Science.gov

Sample records for air support cas

  1. Scorpion: Close Air Support (CAS) aircraft

    NASA Technical Reports Server (NTRS)

    Allen, Chris; Cheng, Rendy; Koehler, Grant; Lyon, Sean; Paguio, Cecilia

    1991-01-01

    The objective is to outline the results of the preliminary design of the Scorpion, a proposed close air support aircraft. The results obtained include complete preliminary analysis of the aircraft in the areas of aerodynamics, structures, avionics and electronics, stability and control, weight and balance, propulsion systems, and costs. A conventional wing, twin jet, twin-tail aircraft was chosen to maximize the desirable characteristics. The Scorpion will feature low speed maneuverability, high survivability, low cost, and low maintenance. The life cycle cost per aircraft will be 17.5 million dollars. The maximum takeoff weight will be 52,760 pounds. Wing loading will be 90 psf. The thrust to weight will be 0.6 lbs/lb. This aircraft meets the specified mission requirements. Some modifications have been suggested to further optimize the design.

  2. Preliminary design of a family of close air support aircraft

    NASA Technical Reports Server (NTRS)

    Tuschhoff, Jeff; Bartel, Rudy; Cox, Brian; Darrah, Paul; Drake, TY; Hendrich, Louis; Hicks, Robin; Holt, Mark; Hoyle, Mark; Kerns, Brian

    1989-01-01

    A family of three Close Air Support (CAS) aircraft is presented. These aircraft are designed with commonality as the main design objective to reduce the life cycle cost. The aircraft are low wing, twin-boom, pusher turbo-prop configurations. The amount of information displayed to the pilot was reduced to a minimum to greatly simplify the cockpit. The aircraft met the mission specifications and the performance and cost characteristics compared well with other CAS aircraft. The concept of a family of CAS aircraft seems viable after preliminary design.

  3. Air Risk Information Support Center

    SciTech Connect

    Shoaf, C.R.; Guth, D.J.

    1990-12-31

    The Air Risk Information Support Center (Air RISC) was initiated in early 1988 by the US Environmental Protection Agency`s (EPA) Office of Health and Environmental Assessment (OHEA) and the Office of Air Quality Planning and Standards (OAQPS) as a technology transfer effort that would focus on providing information to state and local environmental agencies and to EPA Regional Offices in the areas of health, risk, and exposure assessment for toxic air pollutants. Technical information is fostered and disseminated by Air RISCs three primary activities: (1) a {open_quotes}hotline{close_quotes}, (2) quick turn-around technical assistance projects, and (3) general technical guidance projects. 1 ref., 2 figs.

  4. The Guardian: Preliminary design of a close air support aircraft

    NASA Technical Reports Server (NTRS)

    Haag, Jonathan; Huber, David; Mcinerney, Kelly; Mulligan, Greg; Pessin, David; Seelos, Michael

    1991-01-01

    One design is presented of a Close Air Support (CAS) aircraft. It is a canard wing, twin engine, twin vertical tail aircraft that has the capability to cruise at 520 knots. The Guardian contains state of the art flight control systems. Specific highlights of the Guardian include: (1) low cost (the acquisition cost per airplane is $13.6 million for a production of 500 airplanes); (2) low maintenance (it was designed to be easily maintainable in unprepared fields); and (3) high versatility (it can perform a wide range of missions). Along with being a CAS aircraft, it is capable of long ferry missions, battlefield interdiction, maritime attack, and combat rescue. The Guardian is capable of a maximum ferry of 3800 nm, can takeoff in a distance of 1700 ft, land in a ground roll distance of 1644 ft. It has a maximum takeoff weight of 48,753 lbs, and is capable of carrying up to 19,500 lbs of ordinance.

  5. Preliminary design of a family of three close air support aircraft

    NASA Technical Reports Server (NTRS)

    Cox, Brian; Darrah, Paul; Lussier, Wayne; Mills, Nikos

    1989-01-01

    A family of three Close Air Support aircraft is presented. These aircraft are designed with commonality as the main design objective to reduce the life cycle cost. The aircraft are low wing, twin-boom, pusher turbo-prop configurations. The amount of information displayed to the pilot was reduced to a minimum to greatly simplify the cockpit. The aircraft met the mission specifications and the performance and cost characteristics compared well with other CAS aircraft. The concept of a family of CAS aircraft seems viable after preliminary design.

  6. CALM: Complex Adaptive System (CAS)-Based Decision Support for Enabling Organizational Change

    NASA Astrophysics Data System (ADS)

    Adler, Richard M.; Koehn, David J.

    Guiding organizations through transformational changes such as restructuring or adopting new technologies is a daunting task. Such changes generate workforce uncertainty, fear, and resistance, reducing morale, focus and performance. Conventional project management techniques fail to mitigate these disruptive effects, because social and individual changes are non-mechanistic, organic phenomena. CALM (for Change, Adaptation, Learning Model) is an innovative decision support system for enabling change based on CAS principles. CALM provides a low risk method for validating and refining change strategies that combines scenario planning techniques with "what-if" behavioral simulation. In essence, CALM "test drives" change strategies before rolling them out, allowing organizations to practice and learn from virtual rather than actual mistakes. This paper describes the CALM modeling methodology, including our metrics for measuring organizational readiness to respond to change and other major CALM scenario elements: prospective change strategies; alternate futures; and key situational dynamics. We then describe CALM's simulation engine for projecting scenario outcomes and its associated analytics. CALM's simulator unifies diverse behavioral simulation paradigms including: adaptive agents; system dynamics; Monte Carlo; event- and process-based techniques. CALM's embodiment of CAS dynamics helps organizations reduce risk and improve confidence and consistency in critical strategies for enabling transformations.

  7. Development Process of a Praxeology for Supporting the Teaching of Proofs in a CAS Environment Based on Teachers' Experience in a Professional Development Course

    ERIC Educational Resources Information Center

    Zehavi, Nurit; Mann, Giora

    2011-01-01

    This paper presents the development process of a "praxeology" (theory-of-practice) for supporting the teaching of proofs in a CAS environment. The characteristics of the praxeology were elaborated within the frame of a professional development course for teaching analytic geometry with CAS. The theoretical framework draws on Chevallard's…

  8. Proposal for a low cost close air support aircraft for the year 2000: The Raptor

    NASA Technical Reports Server (NTRS)

    Brown, Jerome D.; Dewitt, Ward S.; Mcdonald, Mark; Riley, John W.; Roberts, Anthony E.; Watson, Sean; Whelan, Margaret M.

    1991-01-01

    The Raptor is a proposed low cost Close Air Support (CAS) aircraft for the U.S. Military. The Raptor incorporates a 'cranked arrow' wing planform, and uses canards instead of a traditional horizontal tail. The Raptor is designed to be capable of responsive delivery of effective ordnance in close proximity to friendly ground forces during the day, night, and 'under the weather' conditions. Details are presented of the Raptor's mission, configuration, performance, stability and control, ground support, manufacturing, and overall cost to permit engineering evaluation of the proposed design. A description of the design process and analysis methods used is also provided.

  9. AIRS Mission Support from GES DISC

    NASA Technical Reports Server (NTRS)

    Wei, Jennifer; Hearty, Thomas; Savtchenko, Audrey; Ding, Feng; Esfandiari, Ed; Theobald, Mike; Vollmer, Bruce; Kempler, Steve

    2015-01-01

    This talk will describe the support and distribution of AIRS (Atmospheric Infra Red Sounding) data products that are archived and distributed from the Goddard Earth Sciences Data and Information Services Center. Along with data stewardship, an important mission of GES DISC is to enhance the usability of data and broaden the user base. We will provide a brief summary of the current online archive and distribution metrics for the AIRS v5 and v6 products. We will also describe collaborative data sets and services (e.g., visualization and potential science applications) and solicit feedback for potential future services.

  10. Manx: Close air support aircraft preliminary design

    NASA Technical Reports Server (NTRS)

    Amy, Annie; Crone, David; Hendrickson, Heidi; Willis, Randy; Silva, Vince

    1991-01-01

    The Manx is a twin engine, twin tailed, single seat close air support design proposal for the 1991 Team Student Design Competition. It blends advanced technologies into a lightweight, high performance design with the following features: High sensitivity (rugged, easily maintained, with night/adverse weather capability); Highly maneuverable (negative static margin, forward swept wing, canard, and advanced avionics result in enhanced aircraft agility); and Highly versatile (design flexibility allows the Manx to contribute to a truly integrated ground team capable of rapid deployment from forward sites).

  11. Air support facilities. [interface between air and surface transportation systems

    NASA Technical Reports Server (NTRS)

    1975-01-01

    Airports are discussed in terms of the interface between the ground and air for transportation systems. The classification systems, design, facilities, administration, and operations of airports are described.

  12. Air Systems Provide Life Support to Miners

    NASA Technical Reports Server (NTRS)

    2014-01-01

    Through a Space Act Agreement with Johnson Space Center, Paragon Space Development Corporation, of Tucson, Arizona, developed the Commercial Crew Transport-Air Revitalization System, designed to provide clean air for crewmembers on short-duration space flights. The technology is now being used to help save miners' lives in the event of an underground disaster.

  13. Cyclone: A close air support aircraft for tomorrow

    NASA Technical Reports Server (NTRS)

    Cox, George; Croulet, Donald; Dunn, James; Graham, Michael; Ip, Phillip; Low, Scott; Vance, Gregg; Volckaert, Eric

    1991-01-01

    To meet the threat of the battlefield of the future, the U.S. ground forces will require reliable air support. To provide this support, future aircrews demand a versatile close air support aircraft capable of delivering ordinance during the day, night, or in adverse weather with pin-point accuracy. The Cyclone aircraft meets these requirements, packing the 'punch' necessary to clear the way for effective ground operations. Possessing anti-armor, missile, and precision bombing capability, the Cyclone will counter the threat into the 21st Century. Here, it is shown that the Cyclone is a realistic, economical answer to the demand for a capable close air support aircraft.

  14. A-2000: Close air support aircraft design team

    NASA Technical Reports Server (NTRS)

    Carrannanto, Paul; Lim, Don; Lucas, Evangeline; Risse, Alan; Weaver, Dave; Wikse, Steve

    1991-01-01

    The US Air Force is currently faced with the problem of providing adequate close air support for ground forces. Air response to troops engaged in combat must be rapid and devastating due to the highly fluid battle lines of the future. The A-2000 is the result of a study to design an aircraft to deliver massive fire power accurately. The low cost A-2000 incorporates: large weapons payload; excellent maneuverability; all weather and terrain following capacity; redundant systems; and high survivability.

  15. Controlling UCAVs by JTACs in CAS missions

    NASA Astrophysics Data System (ADS)

    Kumaş, A. E.

    2014-06-01

    By means of evolving technology, capabilities of UAVs (Unmanned Aerial Vehicle)s are increasing rapidly. This development provides UAVs to be used in many different areas. One of these areas is CAS (Close Air Support) mission. UAVs have several advantages compared to manned aircraft, however there are also some problematic areas. The remote controlling of these vehicles from thousands of nautical miles away via satellite may lead to various problems both ethical and tactical aspects. Therefore, CAS missions require a good level of ALI (Air-Land Integration), a high SA (situational awareness) and precision engagement. In fact, there is an aware friendly element in the target area in CAS missions, unlike the other UAV operations. This element is an Airman called JTAC (Joint Terminal Attack Controller). Unlike the JTAC, UAV operators are too far away from target area and use the limited FOV (Field of View) provided by camera and some other sensor data. In this study, target area situational awareness of a UAV operator and a JTAC, in a high-risk mission for friendly ground forces and civilians such as CAS, are compared. As a result of this comparison, answer to the question who should control the UCAV (Unmanned Combat Aerial Vehicle) in which circumstances is sought. A literature review is made in UAV and CAS fields and recent air operations are examined. The control of UCAV by the JTAC is assessed by SWOT analysis and as a result it is deduced that both control methods can be used in different situations within the framework of the ROE (Rules Of Engagement) is reached.

  16. 6. Soft support building, view towards south Ellsworth Air ...

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

    6. Soft support building, view towards south - Ellsworth Air Force Base, Delta Flight, Launch Facility D-6, 4 miles north of Badlands National Park Headquarters, 4.5 miles east of Jackson County line on county road, Interior, Jackson County, SD

  17. High efficiency, down flow air filter sealing and support system

    SciTech Connect

    Mattison, A.H.

    1986-07-15

    An assembly of high efficiency air filter units through which essentially all air entering a clean space below the units must pass to remove particulate matter down to sub-micron size from the air, the assembly comprising: (a) a plurality of air filter units each having a filter core of pleated media sealed in air-tight engagement on four sides to a surrounding, box-like, rigid frame, having side and end members; (b) means for supporting the filter units adjacent the upper surfaces thereof from structure above the space with adjacent units having the side and end members thereof providing adjoining vertical surfaces in closely spaced relation with the lower surfaces of the units in essentially the same horizontal plane to form at least a portion of the top of the space; and (c) a caulking material filling all spaces between the adjoining vertical surfaces of adjacent filter units, effectively sealing the spaces and providing the sole means preventing passage of air around the units.

  18. AIRS Data Support at NASA Goddard Earth Science DISC DAAC

    NASA Astrophysics Data System (ADS)

    Cho, S.; Qin, J.; Sharma, A.

    2002-05-01

    The Atmospheric Infrared Sounder (AIRS) is selected by NASA to fly on the second Earth Observing System (EOS) polar orbiting platform, EOS Aqua, which is launched in April 2002. AIRS, together with Advanced Microwave Sounding Unit (AMSU) and Humidity Sounder for Brazil (HSB), is designed to meet the requirements of the NASA Earth Science Enterprise climate research program and the NOAA operational weather forecasting The data products from the AIRS/AMSU/HSB will be archived and distributed at the Goddard Distributed Active Archive Center (GDAAC) located in the NASA Goddard Earth Sciences Data and Information Services Center (GES DAAC) in later 2002. This new dataset consists of radiances, geo-locations and atmospheric products, such as, temperature, humidity, cloud and ozone, providing measurements for temperature at an accuracy of 1 o C in layers 1 km thick and humidity with an accuracy of 20 % in layers 2 km thick in the troposphere. The data will be freely available via WWW interfaces, or an FTP containing subsetted and reformatted data products. The GES DISC DAAC Search and Order allows users to search for data by following particular paths down the hierarchy. This simple point-and- click navigational web interface shows temporal and spatial coverage, item size, description and browse images for AIRS data and one can customize search using spatial,temporal, attribute and parameter search. The EOS Data Gateway (EDG) is another user interface for searching and ordering the AIRS data together with other data products obtained from EOS instruments. The Atmospheric Dynamics Data Support Team (ADDST) at the GES DISC/DAAC will provide various services to assist users in understanding, accessing, and using AIRS data product. The ADDST has been developing tools to read, visualize and analyze the AIRS data, channel/parameter subsetting of AIRS HDF-EOS data products and supplying documentation and readme et al. Other services provided by the ADDST will contain assistance

  19. Merging Air Quality and Public Health Decision Support Systems

    NASA Astrophysics Data System (ADS)

    Hudspeth, W. B.; Bales, C. L.

    2003-12-01

    The New Mexico Air Quality Mapper (NMAQM) is a Web-based, open source GIS prototype application that Earth Data Analysis Center is developing under a NASA Cooperative Agreement. NMAQM enhances and extends existing data and imagery delivery systems with an existing Public Health system called the Rapid Syndrome Validation Project (RSVP). RSVP is a decision support system operating in several medical and public health arenas. It is evolving to ingest remote sensing data as input to provide early warning of human health threats, especially those related to anthropogenic atmospheric pollutants and airborne pathogens. The NMAQM project applies measurements of these atmospheric pollutants, derived from both remotely sensed data as well as from in-situ air quality networks, to both forecasting and retrospective analyses that influence human respiratory health. NMAQM provides a user-friendly interface for visualizing and interpreting environmentally-linked epidemiological phenomena. The results, and the systems made to provide the information, will be applicable not only to decision-makers in the public health realm, but also to air quality organizations, demographers, community planners, and other professionals in information technology, and social and engineering sciences. As an accessible and interactive mapping and analysis application, it allows environment and health personnel to study historic data for hypothesis generation and trend analysis, and then, potentially, to predict air quality conditions from daily data acquisitions. Additional spin off benefits to such users include the identification of gaps in the distribution of in-situ monitoring stations, the dissemination of air quality data to the public, and the discrimination of local vs. more regional sources of air pollutants that may bear on decisions relating to public health and public policy.

  20. Supporting the Future Air Traffic Control Projection Process

    NASA Technical Reports Server (NTRS)

    Davison, Hayley J.; Hansman, R. John, Jr.

    2002-01-01

    In air traffic control, projecting what the air traffic situation will be over the next 30 seconds to 30 minutes is a key process in identifying conflicts that may arise so that evasive action can be taken upon discovery of these conflicts. A series of field visits in the Boston and New York terminal radar approach control (TRACON) facilities and in the oceanic air traffic control facilities in New York and Reykjavik, Iceland were conducted to investigate the projection process in two different ATC domains. The results from the site visits suggest that two types of projection are currently used in ATC tasks, depending on the type of separation minima and/or traffic restriction and information display used by the controller. As technologies improve and procedures change, care should be taken by designers to support projection through displays, automation, and procedures. It is critical to prevent time/space mismatches between interfaces and restrictions. Existing structure in traffic dynamics could be utilized to provide controllers with useful behavioral models on which to build projections. Subtle structure that the controllers are unable to internalize could be incorporated into an ATC projection aid.

  1. The Eliminator: A design of a close air support aircraft

    NASA Technical Reports Server (NTRS)

    Hendrix, Mandy; Hoang, TY; Kokolios, Alex; Selyem, Sharon; Wardell, Mark; Winterrowd, David

    1991-01-01

    The Eliminator is the answer to the need for an affordable, maintainable, survivable, high performance close air support aircraft primarily for the United States, but with possible export sales to foreign customers. The Eliminator is twin turbofan, fixed wing aircraft with high mounted canards and low mounted wings. It is designed for high subsonic cruise and an attack radius of 250 nautical miles. Primarily it would carry 20 500 pound bombs as its main ordnance , but is versatile enough to carry a variety of weapons configurations to perform several different types of missions. It carries state of the art navigation and targeting systems to deliver its payload with pinpoint precision and is designed for maximum survivability of the crew and aircraft for a safe return and quick turnaround. It can operate from fields as short as 1800 ft. with easy maintenance for dispersed operation during hostile situations. It is designed for exceptional maneuverability and could be used in a variety of roles from air-to-air operations to anti-submarine warfare and maritime patrol duties.

  2. COMMUNICATING RISK INFORMATION TO STATE AND LOCAL AIR POLLUTION CONTROL AGENCIES VIA U.S. EPA'S AIR RISK INFORMATION SUPPORT CENTER (AIR RISC)

    EPA Science Inventory

    The Air Risk Information Support Center (Air RISC) has been organized by U.S. EPA's offices of Air Quality Planning and Standards and Health and Environmental Assessment. The center has been developed in cooperation with the State and Territorial air Pollution Control Program Adm...

  3. 32 CFR 644.535 - Support in clearance of Air Force lands.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 32 National Defense 4 2011-07-01 2011-07-01 false Support in clearance of Air Force lands. 644.535 Section 644.535 National Defense Department of Defense (Continued) DEPARTMENT OF THE ARMY (CONTINUED) REAL... Excess Land and Improvements § 644.535 Support in clearance of Air Force lands. Where Air Force...

  4. 32 CFR 644.535 - Support in clearance of Air Force lands.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 32 National Defense 4 2014-07-01 2013-07-01 true Support in clearance of Air Force lands. 644.535 Section 644.535 National Defense Department of Defense (Continued) DEPARTMENT OF THE ARMY (CONTINUED) REAL... Excess Land and Improvements § 644.535 Support in clearance of Air Force lands. Where Air Force...

  5. 32 CFR 644.535 - Support in clearance of Air Force lands.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 32 National Defense 4 2013-07-01 2013-07-01 false Support in clearance of Air Force lands. 644.535 Section 644.535 National Defense Department of Defense (Continued) DEPARTMENT OF THE ARMY (CONTINUED) REAL... Excess Land and Improvements § 644.535 Support in clearance of Air Force lands. Where Air Force...

  6. 32 CFR 644.535 - Support in clearance of Air Force lands.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 32 National Defense 4 2010-07-01 2010-07-01 true Support in clearance of Air Force lands. 644.535 Section 644.535 National Defense Department of Defense (Continued) DEPARTMENT OF THE ARMY (CONTINUED) REAL... Excess Land and Improvements § 644.535 Support in clearance of Air Force lands. Where Air Force...

  7. 32 CFR 644.535 - Support in clearance of Air Force lands.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 32 National Defense 4 2012-07-01 2011-07-01 true Support in clearance of Air Force lands. 644.535 Section 644.535 National Defense Department of Defense (Continued) DEPARTMENT OF THE ARMY (CONTINUED) REAL... Excess Land and Improvements § 644.535 Support in clearance of Air Force lands. Where Air Force...

  8. Characterizing and Supporting Change in Algebra Students' Representational Fluency in a CAS/Paper-and-Pencil Environment

    ERIC Educational Resources Information Center

    Fonger, Nicole L.

    2012-01-01

    Representational fluency (RF) includes an ability to interpret, create, move within and among, and connect tool-based representations of mathematical objects. Taken as an indicator of conceptual understanding, there is a need to better support school algebra students' RF in learning environments that utilize both computer algebra systems…

  9. Wind Prediction Accuracy for Air Traffic Management Decision Support Tools

    NASA Technical Reports Server (NTRS)

    Cole, Rod; Green, Steve; Jardin, Matt; Schwartz, Barry; Benjamin, Stan

    2000-01-01

    The performance of Air Traffic Management and flight deck decision support tools depends in large part on the accuracy of the supporting 4D trajectory predictions. This is particularly relevant to conflict prediction and active advisories for the resolution of conflicts and the conformance with of traffic-flow management flow-rate constraints (e.g., arrival metering / required time of arrival). Flight test results have indicated that wind prediction errors may represent the largest source of trajectory prediction error. The tests also discovered relatively large errors (e.g., greater than 20 knots), existing in pockets of space and time critical to ATM DST performance (one or more sectors, greater than 20 minutes), are inadequately represented by the classic RMS aggregate prediction-accuracy studies of the past. To facilitate the identification and reduction of DST-critical wind-prediction errors, NASA has lead a collaborative research and development activity with MIT Lincoln Laboratories and the Forecast Systems Lab of the National Oceanographic and Atmospheric Administration (NOAA). This activity, begun in 1996, has focussed on the development of key metrics for ATM DST performance, assessment of wind-prediction skill for state of the art systems, and development/validation of system enhancements to improve skill. A 13 month study was conducted for the Denver Center airspace in 1997. Two complementary wind-prediction systems were analyzed and compared to the forecast performance of the then standard 60 km Rapid Update Cycle - version 1 (RUC-1). One system, developed by NOAA, was the prototype 40-km RUC-2 that became operational at NCEP in 1999. RUC-2 introduced a faster cycle (1 hr vs. 3 hr) and improved mesoscale physics. The second system, Augmented Winds (AW), is a prototype en route wind application developed by MITLL based on the Integrated Terminal Wind System (ITWS). AW is run at a local facility (Center) level, and updates RUC predictions based on an

  10. Attitudes Toward Family Enrichment and Support Programs among Air Force Families.

    ERIC Educational Resources Information Center

    Orthner, Dennis K.; Bowen, Gary L.

