquid Chromatography in 1982.

    ERIC Educational Resources Information Center

    Freeman, David H.

    1982-01-01

    Reviews trends in liquid chromatography including apparatus, factors affecting efficient separation of a mixture (peak sharpness and speed), simplified problem-solving, adsorption, bonded phase chromatography, ion selectivity, and size exclusion. The current trend is to control chemical selectivity by the liquid phase. (Author/JN)

  10. Tunable liquid crystal lasers

    NASA Astrophysics Data System (ADS)

    Woltman, Scott J.

    Liquid crystal lasers are dye-doped distributed feedback lasing systems. Fabricated by coupling the periodic structure of a liquid crystal medium with a fluorescent dye, the emission from these systems is tunable by controlling the liquid crystal system---be it through electric or thermal field effects, photochemical reactions, mechanical deformations, etc. The laser action arises from an extended interaction time between the radiation field, the laser emission, and the matter field, the periodic liquid crystal medium, at the edge of the photonic band gap. In this thesis, several tunable liquid crystal laser systems are investigated: cholesteric liquid crystals, holographic-polymer dispersed liquid crystals and liquid crystal polarization gratings. The primary focus has been to fabricate systems that are tunable through electrical means, as applications requiring mechanical or thermal changes are often difficult to control. Cholesteric liquid crystal lasers are helical Bragg reflectors, with a band gap for circularly polarized light of equivalent handedness to their helix. These materials were doped with a laser dye and laser emission was observed. The use of an in-plane electric field tends to unwind the helical pitch of the film and in doing so tunable emission was demonstrated for ˜15 nm. Holographic-polymer dispersed liquid crystals (H-PDLCs) are grating structures consisting of alternating layers of polymer and liquid crystal, with different indices of refraction. The application of an electric field index matches these layers and switches off the grating. Thus, laser emission can be switched on and off through the use of an electric field. Spatially tunable H-PDLC lasers were fabricated by creating chirped gratings, formed by divergent beams. The emission was shown to tune ˜5 nm as the pump beam was translated across a 1 inch film. Liquid crystal polarization gratings use photo-patterned alignment layers, through a polarization holography exposure, to

  11. Liquid metal enabled pump

    PubMed Central

    Tang, Shi-Yang; Khoshmanesh, Khashayar; Sivan, Vijay; Petersen, Phred; O’Mullane, Anthony P.; Abbott, Derek; Mitchell, Arnan; Kalantar-zadeh, Kourosh

    2014-01-01

    Small-scale pumps will be the heartbeat of many future micro/nanoscale platforms. However, the integration of small-scale pumps is presently hampered by limited flow rate with respect to the input power, and their rather complicated fabrication processes. These issues arise as many conventional pumping effects require intricate moving elements. Here, we demonstrate a system that we call the liquid metal enabled pump, for driving a range of liquids without mechanical moving parts, upon the application of modest electric field. This pump incorporates a droplet of liquid metal, which induces liquid flow at high flow rates, yet with exceptionally low power consumption by electrowetting/deelectrowetting at the metal surface. We present theory explaining this pumping mechanism and show that the operation is fundamentally different from other existing pumps. The presented liquid metal enabled pump is both efficient and simple, and thus has the potential to fundamentally advance the field of microfluidics. PMID:24550485

  12. Liquid liquid phase transition in Stillinger Weber silicon

    NASA Astrophysics Data System (ADS)

    Beaucage, Philippe; Mousseau, Normand

    2005-04-01

    It was recently demonstrated that Stillinger-Weber silicon undergoes a liquid-liquid first-order phase transition deep into the supercooled region (Sastry and Angell 2003 Nat. Mater. 2 739). Here we study the effects of perturbations on this phase transition. We show that the order of the liquid-liquid transition changes with negative pressure. We also find that the liquid-liquid transition disappears when the three-body term of the potential is strengthened by as little as 5%. This implies that the details of the potential could affect strongly the nature and even the existence of the liquid-liquid phase.

  13. Liquid Water, the ``Most Complex'' Liquid: New Results in Bulk, Nanoconfined, and Biological Environments

    NASA Astrophysics Data System (ADS)

    Stanley, H. Eugene

    2010-03-01

    We will introduce some of the 63 anomalies of the most complex of liquids, water. We will demonstrate some recent progress in understanding these anomalies by combining information provided by recent experiments and simulations on water in bulk, nanoconfined, and biological environments. We will interpret evidence from recent experiments designed to test the hypothesis that liquid water may display ``polymorphism'' in that it can exist in two different phases---and discuss recent work on water's transport anomalies [1] as well as the unusual behavior of water in biological environments [2]. Finally, we will discuss how the general concept of liquid polymorphism [3] is proving useful in understanding anomalies in other liquids, such as silicon, silica, and carbon, as well as metallic glasses, which have in common that they are characterized by two characteristic length scales in their interactions. This work was supported by NSF Chemistry Division, and carried out in collaboration with a number of colleagues, chief among whom are C. A. Angell, M. C. Barbosa, M. C. Bellissent, L. Bosio, F. Bruni, S. V. Buldyrev, M. Canpolat, S. -H. Chen, P. G. Debenedetti, U. Essmann,G. Franzese, A. Geiger, N. Giovambattista, S. Han, P. Kumar, E. La Nave,G. Malescio, F. Mallamace, M. G. Mazza, O. Mishima, P. Netz, P. H. Poole, P. J. Rossky, R. Sadr,S. Sastry, A. Scala, F. Sciortino, A. Skibinsky, F. W. Starr, K. C. Stokely J. Teixeira, L. Xu, and Z. Yan.[4pt] [1] L. Xu, F. Mallamace, Z. Yan, F. W. Starr, S. V. Buldyrev, and H. E. Stanley, ``Appearance of a Fractional Stokes-Einstein Relation in Water and a Structural Interpretation of Its Onset,'' Nature Physics 5, 565--569 (2009). [0pt] [2] P. Kumar, Z. Yan, L. Xu, M. G. Mazza, S. V. Buldyrev, S. -H. Chen. S. Sastry, and H. E. Stanley, ``Glass Transition in Biomolecules and the Liquid-Liquid Critical Point of Water,'' Phys. Rev. Lett. 97, 177802 (2006). [0pt] [3] H. E. Stanley, ed. , Liquid Polymorphism [Advances in Chemical Physics

  14. The α1A-adrenoceptor gene is required for the α1L-adrenoceptor-mediated response in isolated preparations of the mouse prostate

    PubMed Central

    Gray, K T; Short, J L; Ventura, S

    2008-01-01

    Background and purpose: This study investigated whether deletion of the α1A-adrenoceptor gene influences contractile responses of mouse prostate to noradrenaline. Responses of mouse prostate to noradrenaline are known to be mediated by α1L-adrenoceptors, which are thought to be a functional phenotype of α1A-adrenoceptor. Experimental approach: Prostate tissues from α1A-adrenoceptor knockout mice which were homozygous (α1A−/−) and heterozygous (α1A+/−) for the disrupted α1A-adrenoceptor gene, as well as wild-type (α1A+/+) littermates were mounted in glass-isolated organ baths. Electrical field stimulation of nerves and exogenous application of noradrenaline were used to investigate the effects of α1A-adrenoceptor disruption on prostate contractility. Key results: Frequency–response curves to electrical field stimulation (0.5 ms pulse duration, 60 V, 0.1–20 Hz) yielded frequency-dependent contractions. At frequencies of 10 and 20 Hz, prostates from α1A−/− mice elicited an approximately 30% decreased response compared with prostates from α1A+/+ mice. Prazosin (0.3 μM) attenuated responses to electrical field stimulation in prostates from α1A+/+ and α1A+/− mice but not from α1A−/− mice. Increasing concentrations of exogenously administered noradrenaline (10 nM–1 mM) produced mean concentration–response curves in prostates from α1A+/+ and α1A+/− mice, which were not different. Maximum responses to noradrenaline were decreased by approximately 80% in prostates from α1A−/− mice compared with α1A+/+ mice. Prazosin attenuated responses to noradrenaline in all genotypes. Conclusions and implications: α1L-Adrenoceptor-mediated responses in mouse prostate are abolished in α1A−/− mice, demonstrating that the α1A-adrenoceptor gene is essential to the manifestation of the prostatic α1L-adrenoceptor phenotype. This implies that α1L-adrenoceptors are indeed a functional phenotype of α1A-adrenoceptor. PMID

  15. Renewable liquid reflecting zone plate

    DOEpatents

    Toor, Arthur; Ryutov, Dmitri D.

    2003-12-09

    A renewable liquid reflecting zone plate. Electrodes are operatively connected to a dielectric liquid in a circular or other arrangement to produce a reflecting zone plate. A system for renewing the liquid uses a penetrable substrate.

  16. Noncavitating Pump For Liquid Helium

    NASA Technical Reports Server (NTRS)

    Hasenbein, Robert; Izenson, Michael; Swift, Walter; Sixsmith, Herbert

    1996-01-01

    Immersion pump features high efficiency in cryogenic service. Simple and reliable centrifugal pump transfers liquid helium with mass-transfer efficiency of 99 percent. Liquid helium drawn into pump by helical inducer, which pressurizes helium slightly to prevent cavitation when liquid enters impeller. Impeller then pressurizes liquid. Purpose of pump to transfer liquid helium from supply to receiver vessel, or to provide liquid helium flow for testing and experimentation.

  17. Nanowire liquid pumps.

    PubMed

    Huang, Jian Yu; Lo, Yu-Chieh; Niu, Jun Jie; Kushima, Akihiro; Qian, Xiaofeng; Zhong, Li; Mao, Scott X; Li, Ju

    2013-04-01

    The ability to form tiny droplets of liquids and control their movements is important in printing or patterning, chemical reactions and biological assays. So far, such nanofluidic capabilities have principally used components such as channels, nozzles or tubes, where a solid encloses the transported liquid. Here, we show that liquids can flow along the outer surface of solid nanowires at a scale of attolitres per second and the process can be directly imaged with in situ transmission electron microscopy. Microscopy videos show that an ionic liquid can be pumped along tin dioxide, silicon or zinc oxide nanowires as a thin precursor film or as beads riding on the precursor film. Theoretical analysis suggests there is a critical film thickness of ∼10 nm below which the liquid flows as a flat film and above which it flows as discrete beads. This critical thickness is the result of intermolecular forces between solid and liquid, which compete with liquid surface energy and Rayleigh-Plateau instability. PMID:23542904

  18. Liquid sampling system

    DOEpatents

    Larson, Loren L.