    1982-01-01

    Drawing upon survey data from a probability sample of 331 U.S. Air Force married couples and 101 Air Force single parents, determined the attitudes of Air Force families toward family enrichment and support programs. States implications for family programing. (Author)

  11. Removal of formaldehyde from air using functionalized silica supports.

    PubMed

    Ewlad-Ahmed, Abdunaser M; Morris, Michael A; Patwardhan, Siddharth V; Gibson, Lorraine T

    2012-12-18

    This paper demonstrates the use of functionalized meso-silica materials (MCM-41 or SBA-15) as adsorbents for formaldehyde (H₂CO) vapor from contaminated air. Additionally new green nanosilica (GNs) materials were prepared via a bioinspired synthesis route and were assessed for removal of H₂CO from contaminated indoor air. These exciting new materials were prepared via rapid, 15 min, environmentally friendly synthesis routes avoiding any secondary pollution. They provided an excellent platform for functionalization and extraction of H₂CO demonstrating similar performance to the conventional meso-silica materials. To the authors' knowledge this is the first reported practical application of this material type. Prior to trapping, all materials were functionalized with amino-propyl groups which led to chemisorption of H₂CO; removing it permanently from air. No retention of H₂CO was achieved with nonfunctionalized material and it was observed that best extraction performance required a dynamic adsorption setup when compared to passive application. These results demonstrate the first application of GNs as potential adsorbents and functionalized meso-silica for use in remediation of air pollution in indoor air. PMID:23181357

  12. Scale Issues in Air Quality Modeling Policy Support

    EPA Science Inventory

    This study examines the issues relating to the use of regional photochemical air quality models for evaluating their performance in reproducing the spatio-temporal features embedded in the observations and for designing emission control strategies needed to achieve compliance wit...

  13. Supporting Air-Conditioning Controller Design Using Evolutionary Computation

    NASA Astrophysics Data System (ADS)

    Kojima, Kazuyuki; Watanuki, Keiichi

    In recent years, as part of the remarkable development of electronic techniques, electronic control has been applied to various systems. Many sensors and actuators have been implemented into those systems, and energy efficiency and performance have been greatly improved. However, these systems have been complicated, and much time has been required to develop system controllers. In this paper, a method of automatic controller design for those systems is described. In order to automate the design of an electronic controller, an evolutionary hardware is applied. First, the framework for applying the genetic algorithm to the automation of controller design is described. In particular, the coding of a chromosome is shown in detail. Then, how to make a fitness function is represented, with an air conditioner as an example, and the controller of the air conditioner is developed automatically using our proposed framework. Finally, an evolutionary simulation is performed to confirm our framework.

  14. Air feed tube support system for a solid oxide fuel cell generator

    DOEpatents

    Doshi, Vinod B.; Ruka, Roswell J.; Hager, Charles A.

    2002-01-01

    A solid oxide fuel cell generator (12), containing tubular fuel cells (36) with interior air electrodes (18), where a supporting member (82) containing a plurality of holes (26) supports oxidant feed tubes (51), which pass from an oxidant plenum (52") into the center of the fuel cells, through the holes (26) in the supporting member (82), where a compliant gasket (86) around the top of the oxidant feed tubes and on top (28) of the supporting member (82) helps support the oxidant feed tubes and center them within the fuel cells, and loosen the tolerance for centering the air feed tubes.

  15. Air and water quality monitor assessment of life support subsystems

    NASA Technical Reports Server (NTRS)

    Whitley, Ken; Carrasquillo, Robyn L.; Holder, D.; Humphries, R.

    1988-01-01

    Preprotype air revitalization and water reclamation subsystems (Mole Sieve, Sabatier, Static Feed Electrolyzer, Trace Contaminant Control, and Thermoelectric Integrated Membrane Evaporative Subsystem) were operated and tested independently and in an integrated arrangement. During each test, water and/or gas samples were taken from each subsystem so that overall subsystem performance could be determined. The overall test design and objectives for both subsystem and integrated subsystem tests were limited, and no effort was made to meet water or gas specifications. The results of chemical analyses for each of the participating subsystems are presented along with other selected samples which were analyzed for physical properties and microbiologicals.

  16. Gold-Palladium nanoparticles supported by mesoporous β-MnO2 air electrode for rechargeable Li-Air battery

    NASA Astrophysics Data System (ADS)

    Thapa, Arjun Kumar; Shin, Tae Ho; Ida, Shintaro; Sumanasekera, Gamini U.; Sunkara, Mahendra K.; Ishihara, Tatsumi

    2012-12-01

    The electrochemical performance and electrode reaction using Au-Pd nanoparticle (NP) supported mesoporous β-MnO2 as a cathode catalyst for rechargeable Lithium-Air (Li-Air) battery is reported here for the first time. In this study, Au-Pd NP-supported mesoporous β-MnO2 was successfully synthesized by hydrothermal process using a silica KIT-6 template. It has an initial discharge capacity of ca. 775 mAh g-1 with high reversible capacity at a current density of 0.13 mA cm-2. The Au-Pd NP-supported mesoporous β-MnO2 cathode catalyst, which enhances the kinetic of oxygen reduction and evolution reactions (ORR/OERs), thereby improves energy and coulombic efficiency of the Li-Air cell. Raman spectroscopy and ex-situ XRD results of the Au-Pd NP-supported mesoporous β-MnO2 air electrode suggest that the observed capacity comes from oxidation of Li+ to form Li2O2 during discharge to 2.0 V.

  17. Air Quality Modeling in Support of the Near-road EXposures and effects of Urban air pollutants Study (NEXUS)

    EPA Science Inventory

    The paper presents the results of the model applications to estimate exposure metrics in support of an epidemiologic study in Detroit, Michigan. The Near-road Exposures to Urban air pollutant Study (NEXUS) design includes determining if children in Detroit, MI with asthma living ...

  18. Studying the Proteomic Composition of Expired Air Condensate in Newborns on Breathing Support.

    PubMed

    Kononikhin, A S; Ryndin, A Yu; Starodubtseva, N L; Chagovets, V V; Burov, A A; Bugrova, A E; Kostyukevich, Yu I; Popov, I A; Frankevich, V E; Ionov, O V; Zubkov, V V; Nikolaev, E N

    2016-04-01

    This study was designed to collect and perform a proteomic analysis of expired air condensate in newborns receiving respiratory support at the Department of Resuscitation and Intensive Care. The proteomic composition of expired air condensate was evaluated in newborns at various stages of development and with different abnormalities. PMID:27165072

  19. [CAS General Standards 2012

    ERIC Educational Resources Information Center

    Council for the Advancement of Standards in Higher Education, 2011

    2011-01-01

    The mission of the Council for the Advancement of Standards in Higher Education (CAS) is to promote the improvement of programs and services to enhance the quality of student learning and development. CAS is a consortium of professional associations who work collaboratively to develop and promulgate standards and guidelines and to encourage…

  20. Local-Scale Air Quality Modeling in Support of Human Health and Exposure Research (Invited)

    NASA Astrophysics Data System (ADS)

    Isakov, V.

    2010-12-01

    Spatially- and temporally-sparse information on air quality is a key concern for air-pollution-related environmental health studies. Monitor networks are sparse in both space and time, are costly to maintain, and are often designed purposely to avoid detecting highly localized sources. Recent studies have shown that more narrowly defining the geographic domain of the study populations and improvements in the measured/estimated ambient concentrations can lead to stronger associations between air pollution and hospital admissions and mortality records. Traditionally, ambient air quality measurements have been used as a primary input to support human health and exposure research. However, there is increasing evidence that the current ambient monitoring network is not capturing sharp gradients in exposure due to the presence of high concentration levels near, for example, major roadways. Many air pollutants exhibit large concentration gradients near large emitters such as major roadways, factories, ports, etc. To overcome these limitations, researchers are now beginning to use air quality models to support air pollution exposure and health studies. There are many advantages to using air quality models over traditional approaches based on existing ambient measurements alone. First, models can provide spatially- and temporally-resolved concentrations as direct input to exposure and health studies and thus better defining the concentration levels for the population in the geographic domain. Air quality models have a long history of use in air pollution regulations, and supported by regulatory agencies and a large user community. Also, models can provide bidirectional linkages between sources of emissions and ambient concentrations, thus allowing exploration of various mitigation strategies to reduce risk to exposure. In order to provide best estimates of air concentrations to support human health and exposure studies, model estimates should consider local-scale features

  1. Development of a large support surface for an air-bearing type zero-gravity simulator

    NASA Technical Reports Server (NTRS)

    Glover, K. E.

    1976-01-01

    The methods used in producing a large, flat surface to serve as the supporting surface for an air-bearing type zero-gravity simulator using low clearance, thrust-pad type air bearings are described. Major problems encountered in the use of self-leveled epoxy coatings in this surface are discussed and techniques are recommended which proved effective in overcoming these problems. Performance requirements of the zero-gravity simulator vehicle which were pertinent to the specification of the air-bearing support surface are also discussed.

  2. Air Quality Modeling in Support of the Near-Road Exposures and Effects of Urban Air Pollutants Study (NEXUS)

    PubMed Central

    Isakov, Vlad; Arunachalam, Saravanan; Batterman, Stuart; Bereznicki, Sarah; Burke, Janet; Dionisio, Kathie; Garcia, Val; Heist, David; Perry, Steve; Snyder, Michelle; Vette, Alan

    2014-01-01

    A major challenge in traffic-related air pollution exposure studies is the lack of information regarding pollutant exposure characterization. Air quality modeling can provide spatially and temporally varying exposure estimates for examining relationships between traffic-related air pollutants and adverse health outcomes. A hybrid air quality modeling approach was used to estimate exposure to traffic-related air pollutants in support of the Near-Road Exposures and Effects of Urban Air Pollutants Study (NEXUS) conducted in Detroit (Michigan, USA). Model-based exposure metrics, associated with local variations of emissions and meteorology, were estimated using a combination of the American Meteorological Society/Environmental Protection Agency Regulatory Model (AERMOD) and Research LINE-source dispersion model for near-surface releases (RLINE) dispersion models, local emission source information from the National Emissions Inventory, detailed road network locations and traffic activity, and meteorological data from the Detroit City Airport. The regional background contribution was estimated using a combination of the Community Multi-scale Air Quality (CMAQ) and the Space-Time Ordinary Kriging (STOK) models. To capture the near-road pollutant gradients, refined “mini-grids” of model receptors were placed around participant homes. Exposure metrics for CO, NOx, PM2.5 and its components (elemental and organic carbon) were predicted at each home location for multiple time periods including daily and rush hours. The exposure metrics were evaluated for their ability to characterize the spatial and temporal variations of multiple ambient air pollutants compared to measurements across the study area. PMID:25166917

  3. Air quality modeling in support of the Near-Road Exposures and Effects of Urban Air Pollutants Study (NEXUS).

    PubMed

    Isakov, Vlad; Arunachalam, Saravanan; Batterman, Stuart; Bereznicki, Sarah; Burke, Janet; Dionisio, Kathie; Garcia, Val; Heist, David; Perry, Steve; Snyder, Michelle; Vette, Alan

    2014-09-01

    A major challenge in traffic-related air pollution exposure studies is the lack of information regarding pollutant exposure characterization. Air quality modeling can provide spatially and temporally varying exposure estimates for examining relationships between traffic-related air pollutants and adverse health outcomes. A hybrid air quality modeling approach was used to estimate exposure to traffic-related air pollutants in support of the Near-Road Exposures and Effects of Urban Air Pollutants Study (NEXUS) conducted in Detroit (Michigan, USA). Model-based exposure metrics, associated with local variations of emissions and meteorology, were estimated using a combination of the American Meteorological Society/Environmental Protection Agency Regulatory Model (AERMOD) and Research LINE-source dispersion model for near-surface releases (RLINE) dispersion models, local emission source information from the National Emissions Inventory, detailed road network locations and traffic activity, and meteorological data from the Detroit City Airport. The regional background contribution was estimated using a combination of the Community Multi-scale Air Quality (CMAQ) and the Space-Time Ordinary Kriging (STOK) models. To capture the near-road pollutant gradients, refined "mini-grids" of model receptors were placed around participant homes. Exposure metrics for CO, NOx, PM2.5 and its components (elemental and organic carbon) were predicted at each home location for multiple time periods including daily and rush hours. The exposure metrics were evaluated for their ability to characterize the spatial and temporal variations of multiple ambient air pollutants compared to measurements across the study area. PMID:25166917

  4. CAS-1 lunar soil simulant

    NASA Astrophysics Data System (ADS)

    Zheng, Yongchun; Wang, Shijie; Ouyang, Ziyuan; Zou, Yongliao; Liu, Jianzhong; Li, Chunlai; Li, Xiongyao; Feng, Junming

    2009-02-01

    Lunar soil simulant is a geochemical reproduction of lunar regolith, and is needed for lunar science and engineering researches. This paper describes a new lunar soil simulant, CAS-1, prepared by the Chinese Academy of Sciences, to support lunar orbiter, soft-landing mission and sample return missions of China’s Lunar Exploration Program, which is scheduled for 2004 2020. Such simulants should match the samples returned from the Moon, all collected from the lunar regolith rather than outcrops. The average mineral and chemical composition of lunar soil sample returned from the Apollo 14 mission, which landed on the Fra Mauro Formation, is chosen as the model for the CAS-1 simulant. Source material for this simulant was a low-Ti basaltic scoria dated at 1600 years from the late Quaternary volcanic area in the Changbai Mountains of northeast China. The main minerals of this rock are pyroxene, olivine, and minor plagioclase, and about 20 40% modal glass. The scoria was analyzed by XRF and found to be chemically similar to Apollo 14 lunar sample 14163. It was crushed in an impact mill with a resulting median particle size 85.9 μm, similar to Apollo soils. Bulk density, shear resistance, complex permittivity, and reflectance spectra were also similar to Apollo 14 soil. We conclude that CAS-1 is an ideal lunar soil simulant for science and engineering research of future lunar exploration program.

  5. A Distributed Simulation Facility to Support Human Factors Research in Advanced Air Transportation Technology

    NASA Technical Reports Server (NTRS)

    Amonlirdviman, Keith; Farley, Todd C.; Hansman, R. John, Jr.; Ladik, John F.; Sherer, Dana Z.

    1998-01-01

    A distributed real-time simulation of the civil air traffic environment developed to support human factors research in advanced air transportation technology is presented. The distributed environment is based on a custom simulation architecture designed for simplicity and flexibility in human experiments. Standard Internet protocols are used to create the distributed environment, linking all advanced cockpit simulator, all Air Traffic Control simulator, and a pseudo-aircraft control and simulation management station. The pseudo-aircraft control station also functions as a scenario design tool for coordinating human factors experiments. This station incorporates a pseudo-pilot interface designed to reduce workload for human operators piloting multiple aircraft simultaneously in real time. The application of this distributed simulation facility to support a study of the effect of shared information (via air-ground datalink) on pilot/controller shared situation awareness and re-route negotiation is also presented.

  6. Design of an air traffic computer simulation system to support investigation of civil tiltrotor aircraft operations

    NASA Technical Reports Server (NTRS)

    Rogers, Ralph V.

    1992-01-01

    This research project addresses the need to provide an efficient and safe mechanism to investigate the effects and requirements of the tiltrotor aircraft's commercial operations on air transportation infrastructures, particularly air traffic control. The mechanism of choice is computer simulation. Unfortunately, the fundamental paradigms of the current air traffic control simulation models do not directly support the broad range of operational options and environments necessary to study tiltrotor operations. Modification of current air traffic simulation models to meet these requirements does not appear viable given the range and complexity of issues needing resolution. As a result, the investigation of systemic, infrastructure issues surrounding the effects of tiltrotor commercial operations requires new approaches to simulation modeling. These models should be based on perspectives and ideas closer to those associated with tiltrotor air traffic operations.

  7. Predicting Human Error in Air Traffic Control Decision Support Tools and Free Flight Concepts

    NASA Technical Reports Server (NTRS)

    Mogford, Richard; Kopardekar, Parimal

    2001-01-01

    The document is a set of briefing slides summarizing the work the Advanced Air Transportation Technologies (AATT) Project is doing on predicting air traffic controller and airline pilot human error when using new decision support software tools and when involved in testing new air traffic control concepts. Previous work in this area is reviewed as well as research being done jointly with the FAA. Plans for error prediction work in the AATT Project are discussed. The audience is human factors researchers and aviation psychologists from government and industry.

  8. Air

    MedlinePlus

    ... do to protect yourself from dirty air . Indoor air pollution and outdoor air pollution Air can be polluted indoors and it can ... this chart to see what things cause indoor air pollution and what things cause outdoor air pollution! Indoor ...

  9. Time Evolution of the Wettability of Supported Graphene under Ambient Air Exposure

    PubMed Central

    2016-01-01

    The wettability of graphene is both fundamental and crucial for interfacing in most applications, but a detailed understanding of its time evolution remains elusive. Here we systematically investigate the wettability of metal-supported, chemical vapor deposited graphene films as a function of ambient air exposure time using water and various other test liquids with widely different surface tensions. The wettability of graphene is not constant, but varies with substrate interactions and air exposure time. The substrate interactions affect the initial graphene wettability, where, for instance, water contact angles of ∼85 and ∼61° were measured for Ni and Cu supported graphene, respectively, after just minutes of air exposure. Analysis of the surface free energy components indicates that the substrate interactions strongly influence the Lewis acid–base component of supported graphene, which is considerably weaker for Ni supported graphene than for Cu supported graphene, suggesting that the classical van der Waals interaction theory alone is insufficient to describe the wettability of graphene. For prolonged air exposure, the effect of physisorption of airborne contaminants becomes increasingly dominant, resulting in an increase of water contact angle that follows a universal linear-logarithmic relationship with exposure time, until saturating at a maximum value of 92–98°. The adsorbed contaminants render all supported graphene samples increasingly nonpolar, although their total surface free energy decreases only by 10–16% to about 37–41 mJ/m2. Our finding shows that failure to account for the air exposure time may lead to widely different wettability values and contradicting arguments about the wetting transparency of graphene. PMID:26900413

  10. 75 FR 48552 - Automatic Dependent Surveillance-Broadcast (ADS-B) Out Performance Requirements To Support Air...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-08-11

    ... (ADS-B) Out Performance Requirements To Support Air Traffic Control (ATC) Service; OMB Approval of..., ``Automatic Dependent Surveillance- Broadcast (ADS-B) Out Performance Requirements To Support Air Traffic... rule, ``Automatic Dependent Surveillance-Broadcast (ADS-B) Out Performance Requirements To Support...

  11. Design of an air traffic computer simulation system to support investigation of civil tiltrotor aircraft operations

    NASA Technical Reports Server (NTRS)

    Rogers, Ralph V.

    1993-01-01

    The TATSS Project's goal was to develop a design for computer software that would support the attainment of the following objectives for the air traffic simulation model: (1) Full freedom of movement for each aircraft object in the simulation model. Each aircraft object may follow any designated flight plan or flight path necessary as required by the experiment under consideration. (2) Object position precision up to +/- 3 meters vertically and +/- 15 meters horizontally. (3) Aircraft maneuvering in three space with the object position precision identified above. (4) Air traffic control operations and procedures. (5) Radar, communication, navaid, and landing aid performance. (6) Weather. (7) Ground obstructions and terrain. (8) Detection and recording of separation violations. (9) Measures of performance including deviations from flight plans, air space violations, air traffic control messages per aircraft, and traditional temporal based measures.

  12. A GIS based decision support system for estimation, visualization and analysis of air pollution for large Turkish cities

    NASA Astrophysics Data System (ADS)

    Elbir, Tolga

    A decision support system has been developed to support local authorities in air quality management for big Turkish cities. The system is based on CALPUFF dispersion model, digital maps and related databases to estimate the emissions and spatial distribution of air pollutants. It applies a geographical information system. The system estimates ambient air pollution levels at high temporal and spatial resolutions. The system enables mapping of emissions and air quality levels. Mapping and scenario results can be compared with air quality limits. Impact assessment of air pollution abatement measures can also be carried out.

  13. The SnoDog: Preliminary design of a close air support aircraft

    NASA Technical Reports Server (NTRS)

    Ashbaugh, Scott; Bartel, Kent; Cavalli, J. R.; Chan, John; Chung, Jason; Dimaranan, Liza; Freese, Mike; Levitt, Rick; Soban, Dani

    1991-01-01

    U.S. military forces are presently searching for the next generation Close Air Support aircraft. The following report presents the SnoDog, a low-cost ($14.8 million) aircraft capable of operating from remote battlefields and unimproved airstrips. The configuration consists of a conventional, low aspect-ratio wing, twin booms, twin canted vertical stabilizers along with a high-mounted joined horizontal tail. A supercritical airfoil for the wing enhances aerodynamic performance, while the SnoDog's instability increases maneuverability over current close air support aircraft. Survivability was incorporated into the design by the use of a titanium tub to protect the cockpit from anti-aircraft artillery, as well as, the twin booms and retracted gear disposition. The booms aid survivability by supplying separated, redundant controls, and the landing gear are slightly exposed when retracted to enable a belly landing in emergencies. Designed to fly at Mach .76, the SnoDog is powered by two low-bypass turbofan engines. Engine accessibility and interchangeable parts make the SnoDog highly maintainable. The SnoDog is adaptable to many different missions, as it is capable of carrying advanced avionics pods, carrying external fuel tanks or refueling in-air, and carrying various types of munitions. This makes the SnoDog a multirole aircraft capable of air-to-air and air-to-ground combat. This combination of features make the SnoDog unique as a close air support aircraft, capable of meeting the U.S. military's future needs.

  14. DEVELOPMENT AND APPLICATIONS OF CFD SIMULATIONS SUPPORTING URBAN AIR QUALITY AND HOMELAND SECURITY

    EPA Science Inventory

    Prior to September 11, 2001 developments of Computational Fluid Dynamics (CFD) were begun to support air quality applications. CFD models are emerging as a promising technology for such assessments, in part due to the advancing power of computational hardware and software. CFD si...

  15. DEVELOPMENT AND APPLICATIONS OF CFD SIMULATIONS IN SUPPORT OF AIR QUALITY STUDIES INVOLVING BUILDINGS

    EPA Science Inventory

    There is a need to properly develop the application of Computational Fluid Dynamics (CFD) methods in support of air quality studies involving pollution sources near buildings at industrial sites. CFD models are emerging as a promising technology for such assessments, in part due ...