    1987-01-01

    A conduit extends from a reservoir through a sampling station and back to the reservoir in a closed loop. A jet ejector in the conduit establishes suction for withdrawing liquid from the reservoir. The conduit has a self-healing septum therein upstream of the jet ejector for receiving one end of a double-ended cannula, the other end of which is received in a serum bottle for sample collection. Gas is introduced into the conduit at a gas bleed between the sample collection bottle and the reservoir. The jet ejector evacuates gas from the conduit and the bottle and aspirates a column of liquid from the reservoir at a high rate. When the withdrawn liquid reaches the jet ejector the rate of flow therethrough reduces substantially and the gas bleed increases the pressure in the conduit for driving liquid into the sample bottle, the gas bleed forming a column of gas behind the withdrawn liquid column and interrupting the withdrawal of liquid from the reservoir. In the case of hazardous and toxic liquids, the sample bottle and the jet ejector may be isolated from the reservoir and may be further isolated from a control station containing remote manipulation means for the sample bottle and control valves for the jet ejector and gas bleed.

  19. Liquid sampling system

    DOEpatents

    Larson, L.L.

    1984-09-17

    A conduit extends from a reservoir through a sampling station and back to the reservoir in a closed loop. A jet ejector in the conduit establishes suction for withdrawing liquid from the reservoir. The conduit has a self-healing septum therein upstream of the jet ejector for receiving one end of a double-ended cannula, the other end of which is received in a serum bottle for sample collection. Gas is introduced into the conduit at a gas bleed between the sample collection bottle and the reservoir. The jet ejector evacuates gas from the conduit and the bottle and aspirates a column of liquid from the reservoir at a high rate. When the withdrawn liquid reaches the jet ejector the rate of flow therethrough reduces substantially and the gas bleed increases the pressure in the conduit for driving liquid into the sample bottle, the gas bleed forming a column of gas behind the withdrawn liquid column and interrupting the withdrawal of liquid from the reservoir. In the case of hazardous and toxic liquids, the sample bottle and the jet ejector may be isolated from the reservoir and may be further isolated from a control station containing remote manipulation means for the sample bottle and control valves for the jet ejector and gas bleed. 5 figs.

  20. Spreading of miscible liquids

    NASA Astrophysics Data System (ADS)

    Walls, Daniel J.; Haward, Simon J.; Shen, Amy Q.; Fuller, Gerald G.

    2016-05-01

    Miscible liquids commonly contact one another in natural and technological situations, often in the proximity of a solid substrate. In the scenario where a drop of one liquid finds itself on a solid surface and immersed within a second, miscible liquid, it will spread spontaneously across the surface. We show experimental findings of the spreading of sessile drops in miscible environments that have distinctly different shape evolution and power-law dynamics from sessile drops that spread in immiscible environments, which have been reported previously. We develop a characteristic time to scale radial data of the spreading sessile drops based on a drainage flow due to gravity. This time scale is effective for a homologous subset of the liquids studied. However, it has limitations when applied to significantly chemically different, yet miscible, liquid pairings; we postulate that the surface energies between each liquid and the solid surface becomes important for this other subset of the liquids studied. Initial experiments performed with pendant drops in miscible environments support the drainage flow observed in the sessile drop systems.

  1. Liquid metal drop ejection

    NASA Technical Reports Server (NTRS)

    Khuri-Yakub, B. T.

    1993-01-01

    The aim of this project was to demonstrate the possibility of ejecting liquid metals using drop on demand printing technology. The plan was to make transducers for operation in the 100 MHz frequency range and to use these transducers to demonstrate the ability to eject drops of liquid metals such as gallium. Two transducers were made by indium bonding piezoelectric lithium niobate to quartz buffer rods. The lithium niobate plates were thinned by mechanical polishing to a thickness of 37 microns for operation at 100 MHz. Hemispherical lenses were polished in the opposite ends of the buffer rods. The lenses, which focus the sound waves in the liquid metal, had an F-number equals 1. A mechanical housing was made to hold the transducers and to allow precise control over the liquid level above the lens. We started by demonstrating the ability to eject drops of water on demand. The drops of water had a diameter of 15 microns which corresponds to the wavelength of the sound wave in the water. A videotape of this ejection was made. We then used a mixture of Gallium and Indium (used to lower the melting temperature of the Gallium) to demonstrate the ejection of liquid metal drops. This proved to be difficult because of the oxide skin which forms on the surface of the liquid. In some instances, we were able to eject metal drops, however, this was not consistent and reproducible. An experiment was set up at NASA-Lewis to stabilize the process of drop on demand liquid metal ejection. The object was to place the transducer and liquid metal in a vacuum station so that no oxide would form on the surface. We were successful in demonstrating that liquid metals could be ejected on demand and that this technology could be used for making sheet metal in space.

  2. Liquid-level detector

    DOEpatents

    Not Available

    1981-01-29

    Aliquid level sensor is described which has a pair of upright conductors spaced by an insulator defining a first high resistance path between the conductors. An electrically conductive path is interposed between the upright conductors at a discrete location at which liquid level is to be measured. It includes a liquid accessible gap of a dimension such that the electrical resistance across the conductor when the gap is filled with the liquid is detectably less than when the gap is emptied. The conductor might also be physically altered by temperature changes to serve also as an indicator of elevated temperature.

  3. Liquid metal electric pump

    DOEpatents

    Abbin, Joseph P.; Andraka, Charles E.; Lukens, Laurance L.; Moreno, James B.

    1992-01-01

    An electrical pump for pumping liquid metals to high pressures in high temperature environments without the use of magnets or moving mechanical parts. The pump employs a non-porous solid electrolyte membrane, typically ceramic, specific to the liquid metal to be pumped. A DC voltage is applied across the thickness of the membrane causing ions to form and enter the membrane on the electrically positive surface, with the ions being neutralized on the opposite surface. This action provides pumping of the liquid metal from one side of the non-porous solid electrolyte membrane to the other.

  4. Liquid thickness gauge

    NASA Technical Reports Server (NTRS)

    Weinstein, Leonard M. (Inventor)

    1988-01-01

    A method and apparatus are developed to measure the thickness of a liquid on a surface independent of liquid conductivity. Two pairs of round, corrosion resistant wires are mounted in an insulating material such that the cross-sectional area of each wire is flush with and normal to the surface. The resistance between each pair of wires is measured using two ac resistance measuring circuits, in which the ratio of the outputs of the two resistance measuring circuits is indicative of the thickness of the liquid on the surface.

  5. Pressurized liquid filled tendons

    SciTech Connect

    Burns, G.E.

    1987-05-12

    This patent describes an apparatus for detecting a leak in a tension leg platform tendon, comprising: a fluid-tight tensioned tubular tendon, the tendon connected on its upper end to a buoyant offshore structure and on its lower end to an anchor means. The anchor means is connected to the sea floor; means for supplying liquid to the tendon; means for pressurizing the liquid in excess of the maximum hydrostatic pressure exerted by the sea water on the tendon; and means for monitoring pressure, the means monitoring variations in liquid pressure to the tendon.

  6. Generation and deposition of Aβ43 by the virtually inactive presenilin-1 L435F mutant contradicts the presenilin loss-of-function hypothesis of Alzheimer's disease.

    PubMed

    Kretner, Benedikt; Trambauer, Johannes; Fukumori, Akio; Mielke, Janina; Kuhn, Peer-Hendrik; Kremmer, Elisabeth; Giese, Armin; Lichtenthaler, Stefan F; Haass, Christian; Arzberger, Thomas; Steiner, Harald

    2016-01-01

    As stated by the prevailing amyloid cascade hypothesis, Alzheimer's disease (AD) is caused by the aggregation and cerebral deposition of long amyloid-β peptide (Aβ) species, which are released from a C-terminal amyloid precursor protein fragment by γ-secretase. Mutations in its catalytic subunit presenilin-1 (PS1) increase the Aβ42 to Aβ40 ratio and are the major cause of familial AD (FAD). An opposing hypothesis states that loss of essential presenilin functions underlies the disease. A major argument for this hypothesis is the observation that the nearly inactive PS1 L435F mutant, paradoxically, causes FAD We now show that the very little Aβ generated by PS1 L435F consists primarily of Aβ43, a highly amyloidogenic species which was overlooked in previous studies of this mutant. We further demonstrate that the generation of Aβ43 is not due to a trans-dominant effect of this mutant on WT presenilin. Furthermore, we found Aβ43-containing plaques in brains of patients with this mutation. The aberrant generation of Aβ43 by this particular mutant provides a direct objection against the presenilin hypothesis. PMID:26988102

  7. Gain of a New Exon by a Lineage-Specific Alu Element-Integration Event in the BCS1L Gene during Primate Evolution

    PubMed Central

    Park, Sang-Je; Kim, Young-Hyun; Lee, Sang-Rae; Choe, Se-Hee; Kim, Myung-Jin; Kim, Sun-Uk; Kim, Ji-Su; Sim, Bo-Woong; Song, Bong-Seok; Jeong, Kang-Jin; Jin, Yeung-Bae; Lee, Youngjeon; Park, Young-Ho; Park, Young Il; Huh, Jae-Won; Chang, Kyu-Tae

    2015-01-01

    BCS1L gene encodes mitochondrial protein and is a member of conserved AAA protein family. This gene is involved in the incorporation of Rieske FeS and Qcr10p into complex III of respiratory chain. In our previous study, AluYRa2-derived alternative transcript in rhesus monkey genome was identified. However, this transcript has not been reported in human genome. In present study, we conducted evolutionary analysis of AluYRa2-exonized transcript with various primate genomic DNAs and cDNAs from humans, rhesus monkeys, and crab-eating monkeys. Remarkably, our results show that AluYRa2 element has only been integrated into genomes of Macaca species. This Macaca lineage-specific integration of AluYRa2 element led to exonization event in the first intron region of BCS1L gene by producing a conserved 3′ splice site. Intriguingly, in rhesus and crab-eating monkeys, more diverse transcript variants by alternative splicing (AS) events, including exon skipping and different 5′ splice sites from humans, were identified. Alignment of amino acid sequences revealed that AluYRa2-exonized transcript has short N-terminal peptides. Therefore, AS events play a major role in the generation of various transcripts and proteins during primate evolution. In particular, lineage-specific integration of Alu elements and species-specific Alu-derived exonization events could be important sources of gene diversification in primates. PMID:26537194