  16. Informational webinar for EPA STAR RFA on "Air, Climate and Energy (ACE) Centers: Science Supporting Solutions"

    EPA Science Inventory

    The purpose of this webinar presentation is to discuss the application process and required elements for the Air, Climate and Energy (ACE) Centers: Science Supporting Solutions RFA. EPA is seeking research on the development of sound science to systematically inform policy makers...

  17. DEVELOPMENT OF A SUB-SLAB AIR SAMPLING PROTOCOL TO SUPPORT ASSESSMENT OF VAPOR INTRUSION

    EPA Science Inventory

    The primary purpose of this research effort is to develop a methodology for sub-slab sampling to support the EPA guidance and vapor intrusion investigations after vapor intrusion has been established at a site. Methodologies for sub-slab air sampling are currently lacking in ref...

  18. Hadoop-Based Distributed System for Online Prediction of Air Pollution Based on Support Vector Machine

    NASA Astrophysics Data System (ADS)

    Ghaemi, Z.; Farnaghi, M.; Alimohammadi, A.

    2015-12-01

    The critical impact of air pollution on human health and environment in one hand and the complexity of pollutant concentration behavior in the other hand lead the scientists to look for advance techniques for monitoring and predicting the urban air quality. Additionally, recent developments in data measurement techniques have led to collection of various types of data about air quality. Such data is extremely voluminous and to be useful it must be processed at high velocity. Due to the complexity of big data analysis especially for dynamic applications, online forecasting of pollutant concentration trends within a reasonable processing time is still an open problem. The purpose of this paper is to present an online forecasting approach based on Support Vector Machine (SVM) to predict the air quality one day in advance. In order to overcome the computational requirements for large-scale data analysis, distributed computing based on the Hadoop platform has been employed to leverage the processing power of multiple processing units. The MapReduce programming model is adopted for massive parallel processing in this study. Based on the online algorithm and Hadoop framework, an online forecasting system is designed to predict the air pollution of Tehran for the next 24 hours. The results have been assessed on the basis of Processing Time and Efficiency. Quite accurate predictions of air pollutant indicator levels within an acceptable processing time prove that the presented approach is very suitable to tackle large scale air pollution prediction problems.

  19. Quantitative Characterizations of Ultrashort Echo (UTE) Images for Supporting Air-Bone Separation in the Head

    PubMed Central

    Hsu, Shu-Hui; Cao, Yue; Lawrence, Theodore S.; Tsien, Christina; Feng, Mary; Grodzki, David M.; Balter, James M.

    2015-01-01

    Accurate separation of air and bone is critical for creating synthetic CT from MRI to support Radiation Oncology workflow. This study compares two different ultrashort echo-time sequences in the separation of air from bone, and evaluates post-processing methods that correct intensity nonuniformity of images and account for intensity gradients at tissue boundaries to improve this discriminatory power. CT and MRI scans were acquired on 12 patients under an institution review board-approved prospective protocol. The two MRI sequences tested were ultra-short TE imaging using 3D radial acquisition (UTE), and using pointwise encoding time reduction with radial acquisition (PETRA). Gradient nonlinearity correction was applied to both MR image volumes after acquisition. MRI intensity nonuniformity was corrected by vendor-provided normalization methods, and then further corrected using the N4itk algorithm. To overcome the intensity-gradient at air-tissue boundaries, spatial dilations, from 0 to 4 mm, were applied to threshold-defined air regions from MR images. Receiver operating characteristic (ROC) analyses, by comparing predicted (defined by MR images) versus “true” regions of air and bone (defined by CT images), were performed with and without residual bias field correction and local spatial expansion. The post-processing corrections increased the areas under the ROC curves (AUC) from 0.944 ± 0.012 to 0.976 ± 0.003 for UTE images, and from 0.850 ± 0.022 to 0.887 ± 0.012 for PETRA images, compared to without corrections. When expanding the threshold-defined air volumes, as expected, sensitivity of air identification decreased with an increase in specificity of bone discrimination, but in a non-linear fashion. A 1-mm air mask expansion yielded AUC increases of 1% and 4% for UTE and PETRA images, respectively. UTE images had significantly greater discriminatory power in separating air from bone than PETRA images. Post-processing strategies improved the

  20. Quantitative characterizations of ultrashort echo (UTE) images for supporting air-bone separation in the head

    NASA Astrophysics Data System (ADS)

    Hsu, Shu-Hui; Cao, Yue; Lawrence, Theodore S.; Tsien, Christina; Feng, Mary; Grodzki, David M.; Balter, James M.

    2015-04-01

    Accurate separation of air and bone is critical for creating synthetic CT from MRI to support Radiation Oncology workflow. This study compares two different ultrashort echo-time sequences in the separation of air from bone, and evaluates post-processing methods that correct intensity nonuniformity of images and account for intensity gradients at tissue boundaries to improve this discriminatory power. CT and MRI scans were acquired on 12 patients under an institution review board-approved prospective protocol. The two MRI sequences tested were ultra-short TE imaging using 3D radial acquisition (UTE), and using pointwise encoding time reduction with radial acquisition (PETRA). Gradient nonlinearity correction was applied to both MR image volumes after acquisition. MRI intensity nonuniformity was corrected by vendor-provided normalization methods, and then further corrected using the N4itk algorithm. To overcome the intensity-gradient at air-tissue boundaries, spatial dilations, from 0 to 4 mm, were applied to threshold-defined air regions from MR images. Receiver operating characteristic (ROC) analyses, by comparing predicted (defined by MR images) versus ‘true’ regions of air and bone (defined by CT images), were performed with and without residual bias field correction and local spatial expansion. The post-processing corrections increased the areas under the ROC curves (AUC) from 0.944 ± 0.012 to 0.976 ± 0.003 for UTE images, and from 0.850 ± 0.022 to 0.887 ± 0.012 for PETRA images, compared to without corrections. When expanding the threshold-defined air volumes, as expected, sensitivity of air identification decreased with an increase in specificity of bone discrimination, but in a non-linear fashion. A 1 mm air mask expansion yielded AUC increases of 1 and 4% for UTE and PETRA images, respectively. UTE images had significantly greater discriminatory power in separating air from bone than PETRA images. Post-processing strategies improved the

  1. Cas9 Functionally Opens Chromatin.

    PubMed

    Barkal, Amira A; Srinivasan, Sharanya; Hashimoto, Tatsunori; Gifford, David K; Sherwood, Richard I

    2016-01-01

    Using a nuclease-dead Cas9 mutant, we show that Cas9 reproducibly induces chromatin accessibility at previously inaccessible genomic loci. Cas9 chromatin opening is sufficient to enable adjacent binding and transcriptional activation by the settler transcription factor retinoic acid receptor at previously unbound motifs. Thus, we demonstrate a new use for Cas9 in increasing surrounding chromatin accessibility to alter local transcription factor binding. PMID:27031353

  2. Cas9 Functionally Opens Chromatin

    PubMed Central

    Barkal, Amira A.; Srinivasan, Sharanya; Hashimoto, Tatsunori; Gifford, David K.; Sherwood, Richard I.

    2016-01-01

    Using a nuclease-dead Cas9 mutant, we show that Cas9 reproducibly induces chromatin accessibility at previously inaccessible genomic loci. Cas9 chromatin opening is sufficient to enable adjacent binding and transcriptional activation by the settler transcription factor retinoic acid receptor at previously unbound motifs. Thus, we demonstrate a new use for Cas9 in increasing surrounding chromatin accessibility to alter local transcription factor binding. PMID:27031353

  3. Air quality impact analysis in support of the new production reactor environmental impact statement

    SciTech Connect

    Hadley, D L

    1991-04-01

    The Pacific Northwest Laboratory (PNL) conducted this air quality impact analysis for the US Department of Energy (DOE). The purpose of this work was to provide Argonne National Laboratory (ANL) with the required estimates of ground-level concentrations of five criteria air pollutants at the Hanford Site boundary from each of the stationary sources associated with the new production reactor (NPR) and its supporting facilities. The DOE proposes to provide new production capacity for the primary production of tritium and secondary production of plutonium to support the US nuclear weapons program. Three alternative reactor technologies are being considered by DOE: the light-water reactor, the low-temperature, heavy-water reactor, and the modular high-temperature, gas-cooled reactor. In this study, PNL provided estimates of the impacts of the proposed action on the ground-level concentration of the criteria air pollutants for each of the alternative technologies. The criteria pollutants were sulfur dioxide, nitrogen dioxide, carbon monoxide, total suspended particulates, and particulates with a diameter of less than 10 microns. Ground-level concentrations were estimated for the peak construction phase activities expected to occur in 1997 and for the operational phase activities beginning in the year 2000. Ground-level concentrations of the primary air pollutants were estimated to be well below any of the applicable national or state ambient air quality standards. 12 refs., 19 tabs.

  4. Addendum to Air Quality: Decision Support Tools, Partner Plans, Working Groups, Committees

    NASA Technical Reports Server (NTRS)

    Holekamp, Kara; Frisbie, Troy; Estep, Lee

    2005-01-01

    In the original report dated February 11, 2005, the utility of NASA Earth science data in the air quality activities of other agencies and organizations was assessed by reviewing strategic and mission plans and by conducting personal interviews with agency experts to identify and investigate agencies with the potential for partnership with NASA. The overarching agency strategic plans were reviewed and commonalities such as the desire for partnerships and technology development were noted. This addendum to the original report contains such information about the Tennessee Valley Authority and will be inserted as Section 2.6 of "Air Quality: Decision Support Tools, Partner Plans, Working Groups, Committees."

  5. Addendum to Air Quality: Decision Support Tools, Partner Plans, Working Groups, Committees

    NASA Technical Reports Server (NTRS)

    Holekamp, Kara; Frisbie, Troy; Estep, Lee

    2005-01-01

    In the original report dated February 11, 2005, the utility of the NASA Earth science data in the air quality activities of other agencies and organizations was assessed by reviewing strategic and mission plans and by conducting personal interviews with agency experts to identify and investigate agencies with the potential for partnership with NASA. The overarching agency strategic plans were reviewed and commonalities such as the desire for partnerships and technology development were noted. The addendum to the original report contains such information about the Tennessee Valley Authority and will be inserted in Section 2.6 of "Air Quality Decision Support Tools, Partner Plans, Working Groups, Committees".

  6. Turboprop aircraft against terrorism: a SWOT analysis of turboprop aircraft in CAS operations

    NASA Astrophysics Data System (ADS)

    Yavuz, Murat; Akkas, Ali; Aslan, Yavuz

    2012-06-01

    Today, the threat perception is changing. Not only for countries but also for defence organisations like NATO, new threat perception is pointing terrorism. Many countries' air forces become responsible of fighting against terorism or Counter-Insurgency (COIN) Operations. Different from conventional warfare, alternative weapon or weapon systems are required for such operatioins. In counter-terrorism operations modern fighter jets are used as well as helicopters, subsonic jets, Unmanned Aircraft Systems (UAS), turboprop aircraft, baloons and similar platforms. Succes and efficiency of the use of these platforms can be determined by evaluating the conditions, the threats and the area together. Obviously, each platform has advantages and disadvantages for different cases. In this research, examples of turboprop aircraft usage against terrorism and with a more general approach, turboprop aircraft for Close Air Support (CAS) missions from all around the world are reviewed. In this effort, a closer look is taken at the countries using turboprop aircraft in CAS missions while observing the fields these aircraft are used in, type of operations, specifications of the aircraft, cost and the maintenance factors. Thus, an idea about the convenience of using these aircraft in such operations can be obtained. A SWOT analysis of turboprop aircraft in CAS operations is performed. This study shows that turboprop aircraft are suitable to be used in counter-terrorism and COIN operations in low threat environment and is cost benefical compared to jets.

  7. Transformation of OODT CAS to Perform Larger Tasks

    NASA Technical Reports Server (NTRS)

    Mattmann, Chris; Freeborn, Dana; Crichton, Daniel; Hughes, John; Ramirez, Paul; Hardman, Sean; Woollard, David; Kelly, Sean

    2008-01-01

    A computer program denoted OODT CAS has been transformed to enable performance of larger tasks that involve greatly increased data volumes and increasingly intensive processing of data on heterogeneous, geographically dispersed computers. Prior to the transformation, OODT CAS (also alternatively denoted, simply, 'CAS') [wherein 'OODT' signifies 'Object-Oriented Data Technology' and 'CAS' signifies 'Catalog and Archive Service'] was a proven software component used to manage scientific data from spaceflight missions. In the transformation, CAS was split into two separate components representing its canonical capabilities: file management and workflow management. In addition, CAS was augmented by addition of a resource-management component. This third component enables CAS to manage heterogeneous computing by use of diverse resources, including high-performance clusters of computers, commodity computing hardware, and grid computing infrastructures. CAS is now more easily maintainable, evolvable, and reusable. These components can be used separately or, taking advantage of synergies, can be used together. Other elements of the transformation included addition of a separate Web presentation layer that supports distribution of data products via Really Simple Syndication (RSS) feeds, and provision for full Resource Description Framework (RDF) exports of metadata.

  8. Soil-based filtration technology for air purification: potentials for environmental and space life support application

    NASA Astrophysics Data System (ADS)

    Nelson, Mark; Bohn, Hinrich

    Soil biofiltration, also known as Soil bed reactor (SBR), technology was originally developed in Germany to take advantage of the diversity in microbial mechanisms to control gases producing malodor in industrial processes. The approach has since gained wider international acceptance and seen numerous improvements, for example, by the use of high-organic compost beds to maximize microbial processes. This paper reviews the basic mechanisms which underlay soil processes involved in air purification, advantages and limitations of the technology and the cur-rent research status of the approach. Soil biofiltration has lower capital and operating/energetic costs than conventional technologies and is well adapted to handle contaminants in moderate concentrations. The systems can be engineered to optimize efficiency though manipulation of temperature, pH, moisture content, soil organic matter and airflow rates. SBR technology was modified for application in the Biosphere 2 project, which demonstrated in preparatory research with a number of closed system testbeds that soil could also support crop plants while also serving as soil filters with air pumps to push air through the soil. This Biosphere 2 research demonstrated in several closed system testbeds that a number of important trace gases could be kept under control and led to the engineering of the entire agricultural soil of Biosphere 2 to serve as a soil filtration unit for the facility. Soil biofiltration, coupled with food crop produc-tion, as a component of bioregenerative space life support systems has the advantages of lower energy use and avoidance of the consumables required for other air purification approaches. Expanding use of soil biofiltration can aid a number of environmental applications, from the mitigation of indoor air pollution, improvement of industrial air emissions and prevention of accidental release of toxic gases.

  9. Development of a GIS-based decision support system for urban air quality management in the city of Istanbul

    NASA Astrophysics Data System (ADS)

    Elbir, Tolga; Mangir, Nizamettin; Kara, Melik; Simsir, Sedef; Eren, Tuba; Ozdemir, Seda

    2010-02-01

    A decision support system has been developed for urban air quality management in the metropolitan area of Istanbul. The system is based on CALMET/CALPUFF dispersion modeling system, digital maps, and related databases to estimate the emissions and spatial distribution of air pollutants with the help of a GIS software. The system estimates ambient air pollution levels at high temporal and spatial resolutions and enables mapping of emissions and air quality levels. Mapping and scenario results can be compared with air quality limits. Impact assessment of air pollution abatement measures can also be carried out.

  10. Nanoscale metal oxide and supported metal catalysts for Li-air battery

    NASA Astrophysics Data System (ADS)

    Huang, Kan

    The dissertation work focuses on research and development of durable nanoscale catalysts and supports for rechargeable Li-air batteries that use aqueous catholytes. Transition metal oxides, TiO2 and Nb2 O5 in particular, were prepared from a sol-gel process in the form of nanocoatings (5˜50 nm) on carbon nanotubes (CNTs) and studied as catalyst supports. Carbon doping in the oxides and post annealing significantly increased their electronic conductivity. Pt catalyst on the support with TiO 2 (Pt/c-TiO2/CNTs) showed a much better oxygen reduction reaction (ORR) activity than a commercial Pt on carbon black (Pt/C). Negligible loss (< 3%) in ORR activity was found in Pt/c-TiO2/CNTs as compared to more than 50% loss in Pt/C, demonstrating a significantly improved durability in the developed catalysts. However, Pt/c-Nb2O5/CNTs was found to be worse in ORR activity and durability, suggesting that c-Nb 2O5/CNTs may not be a good support. CNTs have fibrous shape and would provide a unique porous structure as electrode. Their buckypapers were made and used to support catalysts of Pt and IrO2 in the cathodes of Li-air batteries with sulfuric acid catholyte. At low Pt loading (5 wt.%) without IrO2 on the buckypaper cathode, the Li-air cell achieved a discharging capacity of 306 mAh/g and a specific energy of 1067 Wh/kg at 0.2 mA/cm2. A significant charge overpotential reduction (˜ 0.3 V) was achieved when IrO2 was also used to form a bifunctional catalyst with Pt on the buckypapers. The round trip efficiency was increased from 72% to 81% with the bifunctional cathode, demonstrating a higher energy conversion efficiency.

  11. Multi-Agent Diagnosis and Control of an Air Revitalization System for Life Support in Space

    NASA Technical Reports Server (NTRS)

    Malin, Jane T.; Kowing, Jeffrey; Nieten, Joseph; Graham, Jeffrey s.; Schreckenghost, Debra; Bonasso, Pete; Fleming, Land D.; MacMahon, Matt; Thronesbery, Carroll

    2000-01-01

    An architecture of interoperating agents has been developed to provide control and fault management for advanced life support systems in space. In this adjustable autonomy architecture, software agents coordinate with human agents and provide support in novel fault management situations. This architecture combines the Livingstone model-based mode identification and reconfiguration (MIR) system with the 3T architecture for autonomous flexible command and control. The MIR software agent performs model-based state identification and diagnosis. MIR identifies novel recovery configurations and the set of commands required for the recovery. The AZT procedural executive and the human operator use the diagnoses and recovery recommendations, and provide command sequencing. User interface extensions have been developed to support human monitoring of both AZT and MIR data and activities. This architecture has been demonstrated performing control and fault management for an oxygen production system for air revitalization in space. The software operates in a dynamic simulation testbed.

  12. Closed-loop Habitation Air Revitalization Model for Regenerative Life Support Systems

    NASA Technical Reports Server (NTRS)

    Hart, Maxwell M.

    1991-01-01

    The primary function of any life support system is to keep the crew alive by providing breathable air, potable water, edible food, and for disposal of waste. In a well-balanced or regenerative life support system, the various components are each using what is available and producing what is needed by other components so that there will always be enough chemicals in the form in which they are needed. Humans are not just users, but also one of the participating parts of the system. If a system could continuously recycle the original chemicals, this would make it virtually a Closed-loop Habitation (CH). Some difficulties in trying to create a miniature version of a CH are briefly discussed. In a miniature CH, a minimal structure must be provided and the difference must be made up by artificial parts such as physicochemical systems that perform the conversions that the Earth can achieve naturally. To study the interactions of these parts, a computer model was designed that simulates a miniature CH with emphasis on the air revitalization part. It is called the Closed-loop Habitation Air Revitalization Model (CHARM).

  13. [The main ways of improvement of medical support of the Air Forces in modern conditions].

    PubMed

    Blaginin, A A; Grebeniuk, A N; Lizogub, I N

    2014-02-01

    Blaginin A.A., Grebenyuk A.N., Lizogub LN. - The main ways of improvement of medical support of the Air Forces in modern conditions. Aircrew conducting active hostilities suffers from the whole spectrum of factors and conditions of the combat situation. The main task for the medical service of the Air Force is to carry out preventive and curative action for aviation specialists who are responsible for the combat capability of aircraft formations. The medical service of the Air Force must have forces and facilities for planning, organization and implementation of the treatment of lightly wounded and sick aviation professionals with short periods of recovery, medical rehabilitation of aircrew qfter suffering injuries, diseases, sanatorium therapy of aircrew with partial failure of health, outpatient and inpatient medical examination aircrew - flight commissions, preventive rest of aviation specialists with symptoms of chronic fatigue. Should be trained aviation physicians, including both basic military medical education and in-depth study of the medical aspects of various fields of personnel of the Air Force. PMID:25046924

  14. Air Quality Modeling in Support of the Near-Road Exposures and Effects of Urban Air Pollutants Study (NEXUS)

    EPA Science Inventory

    A major challenge in traffic-related air pollution exposure studies is the lack of information regarding pollutant exposure characterization. Air quality modeling can provide spatially and temporally varying exposure estimates for examining relationships between traffic-related a...

  15. Measuring critical care air support teams' performance during extended periods of duty.

    PubMed

    Lamb, Di

    2010-01-01

    The Royal Air Force (RAF) Critical Care Air Support Teams (CCASTs) aeromedically evacuate seriously injured service personnel. Long casualty evacuation chains create logistical constraints that must be considered when aeromedically evacuating patients. One constraint is the length of a CCAST mission and its potential effect on team member performance. Despite no evidence of patient care compromise, the RAF has commissioned a study to investigate whether CCAST mission length influences performance. Describing and understanding the role of a CCAST enabled fatigue to be defined. Factors essential to studying fatigue were then identified that were used to develop a theoretical model for designing a study to measure the effects of fatigue on CCAST performance. Relevant factors include the patient's clinical condition, team members' cognition and vigilance levels, and the occupational aviation environment. Further factors influencing overall performance include the duration and complexity of patient interventions, mission length, circadian influences, and fatigue countermeasures. PMID:20683231

  16. CRISPR/Cas9 in Genome Editing and Beyond.

    PubMed

    Wang, Haifeng; La Russa, Marie; Qi, Lei S

    2016-06-01

    The Cas9 protein (CRISPR-associated protein 9), derived from type II CRISPR (clustered regularly interspaced short palindromic repeats) bacterial immune systems, is emerging as a powerful tool for engineering the genome in diverse organisms. As an RNA-guided DNA endonuclease, Cas9 can be easily programmed to target new sites by altering its guide RNA sequence, and its development as a tool has made sequence-specific gene editing several magnitudes easier. The nuclease-deactivated form of Cas9 further provides a versatile RNA-guided DNA-targeting platform for regulating and imaging the genome, as well as for rewriting the epigenetic status, all in a sequence-specific manner. With all of these advances, we have just begun to explore the possible applications of Cas9 in biomedical research and therapeutics. In this review, we describe the current models of Cas9 function and the structural and biochemical studies that support it. We focus on the applications of Cas9 for genome editing, regulation, and imaging, discuss other possible applications and some technical considerations, and highlight the many advantages that CRISPR/Cas9 technology offers. PMID:27145843

  17. Local Voltage Support from Distributed Energy Resources to Prevent Air Conditioner Motor Stalling

    SciTech Connect

    Baone, Chaitanya A; Xu, Yan; Kueck, John D

    2010-01-01

    Microgrid voltage collapse often happens when there is a high percentage of low inertia air-conditioning (AC) motors in the power systems. The stalling of the AC motors results in Fault Induced Delayed Voltage Recovery (FIDVR). A hybrid load model including typical building loads, AC motor loads, and other induction motor loads is built to simulate the motoring stalling phenomena. Furthermore, distributed energy resources (DE) with local voltage support capability are utilized to boost the local bus voltage during a fault, and prevent the motor stalling. The simulation results are presented. The analysis of the simulation results show that local voltage support from multiple DEs can effectively and economically solve the microgrid voltage collapse problem.

  18. An Air Revitalization Model (ARM) for Regenerative Life Support Systems (RLSS)

    NASA Technical Reports Server (NTRS)

    Hart, Maxwell M.

    1990-01-01

    The primary objective of the air revitalization model (ARM) is to determine the minimum buffer capacities that would be necessary for long duration space missions. Several observations are supported by the current configuration sizes: the baseline values for each gas and the day to day or month to month fluctuations that are allowed. The baseline values depend on the minimum safety tolerances and the quantities of life support consumables necessary to survive the worst case scenarios within those tolerances. Most, it not all, of these quantities can easily be determined by ARM once these tolerances are set. The day to day fluctuations also require a command decision. It is already apparent from the current configuration of ARM that the tighter these fluctuations are controlled, the more energy used, the more nonregenerable hydrazine consumed, and the larger the required capacities for the various gas generators. All of these relationships could clearly be quantified by one operational ARM.

  19. Use of UAS to Support Management in Precision Agriculture: The AggieAir Experience

    NASA Astrophysics Data System (ADS)

    McKee, M.; Torres-Rua, A. F.; ELarab, M.; Hassan Esfahani, L.; Jensen, A.

    2015-12-01

    Remote sensing applications for precision agriculture depend on acquiring actionable information at high spatial resolution and at a temporal frequency appropriate for timely responses. Unmanned aerial systems (UAS) are capable of providing such imagery for use in various applications for precision agriculture (yield estimation, evapotranspiration, etc.). AggieAirTM, a UAS platform and sensory array, was designed and developed at Utah State University to acquire high-resolution imagery (0.15m -0.6 m) in the visual, near infrared, red edge, and thermal infrared spectra. Spectral data obtained from AggieAir are used to develop soil moisture, plant chlorophyll, leaf nitrogen and actual evapotranspiration estimates to support management in precision agriculture. This presentation will focus on experience in using the AggieAir system to provide information products of possible interest in precision agriculture. The discussion will include information about the direction and rate of development of UAS technology and the current and anticipated future state of the regulatory environment for use of these systems in the U.S.

  20. Barriers and Facilitators to Use of Air Force Family Support Centers (FSCs): Lessons for Civilian and Military Sectors.

    ERIC Educational Resources Information Center

    Albano, Sondra

    This study focused on non-use of Air Force Family Support Centers (FSCs), a global network of programs and services designed to facilitate family adaptation to the mobile military lifestyle and to help commanders and supervisors respond to family needs. The study used the 1993 Air Force Needs Assessment Survey to investigate what factors…

  1. Illustrations and Supporting Texts for Sound Standing Waves of Air Columns in Pipes in Introductory Physics Textbooks

    ERIC Educational Resources Information Center

    Zeng, Liang; Smith, Chris; Poelzer, G. Herold; Rodriguez, Jennifer; Corpuz, Edgar; Yanev, George

    2014-01-01

    In our pilot studies, we found that many introductory physics textbook illustrations with supporting text for sound standing waves of air columns in open-open, open-closed, and closed-closed pipes inhibit student understanding of sound standing wave phenomena due to student misunderstanding of how air molecules move within these pipes. Based on…

  2. Ruthenium-based electrocatalysts supported on reduced graphene oxide for lithium-air batteries.

    PubMed

    Jung, Hun-Gi; Jeong, Yo Sub; Park, Jin-Bum; Sun, Yang-Kook; Scrosati, Bruno; Lee, Yun Jung

    2013-04-23

    Ruthenium-based nanomaterials supported on reduced graphene oxide (rGO) have been investigated as air cathodes in non-aqueous electrolyte Li-air cells using a TEGDME-LiCF3SO3 electrolyte. Homogeneously distributed metallic ruthenium and hydrated ruthenium oxide (RuO2·0.64H2O), deposited exclusively on rGO, have been synthesized with average size below 2.5 nm. The synthesized hybrid materials of Ru-based nanoparticles supported on rGO efficiently functioned as electrocatalysts for Li2O2 oxidation reactions, maintaining cycling stability for 30 cycles without sign of TEGDME-LiCF3SO3 electrolyte decomposition. Specifically, RuO2·0.64H2O-rGO hybrids were superior to Ru-rGO hybrids in catalyzing the OER reaction, significantly reducing the average charge potential to ∼3.7 V at the high current density of 500 mA g(-1) and high specific capacity of 5000 mAh g(-1). PMID:23540570

  3. Fisk-based criteria to support validation of detection methods for drinking water and air.

    SciTech Connect

    MacDonell, M.; Bhattacharyya, M.; Finster, M.; Williams, M.; Picel, K.; Chang, Y.-S.; Peterson, J.; Adeshina, F.; Sonich-Mullin, C.; Environmental Science Division; EPA

    2009-02-18

    This report was prepared to support the validation of analytical methods for threat contaminants under the U.S. Environmental Protection Agency (EPA) National Homeland Security Research Center (NHSRC) program. It is designed to serve as a resource for certain applications of benchmark and fate information for homeland security threat contaminants. The report identifies risk-based criteria from existing health benchmarks for drinking water and air for potential use as validation targets. The focus is on benchmarks for chronic public exposures. The priority sources are standard EPA concentration limits for drinking water and air, along with oral and inhalation toxicity values. Many contaminants identified as homeland security threats to drinking water or air would convert to other chemicals within minutes to hours of being released. For this reason, a fate analysis has been performed to identify potential transformation products and removal half-lives in air and water so appropriate forms can be targeted for detection over time. The risk-based criteria presented in this report to frame method validation are expected to be lower than actual operational targets based on realistic exposures following a release. Note that many target criteria provided in this report are taken from available benchmarks without assessing the underlying toxicological details. That is, although the relevance of the chemical form and analogues are evaluated, the toxicological interpretations and extrapolations conducted by the authoring organizations are not. It is also important to emphasize that such targets in the current analysis are not health-based advisory levels to guide homeland security responses. This integrated evaluation of chronic public benchmarks and contaminant fate has identified more than 200 risk-based criteria as method validation targets across numerous contaminants and fate products in drinking water and air combined. The gap in directly applicable values is

  4. Commentary: Is the Air Pollution Health Research Community Prepared to Support a Multipollutant Air Quality Management Framework?

    EPA Science Inventory

    Ambient air pollution is always encountered as a complex mixture, but past regulatory and research strategies largely focused on single pollutants, pollutant classes, and sources one-at-a-time. There is a trend toward managing air quality in a progressively “multipollutant” manne...

  5. Analysis of Coolant-flow Requirements for an Improved, Internal-strut-supported, Air-cooled Turbine-rotor Blade

    NASA Technical Reports Server (NTRS)

    Schramm, Wilson B; Nachtigall, Alfred J

    1952-01-01

    An analytical evaluation of a new typ An analytical evaluation of a new type of air-cooled turbine-rotor-blade design, based on the principle of submerging the load-carrying element in cooling air within a thin high-temperature sheel, indicates that this principle of blade design permits the load carrying element to be operated at considerably lower temperature than that of the enveloping shell. Comparison with an air-cooled shell-supported air-cooled blade has greater potentiality to withstand increased stresses that can be anticipated in future engines.

  6. Predicting Weight Support Based on Wake Measurements of a Flying Bird in Still Air

    NASA Astrophysics Data System (ADS)

    Gutierrez, Eric; Lentink, David

    2014-11-01

    The wake development of a freely flying Pacific Parrotlet (Forpus coelestis) was examined in still air. The bird was trained to fly from perch to perch through the laser sheet while wearing custom-made laser safety goggles. This enabled a detailed study of the evolution of the vortices shed in its wake using high-speed particle image velocimetry at 1000 Hz in the plane transverse to the flight path. The measurement started when the bird was approximately 0.25 wingbeats in front of the laser sheet and stopped after it traveled 3.5 wingbeats beyond the laser sheet. The instantaneous lift force that supports body weight was calculated based on the velocity field, using both the Kuttta-Joukowski and the actuator disk quasi-steady model. During the first few flaps, both models predict an instantaneous lift that is reasonably close to the weight of the bird. Several flaps away from the laser sheet, however, the models predict that the lift steadily declines to about 50% of the weight of the bird. In contrast to earlier reports for bat wakes in wind tunnels, these findings for bird wakes in still air suggest that the predictive strength of quasi-steady force calculations depends on the distance between the animal and the laser sheet.

  7. OHIO RIVER BASIN ENERGY STUDY: AIR QUALITY AND RELATED IMPACTS. VOLUME I. DOCUMENTATION IN SUPPORT OF KEY ORBES AIR QUALITY FINDINGS

    EPA Science Inventory

    This report was prepared as part of the Ohio River Basin Energy Study (ORBES), a multi-disciplinary research program supported by the Environmental Protection Agency. The extensive air quality analysis undertaken for the ORBES included examination of pollutant emissions and resul...

  8. Harnessing heterologous and endogenous CRISPR-Cas machineries for efficient markerless genome editing in Clostridium

    PubMed Central

    Pyne, Michael E.; Bruder, Mark R.; Moo-Young, Murray; Chung, Duane A.; Chou, C. Perry

    2016-01-01

    Application of CRISPR-Cas9 systems has revolutionized genome editing across all domains of life. Here we report implementation of the heterologous Type II CRISPR-Cas9 system in Clostridium pasteurianum for markerless genome editing. Since 74% of species harbor CRISPR-Cas loci in Clostridium, we also explored the prospect of co-opting host-encoded CRISPR-Cas machinery for genome editing. Motivation for this work was bolstered from the observation that plasmids expressing heterologous cas9 result in poor transformation of Clostridium. To address this barrier and establish proof-of-concept, we focus on characterization and exploitation of the C. pasteurianum Type I-B CRISPR-Cas system. In silico spacer analysis and in vivo interference assays revealed three protospacer adjacent motif (PAM) sequences required for site-specific nucleolytic attack. Introduction of a synthetic CRISPR array and cpaAIR gene deletion template yielded an editing efficiency of 100%. In contrast, the heterologous Type II CRISPR-Cas9 system generated only 25% of the total yield of edited cells, suggesting that native machinery provides a superior foundation for genome editing by precluding expression of cas9 in trans. To broaden our approach, we also identified putative PAM sequences in three key species of Clostridium. This is the first report of genome editing through harnessing native CRISPR-Cas machinery in Clostridium. PMID:27157668

  9. Harnessing heterologous and endogenous CRISPR-Cas machineries for efficient markerless genome editing in Clostridium.

    PubMed

    Pyne, Michael E; Bruder, Mark R; Moo-Young, Murray; Chung, Duane A; Chou, C Perry

    2016-01-01

    Application of CRISPR-Cas9 systems has revolutionized genome editing across all domains of life. Here we report implementation of the heterologous Type II CRISPR-Cas9 system in Clostridium pasteurianum for markerless genome editing. Since 74% of species harbor CRISPR-Cas loci in Clostridium, we also explored the prospect of co-opting host-encoded CRISPR-Cas machinery for genome editing. Motivation for this work was bolstered from the observation that plasmids expressing heterologous cas9 result in poor transformation of Clostridium. To address this barrier and establish proof-of-concept, we focus on characterization and exploitation of the C. pasteurianum Type I-B CRISPR-Cas system. In silico spacer analysis and in vivo interference assays revealed three protospacer adjacent motif (PAM) sequences required for site-specific nucleolytic attack. Introduction of a synthetic CRISPR array and cpaAIR gene deletion template yielded an editing efficiency of 100%. In contrast, the heterologous Type II CRISPR-Cas9 system generated only 25% of the total yield of edited cells, suggesting that native machinery provides a superior foundation for genome editing by precluding expression of cas9 in trans. To broaden our approach, we also identified putative PAM sequences in three key species of Clostridium. This is the first report of genome editing through harnessing native CRISPR-Cas machinery in Clostridium. PMID:27157668

  10. Dual nuclease activity of a Cas2 protein in CRISPR-Cas subtype I-B of Leptospira interrogans.

    PubMed

    Dixit, Bhuvan; Ghosh, Karukriti Kaushik; Fernandes, Gary; Kumar, Pankaj; Gogoi, Prerana; Kumar, Manish

    2016-04-01

    Leptospira interrogans serovar Copenhageni strain Fiocruz L1-130 carries a set of cas genes associated with CRISPR-Cas subtype I-B. Herein, we report for the first time active transcription of a set of cas genes (cas1 to cas8) of L. interrogans where cas4, cas1, cas2 and cas6, cas3, cas8, cas7, cas5 are clustered together in two independent operons. As an initial step toward comprehensive understanding of CRISPR-Cas system in spirochete, the biochemical study of one of the core Leptospira Cas2 proteins (Lep_Cas2) showed nuclease activity on both DNA and RNA in a nonspecific manner. Additionally, unlike other known Cas2 proteins, Lep_Cas2 showed metal-independent RNase activity and preferential activity on RNA over DNA. These results provide insight for understanding Cas2 diversity existing in the prokaryotic adaptive immune system. PMID:26950513

  11. Conceptual air sparging decision tool in support of the development of an air sparging optimization decision tool

    SciTech Connect

    1995-09-01

    The enclosed document describes a conceptual decision tool (hereinafter, Tool) for determining applicability of and for optimizing air sparging systems. The Tool was developed by a multi-disciplinary team of internationally recognized experts in air sparging technology, lead by a group of project and task managers at Parsons Engineering Science, Inc. (Parsons ES). The team included Mr. Douglas Downey and Dr. Robert Hinchee of Parsons ES, Dr. Paul Johnson of Arizona State University, Dr. Richard Johnson of Oregon Graduate Institute, and Mr. Michael Marley of Envirogen, Inc. User Community Panel Review was coordinated by Dr. Robert Siegrist of Colorado School of Mines (also of Oak Ridge National Laboratory) and Dr. Thomas Brouns of Battelle/Pacific Northwest Laboratory. The Tool is intended to provide guidance to field practitioners and environmental managers for evaluating the applicability and optimization of air sparging as remedial action technique.

  12. Compressed air energy storage monitoring to support refrigerated mined rock cavern technology.

    SciTech Connect

    Lee, Moo Yul; Bauer, Stephen J.

    2004-06-01

    This document is the final report for the Compressed Air Energy Storage Monitoring to Support Refrigerated-Mined Rock Cavern Technology (CAES Monitoring to Support RMRCT) (DE-FC26-01NT40868) project to have been conducted by CAES Development Co., along with Sandia National Laboratories. This document provides a final report covering tasks 1.0 and subtasks 2.1, 2.2, and 2.5 of task 2.0 of the Statement of Project Objectives and constitutes the final project deliverable. The proposed work was to have provided physical measurements and analyses of large-scale rock mass response to pressure cycling. The goal was to develop proof-of-concept data for a previously developed and DOE sponsored technology (RMRCT or Refrigerated-Mined Rock Cavern Technology). In the RMRCT concept, a room and pillar mine developed in rock serves as a pressure vessel. That vessel will need to contain pressure of about 1370 psi (and cycle down to 300 psi). The measurements gathered in this study would have provided a means to determine directly rock mass response during cyclic loading on the same scale, under similar pressure conditions. The CAES project has been delayed due to national economic unrest in the energy sector.

  13. NASA Langley's Formal Methods Research in Support of the Next Generation Air Transportation System

    NASA Technical Reports Server (NTRS)

    Butler, Ricky W.; Munoz, Cesar A.

    2008-01-01

    This talk will provide a brief introduction to the formal methods developed at NASA Langley and the National Institute for Aerospace (NIA) for air traffic management applications. NASA Langley's formal methods research supports the Interagency Joint Planning and Development Office (JPDO) effort to define and develop the 2025 Next Generation Air Transportation System (NGATS). The JPDO was created by the passage of the Vision 100 Century of Aviation Reauthorization Act in Dec 2003. The NGATS vision calls for a major transformation of the nation s air transportation system that will enable growth to 3 times the traffic of the current system. The transformation will require an unprecedented level of safety-critical automation used in complex procedural operations based on 4-dimensional (4D) trajectories that enable dynamic reconfiguration of airspace scalable to geographic and temporal demand. The goal of our formal methods research is to provide verification methods that can be used to insure the safety of the NGATS system. Our work has focused on the safety assessment of concepts of operation and fundamental algorithms for conflict detection and resolution (CD&R) and self- spacing in the terminal area. Formal analysis of a concept of operations is a novel area of application of formal methods. Here one must establish that a system concept involving aircraft, pilots, and ground resources is safe. The formal analysis of algorithms is a more traditional endeavor. However, the formal analysis of ATM algorithms involves reasoning about the interaction of algorithmic logic and aircraft trajectories defined over an airspace. These trajectories are described using 2D and 3D vectors and are often constrained by trigonometric relations. Thus, in many cases it has been necessary to unload the full power of an advanced theorem prover. The verification challenge is to establish that the safety-critical algorithms produce valid solutions that are guaranteed to maintain separation

  14. A Method for Making Cross-Comparable Estimates of the Benefits of Decision Support Technologies for Air Traffic Management

    NASA Technical Reports Server (NTRS)

    Lee, David; Long, Dou; Etheridge, Mel; Plugge, Joana; Johnson, Jesse; Kostiuk, Peter

    1998-01-01

    We present a general method for making cross comparable estimates of the benefits of NASA-developed decision support technologies for air traffic management, and we apply a specific implementation of the method to estimate benefits of three decision support tools (DSTs) under development in NASA's advanced Air Transportation Technologies Program: Active Final Approach Spacing Tool (A-FAST), Expedite Departure Path (EDP), and Conflict Probe and Trial Planning Tool (CPTP). The report also reviews data about the present operation of the national airspace system (NAS) to identify opportunities for DST's to reduce delays and inefficiencies.

  15. Gulf of Mexico Air Quality: CALIPSO Support for Gulf of Mexico Air Quality Relating to the Deepwater Horizon Oil Spill

    NASA Technical Reports Server (NTRS)

    Nguyen, Myngoc T.; Lapointe, Stephen; Jennings, Brittney; Zoumplis, Angela

    2011-01-01

    On April 20, 2010, an oil platform belonging to BP exploded and leaked a huge volume of oil into the Gulf of Mexico. In an effort to control the spread of the oil, BP applied dispersants such as Corexit and conducted in-situ burnings of the oil. This catastrophe created a complex chain of events that affected not only the fragile water and land ecosystems, but the humans who breathe the air every day. Thousands of people were exposed to fumes associated with oil vapors from the spill, burning of the oil, and the toxic mixture of dispersants. While aiding in clean-up efforts, local fishermen were directly exposure to fumes when working on the Gulf. A notable amount of Gulf Coast residents were also exposed to the oil fumes as seasonal southeasterly winds blew vapors toward land. The Volatile Organic Compounds (VOC) found in oil vapors include: benzene, toluene, ethyl benzene, xylene, naphthalene, hydrogen sulfide and particulate matter (PM). Increases in water temperature and sunlight due to the summer season allow for these VOCs and PM to evaporate into the air more rapidly. Aside from the VOCs found in oil vapors, the dispersant being used to break up the oil is highly toxic and is thought to be even more toxic than the oil itself (EPA website, 2010). To protect human health, the environment, and to make informed policy decisions relevant to the spill, the EPA Region 6 has continuously monitored the affected areas carefully for levels of pollutants in the outdoor air that are associated with petroleum products and the burning of oil along the coast. In an effort to prevent, prepare for, and respond to future oil spills that occur in and around inland waters of the United States, the EPA has been working with local, state, and federal response partners. Air quality measurements were collected by the EPA at five active monitoring systems stationed along the coast.

  16. Information Requirements for Supervisory Air Traffic Controllers in Support of a Wake Vortex Departure System

    NASA Technical Reports Server (NTRS)

    Lohr, Gary W.; Williams, Daniel M.; Trujillo, Anna C.

    2008-01-01

    Closely Space Parallel Runway (CSPR) configurations are capacity limited for departures due to the requirement to apply wake vortex separation standards from traffic departing on the adjacent parallel runway. To mitigate the effects of this constraint, a concept focusing on wind dependent departure operations has been developed, known as the Wake Turbulence Mitigation for Departures (WTMD). This concept takes advantage of the fact that crosswinds of sufficient velocity blow wakes generated by aircraft departing from the downwind runway away from the upwind runway. Consequently, under certain conditions, wake separations on the upwind runway would not be required based on wakes generated by aircraft on the downwind runway, as is currently the case. It follows that information requirements, and sources for this information, would need to be determined for airport traffic control tower (ATCT) supervisory personnel who would be charged with decisions regarding use of the procedure. To determine the information requirements, data were collected from ATCT supervisors and controller-in-charge qualified individuals at Lambert-St. Louis International Airport (STL) and George Bush Houston Intercontinental Airport (IAH). STL and IAH were chosen as data collection sites based on the implementation of a WTMD prototype system, operating in shadow mode, at these locations. The 17 total subjects (STL: 5, IAH: 12) represented a broad-base of air traffic experience. Results indicated that the following information was required to support the conduct of WTMD operations: current and forecast weather information, current and forecast traffic demand and traffic flow restrictions, and WTMD System status information and alerting. Subjects further indicated that the requisite information is currently available in the tower cab with the exception of the WTMD status and alerting. Subjects were given a demonstration of a display supporting the prototype systems and unanimously stated that the

  17. Workload-Matched Adaptive Automation Support of Air Traffic Controller Information Processing Stages

    NASA Technical Reports Server (NTRS)

    Kaber, David B.; Prinzel, Lawrence J., III; Wright, Melanie C.; Clamann, Michael P.

    2002-01-01

    Adaptive automation (AA) has been explored as a solution to the problems associated with human-automation interaction in supervisory control environments. However, research has focused on the performance effects of dynamic control allocations of early stage sensory and information acquisition functions. The present research compares the effects of AA to the entire range of information processing stages of human operators, such as air traffic controllers. The results provide evidence that the effectiveness of AA is dependent on the stage of task performance (human-machine system information processing) that is flexibly automated. The results suggest that humans are better able to adapt to AA when applied to lower-level sensory and psychomotor functions, such as information acquisition and action implementation, as compared to AA applied to cognitive (analysis and decision-making) tasks. The results also provide support for the use of AA, as compared to completely manual control. These results are discussed in terms of implications for AA design for aviation.

  18. GHG and Air Pollution Co-benefits Analysis to Support Decision Making in Hyderabad, India

    NASA Astrophysics Data System (ADS)

    Guttikunda, S.; Shah, M.