  8. LYRM7/MZM1L is a UQCRFS1 chaperone involved in the last steps of mitochondrial Complex III assembly in human cells.

    PubMed

    Sánchez, Ester; Lobo, Teresa; Fox, Jennifer L; Zeviani, Massimo; Winge, Dennis R; Fernández-Vizarra, Erika

    2013-03-01

    The mammalian Complex III (CIII) assembly process is yet to be completely understood. There is still a lack in understanding of how the structural subunits are put together and which additional factors are involved. Here we describe the identification and characterization of LYRM7, a human protein displaying high sequence homology to the Saccharomyces cerevisiae protein Mzm1, which was recently shown as an assembly factor for Rieske Fe-S protein incorporation into the yeast cytochrome bc(1) complex. We conclude that human LYRM7, which we propose to be renamed MZM1L (MZM1-like), works as a human Rieske Fe-S protein (UQCRFS1) chaperone, binding to this subunit within the mitochondrial matrix and stabilizing it prior to its translocation and insertion into the late CIII dimeric intermediate within the mitochondrial inner membrane. Thus, LYRM7/MZM1L is a novel human CIII assembly factor involved in the UQCRFS1 insertion step, which enables formation of the mature and functional CIII enzyme. PMID:23168492

  9. TERT-CLPTM1L Polymorphism rs401681 Contributes to Cancers Risk: Evidence from a Meta-Analysis Based on 29 Publications

    PubMed Central

    Yin, Ming; Sun, Jingwen; Liu, Li; Qin, Qin; Li, Xiaorong; Long, Lu; Nie, Shaofa; Wei, Sheng

    2012-01-01

    Background Some common genetic variants of TERT-CLPTM1L gene, which encode key protein subunits of telomerase, have been suggested to play a crucial role in tumorigenesis. The TERT-CLPTM1L polymorphism rs401681 was of special interest for cancers risk but with inconclusive results. Methodology/Principal Findings We performed a comprehensive meta-analysis of 29 publications with a total of 91263 cases and 735952 controls. We assessed the strength of the association between rs401681 and overall cancers risk and performed subgroup analyses by cancer type, ethnicity, source of control, sample size and expected power. Rs401681 C allele was found to be associated with marginally increased cancers risk, with per allele OR of 1.04 (95%CI = 1.00–1.08, Pheterogeneity<0.001) and an expected power of 1.000. Following further stratified analyses, the increased cancers risk were discovered in subgroups of lung, bladder, prostate, basal cell carcinomas and Asians, while a declined risk of pancreatic cancer and melanoma were detected. Conclusions/Significance These findings suggested that rs401681 C allele was a low-penetrance risk allele for the development of cancers of lung, bladder, prostate and basal cell carcinoma, but a potential protective allele for melanoma and pancreatic cancer. PMID:23226346

  10. Liquid cooled garments

    NASA Technical Reports Server (NTRS)

    1975-01-01

    Liquid cooled garments employed in several applications in which severe heat is encountered are discussed. In particular, the use of the garments to replace air line cooling units in a variety of industrial processing situations is discussed.

  11. Wettability by Ionic Liquids.

    PubMed

    Liu, Hongliang; Jiang, Lei

    2016-01-01

    Ionic liquids (ILs) have become particularly attractive recently because they have demonstrated themselves to be important construction units in the broad fields of chemistry and materials science, from catalysis and synthesis to analysis and electrochemistry, from functional fluids to clean energy, from nanotechnology to functional materials. One of the greatest issues that determines the performance of ILs is the wettability of correlated surfaces. In this concept article, the key developments and issues in IL wettability are surveyed, including the electrowetting of ILs in gas-liquid-solid systems and liquid-liquid-solid systems, ILs as useful probe fluids, the superwettability of Ils, and future directions in IL wettability. This should generate extensive interest in the field and encourage more scientists to engage in this area to tackle its scientific challenges. PMID:26619157

  12. Liquid metal boiling inception

    NASA Technical Reports Server (NTRS)

    Sabin, C. M.; Poppendiek, H. F.; Mouritzen, G.; Meckel, P. T.; Cloakey, J. E.

    1972-01-01

    An experimental study of the inception of boiling in potassium in forced convection is reported. The boiler consisted of a 0.19-inch inside diameter, niobium-1% zirconium boiler tube approximately six feet long. Heating was accomplished by direct electrical tube wall conduction. Experiments were performed with both all-liquid fill and two-phase fill startup sequences and with a range of flow rates, saturation temperatures, inert gas levels, and fill liquid temperatures. Superheat of the liquid above the equilibrium saturation temperature was observed in all the experiments. Incipient boiling liquid superheat ranged from a few degrees to several hundred. Comparisons of these data with other data and with several analytical treatments are presented.

  13. Liquid level controller

    DOEpatents

    Mangus, J.D.; Redding, A.H.

    1975-07-15

    A system for maintaining two distinct sodium levels within the shell of a heat exchanger having a plurality of J-shaped modular tube bundles each enclosed in a separate shell which extends from a common base portion. A lower liquid level is maintained in the base portion and an upper liquid level is maintained in the shell enwrapping the long stem of the J-shaped tube bundles by utilizing standpipes with a notch at the lower end which decreases in open area the distance from the end of the stand pipe increases and a supply of inert gas fed at a constant rate to produce liquid levels, which will remain generally constant as the flow of liquid through the vessel varies. (auth)

  14. Lacerations - liquid bandage

    MedlinePlus

    ... painless to apply. Skin adhesives, or liquid bandages, seal the cut closed after only one application. There ... scrub the site. Doing so may loosen the seal or even remove the adhesive completely. The seal ...

  15. Diet - full liquid

    MedlinePlus

    ... 7 of the foods you can eat for breakfast, lunch, and dinner. Liquid foods DO NOT include " ... Nonfat dry milk added to your drinks Instant breakfast powder added to milk, puddings, custards, and milkshakes ...

  16. Lacerations - liquid bandage

    MedlinePlus

    ... causes only slight burning when applied. Liquid bandages, seal the cut closed after only 1 application. There ... you can shower or bathe without worry. The seal lasts for 5 to 10 days. It will ...

  17. Safer Liquid Natural Gas

    NASA Technical Reports Server (NTRS)

    1976-01-01

    After the disaster of Staten Island in 1973 where 40 people were killed repairing a liquid natural gas storage tank, the New York Fire Commissioner requested NASA's help in drawing up a comprehensive plan to cover the design, construction, and operation of liquid natural gas facilities. Two programs are underway. The first transfers comprehensive risk management techniques and procedures which take the form of an instruction document that includes determining liquid-gas risks through engineering analysis and tests, controlling these risks by setting up redundant fail safe techniques, and establishing criteria calling for decisions that eliminate or accept certain risks. The second program prepares a liquid gas safety manual (the first of its kind).

  18. Liquid piston Stirling engines

    SciTech Connect

    West, C.D.

    1983-01-01

    This book is a presentation on piston stirling engines. Topics covered include: liquid piston engines; basic design and power calculations; more advanced power calculations; design example; and past research work and some present research needs.

  19. Liquid-Cooled Garment

    NASA Technical Reports Server (NTRS)

    1977-01-01

    A liquid-cooled bra, offshoot of Apollo moon suit technology, aids the cancer-detection technique known as infrared thermography. Water flowing through tubes in the bra cools the skin surface to improve resolution of thermograph image.

  20. Liquid sample processor

    NASA Technical Reports Server (NTRS)

    Jahnsen, V. J.; Campen, C. F., Jr.

    1975-01-01

    Processor is automatic and includes series of extraction tubes packed with fibrous absorbent material of large surface area. When introduced into these tubes, liquid test samples become completely absorbed by packing material as thin film.

  1. 114. WEST SIDE OF LIQUID OXYGEN CONTROL ROOM (205). LIQUID ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    114. WEST SIDE OF LIQUID OXYGEN CONTROL ROOM (205). LIQUID NITROGEN (LN2) SUBCOOLER ON LEFT; SKID 8, LIQUID OXYGEN CONTROLLER FOR SWITCHING BETWEEN RAPID-LOAD AND TOPPING ON RIGHT. LIQUID OXYGEN LINE FROM SKID 9A AT RIGHT EDGE OF PHOTO. - Vandenberg Air Force Base, Space Launch Complex 3, Launch Pad 3 East, Napa & Alden Roads, Lompoc, Santa Barbara County, CA

  2. Applications of ionic liquids.

    PubMed

    Patel, Divia Dinesh; Lee, Jong-Min

    2012-06-01

    Ionic liquids have recently gained popularity in the scientific community owing to their special properties and characteristics. One of the reasons why ionic liquids have been termed "green solvents" is due to their negligible vapour pressure. Their use in electrochemical, biological and metal extraction applications is discussed. Wide research has been carried out for their use in batteries, solar panels, fuel cells, drug deliveries and biomass pretreatments. This work aims to consolidate the various findings from previous works in these areas. PMID:22711528

  3. Excitability in liquid crystal.

    PubMed

    Coullet, P.; Frisch, T.; Gilli, J. M.; Rica, S.

    1994-09-01

    The spiral waves observed in a liquid crystal submitted to a vertical electric field and a horizontal rotating magnetic field are explained in the framework of a purely mechanical description of the liquid crystal. The originality of the experiment described in this paper is the presence of the vertical electric field which allows us to analyze the spiral waves in the framework of a weakly nonlinear theory. PMID:12780124

  4. Liquid cooled helmet

    NASA Technical Reports Server (NTRS)

    Elkins, William (Inventor); Williams, Bill A. (Inventor)

    1979-01-01

    Liquid cooled helmet comprising a cap of flexible material adapted to fit the head of a person, cooling panels mounted inside the cap forming passageways for carrying a liquid coolant, the panels being positioned to engage the cranium and neck of a person wearing the helmet, inlet and outlet lines communicating with the passageways, and releasable straps for securing the helmet about the neck of the wearer.

  5. Compact Liquid Deaerator

    NASA Technical Reports Server (NTRS)

    Yamauchi, S. T.

    1982-01-01

    Gases are removed from liquids by a new deaerator that takes up only 5 inches (12.7 cm) at top of a medium-sized storage tank. Deaerator has a multiple cascading header that exposes more fluid at lower pressures than typical commercial deaerators. Potential applications are in hydraulic systems for aircraft and heavy machinery, in cooling systems where deaerated liquid is needed to prevent cavitation of pump.