    2008-12-01

    The increasing energy demand in the transport and industrial sectors accounts for a high carbon footprint in Hyderabad, India, and consequently to increasing air pollution. Integrated Environmental Strategies program under US EPA supported the analysis of Andhra Pradesh Pollution Control Board (PCB), to identify the major sources of pollution (local and global) and prioritize a series of strategies to better address mitigation in a cost effective manner. In Hyderabad, under the current trends, PM10 and CO2 emissions in 2020 are estimated to increase ~50 percent, compared to 2006 levels to ~43.5 ktons and ~10.3 million tons respectively. A co-benefits framework was implemented in analyzing the future control scenarios for human health benefits and carbon savings. Overall, implementing a series of interventions ranging from urban planning including better transport planning with bus rapid transport and metro rail, relocation of industries, and waste management, are expected to reduce the local and global emissions below the 2006 levels and yield an estimated ~US 196 million and ~US 492 million, in 2010 and 2020 respectively, in combined benefits of health and carbon savings. The PCB is coordinating the efforts for planning and implementation of these strategies. This paper will focus on presenting the methodology utilized for estimating emissions, pollutant dispersion, and impact on local and global environments, evaluated against the business as usual scenarios.

  19. Supported noble metal catalysts in the catalytic wet air oxidation of industrial wastewaters and sewage sludges.

    PubMed

    Besson, M; Descorme, C; Bernardi, M; Gallezot, P; di Gregorio, F; Grosjean, N; Minh, D Pham; Pintar, A

    2010-12-01

    This paper reviews some catalytic wet air oxidation (CWAO) investigations of industrial wastewaters over platinum and ruthenium catalysts supported on TiO2 and ZrO2 formulated to be active and resistant to leaching, with particular focus on the stability of the catalyst. Catalyst recycling experiments were performed in batch reactors and long-term stability tests were conducted in trickle-bed reactors. The catalyst did not leach upon treatment of Kraft bleaching plant and olive oil mill effluents, and could be either recycled or used for long periods of time in continuous reactors. Conversely, these catalysts were rapidly leached when used to treat effluents from the production of polymeric membranes containing N,N-dimethylformamide. The intermediate formation of amines, such as dimethylamine and methylamine with a high complexing capacity for the metal, was shown to be responsible for the metal leaching. These heterogeneous catalysts also deactivated upon CWAO of sewage sludges due to the adsorption of the solid organic matter. Pre-sonication of the sludge to disintegrate the flocs and improve solubility was inefficient. PMID:21214003

  20. NASA GES DISC support of CO2 Data from OCO-2, ACOS, and AIRS

    NASA Technical Reports Server (NTRS)

    Wei, Jennifer C; Vollmer, Bruce E.; Savtchenko, Andrey K.; Hearty, Thomas J; Albayrak, Rustem Arif; Deshong, Barbara E.

    2013-01-01

    NASA Goddard Earth Sciences Data and Information Services Centers (GES DISC) is the data center assigned to archive and distribute current AIRS, ACOS data and data from the upcoming OCO-2 mission. The GES DISC archives and supports data containing information on CO2 as well as other atmospheric composition, atmospheric dynamics, modeling and precipitation. Along with the data stewardship, an important mission of GES DISC is to facilitate access to and enhance the usability of data as well as to broaden the user base. GES DISC strives to promote the awareness of science content and novelty of the data by working with Science Team members and releasing news articles as appropriate. Analysis of events that are of interest to the general public, and that help in understanding the goals of NASA Earth Observing missions, have been among most popular practices.Users have unrestricted access to a user-friendly search interface, Mirador, that allows temporal, spatial, keyword and event searches, as well as an ontology-driven drill down. Variable subsetting, format conversion, quality screening, and quick browse, are among the services available in Mirador. The majority of the GES DISC data are also accessible through OPeNDAP (Open-source Project for a Network Data Access Protocol) and WMS (Web Map Service). These services add more options for specialized subsetting, format conversion, image viewing and contributing to data interoperability.

  1. Illustrations and supporting texts for sound standing waves of air columns in pipes in introductory physics textbooks

    NASA Astrophysics Data System (ADS)

    Zeng, Liang; Smith, Chris; Poelzer, G. Herold; Rodriguez, Jennifer; Corpuz, Edgar; Yanev, George

    2014-12-01

    In our pilot studies, we found that many introductory physics textbook illustrations with supporting text for sound standing waves of air columns in open-open, open-closed, and closed-closed pipes inhibit student understanding of sound standing wave phenomena due to student misunderstanding of how air molecules move within these pipes. Based on the construct of meaningful learning from cognitive psychology and semiotics, a quasiexperimental study was conducted to investigate the comparative effectiveness of two alternative approaches to student understanding: a traditional textbook illustration approach versus a newly designed air molecule motion illustration approach. Thirty volunteer students from introductory physics classes were randomly assigned to two groups of 15 each. Both groups were administered a presurvey. Then, group A read the air molecule motion illustration handout, and group B read a traditional textbook illustration handout; both groups were administered postsurveys. Subsequently, the procedure was reversed: group B read the air molecule motion illustration handout and group A read the traditional textbook illustration handout. This was followed by a second postsurvey along with an exit research questionnaire. The study found that the majority of students experienced meaningful learning and stated that they understood sound standing wave phenomena significantly better using the air molecule motion illustration approach. This finding provides a method for physics education researchers to design illustrations for abstract sound standing wave concepts, for publishers to improve their illustrations with supporting text, and for instructors to facilitate deeper learning in their students on sound standing waves.

  2. 75 FR 37711 - Automatic Dependent Surveillance-Broadcast (ADS-B) Out Performance Requirements To Support Air...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-06-30

    ... Support Air Traffic Control (ATC) Service'' (75 FR 30160). There are three footnotes in the preamble for... Amendments,'' published October 16, 2009 (74 FR 53368), the FAA revised part 21 subpart O, and Sec. 21.609... correction to that rule on March 1, 2010 (75 FR 9095) changing the effective date for the revision of...

  3. DEVELOPMENT OF AN AGGREGATION AND EPISODE SELECTION SCHEME TO SUPPORT THE MODELS-3 COMMUNITY MULTISCALE AIR QUALITY MODEL

    EPA Science Inventory

    The development of an episode selection and aggregation approach, designed to support distributional estimation of use with the Models-3 Community Multiscale Air Quality (CMAQ) model, is described. The approach utilized cluster analysis of the 700-hPa east-west and north-south...

  4. Thirsty Walls: A New Paradigm for Air Revitalization in Life Support

    NASA Technical Reports Server (NTRS)

    Graf, John; Brennecke, Joan; Weislogel, Mark

    2015-01-01

    Carbon Dioxide removal systems on submarines are compact and reliable. They use solubility chemistry. They spray a Carbon Dioxide adsorbing chemical directly into the air stream, and allow the liquid to settle. Carbon Dioxide removal systems on ISS are large and need repair. They use adsorption chemistry. They force air through a bed packed with granular zeolite, and heat the bed to desorb the Carbon Dioxide. The thermal cycles cause the zeolite to dust. New advances in additive manufacturing, and a better understanding of uid behavior in microgravity make it possible to expose a liquid directly to air in a microgravity environment. It is now practical to use submarine style solubility chemistry for atmosphere revitalization in space. It is now possible to develop space systems that achieve submarine levels of reliability. New developments in Ionic Liquid research make it possible to match the solubility performance characteristics of MEA used on submarines - with Ionic Liquids that do not release chemical vapors into the air. "Thirsty Walls" provide gentle, passive contact between ventilation air and Air Revitalization functions of temperature control, relative humidity control, and Carbon Dioxide removal. "Thirsty Walls" eliminates the need of large blowers and compressors that need to force air at high velocities through restrictive Air Revitalization hardware.

  5. “Applying Multi-scale Air Quality Models to Support Epidemiologic Studies”.

    EPA Science Inventory

    The Near-road Exposures and Effects of Urban air pollutant Study (NEXUS) investigating whether children with asthma living near major roadways in Detroit, MI have greater health impacts from air pollutants than those living farther away, particularly near roadways with high diese...

  6. Exploiting CRISPR/Cas systems for biotechnology

    PubMed Central

    Sampson, Timothy R.; Weiss, David S.

    2015-01-01

    The Cas9 endonuclease is the central component of the Type II CRISPR/Cas system, a prokaryotic adaptive restriction system against invading nucleic acids, such as those originating from bacteriophages and plasmids. Recently, this RNA-directed DNA endonuclease has been harnessed to target DNA sequences of interest. Here, we review the development of Cas9 as an important tool to not only edit the genomes of a number of different prokaryotic and eukaryotic species, but also as an efficient system for site-specific transcriptional repression or activation. Additionally, a specific Cas9 protein has been observed to target an RNA substrate, suggesting that Cas9 may have the ability to be programmed to target RNA as well. Cas proteins from other CRISPR/Cas subtypes may also be exploited in this regard. Thus, CRISPR/Cas systems represent an effective and versatile biotechnological tool, which will have significant impact on future advancements in genome engineering. PMID:24323919

  7. A female pelvic bone shape model for air/bone separation in support of synthetic CT generation for radiation therapy

    NASA Astrophysics Data System (ADS)

    Liu, Lianli; Cao, Yue; Fessler, Jeffrey A.; Jolly, Shruti; Balter, James M.

    2016-01-01

    Separating bone from air in MR data is one of the major challenges in using MR images to derive synthetic CT. The problem is further complicated when the anatomic regions filled with air are altered across scans due to air mobility, for instance, in pelvic regions, thereby the air regions estimated using an ultrashort echo time (UTE) sequence are invalid in other image series acquired for multispectral classification. This study aims to develop and investigate a female pelvic bone shape model to identify low intensity regions in MRI where air is unlikely to be present in support of synthetic CT generation without UTE imaging. CT scans of 30 patients were collected for the study, 17 of them also have corresponding MR scans. The shape model was built from the CT dataset, where the reference image was aligned to each of the training images using B-spline deformable registration. Principal component analysis was performed on B-spline coefficients for a compact model where shape variance was described by linear combination of principal modes. The model was applied to identify pelvic bone in MR images by deforming the corresponding MR data of the reference image to target MR images, where the search space of the deformation process was constrained within the subspace spanned by principal modes. The local minima in the search space were removed effectively by the shape model, thus supporting an efficient binary search for the optimal solution. We evaluated the model by its efficacy in identifying bone voxels and excluding air regions. The model was tested across the 17 patients that have corresponding MR scans using a leave-one-out cross validation. A simple model using the first leading principal mode only was found to achieve reasonable accuracy, where an averaged 87% of bone voxels were correctly identified. Finally dilation of the optimally fit bone mask by 5 mm was found to cover 96% of bone voxels while minimally impacting the overlap with air (below 0.4%).

  8. CAS-Induced Difficulties in Learning Mathematics?

    ERIC Educational Resources Information Center

    Jankvist, Uffe Thomas; Misfeldt, Morten

    2015-01-01

    In recent years computer algebra systems (CAS) have become an integrated part of the upper secondary school mathematics program. Despite the many positive possibilities of CAS, there also seems to be a flip side of the coin in relation to actual difficulties in learning mathematics, not least because a strong dependence on CAS for mathematical…

  9. "CAS" Statement of Shared Ethical Principles

    ERIC Educational Resources Information Center

    Council for the Advancement of Standards in Higher Education, 2006

    2006-01-01

    The Council for the Advancement of Standards in Higher Education (CAS) has served as a voice for quality assurance and promulgation of standards in higher education for over 25 years. CAS was established to promote inter-association efforts to address quality assurance, student learning, and professional integrity. CAS includes membership of over…

  10. Field validation of sound mitigation models and air pollutant emission testing in support of missile motor disposal activities.

    PubMed

    McFarland, Michael J; Palmer, Glenn R; Kordich, Micheal M; Pollet, Dean A; Jensen, James A; Lindsay, Mitchell H

    2005-08-01

    The U.S. Department of Defense approved activities conducted at the Utah Test and Training Range (UTTR) include both operational readiness test firing of intercontinental ballistic missile motors as well as the destruction of obsolete or otherwise unusable intercontinental ballistic missile motors through open burn/open detonation (OB/ OD). Within the Utah Division of Air Quality, these activities have been identified as having the potential to generate unacceptable noise levels, as well as significant amounts of hazardous air pollutants. Hill Air Force Base, UT, has completed a series of field tests at the UTTR in which sound-monitoring surveillance of OB/OD activities was conducted to validate the Sound Intensity Prediction System (SIPS) model. Using results generated by the SIPS model to support the decision to detonate, the UTTR successfully disposed of missile motors having an aggregate net explosive weight (NEW) of 56,500 lbs without generating adverse noise levels within populated areas. These results suggest that, under appropriate conditions, missile motors of even larger NEW may be detonated without exceeding regulatory noise limits. In conjunction with collecting noise monitoring data, air quality data was collected to support the development of air emission factors for both static missile motor firings and OB/OD activities. Through the installation of 15 ground-based air samplers, the generation of combustion fixed gases, hazardous air pollutants, and chlorides were monitored during the 56,500-lb NEW detonation event. Comparison of field measurements to predictions generated from the U.S. Navy's energetic combustion pollutant formation model, POLU4WN, indicated that, as the detonation fireball expanded from ground zero, organic compounds as well as carbon monoxide continued to oxidize as the hot gases reacted with ambient air. Hazardous air pollutant analysis of air samplers confirmed the presence of chloromethane, benzene, toluene, 1,2-propadiene, and

  11. Monitoring air pollution effects on children for supporting public health policy: the protocol of the prospective cohort MAPEC study

    PubMed Central

    Feretti, D; Ceretti, E; De Donno, A; Moretti, M; Carducci, A; Bonetta, S; Marrese, M R; Bonetti, A; Covolo, L; Bagordo, F; Villarini, M; Verani, M; Schilirò, T; Limina, R M; Grassi, T; Monarca, S; Casini, B; Carraro, E; Zani, C; Mazzoleni, G; Levaggi, R; Gelatti, U

    2014-01-01

    Introduction Genotoxic biomarkers have been studied largely in adult population, but few studies so far have investigated them in children exposed to air pollution. Children are a high-risk group as regards the health effects of air pollution and some studies suggest that early exposure during childhood can play an important role in the development of chronic diseases in adulthood. The objective of the project is to evaluate the associations between the concentration of urban air pollutants and biomarkers of early biological effect in children, and to propose a model for estimating the global risk of early biological effects due to air pollutants and other factors in children. Methods and analysis Two biomarkers of early biological effects, DNA damage by the comet assay and the micronuclei (MN) test, will be investigated in oral mucosa cells of 6–8-year-old children. Concurrently, some toxic airborne pollutants (polycyclic aromatic hydrocarbon (PAH) and nitro-PAH) and in vitro air mutagenicity and toxicity in ultra-fine air particulates (PM0.5) will be evaluated. Furthermore, demographic and socioeconomic variables, other sources of exposures to air pollutants and lifestyle variables will be assessed by a structured questionnaire. The associations between sociodemographic, environmental and other exposure variables and biomarkers of early biological effect using univariate and multivariate models will be analysed. A tentative model for calculating the global absolute risk of having early biological effects caused by air pollution and other variables will be proposed. Ethics and dissemination The project has been approved by the Ethics Committees of the local Health Authorities. The results will be communicated to local Public Health Agencies, for supporting educational programmes and health policy strategies. LIFE+2012 Environment Policy and Governance. LIFE12 ENV/IT/000614. PMID:25227631

  12. DEVELOPMENT AND APPLICATIONS OF CFD IN SUPPORT OF AIR QUALITY STUDIES OF ROADWAY AND BUILDING MICROENVIRONMENTS

    EPA Science Inventory

    There is a need to develop modeling and data analysis tools to increase our understanding of human exposures to air pollutants beyond what can be explained by "limited" field data. Modeling simulations of complex distributions of pollutant concentrations within roadw...

  13. Engineering Task Plan Supporting Rupture of Compressed Air Lines in Contaminated Areas USQ

    SciTech Connect

    JANICEK, G.P.

    2001-11-30

    This Engineering Task Plan (ETP) defines the Tasks and organizational responsibilities for obtaining Tank Farm data relative to pit structures and soil contamination levels for purposes of resolving the rupture of compressed air/gas lines unreviewed safety question.

  14. Integrating Sensor Monitoring Technology into the Current Air Pollution Regulatory Support Paradigm: Practical Considerations

    EPA Science Inventory

    The US Environmental Protection Agency (EPA) along with state, local, and tribal governments operate Federal Reference Method (FRM) and Federal Equivalent Method (FEM) instruments to assess compliance with US air pollution standards designed to protect human and ecosystem health....

  15. Data Quality Objectives Summary Report Supporting Radiological Air Surveillance Monitoring for the INL Site

    SciTech Connect

    Haney, Thomas Jay

    2015-05-01

    This report documents the Data Quality Objectives (DQOs) developed for the Idaho National Laboratory (INL) Site ambient air surveillance program. The development of the DQOs was based on the seven-step process recommended “for systematic planning to generate performance and acceptance criteria for collecting environmental data” (EPA 2006). The process helped to determine the type, quantity, and quality of data needed to meet current regulatory requirements and to follow U.S. Department of Energy guidance for environmental surveillance air monitoring design. It also considered the current air monitoring program that has existed at INL Site since the 1950s. The development of the DQOs involved the application of the atmospheric dispersion model CALPUFF to identify likely contamination dispersion patterns at and around the INL Site using site-specific meteorological data. Model simulations were used to quantitatively assess the probable frequency of detection of airborne radionuclides released by INL Site facilities using existing and proposed air monitors.

  16. Demonstration of Isothermal Compressed Air Energy Storage to Support Renewable Energy Production

    SciTech Connect

    Bollinger, Benjamin

    2015-01-02

    This project develops and demonstrates a megawatt (MW)-scale Energy Storage System that employs compressed air as the storage medium. An isothermal compressed air energy storage (ICAESTM) system rated for 1 MW or more will be demonstrated in a full-scale prototype unit. Breakthrough cost-effectiveness will be achieved through the use of proprietary methods for isothermal gas cycling and staged gas expansion implemented using industrially mature, readily-available components.The ICAES approach uses an electrically driven mechanical system to raise air to high pressure for storage in low-cost pressure vessels, pipeline, or lined-rock cavern (LRC). This air is later expanded through the same mechanical system to drive the electric motor as a generator. The approach incorporates two key efficiency-enhancing innovations: (1) isothermal (constant temperature) gas cycling, which is achieved by mixing liquid with air (via spray or foam) to exchange heat with air undergoing compression or expansion; and (2) a novel, staged gas-expansion scheme that allows the drivetrain to operate at constant power while still allowing the stored gas to work over its entire pressure range. The ICAES system will be scalable, non-toxic, and cost-effective, making it suitable for firming renewables and for other grid applications.

  17. Surveillance and Processing of Foreign DNA by the Escherichia coli CRISPR-Cas System.

    PubMed

    Redding, Sy; Sternberg, Samuel H; Marshall, Myles; Gibb, Bryan; Bhat, Prashant; Guegler, Chantal K; Wiedenheft, Blake; Doudna, Jennifer A; Greene, Eric C

    2015-11-01

    CRISPR-Cas adaptive immune systems protect bacteria and archaea against foreign genetic elements. In Escherichia coli, Cascade (CRISPR-associated complex for antiviral defense) is an RNA-guided surveillance complex that binds foreign DNA and recruits Cas3, a trans-acting nuclease helicase for target degradation. Here, we use single-molecule imaging to visualize Cascade and Cas3 binding to foreign DNA targets. Our analysis reveals two distinct pathways dictated by the presence or absence of a protospacer-adjacent motif (PAM). Binding to a protospacer flanked by a PAM recruits a nuclease-active Cas3 for degradation of short single-stranded regions of target DNA, whereas PAM mutations elicit an alternative pathway that recruits a nuclease-inactive Cas3 through a mechanism that is dependent on the Cas1 and Cas2 proteins. These findings explain how target recognition by Cascade can elicit distinct outcomes and support a model for acquisition of new spacer sequences through a mechanism involving processive, ATP-dependent Cas3 translocation along foreign DNA. PMID:26522594

  18. Association between tensin 1 and p130Cas at focal adhesions links actin inward flux to cell migration

    PubMed Central

    Zhao, Zhihai; Tan, Song Hui; Machiyama, Hiroaki; Kawauchi, Keiko; Araki, Keigo; Hirata, Hiroaki; Sawada, Yasuhiro

    2016-01-01

    ABSTRACT Cell migration is a highly dynamic process that plays pivotal roles in both physiological and pathological processes. We have previously reported that p130Cas supports cell migration through the binding to Src as well as phosphorylation-dependent association with actin retrograde flow at focal adhesions. However, it remains elusive how phosphorylated Cas interacts with actin cytoskeletons. We observe that the actin-binding protein, tensin 1, co-localizes with Cas, but not with its phosphorylation-defective mutant, at focal adhesions in leading regions of migrating cells. While a truncation mutant of tensin 1 that lacks the phosphotyrosine-binding PTB and SH2 domains (tensin 1-SH2PTB) poorly co-localizes or co-immunoprecitates with Cas, bacterially expressed recombinant tensin 1-SH2PTB protein binds to Cas in vitro in a Cas phosphorylation-dependent manner. Furthermore, exogenous expression of tensin 1-SH2PTB, which is devoid of the actin-interacting motifs, interferes with the Cas-driven cell migration, slows down the inward flux of Cas molecules, and impedes the displacement of Cas molecules from focal adhesions. Taken together, our results show that tensin 1 links inwardly moving actin cytoskeletons to phosphorylated Cas at focal adhesions, thereby driving cell migration. PMID:27029899

  19. Experience Of A US Air Force Surgical And Critical Care Team Deployed In Support Of Special Operations Command Africa.

    PubMed

    Delmonaco, Brian L; Baker, Aaron; Clay, Jared; Kilbourn, James

    2016-01-01

    An eight-person team of conventional US Air Force (USAF) medical providers deployed to support US Special Operations Forces (SOF) in North and West Africa for the first time in November 2014. The predeployment training, operations while deployed, and lessons learned from the challenges of performing surgery and medical evacuations in the remote desert environment of Chad and Niger on the continent of Africa are described. The vast area of operations and far-forward posture of these teams requires cooperation between partner African nations, the French military, and SOF to make these medical teams effective providers of surgical and critical care in Africa. The continuous deployment of conventional USAF medical providers since 2014 in support of US Special Operations Command Africa is challenging and will benefit from more medical teams and effective air assets to provide casualty evacuation across the vast area of operations. PMID:27045506

  20. On the nonlinear steady-state response of rigid rotors supported by air foil bearings-Theory and experiments

    NASA Astrophysics Data System (ADS)

    Larsen, Jon S.; Santos, Ilmar F.

    2015-06-01

    The demand for oil-free turbo compressors is increasing. Current trends are divided between active magnetic bearings and air foil bearings (AFB), the latter being important due to mechanical simplicity. AFB supported rotors are sensitive to unbalance due to low damping and nonlinear characteristics, hence accurate prediction of their response is important. This paper gives theoretical and experimental contributions by implementing and validating a new method to simulate the nonlinear steady-state response of a rotor supported by three pads segmented AFBs. The fluid film pressures, foil deflections and rotor movements are simultaneously solved, considering foil stiffness and damping coefficients estimated using a structural model, previously described and validated against experiments.

  1. NASA Earth Observation Systems and Applications for Public Health and Air Quality Models and Decisions Support

    NASA Astrophysics Data System (ADS)

    Estes, S. M.; Haynes, J. A.; Omar, A. H.

    2012-12-01

    Health and Air Quality providers and researchers need environmental data to study and understand the geographic, environmental, and meteorological differences in disease. Satellite remote sensing of the environment offers a unique vantage point that can fill in the gaps of environmental, spatial, and temporal data for tracking disease. This presentation will demonstrate the need for collaborations between multi-disciplinary research groups to develop the full potential of utilizing Earth Observations in studying health. Satellite earth observations present a unique vantage point of the earth's environment from space, which offers a wealth of health applications for the imaginative investigator. The presentation is directly related to Earth Observing systems and Global Health Surveillance and will present research results of the remote sensing environmental observations of earth and health applications, which can contribute to the public health and air quality research. As part of NASA approach and methodology they have used Earth Observation Systems and Applications for Public Health and Air Quality Models to provide a method for bridging gaps of environmental, spatial, and temporal data for tracking disease. This presentation will provide an overview of projects dealing with infectious diseases, water borne diseases and air quality and how many environmental variables effect human health. This presentation will provide a venue where the results of both research and practice using satellite earth observations to study weather and it's role in public health research.

  2. NASA Earth Observation Systems and Applications for Public Health and Air Quality Models and Decisions Support

    NASA Technical Reports Server (NTRS)

    Estes, Sue; Haynes, John; Omar, Ali

    2013-01-01

    Health and Air Quality providers and researchers need environmental data to study and understand the geographic, environmental, and meteorological differences in disease. Satellite remote sensing of the environment offers a unique vantage point that can fill in the gaps of environmental, spatial, and temporal data for tracking disease. This presentation will demonstrate the need for collaborations between multi-disciplinary research groups to develop the full potential of utilizing Earth Observations in studying health. Satellite earth observations present a unique vantage point of the earth's environment from space, which offers a wealth of health applications for the imaginative investigator. The presentation is directly related to Earth Observing systems and Global Health Surveillance and will present research results of the remote sensing environmental observations of earth and health applications, which can contribute to the public health and air quality research. As part of NASA approach and methodology they have used Earth Observation Systems and Applications for Public Health and Air Quality Models to provide a method for bridging gaps of environmental, spatial, and temporal data for tracking disease. This presentation will provide an overview of projects dealing with infectious diseases, water borne diseases and air quality and how many environmental variables effect human health. This presentation will provide a venue where the results of both research and practice using satellite earth observations to study weather and it's role in public health research.

  3. NASA Earth Observation Systems and Applications for Public Health and Air Quality Models and Decisions Support

    NASA Technical Reports Server (NTRS)

    Estes, Sue; Haynes, John; Omar, Ali

    2012-01-01

    Health and Air Quality providers and researchers need environmental data to study and understand the geographic, environmental, and meteorological differences in disease. Satellite remote sensing of the environment offers a unique vantage point that can fill in the gaps of environmental, spatial, and temporal data for tracking disease. This presentation will demonstrate the need for collaborations between multi-disciplinary research groups to develop the full potential of utilizing Earth Observations in studying health. Satellite earth observations present a unique vantage point of the earth's environment from space, which offers a wealth of health applications for the imaginative investigator. The presentation is directly related to Earth Observing systems and Global Health Surveillance and will present research results of the remote sensing environmental observations of earth and health applications, which can contribute to the public health and air quality research. As part of NASA approach and methodology they have used Earth Observation Systems and Applications for Public Health and Air Quality Models to provide a method for bridging gaps of environmental, spatial, and temporal data for tracking disease. This presentation will provide an overview of projects dealing with infectious diseases, water borne diseases and air quality and how many environmental variables effect human health. This presentation will provide a venue where the results of both research and practice using satellite earth observations to study weather and it's role in public health research.

  4. AIR TOXICS MODELING FROM LOCAL TO REGIONAL SCALES TO SUPPORT THE 2002 MULTIPOLLUTANT ASSESSMENT

    EPA Science Inventory

    This research focuses on developing models that can describe the chemical and physical processes affecting concentrations of toxic air pollutants in the atmosphere, at spatial scales, ranging from local (< 1 km) to regional (36 km). One objective of this task is to extend the ca...

  5. NASA Controller Acceptability Study 1(CAS-1) Experiment Description and Initial Observations

    NASA Technical Reports Server (NTRS)

    Chamberlain, James P.; Consiglio, Maria C.; Comstock, James R., Jr.; Ghatas, Rania W.; Munoz, Cesar

    2015-01-01

    This paper describes the Controller Acceptability Study 1 (CAS-1) experiment that was conducted by NASA Langley Research Center personnel from January through March 2014 and presents partial CAS-1 results. CAS-1 employed 14 air traffic controller volunteers as research subjects to assess the viability of simulated future unmanned aircraft systems (UAS) operating alongside manned aircraft in moderate-density, moderate-complexity Class E airspace. These simulated UAS were equipped with a prototype pilot-in-the-loop (PITL) Detect and Avoid (DAA) system, specifically the Self-Separation (SS) function of such a system based on Stratway+ software to replace the see-and-avoid capabilities of manned aircraft pilots. A quantitative CAS-1 objective was to determine horizontal miss distance (HMD) values for SS encounters that were most acceptable to air traffic controllers, specifically HMD values that were assessed as neither unsafely small nor disruptively large. HMD values between 0.5 and 3.0 nautical miles (nmi) were assessed for a wide array of encounter geometries between UAS and manned aircraft. The paper includes brief introductory material about DAA systems and their SS functions, followed by descriptions of the CAS-1 simulation environment, prototype PITL SS capability, and experiment design, and concludes with presentation and discussion of partial CAS-1 data and results.

  6. Is It Worth Using CAS for Symbolic Algebra Manipulation in the Middle Secondary Years? Some Teachers' Views

    ERIC Educational Resources Information Center

    Pierce, Robyn; Ball, Lynda; Stacey, Kaye

    2009-01-01

    The use of Computer Algebra Systems (CAS) in years 9 and 10 classrooms as a tool to support learning or in preparation for senior secondary mathematics is controversial. This paper presents an analysis of the positive and negative aspects of using CAS identified in the literature related to these year levels, along with the perceptions of 12…

  7. A female pelvic bone shape model for air/bone separation in support of synthetic CT generation for radiation therapy.

    PubMed

    Liu, Lianli; Cao, Yue; Fessler, Jeffrey A; Jolly, Shruti; Balter, James M

    2016-01-01

    Separating bone from air in MR data is one of the major challenges in using MR images to derive synthetic CT. The problem is further complicated when the anatomic regions filled with air are altered across scans due to air mobility, for instance, in pelvic regions, thereby the air regions estimated using an ultrashort echo time (UTE) sequence are invalid in other image series acquired for multispectral classification. This study aims to develop and investigate a female pelvic bone shape model to identify low intensity regions in MRI where air is unlikely to be present in support of synthetic CT generation without UTE imaging. CT scans of 30 patients were collected for the study, 17 of them also have corresponding MR scans. The shape model was built from the CT dataset, where the reference image was aligned to each of the training images using B-spline deformable registration. Principal component analysis was performed on B-spline coefficients for a compact model where shape variance was described by linear combination of principal modes. The model was applied to identify pelvic bone in MR images by deforming the corresponding MR data of the reference image to target MR images, where the search space of the deformation process was constrained within the subspace spanned by principal modes. The local minima in the search space were removed effectively by the shape model, thus supporting an efficient binary search for the optimal solution. We evaluated the model by its efficacy in identifying bone voxels and excluding air regions. The model was tested across the 17 patients that have corresponding MR scans using a leave-one-out cross validation. A simple model using the first leading principal mode only was found to achieve reasonable accuracy, where an averaged 87% of bone voxels were correctly identified. Finally dilation of the optimally fit bone mask by 5 mm was found to cover 96% of bone voxels while minimally impacting the overlap with air (below 0.4%). PMID

  8. Spouse Support and the Retention Intentions of Air Force Members: A Basis for Program Development.

    ERIC Educational Resources Information Center

    Bowen, Gary L.

    In recent years, the military services have become interested in developing personnel policies and support programs which enable military personnel to meet military objectives and still maintain a viable personal and family life. A study was undertaken to evaluate the direct and indirect impact of spouse support on the retention intention…

  9. Data Quality Objectives Supporting Radiological Air Emissions Monitoring for the PNNL Site

    SciTech Connect

    Barnett, J. M.; Meier, Kirsten M.; Snyder, Sandra F.; Fritz, Brad G.; Poston, Ted M.; Rhoads, Kathleen

    2010-05-25

    This document of Data Quality Objectives (DQOs) was prepared based on the U.S. Environmental Protection Agency (EPA) Guidance on Systematic Planning Using the Data Quality Objectives Process, EPA, QA/G4, 2/2006 (EPA 2006) as well as several other published DQOs. Pacific Northwest National Laboratory (PNNL) is in the process of developing a radiological air monitoring program for the PNNL Site that is distinct from that of the nearby Hanford Site. Radiological emissions at the PNNL Site result from Physical Sciences Facility (PSF) major emissions units. A team was established to determine how the PNNL Site would meet federal regulations and address guidelines developed to monitor and estimate offsite air emissions of radioactive materials. The result is a program that monitors the impact to the public from the PNNL Site.

  10. Phase 2: HGM air flow tests in support of HEX vane investigation

    NASA Technical Reports Server (NTRS)

    Cox, G. B., Jr.; Steele, L. L.; Eisenhart, D. W.

    1993-01-01

    Following the start of SSME certification testing for the Pratt and Whitney Alternate Turbopump Development (ATD) High Pressure Oxidizer Turbopump (HPOTP), cracking of the leading edge of the inner HEX vane was experienced. The HEX vane, at the inlet of the oxidizer bowl in the Hot Gas Manifold (HGM), accepts the HPOTP turbine discharge flow and turns it toward the Gaseous Oxidizer Heat Exchanger (GOX HEX) coil. The cracking consistently initiated over a specific circumferential region of the hex vane, with other circumferential locations appearing with increased run time. Since cracking had not to date been seen with the baseline HPOTP, a fluid-structural interaction involving the ATD HPOTP turbine exit flowfield and the HEX inner vane was suspected. As part of NASA contract NAS8-36801, Pratt and Whitney conducted air flow tests of the ATD HPOTP turbine turnaround duct flowpath in the MSFC Phase 2 HGM air flow model. These tests included HEX vane strain gages and additional fluctuating pressure gages in the turnaround duct and HEX vane flowpath area. Three-dimensional flow probe measurements at two stations downstream of the turbine simulator exit plane were also made. Modifications to the HPOTP turbine simulator investigated the effects on turbine exit flow profile and velocity components, with the objective of reproducing flow conditions calculated for the actual ATD HPOTP hardware. Testing was done at the MSFC SSME Dynamic Fluid Air Flow (Dual-Leg) Facility, at air supply pressures between 50 and 250 psia. Combinations of turbine exit Mach number and pressure level were run to investigate the effect of flow regime. Information presented includes: (1) Descriptions of turbine simulator modifications to produce the desired flow environment; (2) Types and locations for instrumentation added to the flow model for improved diagnostic capability; (3) Evaluation of the effect of changes to the turbine simulator flowpath on the turbine exit flow environment; and (4

  11. Phase 2: HGM air flow tests in support of HEX vane investigation

    NASA Astrophysics Data System (ADS)

    Cox, G. B., Jr.; Steele, L. L.; Eisenhart, D. W.

    1993-07-01

    Following the start of SSME certification testing for the Pratt and Whitney Alternate Turbopump Development (ATD) High Pressure Oxidizer Turbopump (HPOTP), cracking of the leading edge of the inner HEX vane was experienced. The HEX vane, at the inlet of the oxidizer bowl in the Hot Gas Manifold (HGM), accepts the HPOTP turbine discharge flow and turns it toward the Gaseous Oxidizer Heat Exchanger (GOX HEX) coil. The cracking consistently initiated over a specific circumferential region of the hex vane, with other circumferential locations appearing with increased run time. Since cracking had not to date been seen with the baseline HPOTP, a fluid-structural interaction involving the ATD HPOTP turbine exit flowfield and the HEX inner vane was suspected. As part of NASA contract NAS8-36801, Pratt and Whitney conducted air flow tests of the ATD HPOTP turbine turnaround duct flowpath in the MSFC Phase 2 HGM air flow model. These tests included HEX vane strain gages and additional fluctuating pressure gages in the turnaround duct and HEX vane flowpath area. Three-dimensional flow probe measurements at two stations downstream of the turbine simulator exit plane were also made. Modifications to the HPOTP turbine simulator investigated the effects on turbine exit flow profile and velocity components, with the objective of reproducing flow conditions calculated for the actual ATD HPOTP hardware. Testing was done at the MSFC SSME Dynamic Fluid Air Flow (Dual-Leg) Facility, at air supply pressures between 50 and 250 psia. Combinations of turbine exit Mach number and pressure level were run to investigate the effect of flow regime. Information presented includes: (1) Descriptions of turbine simulator modifications to produce the desired flow environment; (2) Types and locations for instrumentation added to the flow model for improved diagnostic capability; (3) Evaluation of the effect of changes to the turbine simulator flowpath on the turbine exit flow environment; and (4

  12. Airborne lidar mapping of vertical ozone distributions in support of the 1990 Clean Air Act Amendments

    NASA Technical Reports Server (NTRS)

    Uthe, Edward E.; Nielsen, Norman B.; Livingston, John M.

    1992-01-01

    The 1990 Clean Air Act Amendments mandated attainment of the ozone standard established by the U.S. Environmental Protection Agency. Improved photochemical models validated by experimental data are needed to develop strategies for reducing near surface ozone concentrations downwind of urban and industrial centers. For more than 10 years, lidar has been used on large aircraft to provide unique information on ozone distributions in the atmosphere. However, compact airborne lidar systems are needed for operation on small aircraft of the type typically used on regional air quality investigations to collect data with which to develop and validate air quality models. Data presented in this paper will consist of a comparison between airborne differential absorption lidar (DIAL) and airborne in-situ ozone measurements. Also discussed are future plans to improve the airborne ultraviolet-DIAL for ozone and other gas observations and addition of a Fourier Transform Infrared (FTIR) emission spectrometer to investigate the effects of other gas species on vertical ozone distribution.

  13. Expanding CRISPR/Cas9 Genome Editing Capacity in Zebrafish Using SaCas9

    PubMed Central

    Feng, Yan; Chen, Cheng; Han, Yuxiang; Chen, Zelin; Lu, Xiaochan; Liang, Fang; Li, Song; Qin, Wei; Lin, Shuo

    2016-01-01

    The type II CRISPR/Cas9 system has been used widely for genome editing in zebrafish. However, the requirement for the 5′-NGG-3′ protospacer-adjacent motif (PAM) of Cas9 from Streptococcus pyogenes (SpCas9) limits its targeting sequences. Here, we report that a Cas9 ortholog from Staphylococcus aureus (SaCas9), and its KKH variant, successfully induced targeted mutagenesis with high frequency in zebrafish. Confirming previous findings, the SpCas9 variant, VQR, can also induce targeted mutations in zebrafish. Bioinformatics analysis of these new Cas targets suggests that the number of available target sites in the zebrafish genome can be greatly expanded. Collectively, the expanded target repertoire of Cas9 in zebrafish should further facilitate the utility of this organism for genetic studies of vertebrate biology. PMID:27317783

  14. Expanding CRISPR/Cas9 Genome Editing Capacity in Zebrafish Using SaCas9.

    PubMed

    Feng, Yan; Chen, Cheng; Han, Yuxiang; Chen, Zelin; Lu, Xiaochan; Liang, Fang; Li, Song; Qin, Wei; Lin, Shuo

    2016-01-01

    The type II CRISPR/Cas9 system has been used widely for genome editing in zebrafish. However, the requirement for the 5'-NGG-3' protospacer-adjacent motif (PAM) of Cas9 from Streptococcus pyogenes (SpCas9) limits its targeting sequences. Here, we report that a Cas9 ortholog from Staphylococcus aureus (SaCas9), and its KKH variant, successfully induced targeted mutagenesis with high frequency in zebrafish. Confirming previous findings, the SpCas9 variant, VQR, can also induce targeted mutations in zebrafish. Bioinformatics analysis of these new Cas targets suggests that the number of available target sites in the zebrafish genome can be greatly expanded. Collectively, the expanded target repertoire of Cas9 in zebrafish should further facilitate the utility of this organism for genetic studies of vertebrate biology. PMID:27317783

  15. Benchmarking, Research, Development, and Support for ORNL Automated Image and Signature Retrieval (AIR/ASR) Technologies

    SciTech Connect

    Tobin, K.W.

    2004-06-01

    This report describes the results of a Cooperative Research and Development Agreement (CRADA) with Applied Materials, Inc. (AMAT) of Santa Clara, California. This project encompassed the continued development and integration of the ORNL Automated Image Retrieval (AIR) technology, and an extension of the technology denoted Automated Signature Retrieval (ASR), and other related technologies with the Defect Source Identification (DSI) software system that was under development by AMAT at the time this work was performed. In the semiconductor manufacturing environment, defect imagery is used to diagnose problems in the manufacturing line, train yield management engineers, and examine historical data for trends. Image management in semiconductor data systems is a growing cause of concern in the industry as fabricators are now collecting up to 20,000 images each week. In response to this concern, researchers at the Oak Ridge National Laboratory (ORNL) developed a semiconductor-specific content-based image retrieval method and system, also known as AIR. The system uses an image-based query-by-example method to locate and retrieve similar imagery from a database of digital imagery using visual image characteristics. The query method is based on a unique architecture that takes advantage of the statistical, morphological, and structural characteristics of image data, generated by inspection equipment in industrial applications. The system improves the manufacturing process by allowing rapid access to historical records of similar events so that errant process equipment can be isolated and corrective actions can be quickly taken to improve yield. The combined ORNL and AMAT technology is referred to hereafter as DSI-AIR and DSI-ASR.

  16. Advanced Regenerative Environmental Control and Life Support Systems: Air and Water Regeneration

    NASA Technical Reports Server (NTRS)

    Schubert, F. H.; Wynveen, R. A.; Quattrone, P. D.

    1985-01-01

    Extended manned space missions will require regenerative life support techniques. Past manned missions used nonregenerative expendables, except for a molecular sieve based carbon dioxide removal system aboard Skylab. The resupply penalties associated with expendables becomes prohibitive as crew size and mission duration increase. The Space Station scheduled to be operational in the 1990's is based on a crew of four to sixteen and a resupply period of 90 days or greater. It will be the first major spacecraft to employ regenerable techniques for life support. The techniques to be used in the requirements for the space station are addressed.

  17. Using Course Load Matrix Analysis To Support Departmental Planning for Enrollment Expansion. AIR 2002 Forum Paper.

    ERIC Educational Resources Information Center

    Harter, Elizabeth A.; England, Martin D.

    The planning office of a large urban university produced an induced course load matrix (ICLM) analysis to support the university's plans for undergraduate enrollment growth at its three campuses. The ICLM tables, based on the complete course histories of the 1993 entering cohort, summarize the program and course selections of a cohort of students…

  18. Decision Support Systems: An Institutional Research Perspective. AIR Forum 1982 Paper.

    ERIC Educational Resources Information Center

    Sheehan, Bernard S.

    Ideas that have been developed in the decision support systems (DSS) literature that seem particularly relevant to institutional research, planning, and analysis (IRPA) are reviewed. In addition, a survey of practitioners in Canadian universities that provides new information on the rapidly changing current state of information technology,…

  19. Designing a Decision-Support System for Enrollment Management. AIR 1985 Annual Forum Paper.

    ERIC Educational Resources Information Center

    Glover, Robert H.

    University of Hartford's decision-support system for enrollment management, which uses fourth-generation software tools, is described, with attention to the conceptual framework, design and implementation plan, and progress to date. The university's planners, institutional researchers, and admissions and financial aid officers are cooperating in…

  20. Data Quality Objectives Supporting Radiological Air Emissions Monitoring for the PNNL Site

    SciTech Connect

    Barnett, J. Matthew; Meier, Kirsten M.; Snyder, Sandra F.; Fritz, Brad G.; Poston, Theodore M.; Antonio, Ernest J.

    2012-11-12

    Pacific Northwest National Laboratory (PNNL) is in the process of developing a radiological air monitoring program for the PNNL Site that is distinct from that of the nearby Hanford Site. The original DQO (PNNL-19427) considered radiological emissions at the PNNL Site from Physical Sciences Facility (PSF) major emissions units. This first revision considers PNNL Site changes subsequent to the implementation of the original DQO. A team was established to determine how the PNNL Site changes would continue to meet federal regulations and address guidelines developed to monitor air emissions and estimate offsite impacts of radioactive material operations. The result is an updated program to monitor the impact to the public from the PNNL Site. The team used the emission unit operation parameters and local meteorological data as well as information from the PSF Potential-to-Emit documentation and Notices of Construction submitted to the Washington State Department of Health (WDOH). The locations where environmental monitoring stations would most successfully characterize the maximum offsite impacts of PNNL Site emissions from the three PSF buildings with major emission units were determined from these data. Three monitoring station locations were determined during the original revision of this document. This first revision considers expanded Department of Energy operations south of the PNNL Site and relocation of the two offsite, northern monitoring stations to sites near the PNNL Site fenceline. Inclusion of the southern facilities resulted in the proposal for a fourth monitoring station in the southern region. The southern expansion added two minor emission unit facilities and one diffuse emission unit facility. Relocation of the two northern stations was possible due to the use of solar power, rather than the previous limitation of the need for access to AC power, at these more remote locations. Addendum A contains all the changes brought about by the revision 1

  1. A Life-Cycle Cost Estimating Methodology for NASA-Developed Air Traffic Control Decision Support Tools

    NASA Technical Reports Server (NTRS)

    Wang, Jianzhong Jay; Datta, Koushik; Landis, Michael R. (Technical Monitor)

    2002-01-01

    This paper describes the development of a life-cycle cost (LCC) estimating methodology for air traffic control Decision Support Tools (DSTs) under development by the National Aeronautics and Space Administration (NASA), using a combination of parametric, analogy, and expert opinion methods. There is no one standard methodology and technique that is used by NASA or by the Federal Aviation Administration (FAA) for LCC estimation of prospective Decision Support Tools. Some of the frequently used methodologies include bottom-up, analogy, top-down, parametric, expert judgement, and Parkinson's Law. The developed LCC estimating methodology can be visualized as a three-dimensional matrix where the three axes represent coverage, estimation, and timing. This paper focuses on the three characteristics of this methodology that correspond to the three axes.