  6. Liquid Level Sensing System

    NASA Technical Reports Server (NTRS)

    Korman, Valentin (Inventor); Wiley, John T. (Inventor); Duffell, Amanda G. (Inventor)

    2014-01-01

    A liquid level sensing system includes waveguides disposed in a liquid and distributed along a path with a gap between adjacent waveguides. A source introduces electromagnetic energy into the waveguides at a first end of the path. A portion of the electromagnetic energy exits the waveguides at a second end of the path. A detector measures the portion of the electromagnetic energy exiting the second end of the path.

  7. Electrically actuated liquid iris.

    PubMed

    Xu, Miao; Ren, Hongwen; Lin, Yi-Hsin

    2015-03-01

    We report an adaptive iris using dielectric liquids and a radial-interdigitated electrode. A black liquid is confined by a circular gasket with a donut shape. The surrounding of the black liquid is filled with an immiscible liquid. In the relaxing state, the black liquid obtains the largest clear aperture. By applying a voltage, the surface of the black liquid is stretched by the generated dielectric force, resulting in a reduction of its aperture. For the demonstrated iris, the diameter of the aperture can be changed from ∼4.7  mm to ∼1.2  mm when the voltage is applied from 0 to 70  V(rms). The aperture ratio is ∼94%. Owing to the radial-interdigitated electrode, the aperture size of the iris can be effectively switched with a reasonably fast response time. The optical switch is polarization-insensitive. The potential applications of our iris are light shutters, optical attenuators, biomimicry, and wearable devices. PMID:25723444

  8. Liquid-liquid critical point: an analytical approach

    NASA Astrophysics Data System (ADS)

    Daanoun, A.

    2006-09-01

    Theoretical simulations and experimental studies have showed that many systems (like liquid metals) can exhibit two phase transitions: gas-liquid and liquid-liquid. Consequently the fluid phase of these systems presents two critical points, namely the usual gas-liquid (G-L) critical point and the liquid-liquid critical point that results from a phase transition between two liquids of different densities: a low density liquid (LDL) and a high density liquid (HDL). The van der Waals theory for simple fluids [Phys. Rev. E 50, 2913 (1994)] is based on taking a system with purely repulsive forces as a reference, is able to describe two stable first-order phase transitions between fluids of different densities. The particles in our system interact via a total pair potential, which splits into a repulsive VR and a density-dependent attractive VA part.

  9. Liquid crystalline composites containing phyllosilicates

    DOEpatents

    Chaiko, David J.

    2004-07-13

    The present invention provides phyllosilicate-polymer compositions which are useful as liquid crystalline composites. Phyllosilicate-polymer liquid crystalline compositions of the present invention can contain a high percentage of phyllosilicate while at the same time be transparent. Because of the ordering of the particles liquid crystalline composite, liquid crystalline composites are particularly useful as barriers to gas transport.

  10. Transmitting Electromagnetic Energy into Liquids

    NASA Technical Reports Server (NTRS)

    Johnston, E. J.

    1984-01-01

    Rough liquid surface enhances coupling. Agitating surface of liquid nitrogen bath with periodic or aperiodic excitation enhances electromagnetic coupling between microwave horn and blackbody temperature standard immersed in liquid. Useful in interfaces between electromagnetic radiation and liquids. Biomedical, radar, and meteorological applications.

  11. Multigene pathway-based analyses identify nasopharyngeal carcinoma risk associations for cumulative adverse effects of TERT-CLPTM1L and DNA double-strand breaks repair.

    PubMed

    Yee Ko, Josephine Mun; Dai, Wei; Wun Wong, Elibe Hiu; Kwong, Dora; Tong Ng, Wai; Lee, Anne; Kai Cheong Ngan, Roger; Chung Yau, Chun; Tung, Stewart; Li Lung, Maria

    2014-10-01

    The genetic etiology of nasopharyngeal carcinoma (NPC) and mechanisms for inherited susceptibility remain unclear. To examine genetic risk factors for NPC, we hypothesized that heritable risk is attributable to cumulative effects of multiple common low-risk variants. With the premise that individual SNPs only confer subtle effects for cancer risk, a multigenic pathway-based approach was used to systematically examine associations between NPC genetic susceptibility with SNPs in genes in DNA repair pathways and from previously identified cancer genome-wide association study analyses. This case-control study covers 161 genes/loci and focuses on pathway-based analyses in 2,349 Hong Kong individuals, allowing stratification according to NPC familial status for meaningful association analysis. Three SNPs (rs401681, rs6774494 and rs3757318) corresponding to TERT/CLPTM1L (OR 95% CI = 0.77, 0.68-0.88), MDS1-EVI1 (OR 95% CI=0.79 0.69-0.89) and CCDC170 (OR 95% CI = 0.76, 0.66-0.86) conferred modest protective effects individually for NPC risk by the logistic regression analysis after multiple testing adjustment (p(Bonferroni)  < 0.05). Stratification of NPC according to familial status identified rs2380165 in BLM (OR 95% CI = 1.49, 1.20-1.86, p(Bonferroni)  < 0.05) association with family history-positive NPC (FH+ NPC) patients. Multiple SNPs pathway-based analysis revealed that the combined gene dosage effects for increasing numbers of unfavorable genotypes in TERT-CLPTM1L and double-strand break repair (DSBR) conferred elevated risk in FH+ and sporadic NPC patients (ORs per allele, 95% CIs = 1.37, 1.22-1.55, p(Bonferroni)  = 5.00 × 10(-6); 1.17, 1.09-1.26, p(Bonferroni)  = 4.58 × 10(-4) , respectively, in TERT/NHEJ pathways). Our data suggested cumulative increased NPC risk associations with TERT-CLPTM1L and DSBR pathways contribute to genetic susceptibility to NPC and have potential translational relevance for patient stratification and therapeutics. PMID

  12. A SEL1L Mutation Links a Canine Progressive Early-Onset Cerebellar Ataxia to the Endoplasmic Reticulum–Associated Protein Degradation (ERAD) Machinery

    PubMed Central

    Kyöstilä, Kaisa; Cizinauskas, Sigitas; Seppälä, Eija H.; Suhonen, Esko; Jeserevics, Janis; Sukura, Antti; Syrjä, Pernilla; Lohi, Hannes

    2012-01-01

    Inherited ataxias are characterized by degeneration of the cerebellar structures, which results in progressive motor incoordination. Hereditary ataxias occur in many species, including humans and dogs. Several mutations have been found in humans, but the genetic background has remained elusive in dogs. The Finnish Hound suffers from an early-onset progressive cerebellar ataxia. We have performed clinical, pathological, and genetic studies to describe the disease phenotype and to identify its genetic cause. Neurological examinations on ten affected dogs revealed rapidly progressing generalized cerebellar ataxia, tremors, and failure to thrive. Clinical signs were present by the age of 3 months, and cerebellar shrinkage was detectable through MRI. Pathological and histological examinations indicated cerebellum-restricted neurodegeneration. Marked loss of Purkinje cells was detected in the cerebellar cortex with secondary changes in other cortical layers. A genome-wide association study in a cohort of 31 dogs mapped the ataxia gene to a 1.5 Mb locus on canine chromosome 8 (praw = 1.1×10−7, pgenome = 7.5×10−4). Sequencing of a functional candidate gene, sel-1 suppressor of lin-12-like (SEL1L), revealed a homozygous missense mutation, c.1972T>C; p.Ser658Pro, in a highly conserved protein domain. The mutation segregated fully in the recessive pedigree, and a 10% carrier frequency was indicated in a population cohort. SEL1L is a component of the endoplasmic reticulum (ER)–associated protein degradation (ERAD) machinery and has not been previously associated to inherited ataxias. Dysfunctional protein degradation is known to cause ER stress, and we found a significant increase in expression of nine ER stress responsive genes in the cerebellar cortex of affected dogs, supporting the pathogenicity of the mutation. Our study describes the first early-onset neurodegenerative ataxia mutation in dogs, establishes an ERAD–mediated neurodegenerative disease model

  13. Beyond dispersive liquid-liquid microextraction.

    PubMed

    Leong, Mei-I; Fuh, Ming-Ren; Huang, Shang-Da

    2014-03-28

    Dispersive liquid-liquid microextraction (DLLME) and other dispersion liquid-phase microextraction (LPME) methods have been developed since the first DLLME method was reported in 2006. DLLME is simple, rapid, and affords high enrichment factor, this is due to the large contact surface area of the extraction solvent. DLLME is a method suitable for the extraction in many different water samples, but it requires using chlorinated solvents. In recent years, interest in DLLME or dispersion LPME has been focused on the use of low-toxicity solvents and more conveniently practical procedures. This review examines some of the most interesting developments in the past few years. In the first section, DLLME methods are separated in two categories: DLLME with low-density extraction solvent and DLLME with high-density extraction solvent. Besides these methods, many novel special devices for collecting low-density extraction solvent are also mentioned. In addition, various dispersion techniques with LPME, including manual shaking, air-assisted LPME (aspirating and injecting the extraction mixture by syringe), ultrasound-assisted emulsification, vortex-assisted emulsification, surfactant-assisted emulsification, and microwave-assisted emulsification are described. Besides the above methods, combinations of DLLME with other extraction techniques (solid-phase extraction, stir bar sorptive extraction, molecularly imprinted matrix solid-phase dispersion and supercritical fluid extraction) are introduced. The combination of nanotechnique with DLLME is also introduced. Furthermore, this review illustrates the application of DLLME or dispersion LPME methods to separate and preconcentrate various organic analytes, inorganic analytes, and samples. PMID:24582396

  14. Functional dissection of an enhancer-like element located within the second intron of the human U2AF1L4 gene.

    PubMed

    Didych, D A; Smirnov, N A; Kotova, E S; Akopov, S B; Nikolaev, L G; Sverdlov, E D

    2011-08-01

    A detailed functional and evolutionary analysis of an enhancer element of the human genome (enhancer 12) located in the second intron of the U2AF1L4 gene, which we identified earlier, is presented. Overlapping fragments of the studied genome region were analyzed for enhancer activity, and the site responsible for the activity of this element was identified using transient transfections of HeLa cells. Comparison of the enhancer 12 sequence with orthologous sequences from seven primate species revealed the existence of evolutionarily conserved sequences within this element. One of the identified conservative regions is likely responsible for the enhancer activity and is able to specifically interact in vitro with proteins of HeLa cell nuclear extract. The ability of orthologous primate sequences to compete with enhancer 12 for binding with HeLa cell nuclear extract proteins and to enhance the activity of the reporter gene in transient transfection of HeLa cells is demonstrated. PMID:22022969

  15. Visualization of lipid metabolism in the larval zebrafish intestine reveals a relationship between NPC1L1 mediated cholesterol uptake and dietary fatty acids

    PubMed Central

    Walters, James W.; Anderson, Jennifer L.; Bittman, Robert; Pack, Michael; Farber, Steven A.

    2012-01-01

    SUMMARY The small intestine is the primary site of dietary lipid absorption in mammals. The balance of nutrients, microorganisms, bile, and mucus that determine intestinal luminal environment cannot be recapitulated ex vivo, thus complicating studies of lipid absorption. We show that fluorescently labeled lipids can be used to visualize and study lipid absorption in live zebrafish larvae. We demonstrate that the addition of BODIPY-fatty acid to a diet high in atherogenic lipids enables imaging of enterocyte lipid droplet dynamics in real time. We find that a lipid-rich meal promotes BODIPY-cholesterol absorption into an endosomal compartment distinguishable from lipid droplets. We also show that dietary fatty acids promote intestinal cholesterol absorption by rapid relocalization of NPC1L1 to intestinal brush border. These data illustrate the power of the zebrafish system to address longstanding questions in vertebrate digestive physiology. PMID:22749558

  16. Operation with three liquid phases in a staged liquid-liquid contactor

    SciTech Connect

    Leonard, R.A.; Ziegler, A.A.; Wigeland, R.A.; Bane, R.W.; Steindler, M.J.

    1983-03-01

    Operation with three liquid phases was demonstrated in a staged liquid-liquid contactor. The possibility that three liquid phases could be handled in a liquid-liquid contactor normally used with two liquid phases was initially established using a laboratory batch test. Tht three liquid phases were obtained using a thorium flow sheet having high concentrations of both acid and thorium. To analyze the batch test, the concept of a dimensionless dispersion number for use with two liquid phases was extended so that it could be applied to three liquid phases. Based on the batch tests, continuous flow tests were run in a staged liquid-liquid contactor used for solvent extraction. A critical factor in the success of these tests was determining the position of the liquid-liquid interface in the contactor. Thus, a contactor was used which allows the position of the liquid-liquid interface to be adjusted. Actual three-phase operation was demonstrated using a 4-cm annular centrifugal contactor, albeit with a somewhat greater (3 to 4 vol. %) aqueous-phase contamination of the organic exit stream than normal (< 1 vol. %).

  17. Integrated pathway and epistasis analysis reveals interactive effect of genetic variants at TERF1 and AFAP1L2 loci on melanoma risk.

    PubMed

    Brossard, Myriam; Fang, Shenying; Vaysse, Amaury; Wei, Qingyi; Chen, Wei V; Mohamdi, Hamida; Maubec, Eve; Lavielle, Nolwenn; Galan, Pilar; Lathrop, Mark; Avril, Marie-Françoise; Lee, Jeffrey E; Amos, Christopher I; Demenais, Florence

    2015-10-15

    Genome-wide association studies (GWASs) have characterized 13 loci associated with melanoma, which only account for a small part of melanoma risk. To identify new genes with too small an effect to be detected individually but which collectively influence melanoma risk and/or show interactive effects, we used a two-step analysis strategy including pathway analysis of genome-wide SNP data, in a first step, and epistasis analysis within significant pathways, in a second step. Pathway analysis, using the gene-set enrichment analysis (GSEA) approach and the gene ontology (GO) database, was applied to the outcomes of MELARISK (3,976 subjects) and MDACC (2,827 subjects) GWASs. Cross-gene SNP-SNP interaction analysis within melanoma-associated GOs was performed using the INTERSNP software. Five GO categories were significantly enriched in genes associated with melanoma (false discovery rate ≤ 5% in both studies): response to light stimulus, regulation of mitotic cell cycle, induction of programmed cell death, cytokine activity and oxidative phosphorylation. Epistasis analysis, within each of the five significant GOs, showed significant evidence for interaction for one SNP pair at TERF1 and AFAP1L2 loci (pmeta-int  = 2.0 × 10(-7) , which met both the pathway and overall multiple-testing corrected thresholds that are equal to 9.8 × 10(-7) and 2.0 × 10(-7) , respectively) and suggestive evidence for another pair involving correlated SNPs at the same loci (pmeta-int  = 3.6 × 10(-6) ). This interaction has important biological relevance given the key role of TERF1 in telomere biology and the reported physical interaction between TERF1 and AFAP1L2 proteins. This finding brings a novel piece of evidence for the emerging role of telomere dysfunction into melanoma development. PMID:25892537

  18. Microgravity liquid propellant management

    NASA Technical Reports Server (NTRS)

    Hung, R. J.

    1990-01-01

    The requirement to settle or to position liquid fluid over the outlet end of a spacecraft propellant tank prior to main engine restart, poses a microgravity fluid behavior problem. Resettlement or reorientation of liquid propellant can be accomplished by providing optimal acceleration to the spacecraft such that the propellant is reoriented over the tank outlet without any vapor entrainment, any excessive geysering, or any other undersirable fluid motion for the space fluid management under microgravity environment. The most efficient technique is studied for propellant resettling through the minimization of propellant usage and weight penalties. Both full scale and subscale liquid propellant tank of Space Transfer Vehicle were used to simulate flow profiles for liquid hydrogen reorientation over the tank outlet. In subscale simulation, both constant and impulsive resettling acceleration were used to simulate the liquid flow reorientation. Comparisons between the constant reverse gravity acceleration and impulsive reverse gravity acceleration to be used for activation of propellant resettlement shows that impulsive reverse gravity thrust is superior to constant reverse gravity thrust.

  19. Carbenes from ionic liquids.

    PubMed

    Hollóczki, Oldamur; Nyulászi, László

    2014-01-01

    In the last decade an explosive development has been observed in the fields of both ionic liquids (ILs) as potential chemically inert solvents with many possible technical applications, and N-heterocyclic carbenes (NHCs) as catalysts with superb performance. Since the cations of many ILs can be deprotonated by strong bases yielding NHCs, this two fields are inherently connected. It has only recently been recognized that some of the commonly used basic anions of the ILs (such as acetate) are able to deprotonate azolium cations. While the resulting NHC could clearly be observed in the vapor phase, in the liquid - where the mutual electrostatic interactions within the ion network stabilize the ion pairs - the neutral NHC cannot be detected by commonly used analytical techniques; however, from these ionic liquids NHCs can be trapped, e.g., by complex formation, or more importantly these ILs can be directly used as catalysts, since the NHC content is sufficiently large for these applications. Apart from imidazole-2-ylidenes, the formation of other highly reactive neutral species ("abnormal carbenes," 2-alkylideneimidazoles, pyridine-ylidenes or pyridinium-ylides) is feasible in highly basic ionic liquids. The cross-fertilizing overlap between the two fields may provide access to a great advance in both areas, and we give an overview here on the results published so far, and also on the remaining possibilities and challenges in the concept of "carbenes from ionic liquids." PMID:23539381

  20. Characterization of VvSERK1, VvSERK2, VvSERK3 and VvL1L genes and their expression during somatic embryogenesis of grapevine (Vitis vinifera L.).

    PubMed

    Schellenbaum, Paul; Jacques, Alban; Maillot, Pascale; Bertsch, Christophe; Mazet, Flore; Farine, Sibylle; Walter, Bernard

    2008-12-01

    Little is known about the genes expressed during grapevine somatic embryogenesis. Both groups of Somatic Embryogenesis Receptor Kinase (SERK) and Leafy Cotyledon (LEC and L1L) genes seem to play key roles during somatic embryogenesis in various plant species. Therefore, we identified and analysed the sequences of VvSERK and VvL1L (Leafy cotyledon1-Like) genes. The deduced amino acid sequences of VvSERK1, VvSERK2 and VvSERK3 are very similar to that of registered SERK proteins, with highest homologies for the kinase domain in the C-terminal region. The amino acid sequence of VvL1L presents all the domains that are characteristic for LEC1 and L1L proteins, particularly, the 16 amino acid residues that serve as signature of the B-domain. Phylogenetic analysis distinguishes members of subclass LEC1 and subclass L1L, and VvL1L is closely related to L1L proteins. Using semi-quantitative RT-PCR, we studied gene expression of VvSERK1, VvSERK2, VvSERK3 and VvL1L in calli and somatic embryos obtained from anther culture of Vitis vinifera L. cv Chardonnay. Expression of VvSERK2 is relatively stable during in vitro culture. In contrast, VvSERK1, VvSERK3 and VvL1L are expressed more 4 to 6 weeks after transfer of the calli onto embryo induction medium, before the visible appearance of embryos on the calli as seen by environmental scanning electron microscopy. Later on (8 weeks after transfer) VvSERK1 expression is maintained in the embryogenic calli and VvSERK3 in the embryos, whereas VvL1L expression is very low. All together, these data suggest the involvement of VvSERK and VvL1L genes in grapevine somatic embryogenesis. PMID:18766346

  1. Thermohydraulics in liquid metals

    NASA Astrophysics Data System (ADS)

    Kottowski, H. M.

    Heat transfer problems in single-phase and two-phase liquid metal forced convection flow are reviewed. Liquid metal boiling heat transfer in pool flow; and dry out heat fluxes are considered. It is shown that in technological plants working with liquid metals, superheating up to 150 C occurs, and can lead to nonstationary hydraulic transition between the single-phase and established two-phase flows. Boiling phases relative to subcooled boiling and bubble boiling have no importance for technological processes. Piston, slug and annular flow patterns dominate. On the basis of the flow patterns observed during boiling, the separate flow model principle is the only one suitable for calculating the two-phase flow pressure drop. Using this model and total pressure drop measurements, a relationship for the two-phase frictional pressure characteristic, valid for tubular and annular geometry, can be determined.

  2. Liquid lubrication in space

    NASA Technical Reports Server (NTRS)

    Zaretsky, Erwin V.

    1990-01-01

    The requirement for long-term, reliable operation of aerospace mechanisms has, with a few exceptions, pushed the state of the art in tribology. Space mission life requirements in the early 1960s were generally 6 months to a year. The proposed U.S. space station schedule to be launched in the 1990s must be continuously usable for 10 to 20 years. Liquid lubrication systems are generally used for mission life requirements longer than a year. Although most spacecraft or satellites have reached their required lifetimes without a lubrication-related failure, the application of liquid lubricants in the space environment presents unique challenges. The state of the art of liquid lubrication in space as well as the problems and their solutions are reviewed.