  2. Spouse abuse among United States Air Force personnel who deployed in support of Operation Iraqi Freedom/Operation Enduring Freedom.

    PubMed

    Rabenhorst, Mandy M; McCarthy, Randy J; Thomsen, Cynthia J; Milner, Joel S; Travis, Wendy J; Foster, Rachel E; Copeland, Carol W

    2013-10-01

    The authors examined spouse abuse perpetration among all married U.S. Air Force personnel who deployed in support of Operation Iraqi Freedom/Operation Enduring Freedom. Using Poisson and conditional Poisson regression, they compared rates of spouse abuse perpetration predeployment and postdeployment in the population of married U.S. Air Force personnel who had a combat-related deployment between October 1, 2001 and October 31, 2008 (N = 156,296). Just over 2% (n = 3,524) of deployers perpetrated at least one substantiated incident of spouse physical or emotional abuse within the 308,197,653 days at risk for abuse during the study period. Male deployers perpetrated spouse abuse at approximately twice the rate of female deployers. Regarding changes in rates of spouse abuse perpetration postdeployment versus predeployment among all deployers, the authors found no differences overall; however, several deployer and incident-related characteristics moderated this effect. Rates of emotional abuse, mild abuse, and abuse not involving alcohol were significantly lower postdeployment, whereas rates of moderate/severe abuse and abuse involving alcohol were significantly higher postdeployment. Although the majority of U.S. Air Force deployers did not perpetrate any substantiated incidents of spouse abuse, there was variability in the impact of deployment on spouse abuse rates before versus after deployment. The finding that rates of moderate/severe spouse abuse incidents involving alcohol were higher postdeployment suggests a need for focused prevention/intervention efforts. PMID:24015706

  3. Data Quality Objectives Supporting Radiological Air Emissions Monitoring for the Marine Sciences Laboratory, Sequim Site

    SciTech Connect

    Barnett, J. Matthew; Meier, Kirsten M.; Snyder, Sandra F.; Antonio, Ernest J.; Fritz, Brad G.; Poston, Theodore M.

    2012-12-27

    This document of Data Quality Objectives (DQOs) was prepared based on the U.S. Environmental Protection Agency (EPA) Guidance on Systematic Planning Using the Data Quality Objectives Process, EPA, QA/G4, 2/2006 (EPA 2006), as well as several other published DQOs. The intent of this report is to determine the necessary steps required to ensure that radioactive emissions to the air from the Marine Sciences Laboratory (MSL) headquartered at the Pacific Northwest National Laboratory’s Sequim Marine Research Operations (Sequim Site) on Washington State’s Olympic Peninsula are managed in accordance with regulatory requirements and best practices. The Sequim Site was transitioned in October 2012 from private operation under Battelle Memorial Institute to an exclusive use contract with the U.S. Department of Energy, Office of Science, Pacific Northwest Site Office.

  4. Testing an advanced satellite technique for dust detection as a decision support system for the air quality assessment

    NASA Astrophysics Data System (ADS)

    Falconieri, Alfredo; Filizzola, Carolina; Femiano, Rossella; Marchese, Francesco; Sannazzaro, Filomena; Pergola, Nicola; Tramutoli, Valerio; Di Muro, Ersilia; Divietri, Mariella; Crisci, Anna Maria; Lovallo, Michele; Mangiamele, Lucia; Vaccaro, Maria Pia; Palma, Achille

    2014-05-01

    In order to correctly apply the European directive for air quality (2008/50/CE), local Authorities are often requested to discriminate the possible origin (natural/anthropic) of anomalous concentration of pollutants in the air (art.20 Directive 2008/50/CE). In this framework, it's been focused on PM10 and PM2,5 concentrations and sources. In fact, depending on their origin, appropriate counter-measures can be taken devoted to prevent their production (e.g. by traffic restriction) or simply to reduce their impact on citizen health (e.g. information campaigns). In this context suitable satellite techniques can be used in order to identify natural sources (particularly Saharan dust, but also volcanic ash or forest fire smoke) that can be responsible of over-threshold concentration of PM10/2,5 in populated areas. In the framework of the NIBS (Networking and Internationalization of Basilicata Space Technologies) project, funded by the Basilicata Region within the ERDF 2007-2013 program, the School of Engineering of University of Basilicata, the Institute of Methodologies for Environmental Analysis of National Research Council (IMAA-CNR) and the Regional Agency for the Protection of the Environment of Basilicata Region (ARPAB) have started a collaboration devoted to assess the potential of the use of advanced satellite techniques for Saharan dust events identification to support ARPAB activities related to the application of the European directive for air quality (2008/50/CE) in Basilicata region. In such a joint activity, the Robust Satellite Technique (RST) approach has been assessed and tested as a decision support system for monitoring and evaluating air quality at local and regional level. In particular, RST-DUST products, derived by processing high temporal resolution data provided by SEVIRI (Spinning Enhanced Visible and Infrared Imager) sensor on board Meteosat Second Generation platforms, have been analysed together with PM10 measurements performed by the ground

  5. Workshop summary: 'Integrating air quality and climate mitigation - is there a need for new metrics to support decision making?'

    NASA Astrophysics Data System (ADS)

    von Schneidemesser, E.; Schmale, J.; Van Aardenne, J.

    2013-12-01

    Air pollution and climate change are often treated at national and international level as separate problems under different regulatory or thematic frameworks and different policy departments. With air pollution and climate change being strongly linked with regard to their causes, effects and mitigation options, the integration of policies that steer air pollutant and greenhouse gas emission reductions might result in cost-efficient, more effective and thus more sustainable tackling of the two problems. To support informed decision making and to work towards an integrated air quality and climate change mitigation policy requires the identification, quantification and communication of present-day and potential future co-benefits and trade-offs. The identification of co-benefits and trade-offs requires the application of appropriate metrics that are well rooted in science, easy to understand and reflect the needs of policy, industry and the public for informed decision making. For the purpose of this workshop, metrics were loosely defined as a quantified measure of effect or impact used to inform decision-making and to evaluate mitigation measures. The workshop held on October 9 and 10 and co-organized between the European Environment Agency and the Institute for Advanced Sustainability Studies brought together representatives from science, policy, NGOs, and industry to discuss whether current available metrics are 'fit for purpose' or whether there is a need to develop alternative metrics or reassess the way current metrics are used and communicated. Based on the workshop outcome the presentation will (a) summarize the informational needs and current application of metrics by the end-users, who, depending on their field and area of operation might require health, policy, and/or economically relevant parameters at different scales, (b) provide an overview of the state of the science of currently used and newly developed metrics, and the scientific validity of these

  6. Eastern Texas Air Quality Forecasting System to Support TexAQS-II and 8-hour Ozone Modeling

    NASA Astrophysics Data System (ADS)

    Byun, D. W.

    2005-12-01

    The main objective of the Second Texas Air Quality Study (TexAQS-II) for 2005 and 2006 is to understand emissions and processes associated with the formation and transport of ozone and regional haze in Texas. The target research area is the more populated eastern half of the state, roughly from Interstate 35 eastward. Accurate meteorological and photochemical modeling efforts are essential to support this study and further enhance modeling efforts for establishing the State Implementation Plan (SIP) by Texas Commission on Environmental Quality (TCEQ). An air quality forecasting (AQF) system for Eastern Texas has been developed to provide these data and to further facilitate retrospective simulations to allow for model improvement and increased understanding of ozone episodes and emissions. We perform two-day air quality forecasting simulations with the 12-km Eastern Texas regional domain, and the 4-km Houston-Galveston area (HGA) domain utilizing a 48-CPU Beowulf Linux computer system. The dynamic boundary conditions are provided by the 36-km resolution conterminous US (CONUS) domain CMAQ simulations. Initial meteorological conditions are provided by the daily ETA forecast results. The results of individual runs are stored and made available to researchers and state and local officials via internet to study the patterns of air quality and its relationship to weather conditions and emissions. The data during the pre- and post-processing stages are in tens of gigabytes and must be managed efficiently during both the actual real-time and the subsequent computation periods. The nature of these forecasts and the time at which the initial data is available necessitates that models be executed within tight deadlines. A set of complex operational scripts is used to allow automatic operation of the data download, sequencing processors, performing graphical analysis, building database archives, and presenting on the web.

  7. Impact of crop production on air quality in life support dynamics in closed habitats

    SciTech Connect

    Volk, T.

    1987-01-01

    Interest in human-designed closed habitats - where the substances needed for human life support are continuously regenerated from waste products - is growing, as apparent from the National Aeronautics and Space Administration's Closed Ecological Life Support Systems Program, the Soviet Union's Bios experiments, and the Biosphere II Project in Arizona. Nuclear-powered bases on the moon and Mars will have food-growing capabilities, and through gas-exchange processes these crops will alter the atmospheric composition. This study focuses on major gases tied to human life support: CO/sub 2/, O/sub 2/, and water vapor. Since actual systems are years and likely decades away, simulation studies can indicate necessary further research and provide instruction about the predicted behavior of such systems. To look at the first-order plant dynamics, i.e., the production of O/sub 2/ and water vapor and the consumption of CO/sub 2/, a simulation model is constructed with crop, human, and waste subsystems. The plant can either share an atmosphere with the humans or be separate, linked by osmotic or mechanical gas exchangers. The crop subsystem is sketched. Stoichiometric equations for the biosynthesis of protein, carbohydrates, and lipids in the edible portion and carbohydrates, fiber, and lignin in the inedible portion govern growth, mimicking that currently observed in the latest hydroponic wheat experiments.

  8. Active carbon-ceramic sphere as support of ruthenium catalysts for catalytic wet air oxidation (CWAO) of resin effluent.

    PubMed

    Liu, Wei-Min; Hu, Yi-Qiang; Tu, Shan-Tung

    2010-07-15

    Active carbon-ceramic sphere as support of ruthenium catalysts were evaluated through the catalytic wet air oxidation (CWAO) of resin effluent in a packed-bed reactor. Active carbon-ceramic sphere and ruthenium catalysts were characterized by N(2) adsorption and chemisorption measurements. BET surface area and total pore volume of active carbon (AC) in the active carbon-ceramic sphere increase with increasing KOH-to-carbon ratio, and AC in the sample KC-120 possesses values as high as 1100 m(2) g(-1) and 0.69 cm(3) g(-1) (carbon percentage: 4.73 wt.%), especially. Active carbon-ceramic sphere supported ruthenium catalysts were prepared using the RuCl(3) solution impregnation onto these supports, the ruthenium loading was fixed at 1-5 wt.% of AC in the support. The catalytic activity varies according to the following order: Ru/KC-120>Ru/KC-80>Ru/KC-60>KC-120>without catalysts. It is found that the 3 wt.% Ru/KC-120 catalyst displays highest stability in the CWAO of resin effluent during 30 days. Chemical oxygen demand (COD) and phenol removal were about 92% and 96%, respectively at the reaction temperature of 200 degrees C, oxygen pressure of 1.5 MPa, the water flow rate of 0.75 L h(-1) and the oxygen flow rate of 13.5 L h(-1). PMID:20362394

  9. CAS as Environments for Implementing Mathematical Microworlds.

    ERIC Educational Resources Information Center

    Alpers, Burkhard

    2002-01-01

    Investigates whether computer algebra systems (CAS) are suitable environments for implementing mathematical microworlds. Recalls what constitutes a microworld and explores how CAS can be used for implementation, stating potentials as well as limitations. Provides as an example the microworld "Formula 1", implemented in Maple Software. (Author/KHR)

  10. CAS

    SciTech Connect

    Martinez, B.; Pomeroy, G. )

    1989-12-02

    The Security Alarm System is a data acquisition and control system which collects data from intrusion sensors and displays the information in a real-time environment for operators. The Access Control System monitors and controls the movement of personnel with the use of card readers and biometrics hand readers.

  11. Precursor ionization ahead of laser-supported detonation wave in air and argon

    NASA Astrophysics Data System (ADS)

    Shimamura, Kohei; Komurasaki, Kimiya; Koizumi, Hiroyuki; Arakawa, Yoshihiro

    2012-10-01

    Laser-produced plasma in a gaseous form is considered, which has attracted great interest for use in many devices. After breakdown one of possible mechanisms of occurrence of this process is noted as laser-supported detonation wave. This wave consisting of the shock wave and the beam absorbing plasma travels at several kilometers per second along the laser beam channel in the direction opposite to the beam incidence. A Nd: Glass laser and a TEA CO2 laser were utilized. According to shadowgraph and spectroscopic studies, the wave has a velocity of 1-10 km/s, an electron temperature of 2-5 eV and an electron density of 10^24 m-3 after breakdown. For simplicity, the discussion is restricted to one-dimensional flows that considers the radiation from plasma and the collisional ionization by laser irradiation. Assuming that UV photons radiating from laser plasma induce photoionization ahead of ionization front, this ionization frequency fp at the distance lp (mean free path of photon) from the wave front corresponds to 10^10 s-1. This is higher than the collisional ionization frequency (10^5-6 s-1). Analytical velocities (fplp) describing the avalanche ionization in the pre-ionization layer agree with the experimentally observed velocities. These results does not depend on background gas and laser-wavelength.

  12. Calculation Package: Derivation of Facility-Specific Derived Air Concentration (DAC) Values in Support of Spallation Neutron Source Operations

    SciTech Connect

    McLaughlin, David A

    2009-12-01

    Derived air concentration (DAC) values for 175 radionuclides* produced at the Oak Ridge National Laboratory (ORNL) Spallation Neutron Source (SNS), but not listed in Appendix A of 10 CFR 835 (01/01/2009 version), are presented. The proposed DAC values, ranging between 1 E-07 {micro}Ci/mL and 2 E-03 {micro}Ci/mL, were calculated in accordance with the recommendations of the International Commission on Radiological Protection (ICRP), and are intended to support an exemption request seeking regulatory relief from the 10 CFR 835, Appendix A, requirement to apply restrictive DACs of 2E-13 {micro}Ci/mL and 4E-11 {micro}Ci/mL and for non-listed alpha and non-alpha-emitting radionuclides, respectively.

  13. RXTE Observations of Cas A

    NASA Technical Reports Server (NTRS)

    Rothschild, R. E.; Lingenfelter, R. E.; Heindl, W. A.; Blanco, P. R.; Pelling, M. R.; Gruber, D. E.; Allen, G. E.; Jahoda, K.; Swank, J. H.; Woosley, S. E.; Nomoto, K.; Higdon, J. C.; Dermer, Charles D. (Editor); Strickman, Mark S. (Editor); Kurfess, James D. (Editor)

    1997-01-01

    The exciting detection by the COMPTEL instrument of the 1157 keV Ti-44 line from the supernova remnant Cas A sets important new constraints on supernova dynamics and nucleosynthesis. The Ti-44 decay also produces x-ray lines at 68 and 78 keV, whose flux should be essentially the same as that of the gamma ray line. The revised COMPTEL flux of 4 x l0(exp -5) cm(exp -2)s(exp -1) is very near the sensitivity limit for line detection by the HEXTE instrument on RXTE. We report on the results from two RXTE observations - 20 ks during In Orbit Checkout in January 1996 and 200 ks in April 1996. We also find a strong continuum emission suggesting cosmic ray electron acceleration in the remnant.

  14. United States Air Force 611th Air Support Group/Civil Engineering Squadron Elmendorf AFB, Alaska. Remedial investigation and feasibility study. Bullen Point Radar Installation, Alaska. Final report

    SciTech Connect

    Karmi, S.

    1996-03-18

    The United States Air Force (Air Force) has prepared this Remedial investigation/Feasibility Study (RI/FS) report as part of the Installation Restoration Program (IRP) to present results of RI/FS activities at five sites at the Bullen Point radar installation. The IRP provides for investigating, quantifying, and remediating environmental contamination from past waste management activities at Air Force installations throughout the United States.

  15. A Non-Stem-Loop CRISPR RNA Is Processed by Dual Binding Cas6.

    PubMed

    Shao, Yaming; Richter, Hagen; Sun, Shengfang; Sharma, Kundan; Urlaub, Henning; Randau, Lennart; Li, Hong

    2016-04-01

    A subclass of recently discovered CRISPR repeat RNA in bacteria contains minimally recognizable structural features that facilitate an unknown mechanism of recognition and processing by the Cas6 family of endoribonucleases. Cocrystal structures of Cas6 from Methanococcus maripaludis (MmCas6b) bound with its repeat RNA revealed a dual site binding structure and a cleavage site conformation poised for phosphodiester bond breakage. Two non-interacting MmCas6b bind to two separate AAYAA motifs within the same repeat, one distal and one adjacent to the cleavage site. This bound structure potentially competes with a stable but non-productive RNA structure. At the cleavage site, MmCas6b supplies a base pair mimic to stabilize a short 2 base pair stem immediately upstream of the scissile phosphate. Complementary biochemical analyses support the dual-AAYAA binding model and a critical role of the protein-RNA base pair mimic. Our results reveal a previously unknown method of processing non-stem-loop CRISPR RNA by Cas6. PMID:26996962

  16. 48 CFR 9903.201-2 - Types of CAS coverage.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 48 Federal Acquisition Regulations System 7 2014-10-01 2014-10-01 false Types of CAS coverage... ACCOUNTING STANDARDS CONTRACT COVERAGE CAS Program Requirements 9903.201-2 Types of CAS coverage. (a) Full coverage. Full coverage requires that the business unit comply with all of the CAS specified in part...

  17. 48 CFR 9903.201-2 - Types of CAS coverage.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 48 Federal Acquisition Regulations System 7 2010-10-01 2010-10-01 false Types of CAS coverage... ACCOUNTING STANDARDS CONTRACT COVERAGE CAS Program Requirements 9903.201-2 Types of CAS coverage. (a) Full coverage. Full coverage requires that the business unit comply with all of the CAS specified in part...

  18. 48 CFR 9903.201-2 - Types of CAS coverage.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 48 Federal Acquisition Regulations System 7 2013-10-01 2012-10-01 true Types of CAS coverage. 9903... ACCOUNTING STANDARDS CONTRACT COVERAGE CAS Program Requirements 9903.201-2 Types of CAS coverage. (a) Full coverage. Full coverage requires that the business unit comply with all of the CAS specified in part...

  19. Assisting Students' Cognitive Strategies with the Use of CAS

    ERIC Educational Resources Information Center

    Sarvari, Csaba; Lavicza, Zsolt; Klincsik, Mihaly

    2010-01-01

    This paper examines various cognitive strategies applied while CAS (Computer Algebra System) are used in undergraduate-level engineering mathematics teaching and learning. We posed some questions in relation to such CAS use: What kind of tools can CAS offer to enhance different cognitive strategies of students? How can the use of CAS widen the…

  20. 48 CFR 9903.201-2 - Types of CAS coverage.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 48 Federal Acquisition Regulations System 7 2012-10-01 2012-10-01 false Types of CAS coverage... ACCOUNTING STANDARDS CONTRACT COVERAGE CAS Program Requirements 9903.201-2 Types of CAS coverage. (a) Full coverage. Full coverage requires that the business unit comply with all of the CAS specified in part...

  1. Incidence of Type II CRISPR1-Cas Systems in Enterococcus Is Species-Dependent

    PubMed Central

    Lyons, Casandra; Raustad, Nicole; Bustos, Mario A.; Shiaris, Michael

    2015-01-01

    CRISPR-Cas systems, which obstruct both viral infection and incorporation of mobile genetic elements by horizontal transfer, are a specific immune response common to prokaryotes. Antiviral protection by CRISPR-Cas comes at a cost, as horizontally-acquired genes may increase fitness and provide rapid adaptation to habitat change. To date, investigations into the prevalence of CRISPR have primarily focused on pathogenic and clinical bacteria, while less is known about CRISPR dynamics in commensal and environmental species. We designed PCR primers and coupled these with DNA sequencing of products to detect and characterize the presence of cas1, a universal CRISPR-associated gene and proxy for the Type II CRISPR1-Cas system, in environmental and non-clinical Enterococcus isolates. CRISPR1-cas1 was detected in approximately 33% of the 275 strains examined, and differences in CRISPR1 carriage between species was significant. Incidence of cas1 in E. hirae was 73%, nearly three times that of E. faecalis (23.6%) and 10 times more frequent than in E. durans (7.1%). Also, this is the first report of CRISPR1 presence in E. durans, as well as in the plant-associated species E. casseliflavus and E. sulfureus. Significant differences in CRISPR1-cas1 incidence among Enterococcus species support the hypothesis that there is a tradeoff between protection and adaptability. The differences in the habitats of enterococcal species may exert varying selective pressure that results in a species-dependent distribution of CRISPR-Cas systems. PMID:26600384

  2. CRISPR-Cas Adaptive Immune Systems of the Sulfolobales: Unravelling Their Complexity and Diversity

    PubMed Central

    Garrett, Roger A.; Shah, Shiraz A.; Erdmann, Susanne; Liu, Guannan; Mousaei, Marzieh; León-Sobrino, Carlos; Peng, Wenfang; Gudbergsdottir, Soley; Deng, Ling; Vestergaard, Gisle; Peng, Xu; She, Qunxin

    2015-01-01

    The Sulfolobales have provided good model organisms for studying CRISPR-Cas systems of the crenarchaeal kingdom of the archaea. These organisms are infected by a wide range of exceptional archaea-specific viruses and conjugative plasmids, and their CRISPR-Cas systems generally exhibit extensive structural and functional diversity. They carry large and multiple CRISPR loci and often multiple copies of diverse Type I and Type III interference modules as well as more homogeneous adaptation modules. These acidothermophilic organisms have recently provided seminal insights into both the adaptation process, the diverse modes of interference, and their modes of regulation. The functions of the adaptation and interference modules tend to be loosely coupled and the stringency of the crRNA-DNA sequence matching during DNA interference is relatively low, in contrast to some more streamlined CRISPR-Cas systems of bacteria. Despite this, there is evidence for a complex and differential regulation of expression of the diverse functional modules in response to viral infection. Recent work also supports critical roles for non-core Cas proteins, especially during Type III-directed interference, and this is consistent with these proteins tending to coevolve with core Cas proteins. Various novel aspects of CRISPR-Cas systems of the Sulfolobales are considered including an alternative spacer acquisition mechanism, reversible spacer acquisition, the formation and significance of antisense CRISPR RNAs, and a novel mechanism for avoidance of CRISPR-Cas defense. Finally, questions regarding the basis for the complexity, diversity, and apparent redundancy, of the intracellular CRISPR-Cas systems are discussed. PMID:25764276

  3. Computational Assessment of the GT-MHR Graphite Core Support Structural Integrity in Air-Ingress Accident Condition

    SciTech Connect

    Jong B. Lim; Eung S. Kim; Chang H. Oh; Richard R. Schultz; David A. Petti

    2008-10-01

    The objective of this project was to perform stress analysis for graphite support structures of the General Atomics’ 600 MWth GT-MHR prismatic core design using ABAQUS ® (ver. 6.75) to assess their structural integrity in air-ingress accident conditions where the structure weakens over time due to oxidation damages. The graphite support structures of prismatic type GT-MHR was analyzed based on the change of temperature, burn-off and corrosion depth during the accident period predicted by GAMMA, a multi-dimensional gas multi-component mixture analysis code developed in the Republic of Korea (ROK)/United States (US) International –Nuclear Engineering Research Initiative (I-NERI) project. Both the loading and thermal stresses were analyzed, but the thermal stress was not significant, leaving the loading stress to be the major factor. The mechanical strengths are exceeded between 11 to 11.5 days after loss-of-coolant-accident (LOCA), corresponding to 5.5 to 6 days after the start of natural convection.