  3. Two new vortex liquids

    NASA Astrophysics Data System (ADS)

    Anderson, Philip W.

    2007-03-01

    In 1967, Reatto and Chester proposed that solid helium-4 might exhibit superfluidity, and in 1970, Leggett suggested what was thought to be a definitive experimental test: to find non-classical rotational inertia in a toroidal sample. More than three decades later, the observation by Kim and Chan of exactly that effect generated great interest and has been repeated and confirmed by a number of groups. However, many attempts to find actual superflow in truly solid samples have failed. Here, I draw an analogy with a second example of anomalous response to vorticity in a dissipative fluid, the vortex liquid phase in the pseudogap region of high-temperature superconductors, and propose that the solid helium experiments have been mischaracterized: what is observed is not supersolidity but an incompressible vortex liquid. This state is distinct from a conventional liquid in that its properties are dominated by conserved supercurrents flowing around a thermally fluctuating tangle of vortices.

  4. Degree of Recruitment of DOT1L to MLL-AF9 Defines Level of H3K79 Di- and Tri-methylation on Target Genes and Transformation Potential

    PubMed Central

    Kuntimaddi, Aravinda; Achille, Nicholas J.; Thorpe, Jeremy; Lokken, Alyson A.; Singh, Ritambhara; Hemenway, Charles S.; Adli, Mazhar; Zeleznik-Le, Nancy J.; Bushweller, John H.

    2015-01-01

    SUMMARY The MLL gene is a common target of chromosomal translocations found in human leukemia. MLL-fusion leukemia has a consistently poor outcome. One of the most common translocation partners is AF9 (MLLT3). MLL-AF9 recruits DOT1L, a histone 3 lysine 79 methyltransferase (H3K79me1/me2/me3), leading to aberrant gene transcription. We show that DOT1L has three AF9 binding sites, and present the NMR solution structure of a DOT1L-AF9 complex. We generate structure-guided point mutations and find they have graded effects on recruitment of DOT1L to MLL-AF9. ChIP-Seq analyses of H3K79me2 and H3K79me3 show that graded reduction of the DOT1L interaction with MLL-AF9 results in differential loss of H3K79me2 and me3 at MLL-AF9 target genes. Furthermore, the degree of DOT1L recruitment is linked to the level of MLL-AF9 hematopoietic transformation. PMID:25921540

  5. Liquid air cycle engines

    NASA Technical Reports Server (NTRS)

    Rosevear, Jerry

    1992-01-01

    Given here is a definition of Liquid Air Cycle Engines (LACE) and existing relevant technologies. Heat exchanger design and fabrication techniques, the handling of liquid hydrogen to achieve the greatest heat sink capabilities, and air decontamination to prevent heat exchanger fouling are discussed. It was concluded that technology needs to be extended in the areas of design and fabrication of heat exchangers to improve reliability along with weight and volume reductions. Catalysts need to be improved so that conversion can be achieved with lower quantities and lower volumes. Packaging studies need to be investigated both analytically and experimentally. Recycling with slush hydrogen needs further evaluation with experimental testing.

  6. Electroplating Using Ionic Liquids

    NASA Astrophysics Data System (ADS)

    Abbott, Andrew P.; Frisch, Gero; Ryder, Karl S.

    2013-07-01

    Electroplating is a key technology in many large-scale industrial applications such as corrosion-resistant and decorative coatings. Issues with current aqueous processes, such as toxicity of reagents and low current efficiencies, can often be overcome by using ionic liquids, and this approach has turned ionometallurgy into a fast-growing area of research. This review outlines the interactions in ionic liquids that are responsible for the advantageous properties of these solvents in electroplating. It summarizes recent research in which these properties have been analyzed or exploited and highlights fundamental issues in research and technology that need to be addressed.

  7. Liquid metal pump

    DOEpatents

    Pennell, William E.

    1982-01-01

    The liquid metal pump comprises floating seal rings and attachment of the pump diffuser to the pump bowl for isolating structural deflections from the pump shaft bearings. The seal rings also eliminate precision machining on large assemblies by eliminating the need for a close tolerance fit between the mounting surfaces of the pump and the seals. The liquid metal pump also comprises a shaft support structure that is isolated from the pump housing for better preservation of alignment of shaft bearings. The shaft support structure also allows for complete removal of pump internals for inspection and repair.

  8. Solar liquid heating system

    SciTech Connect

    Finn, D.J.

    1990-05-08

    This patent describes a solar heater for heating liquids. It comprises: a heatable bag, a support means supporting the heatable bag, a heatable body of liquid in the heatable bag, the heatable bag being disposed in sunlight so as to become heated thereby, a topside gas bag above the heatable bag, the topside gas bag containing a gas for serving as insulation, a topside fluid bag disposed above the topside gas bag and containing a fluid for further insulation. The bags being substantially gasproof and waterproof and also being flexible whereby the gravity pull on the bags and the flexibility thereof causes the upper sides of the bags to seek horizontal levels.

  9. Improved Capacitive Liquid Sensor

    NASA Technical Reports Server (NTRS)

    Waldman, Francis A.

    1992-01-01

    Improved capacitive sensor used to detect presence and/or measure thickness of layer of liquid. Electrical impedance or admittance of sensor measured at prescribed frequency, and thickness of liquid inferred from predetermined theoretical or experimental relationship between impedance and thickness. Sensor is basically a three-terminal device. Features interdigitated driving and sensing electrodes and peripheral coplanar ground electrode that reduces parasitic effects. Patent-pending because first to utilize ground plane as "shunting" electrode. System less expensive than infrared, microwave, or refractive-index systems. Sensor successfully evaluated in commercial production plants to characterize emulsions, slurries, and solutions.

  10. Turbine meters for liquid measurement

    SciTech Connect

    Wass, D.J.; Allen, C.R.

    1995-12-01

    Liquid turbine meters operate in response to fundamental engineering principles, Operation with a single moving part produces excellent longevity and reliability. Liquid turbine meters display wide rangeability, high accuracy, excellent repeatability, low pressure drop and moderate cost. Liquid turbine meters may be applied to many different fluids with different physical properties and corrosive tendencies. The marriage of liquid turbine meters to electronic instruments allows instantaneous flow calculations and produces the flexibility to display data, store data, transmit data in the most convenient form. Liquid turbine meters should be the first flow measurement instrument considered for liquid measurement applications.

  11. Mixed Stationary Liquid Phases for Gas-Liquid Chromatography.

    ERIC Educational Resources Information Center

    Koury, Albert M.; Parcher, Jon F.

    1979-01-01

    Describes a laboratory technique for use in an undergraduate instrumental analysis course that, using the interpretation of window diagrams, prepares a mixed liquid phase column for gas-liquid chromatography. A detailed procedure is provided. (BT)

  12. 138. LIQUID NITROGEN INSTRUMENT PANEL ON EAST WALL OF LIQUID ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    138. LIQUID NITROGEN INSTRUMENT PANEL ON EAST WALL OF LIQUID NITROGEN CONTROL ROOM (115), LSB (BLDG. 770) - Vandenberg Air Force Base, Space Launch Complex 3, Launch Pad 3 West, Napa & Alden Roads, Lompoc, Santa Barbara County, CA

  13. Liquid propellant densification

    NASA Technical Reports Server (NTRS)

    Lak, Tibor I. (Inventor); Petrilla, Steve P. (Inventor); Lozano, Martin E. (Inventor)

    1997-01-01

    Super cooling the cryogenic liquid propellant in a vehicle propellant tank densities the propellant allowing the vehicle propellant tank to carry more fuel in the same volume tank while lowering the vapor pressure and thus the tank operating pressure. Lowering the tank operating pressure reduces the stress and therefore allows the walls of the tank to be thinner. Both the smaller tank volume and thinner tank wall results in an overall smaller and lighter vehicle with increased payload capability. The cryogenic propellant can be supercooled well below the normal boiling point temperature level by transporting the liquid propellant from the vehicle tanks to a ground based cooling unit which utilizes a combination of heat exchanger and compressor. The compressor lowers the coolant fluid bath pressure resulting in a low temperature boiling liquid which is subsequently used to cool the recirculating liquid. The cooled propellant is then returned to the vehicle propellant tank. In addition to reducing the vehicle size and weight the invention also allows location of the vent valve on the ground, elimination of on-board recirculation pumps or bleed systems, smaller and lighter engine pumps and valves, lighter and more stable ullage gas, and significant reduction in tank fill operation. All of these mentioned attributes provide lower vehicle weight and cost.

  14. Properties of Liquid Plutonium

    SciTech Connect

    Freibert, Franz J.; Mitchell, Jeremy N.; Schwartz, Daniel S.; Saleh, Tarik A.; Migliori, Albert

    2012-08-02

    Unalloyed polycrystalline Pu displays extreme thermal expansion behavior, i.e., {alpha} {yields} {beta} {yields} {gamma} {yields} {delta} increases by 25% in volume and {delta} {yields} {var_epsilon} {yields} liquid decreases by 4.5% in volume. Thus, making it difficult to measure density into the liquid state. Dilatometer outfitted with CaF molten metal cell offers a proven capability to measure thermal expansion in molten metals, but has yet to be proven for Pu. Historic data from the liquid nuclear fuels program will prove extremely useful as a guide to future measurements. 3.3at% Ga changes Pu molten metal properties: 50% increase in viscosity and {approx}3% decrease in density. Fe may decrease the density by a small amount assuming an averaging of densities for Pu-Ga and Pu-Fe liquids. More recent Boivineau (2009) work needs some interpretation, but technique is being employed in (U,Pu)O{sub 2} nuclear fuels program (Pu Futures, 2012).

  15. Liquid laser cavities

    NASA Technical Reports Server (NTRS)

    Bjorklund, S.; Filipescu, N.; Kellermeyer, G. L.; Mc Avoy, N.

    1969-01-01

    Liquid laser cavities have plenum chambers at the ends of the capillary cell which are terminated in transparent optical flats. By use of these cavities, several new europium chelates and a terbium chelate can provide laser action in solution at room temperature.

  16. Clean room wiping liquids

    SciTech Connect

    Harding, W.B.

    1991-12-01

    A water-based liquid containing isopropyl alcohol, ammonium hydroxide, and surfactants was developed to replace 1,1,2-trichlorotrifluoroethane for the dampening of clean room wiping cloths used to wipe clean benches, clean room equipment, and latex finger cots and gloves.

  17. ELECTRONS IN NONPOLAR LIQUIDS.

    SciTech Connect

    HOLROYD,R.A.

    2002-10-22

    Excess electrons can be introduced into liquids by absorption of high energy radiation, by photoionization, or by photoinjection from metal surfaces. The electron's chemical and physical properties can then be measured, but this requires that the electrons remain free. That is, the liquid must be sufficiently free of electron attaching impurities for these studies. The drift mobility as well as other transport properties of the electron are discussed here as well as electron reactions, free-ion yields and energy levels, Ionization processes typically produce electrons with excess kinetic energy. In liquids during thermalization, where this excess energy is lost to bath molecules, the electrons travel some distance from their geminate positive ions. In general the electrons at this point are still within the coulombic field of their geminate ions and a large fraction of the electrons recombine. However, some electrons escape recombination and the yield that escapes to become free electrons and ions is termed G{sub fi}. Reported values of G{sub fi} for molecular liquids range from 0.05 to 1.1 per 100 eV of energy absorbed. The reasons for this 20-fold range of yields are discussed here.

  18. Liquid Rocket Engine Testing

    NASA Technical Reports Server (NTRS)

    Rahman, Shamim

    2005-01-01

    Comprehensive Liquid Rocket Engine testing is essential to risk reduction for Space Flight. Test capability represents significant national investments in expertise and infrastructure. Historical experience underpins current test capabilities. Test facilities continually seek proactive alignment with national space development goals and objectives including government and commercial sectors.

  19. Thermoelectricity in liquid crystals

    NASA Astrophysics Data System (ADS)

    Mohd Said, Suhana; Nordin, Abdul Rahman; Abdullah, Norbani; Balamurugan, S.

    2015-09-01

    The thermoelectric effect, also known as the Seebeck effect, describes the conversion of a temperature gradient into electricity. A Figure of Merit (ZT) is used to describe the thermoelectric ability of a material. It is directly dependent on its Seebeck coefficient and electrical conductivity, and inversely dependent on its thermal conductivity. There is usually a compromise between these parameters, which limit the performance of thermoelectric materials. The current achievement for ZT~2.2 falls short of the expected threshold of ZT=3 to allow its viability in commercial applications. In recent times, advances in organic thermoelectrics been significant, improving by over 3 orders of magnitude over a period of about 10 years. Liquid crystals are newly investigated as candidate thermoelectric materials, given their low thermal conductivity, inherent ordering, and in some cases, reasonable electrical conductivity. In this work the thermoelectric behaviour of a discotic liquid crystal, is discussed. The DLC was filled into cells coated with a charge injector, and an alignment of the columnar axis perpendicular to the substrate was allowed to form. This thermoelectric behavior can be correlated to the order-disorder transition. A reasonable thermoelectric power in the liquid crystal temperature regime was noted. In summary, thermoelectric liquid crystals may have the potential to be utilised in flexible devices, as a standalone power source.

  20. Liquid metal hydrogen barriers

    DOEpatents

    Grover, George M.; Frank, Thurman G.; Keddy, Edward S.

    1976-01-01

    Hydrogen barriers which comprise liquid metals in which the solubility of hydrogen is low and which have good thermal conductivities at operating temperatures of interest. Such barriers are useful in nuclear fuel elements containing a metal hydride moderator which has a substantial hydrogen dissociation pressure at reactor operating temperatures.

  1. Liquid rocket valve components

    NASA Technical Reports Server (NTRS)

    1973-01-01

    A monograph on valves for use with liquid rocket propellant engines is presented. The configurations of the various types of valves are described and illustrated. Design criteria and recommended practices for the various valves are explained. Tables of data are included to show the chief features of valve components in use on operational vehicles.

  2. Liquid rocket valve assemblies

    NASA Technical Reports Server (NTRS)

    1973-01-01

    The design and operating characteristics of valve assemblies used in liquid propellant rocket engines are discussed. The subjects considered are as follows: (1) valve selection parameters, (2) major design aspects, (3) design integration of valve subassemblies, and (4) assembly of components and functional tests. Information is provided on engine, stage, and spacecraft checkout procedures.

  3. Liquid White Enamel.

    ERIC Educational Resources Information Center

    Widmar, Marge

    1985-01-01

    A secondary teacher describes how she has her students use liquid white enamel. With the enameling process, students can create lasting, exciting artwork. They can exercise an understanding of design and color while learning the value of careful, sustained craft skills. (RM)

  4. Liquid Cooled Garments

    NASA Technical Reports Server (NTRS)

    1979-01-01

    Astronauts working on the surface of the moon had to wear liquid-cooled garments under their space suits as protection from lunar temperatures which sometimes reach 250 degrees Fahrenheit. In community service projects conducted by NASA's Ames Research Center, the technology developed for astronaut needs has been adapted to portable cooling systems which will permit two youngsters to lead more normal lives.

  5. Unusually stable liquid foams.

    PubMed

    Rio, Emmanuelle; Drenckhan, Wiebke; Salonen, Anniina; Langevin, Dominique

    2014-03-01

    Obtaining stable liquid foams is an important issue in view of their numerous applications. In some of these, the liquid foam in itself is of interest, in others, the liquid foam acts as a precursor for the generation of solid foam. In this short review, we will make a survey of the existing results in the area. This will include foams stabilised by surfactants, proteins and particles. The origin of the stability is related to the slowing down of coarsening, drainage or coalescence, and eventually to their arrest. The three effects are frequently coupled and in many cases, they act simultaneously and enhance one another. Drainage can be arrested if the liquid of the foam either gels or solidifies. Coalescence is slowed down by gelified foam films, and it can be arrested if the films become very thick and/or rigid. These mechanisms are thus qualitatively easy to identify, but they are less easy to model in order to obtain quantitative predictions. The slowing down of coarsening requests either very thick or small films, and its arrest was observed in cases where the surface compression modulus was large. The detail of the mechanisms at play remains unclear. PMID:24342735

  6. Spontaneous liquid-liquid phase separation of water.

    PubMed

    Yagasaki, Takuma; Matsumoto, Masakazu; Tanaka, Hideki

    2014-02-01

    We report a molecular dynamics simulation demonstrating a fast spontaneous liquid-liquid phase separation of water and a subsequent slow crystallization to ice. It is found that supercooled water separates rapidly into low- and high-density domains so as to reduce the surface energy in the rectangular simulation cell at certain thermodynamic states. The liquid-liquid phase separation, which is about two orders of magnitude faster than the crystallization, suggests a possibility to observe this phenomenon experimentally. PMID:25353404

  7. 49 CFR 173.150 - Exceptions for Class 3 (flammable and combustible liquids).

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... CFR 4.10 and 5.11) is not subject to the requirements of this subchapter if it— (1) Contains 24..., see the List of CFR Sections Affected which appears in the Finding Aids section of the printed volume... SHIPPERS-GENERAL REQUIREMENTS FOR SHIPMENTS AND PACKAGINGS Definitions Classification, Packing...

  8. 3-Methylpiperidinium ionic liquids.

    PubMed

    Belhocine, Tayeb; Forsyth, Stewart A; Gunaratne, H Q Nimal; Nieuwenhuyzen, Mark; Nockemann, Peter; Puga, Alberto V; Seddon, Kenneth R; Srinivasan, Geetha; Whiston, Keith

    2015-04-28

    A wide range of room temperature ionic liquids based on the 3-methylpiperdinium cation core were produced from 3-methylpiperidine, which is a derivative of DYTEK® A amine. First, reaction with 1-bromoalkanes or 1-bromoalkoxyalkanes generated the corresponding tertiary amines (Rmβpip, R = alkyl or alkoxyalkyl); further quaternisation reactions with the appropriate methylating agents yielded the quaternary [Rmmβpip]X salts (X(-) = I(-), [CF3CO2](-) or [OTf](-); Tf = -SO2CF3), and [Rmmβpip][NTf2] were prepared by anion metathesis from the corresponding iodides. All [NTf2](-) salts are liquids at room temperature. [Rmmβpip]X (X(-) = I(-), [CF3CO2](-) or [OTf](-)) are low-melting solids when R = alkyl, but room temperature liquids upon introduction of ether functionalities on R. Neither of the 3-methylpiperdinium ionic liquids showed any signs of crystallisation, even well below 0 °C. Some related non-C-substituted piperidinium and pyrrolidinium analogues were prepared and studied for comparison. Crystal structures of 1-hexyl-1,3-dimethylpiperidinium tetraphenylborate, 1-butyl-3-methylpiperidinium bromide, 1-(2-methoxyethyl)-1-methylpiperidinium chloride and 1-(2-methoxyethyl)-1-methylpyrrolidinium bromide are reported. Extensive structural and physical data are collected and compared to literature data, with special emphasis on the systematic study of the cation ring size and/or asymmetry effects on density, viscosity and ionic conductivity, allowing general trends to be outlined. Cyclic voltammetry shows that 3-methylpiperidinium ionic liquids, similarly to azepanium, piperidinium or pyrrolidinium counterparts, are extremely electrochemically stable; the portfolio of useful alternatives for safe and high-performing electrolytes is thus greatly extended. PMID:25669485

  9. Rockets using Liquid Oxygen

    NASA Technical Reports Server (NTRS)

    Busemann, Adolf

    1947-01-01

    It is my task to discuss rocket propulsion using liquid oxygen and my treatment must be highly condensed for the ideas and experiments pertaining to this classic type of rocket are so numerous that one could occupy a whole morning with a detailed presentation. First, with regard to oxygen itself as compared with competing oxygen carriers, it is known that the liquid state of oxygen, in spite of the low boiling point, is more advantageous than the gaseous form of oxygen in pressure tanks, therefore only liquid oxygen need be compared with the oxygen carriers. The advantages of liquid oxygen are absolute purity and unlimited availability at relatively small cost in energy. The disadvantages are those arising from the impossibility of absolute isolation from heat; consequently, allowance must always be made for a certain degree of vaporization and only vented vessels can be used for storage and transportation. This necessity alone eliminates many fields of application, for example, at the front lines. In addition, liquid oxygen has a lower specific weight than other oxygen carriers, therefore many accessories become relatively larger and heavier in the case of an oxygen rocket, for example, the supply tanks and the pumps. The advantages thus become effective only in those cases where definitely scheduled operation and a large ground organization are possible and when the flight requires a great concentration of energy relative to weight. With the aim of brevity, a diagram of an oxygen rocket will be presented and the problem of various component parts that receive particularly thorough investigation in this classic case but which are also often applicable to other rocket types will be referred to.

  10. Microfabricated Liquid Rocket Motors

    NASA Technical Reports Server (NTRS)

    Epstein, Alan H.; Joppin, C.; Kerrebrock, J. L.; Schneider, Steven J. (Technical Monitor)

    2003-01-01

    Under NASA Glenn Research Center sponsorship, MIT has developed the concept of micromachined, bipropellant, liquid rocket engines. This is potentially a breakthrough technology changing the cost-performance tradeoffs for small propulsion systems, enabling new applications, and redefining the meaning of the term low-cost-access-to-space. With this NASA support, a liquid-cooled, gaseous propellant version of the thrust chamber and nozzle was designed, built, and tested as a first step. DARPA is currently funding MIT to demonstrate turbopumps and controls. The work performed herein was the second year of a proposed three-year effort to develop the technology and demonstrate very high power density, regeneratively cooled, liquid bipropellant rocket engine thrust chamber and nozzles. When combined with the DARPA turbopumps and controls, this work would enable the design and demonstration of a complete rocket propulsion system. The original MIT-NASA concept used liquid oxygen-ethanol propellants. The military applications important to DARPA imply that storable liquid propellants are needed. Thus, MIT examined various storable propellant combinations including N2O4 and hydrazine, and H2O2 and various hydrocarbons. The latter are preferred since they do not have the toxicity of N2O4 and hydrazine. In reflection of the newfound interest in H2O2, it is once again in production and available commercially. A critical issue for the microrocket engine concept is cooling of the walls in a regenerative design. This is even more important at microscale than for large engines due to cube-square scaling considerations. Furthermore, the coolant behavior of rocket propellants has not been characterized at microscale. Therefore, MIT designed and constructed an apparatus expressly for this purpose. The report details measurements of two candidate microrocket fuels, JP-7 and JP-10.

  11. Liquid metal thermoacoustic engine

    SciTech Connect

    Swift, G.W.; Migliori, A.; Wheatley, J.C.

    1986-01-01

    We are studying a liquid metal thermoacoustic engine both theoretically and experimentally. This type of engine promises to produce large quantities of electrical energy from heat at modest efficiency with no moving parts. A sound wave is usually thought of as consisting of pressure oscillations, but always attendant to the pressure oscillation are temperature oscillations. The combination produces a rich variety of ''thermoacoustic'' effects. These effects are usually so small that they are never noticed in everyday life; nevertheless under the right circumstances they can be harnessed to produce powerful heat engines, heat pumps, and refrigerators. In our liquid metal thermoacoustic engine, heat flow from a high temperature source to a low temperature sink generates a high-amplitude standing acoustic wave in liquid sodium. This acoustic power is converted to electric power by a simple magnetohydrodynamic effect at the acoustic oscillation frequency. We have developed a detailed thermoacoustic theory applicable to this engine, and find that a reasonably designed liquid sodium engine operating between 700/sup 0/C and 100/sup 0/C should generate about 60 W/cm/sup 2/ of acoustic power at about 1/3 of Carnot's efficiency. Construction of a 3000 W-thermal laboratory model engine has just been completed, and we have exciting preliminary experimental results as of the time of preparation of this manuscript showing, basically, that the engine works. We have also designed and built a 1 kHz liquid sodium magnetohydrodynamic generator and have extensive measurements on it. It is now very well characterized both experimentally and theoretically. The first generator of its kind, it already converts acoustic power to electric power with 40% efficiency. 16 refs., 5 figs.

  12. Models for a liquid-liquid phase transition

    NASA Astrophysics Data System (ADS)

    Buldyrev, S. V.; Franzese, G.; Giovambattista, N.; Malescio, G.; Sadr-Lahijany, M. R.; Scala, A.; Skibinsky, A.; Stanley, H. E.

    2002-02-01

    We use molecular dynamics simulations to study two- and three-dimensional models with the isotropic double-step potential which in addition to the hard core has a repulsive soft core of larger radius. Our results indicate that the presence of two characteristic repulsive distances (hard core and soft core) is sufficient to explain liquid anomalies and a liquid-liquid phase transition, but these two phenomena may occur independently. Thus liquid-liquid transitions may exist in systems like liquid metals, regardless of the presence of the density anomaly. For 2D, we propose a model with a specific set of hard core and soft core parameters, that qualitatively reproduces the phase diagram and anomalies of liquid water. We identify two solid phases: a square crystal (high density phase), and a triangular crystal (low density phase) and discuss the relation between the anomalies of liquid and the polymorphism of the solid. Similarly to real water, our 2D system may have the second critical point in the metastable liquid phase beyond the freezing line. In 3D, we find several sets of parameters for which two fluid-fluid phase transition lines exist: the first line between gas and liquid and the second line between high-density liquid (HDL) and low-density liquid (LDL). In all cases, the LDL phase shows no density anomaly in 3D. We relate the absence of the density anomaly with the positive slope of the LDL-HDL phase transition line.

  13. Polymorphism in glassy silicon: Inherited from liquid-liquid phase transition in supercooled liquid

    NASA Astrophysics Data System (ADS)

    Zhang, Shiliang; Wang, Li-Min; Zhang, Xinyu; Qi, Li; Zhang, Suhong; Ma, Mingzhen; Liu, Riping

    2015-02-01

    Combining molecular dynamics (MD) simulation and Voronoi polyhedral analyses, we discussed the microstructure evolution in liquid and glassy silicon during cooling by focusing on the fraction of various clusters. Liquid-liquid phase transition (LLPT) is detected in supercooled liquid silicon However, freezing the high-density liquid (HDL) to the glassy state is not achieved as the quenching rate goes up to 1014 K/s. The polyamorphism in glassy silicon is found to be mainly associated with low-density liquid (LDL).

  14. Polymorphism in glassy silicon: Inherited from liquid-liquid phase transition in supercooled liquid

    PubMed Central

    Zhang, Shiliang; Wang, Li-Min; Zhang, Xinyu; Qi, Li; Zhang, Suhong; Ma, Mingzhen; Liu, Riping

    2015-01-01

    Combining molecular dynamics (MD) simulation and Voronoi polyhedral analyses, we discussed the microstructure evolution in liquid and glassy silicon during cooling by focusing on the fraction of various clusters. Liquid-liquid phase transition (LLPT) is detected in supercooled liquid silicon However, freezing the high-density liquid (HDL) to the glassy state is not achieved as the quenching rate goes up to 1014 K/s. The polyamorphism in glassy silicon is found to be mainly associated with low-density liquid (LDL). PMID:25716054

  15. Black Liquid Solar Collector Demonstrator.

    ERIC Educational Resources Information Center

    Weichman, F. L.; Austen, D. J.

    1979-01-01

    Describes the details of constructing, and use of, a solar collector. Uses a black liquid to absorb the energy, the thermosyphon effect to drive the liquid through the collector, and a floodlamp as a surrogate sun. (GA)

  16. Demonstrating Paramagnetism Using Liquid Nitrogen.

    ERIC Educational Resources Information Center

    Simmonds, Ray; And Others

    1994-01-01

    Describes how liquid nitrogen is attracted to the poles of neodymium magnets. Nitrogen is not paramagnetic, so the attraction suggests that the liquid nitrogen contains a small amount of oxygen, which causes the paramagnetism. (MVL)

  17. Predicting Polymer/Liquide Interactions

    NASA Technical Reports Server (NTRS)

    Fedors, R. F.

    1985-01-01

    Calculations of liquid absorption by rubber vulcanizates agree with experimental results. Equation allows calculation of swelling of rubber vulcanizates by liquids, based on knowledge of chemical structure of polymer and solvent. Calculated values agree favorably with experimental data.

  18. Liquid metal thermal electric converter

    DOEpatents

    Abbin, Joseph P.; Andraka, Charles E.; Lukens, Laurance L.; Moreno, James B.

    1989-01-01

    A liquid metal thermal electric converter which converts heat energy to electrical energy. The design of the liquid metal thermal electric converter incorporates a unique configuration which directs the metal fluid pressure to the outside of the tube which results in the structural loads in the tube to be compressive. A liquid metal thermal electric converter refluxing boiler with series connection of tubes and a multiple cell liquid metal thermal electric converter are also provided.

  19. Liquid crystalline composites containing phyllosilicates

    DOEpatents

    Chaiko; David J.

    2007-05-08

    The present invention provides barrier films having reduced gas permeability for use in packaging and coating applications. The barrier films comprise an anisotropic liquid crystalline composite layer formed from phyllosilicate-polymer compositions. Phyllosilicate-polymer liquid crystalline compositions of the present invention can contain a high percentage of phyllosilicate while remaining transparent. Because of the ordering of the particles in the liquid crystalline composite, barrier films comprising liquid crystalline composites are particularly useful as barriers to gas transport.

  20. The human 18S rRNA base methyltransferases DIMT1L and WBSCR22-TRMT112 but not rRNA modification are required for ribosome biogenesis

    PubMed Central

    Zorbas, Christiane; Nicolas, Emilien; Wacheul, Ludivine; Huvelle, Emmeline; Heurgué-Hamard, Valérie; Lafontaine, Denis L. J.

    2015-01-01

    At the heart of the ribosome lie rRNAs, whose catalytic function in translation is subtly modulated by posttranscriptional modifications. In the small ribosomal subunit of budding yeast, on the 18S rRNA, two adjacent adenosines (A1781/A1782) are N6-dimethylated by Dim1 near the decoding site, and one guanosine (G1575) is N7-methylated by Bud23-Trm112 at a ridge between the P- and E-site tRNAs. Here we establish human DIMT1L and WBSCR22-TRMT112 as the functional homologues of yeast Dim1 and Bud23-Trm112. We report that these enzymes are required for distinct pre-rRNA processing reactions leading to synthesis of 18S rRNA, and we demonstrate that in human cells, as in budding yeast, ribosome biogenesis requires the presence of the modification enzyme rather than its RNA-modifying catalytic activity. We conclude that a quality control mechanism has been conserved from yeast to human by which binding of a methyltransferase to nascent pre-rRNAs is a prerequisite to processing, so that all cleaved RNAs are committed to faithful modification. We further report that 18S rRNA dimethylation is nuclear in human cells, in contrast to yeast, where it is cytoplasmic. Yeast and human ribosome biogenesis thus have both conserved and distinctive features. PMID:25851604