  4. A newly discovered Bordetella species carries a transcriptionally active CRISPR-Cas with a small Cas9 endonuclease

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Cas9 endonuclease of the Type II-a clustered regularly interspersed short palindromic repeats (CRISPR), of Streptococcus pyogenes (SpCas9) has been adapted as a widely used tool for genome editing and genome engineering. Herein, we describe a gene encoding a novel Cas9 ortholog (BpsuCas9) and th...

  5. Discharge/charge characteristic of Li-air cells using carbon-supported LaMn0.6Fe0.4O3 as an electrocatalyst

    NASA Astrophysics Data System (ADS)

    Yuasa, Masayoshi; Matsuyoshi, Tsubasa; Kida, Tetsuya; Shimanoe, Kengo

    2013-11-01

    The discharge/charge performance of Li-air cell using the carbon-supported LaMn0.6Fe0.4O3 nanoparticle as a cathode catalyst was investigated in this study. The carbon-supported LaMn0.6Fe0.4O3 nanoparticle was prepared via a reverse homogeneous precipitation method, and fabricated to air electrode. Li-air cell was constructed using air electrode, Li metal foil and 1.0 M LiPF6 in propylene carbonate as a cathode, anode and electrolyte, respectively. As the result, the carbon-supported LaMn0.6Fe0.4O3 nanoparticle exhibited both the oxygen evolution activity and the oxygen reduction activity in the non-aqueous electrolyte. The investigation about the presence and absence of the catalytic layer and the gas diffusion layer revealed that each layer is indispensable for the excellent electrode performance, and that the catalytic layer and the gas diffusion layer has a important role to supply the electrolyte and the oxygen gas, respectively. The investigation about the amount of the catalytic layer and the effect of the oxygen concentration revealed that the oxygen diffusability into the air electrode strongly affects to the discharge capacity of Li-air cells.

  6. A Method for Estimating Urban Background Concentrations in Support of Hybrid Air Pollution Modeling for Environmental Health Studies

    PubMed Central

    Arunachalam, Saravanan; Valencia, Alejandro; Akita, Yasuyuki; Serre, Marc L.; Omary, Mohammad; Garcia, Valerie; Isakov, Vlad

    2014-01-01

    Exposure studies rely on detailed characterization of air quality, either from sparsely located routine ambient monitors or from central monitoring sites that may lack spatial representativeness. Alternatively, some studies use models of various complexities to characterize local-scale air quality, but often with poor representation of background concentrations. A hybrid approach that addresses this drawback combines a regional-scale model to provide background concentrations and a local-scale model to assess impacts of local sources. However, this approach may double-count sources in the study regions. To address these limitations, we carefully define the background concentration as the concentration that would be measured if local sources were not present, and to estimate these background concentrations we developed a novel technique that combines space-time ordinary kriging (STOK) of observations with outputs from a detailed chemistry-transport model with local sources zeroed out. We applied this technique to support an exposure study in Detroit, Michigan, for several pollutants (including NOx and PM2.5), and evaluated the estimated hybrid concentrations (calculated by combining the background estimates that addresses this issue of double counting with local-scale dispersion model estimates) using observations. Our results demonstrate the strength of this approach specifically by eliminating the problem of double-counting reported in previous hybrid modeling approaches leading to improved estimates of background concentrations, and further highlight the relative importance of NOx vs. PM2.5 in their relative contributions to total concentrations. While a key limitation of this approach is the requirement for another detailed model simulation to avoid double-counting, STOK improves the overall characterization of background concentrations at very fine spatial scales. PMID:25321872

  7. A method for estimating urban background concentrations in support of hybrid air pollution modeling for environmental health studies.

    PubMed

    Arunachalam, Saravanan; Valencia, Alejandro; Akita, Yasuyuki; Serre, Marc L; Omary, Mohammad; Garcia, Valerie; Isakov, Vlad

    2014-01-01

    Exposure studies rely on detailed characterization of air quality, either from sparsely located routine ambient monitors or from central monitoring sites that may lack spatial representativeness. Alternatively, some studies use models of various complexities to characterize local-scale air quality, but often with poor representation of background concentrations. A hybrid approach that addresses this drawback combines a regional-scale model to provide background concentrations and a local-scale model to assess impacts of local sources. However, this approach may double-count sources in the study regions. To address these limitations, we carefully define the background concentration as the concentration that would be measured if local sources were not present, and to estimate these background concentrations we developed a novel technique that combines space-time ordinary kriging (STOK) of observations with outputs from a detailed chemistry-transport model with local sources zeroed out. We applied this technique to support an exposure study in Detroit, Michigan, for several pollutants (including NOx and PM2.5), and evaluated the estimated hybrid concentrations (calculated by combining the background estimates that addresses this issue of double counting with local-scale dispersion model estimates) using observations. Our results demonstrate the strength of this approach specifically by eliminating the problem of double-counting reported in previous hybrid modeling approaches leading to improved estimates of background concentrations, and further highlight the relative importance of NOx vs. PM2.5 in their relative contributions to total concentrations. While a key limitation of this approach is the requirement for another detailed model simulation to avoid double-counting, STOK improves the overall characterization of background concentrations at very fine spatial scales. PMID:25321872

  8. V723 Cas a borderline classical nova

    NASA Astrophysics Data System (ADS)

    Friedjung, M.; Iijima, T.

    2002-11-01

    V723 Cas had a light curve similar to that of HR Del before maximum, with a very slow pre-maximum rise, explained according to [2] by the presence of an optically thin wind before maximum unlike the optically thick wind generally seen for classical novae after maximum. Examination of the Fe II emission lines by the SAC method, is compatible with this also having been the case for V723 Cas.

  9. Characterization of Cas9-Guide RNA Orthologs.

    PubMed

    Braff, Jonathan L; Yaung, Stephanie J; Esvelt, Kevin M; Church, George M

    2016-01-01

    In light of the multitude of new Cas9-mediated functionalities, the ability to carry out multiple Cas9-enabled processes simultaneously and in a single cell is becoming increasingly valuable. Accomplishing this aim requires a set of Cas9-guide RNA (gRNA) pairings that are functionally independent and insulated from one another. For instance, two such protein-gRNA complexes would allow for concurrent activation and editing at independent target sites in the same cell. The problem of establishing orthogonal CRISPR systems can be decomposed into three stages. First, putatively orthogonal systems must be identified with an emphasis on minimizing sequence similarity of the Cas9 protein and its associated RNAs. Second, the systems must be characterized well enough to effectively express and target the systems using gRNAs. Third, the systems should be established as orthogonal to one another by testing for activity and cross talk. Here, we describe the value of these orthogonal CRISPR systems, outline steps for selecting and characterizing potentially orthogonal Cas9-gRNA pairs, and discuss considerations for the desired specificity in Cas9-coupled functions. PMID:27140923

  10. Linking Meteorology, Air Quality Models and Observations to Characterize Human Exposures in Support of the Environmental Health Studies

    EPA Science Inventory

    Epidemiologic studies are critical in establishing the association between exposure to air pollutants and adverse health effects. Results of epidemiologic studies are used by U.S. EPA in developing air quality standards to protect the public from the health effects of air polluta...

  11. A Review and Analysis of Remote Sensing Capability for Air Quality Measurements as a Potential Decision Support Tool Conducted by the NASA DEVELOP Program

    NASA Technical Reports Server (NTRS)

    Ross, A.; Richards, A.; Keith, K.; Frew, C.; Boseck, J.; Sutton, S.; Watts, C.; Rickman, D.

    2007-01-01

    This project focused on a comprehensive utilization of air quality model products as decision support tools (DST) needed for public health applications. A review of past and future air quality measurement methods and their uncertainty, along with the relationship of air quality to national and global public health, is vital. This project described current and future NASA satellite remote sensing and ground sensing capabilities and the potential for using these sensors to enhance the prediction, prevention, and control of public health effects that result from poor air quality. The qualitative uncertainty of current satellite remotely sensed air quality, the ground-based remotely sensed air quality, the air quality/public health model, and the decision making process is evaluated in this study. Current peer-reviewed literature suggests that remotely sensed air quality parameters correlate well with ground-based sensor data. A satellite remote-sensed and ground-sensed data complement is needed to enhance the models/tools used by policy makers for the protection of national and global public health communities

  12. Evaluating the sustainability of space life support systems: case study on air revitalisation systems ARES and BIORAT

    NASA Astrophysics Data System (ADS)

    Suomalainen, Emilia; Erkman, Suren

    Space life support systems can be taken as kinds of miniature models of industrial systems found on Earth. The term "industrial" is employed here in a generic sense, referring to all human technological activities. The time scale as well as the physical scope of space life support systems is reduced compared to most terrestrial systems and so is consequently their complexity. These systems can thus be used as a kind of a "laboratory of sustainability" to examine concerns related to the environmental sustainability of industrial systems and in particular to their resource use. Two air revitalisation systems, ARES and BIORAT, were chosen as the test cases of our study. They represent respectively a physico-chemical and a biological life support system. In order to analyse the sustainability of these systems, we began by constructing a generic system representation applicable to both these systems (and to others). The metabolism of the systems was analysed by performing Material Flow Analyses—MFA is a tool frequently employed on terrestrial systems in the field of industrial ecology. Afterwards, static simulation models were developed for both ARES and BIORAT, focusing, firstly, on the oxygen balances of the systems and, secondly, on the total mass balances. It was also necessary to define sustainability indicators adapted to space life support systems in order to evaluate and to compare the performances of ARES and BIORAT. The defined indicators were partly inspired from concepts used in Material Flow Accounting and they were divided into four broad categories: 1. recycling and material use efficiency, 2. autarky and coverage time, 3. resource use and waste creation, and 4. system mass and energy consumption. The preliminary results of our analyses show that the performance of BIORAT is superior compared to ARES in terms of the defined resource use indicators. BIORAT seems especially effective in reprocessing carbon dioxide created by human metabolism. The

  13. Novel strategy to mitigate cathode catalyst degradation during air/air startup cycling via the atmospheric resistive switching mechanism of a hydrogen anode with a platinum catalyst supported on tantalum-doped titanium dioxide

    NASA Astrophysics Data System (ADS)

    Shintani, Haruhiko; Kojima, Yuya; Kakinuma, Katsuyoshi; Watanabe, Masahiro; Uchida, Makoto

    2015-10-01

    We propose a new strategy for alleviating the reverse current phenomenon using a unique "atmospheric resistive switching mechanism" (ARSM) of a metal oxide semiconductor support, such that the electrical resistivity changes depending on the gas atmosphere. The membrane-electrode assembly (MEA) using Ta-doped TiO2-supported platinum (Pt/Ta-TiO2) as the anode catalyst showed approximately one order of magnitude greater resistance in air than in hydrogen. The overpotential of the hydrogen oxidation reaction was negligible up to at least 1.5 A cm-2. The losses of electrochemically active surface area and carbon corrosion of the cathode catalyst during air/air startup cycling were significantly suppressed by the use of the Pt/Ta-TiO2 anode. The decrease in the degradation is attributed to a reduction of the reverse current due to a low oxygen reduction reaction rate at the anode, which showed high resistivity in air. These results demonstrate the effectiveness of the ARSM in mitigating cathode catalyst degradation during air/air startup cycling.

  14. Child maltreatment among U.S. Air Force parents deployed in support of Operation Iraqi Freedom/Operation Enduring Freedom.

    PubMed

    Rabenhorst, Mandy M; McCarthy, Randy J; Thomsen, Cynthia J; Milner, Joel S; Travis, Wendy J; Colasanti, Marie P

    2015-02-01

    This study examined child maltreatment perpetration among 99,697 active-duty U.S. Air Force parents who completed a combat deployment. Using the deploying parent as the unit of analysis, we analyzed whether child maltreatment rates increased postdeployement relative to predeployment. These analyses extend previous research that used aggregate data and extend our previous work that used data from the same period but used the victim as the unit of analysis and included only deploying parents who engaged in child maltreatment. In this study, 2% (n = 1,746) of deploying parents perpetrated child maltreatment during the study period. Although no overall differences were found in child maltreatment rates postdeployment compared to predeployment, several maltreatment-related characteristics qualified this finding. Rates for emotional abuse and mild maltreatment were lower following deployment, whereas child maltreatment rates for severe maltreatment were higher following deployment. The finding that rates of severe child maltreatment, including incidents involving alcohol use, were higher postdeployment suggests a need for additional support services for parents following their return from combat deployment, with a focus on returning parents who have an alcohol use problem. PMID:25424846

  15. Al 2O 3 supported Ru catalysts prepared by thermolysis of Ru 3(CO) 12 for catalytic wet air oxidation

    NASA Astrophysics Data System (ADS)

    Yu, Chaoying; Zhao, Peiqing; Chen, Gexin; Hu, Bin

    2011-06-01

    Low loading catalysts Ru/γ-Al 2O 3 and Ru-Ce/γ-Al 2O 3 were prepared by thermolysis of Ru 3(CO) 12 on γ-Al 2O 3. The catalysts were characterized by XPS, XRD and SEM. Two new Ru species (Ru A and Ru B) were detected during the Ru 3(CO) 12 decomposition process due to chemical interaction with the active OH groups on the surface of Al 2O 3 support, and the reduction of them can lead to more dispersed metallic phases. The sample was completely decomposed at 673 K in H 2, and RuO 2 was formed with minor amounts of Ru 0. When the temperature was increased to 773 K to heat the sample, the ratio of Ru 0 to RuO 2 increased. However, after the addition of CeO 2, only RuO 2 was detected on surface. The catalysts exhibited high activities in Catalytic Wet Air Oxidation (CWAO) of different organic compounds at high concentration such as isopropyl alcohol, phenol, acetic acids and N,N-dimethylformamide, which is attributed to the better dispersion of Ru particles and the addition of CeO 2 further enhanced number of effectively active sites on the cluster-derived catalyst surface.

  16. MWCNT-supported phthalocyanine cobalt as air-breathing cathodic catalyst in glucose/O2 fuel cells

    NASA Astrophysics Data System (ADS)

    Elouarzaki, Kamal; Haddad, Raoudha; Holzinger, Michael; Le Goff, Alan; Thery, Jessica; Cosnier, Serge

    2014-06-01

    Simple and highly efficient glucose fuel cells using abiotic catalysts and different ion exchange membranes were designed. The glucose fuel cells are based on a multi-walled carbon nanotube (MWCNT)-supported cobalt phthalocyanine (CoPc) cathode and a carbon black/platinum (C/Pt) anode. The electrocatalytic activity of the MWCNT/CoPc electrode for oxygen reduction was investigated by cyclic and linear sweep voltammetry. The electrochemical experiments show that CoPc exhibits promising catalytic properties for oxygen reduction due to its high overpotential and efficiency at reduced metal load. The MWCNT/CoPc electrodes were applied to the oxygen reduction reaction as air-breathing cathode in a single-chambered glucose fuel cell. This cathode was associated with a C/Pt anode in fuel cell configurations using either an anion (Nafion®) or a cation (Tokuyama) exchange membrane. The best fuel cell configuration delivered a maximum power density of 2.3 mW cm-2 and a cell voltage of 0.8 V in 0.5 M KOH solution containing 0.5 M glucose using the Tokuyama membrane at ambient conditions. Beside the highest power density per cathodic catalyst mass (383 W g-1), these glucose fuel cells exhibit a high operational stability, delivering 0.3 mW cm-2 after 50 days.

  17. Putting the CAS Standards to Work. Training Manual for the CAS Self Assessment Guides.

    ERIC Educational Resources Information Center

    Yerian, Jean M.; Miller, Theodore K., Ed.

    These 18 self-assessment guides and training manual from the Council for the Advancement of Standards (CAS) for Student Services/Development Programs translate the CAS Standards and Guidelines of 1986 into a format for self-study purposes. These self-study guides allow an institution to assure compliance with minimally-acceptable practice, gain an…

  18. Condition Assessment Survey (CAS) Program. Deficiency standards and inspections methods manual: Volume 12, 0.12 Sitework

    SciTech Connect

    Not Available

    1993-05-01

    General information is presented for asset determinant factor/CAS repair codes/CAS cost factors; guide sheet tool & material listing; testing methods; inspection frequency; standard system design life tables; system work breakdown structure; and general system/material data. Deficiency standards and inspection methods are given for utility distribution systems, central heating, central cooling, electrical, utility support structures, paving roadways/walkways, and tunnels.

  19. United States Air Force 611th Air Support Group Civil Engineering Squadron, Elmendorf AFB, Alaska. Remedial investigation and feasibility study Point Lay Radar Installation, Alaska. Final report

    SciTech Connect

    Karmi, S.

    1996-03-04

    The United States Air Force (Air Force) has prepared this Remedial Investigation/Feasibility Study (RI/FS) report to present the results of RI/FS activities at four sites located at the Point Lay radar installation. The remedial investigation (RI) field activities were conducted at the Point Lay radar installation during the summer of 1993. The four sites at Point Lay were investigated because they were suspected of being contaminated with hazardous substances. RI activities were conducted using methods and procedures specified in the RI/FS Work Plan, Sampling and Analysis Plan (SAP), and Health and Safety Plan.

  20. Air gap winding method and support structure for a super conducting generator and method for forming the same

    DOEpatents

    Hopeck, James Frederick

    2003-11-25

    A method of forming a winding support structure for use with a superconducting rotor wherein the method comprises providing an inner support ring, arranging an outer support ring around the inner support ring, coupling first and second support blocks to the outer support ring and coupling a lamination to the first and second support blocks. A slot is defined between the support blocks and between the outer support ring and the lamination to receive a portion of a winding. An RTV fills any clearance space in the slot.

  1. The Future of Online Services to Faculty: A Pilot Project with CAS Online.

    ERIC Educational Resources Information Center

    Culotta, Wendy A.

    Lottery money in California, part of which is required by law to be distributed to educational institutions, provides avenues for innovative services. A pilot program was supported by the administration of California State University, Long Beach, to provide faculty access to CAS (Chemical Abstracts Service) ONLINE (a remote database), which could…

  2. Rapid characterization of CRISPR-Cas9 protospacer adjacent motif sequence elements.

    PubMed

    Karvelis, Tautvydas; Gasiunas, Giedrius; Young, Joshua; Bigelyte, Greta; Silanskas, Arunas; Cigan, Mark; Siksnys, Virginijus

    2015-01-01

    To expand the repertoire of Cas9s available for genome targeting, we present a new in vitro method for the simultaneous examination of guide RNA and protospacer adjacent motif (PAM) requirements. The method relies on the in vitro cleavage of plasmid libraries containing a randomized PAM as a function of Cas9-guide RNA complex concentration. Using this method, we accurately reproduce the canonical PAM preferences for Streptococcus pyogenes, Streptococcus thermophilus CRISPR3 (Sth3), and CRISPR1 (Sth1). Additionally, PAM and sgRNA solutions for a novel Cas9 protein from Brevibacillus laterosporus are provided by the assay and are demonstrated to support functional activity in vitro and in plants. PMID:26585795

  3. Massive Systemic Air Embolism during Extracorporeal Membrane Oxygenation Support of a Neonate with Acute Respiratory Distress Syndrome after Cardiac Surgery

    PubMed Central

    Timpa, Joseph G.; O’Meara, Carlisle; McILwain, R. Britt; Dabal, Robert J.; Alten, Jeffrey A.

    2011-01-01

    Abstract: Extracorporeal membrane oxygenation (ECMO) is universally accepted as a potential lifesaving therapy for neonates suffering severe cardiorespiratory failure, with survival reported as 81% weaning off ECMO and 69% to hospital discharge in this population. Although ECMO may reduce mortality in certain neonatal patients, it is associated with significant complications. Air in the circuit complicates 4.9% of neonatal ECMO runs, and it is crucial that all ECMO caregivers are trained in the prevention of air embolism and possess the knowledge necessary to efficiently identify and remove air from the ECMO circuit to prevent life threatening consequences. We present a fatal case of neonatal systemic air embolism leading to massive entrainment of air into the ECMO venous return cannula of a neonatal patient with acute respiratory distress syndrome following repair of obstructed total anomalous pulmonary venous connection. We describe the pathophysiology and presentation of this rare condition and the importance of early recognition, due to its high mortality rate. PMID:21848179

  4. 48 CFR 970.3002 - CAS program requirements.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 48 Federal Acquisition Regulations System 5 2012-10-01 2012-10-01 false CAS program requirements. 970.3002 Section 970.3002 Federal Acquisition Regulations System DEPARTMENT OF ENERGY AGENCY....3002 CAS program requirements....

  5. 48 CFR 970.3002 - CAS program requirements.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 48 Federal Acquisition Regulations System 5 2013-10-01 2013-10-01 false CAS program requirements. 970.3002 Section 970.3002 Federal Acquisition Regulations System DEPARTMENT OF ENERGY AGENCY....3002 CAS program requirements....

  6. 48 CFR 970.3002 - CAS program requirements.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 48 Federal Acquisition Regulations System 5 2011-10-01 2011-10-01 false CAS program requirements. 970.3002 Section 970.3002 Federal Acquisition Regulations System DEPARTMENT OF ENERGY AGENCY....3002 CAS program requirements....

  7. 48 CFR 970.3002 - CAS program requirements.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 48 Federal Acquisition Regulations System 5 2014-10-01 2014-10-01 false CAS program requirements. 970.3002 Section 970.3002 Federal Acquisition Regulations System DEPARTMENT OF ENERGY AGENCY....3002 CAS program requirements....

  8. 48 CFR 970.3002 - CAS program requirements.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 48 Federal Acquisition Regulations System 5 2010-10-01 2010-10-01 false CAS program requirements. 970.3002 Section 970.3002 Federal Acquisition Regulations System DEPARTMENT OF ENERGY AGENCY....3002 CAS program requirements....

  9. CRISPR-Cas: Revolutionising genome engineering.

    PubMed

    Nicholson, Samantha Anne; Pepper, Michael Sean

    2016-09-01

    The ability to permanently alter or repair the human genome has been the subject of a number of science fiction films, but with the recent advent of several customisable sequence-specific endonuclease technologies, genome engineering looks set to become a clinical reality in the near future. This article discusses recent advancements in the technology called 'clustered regularly interspaced palindromic repeat (CRISPR)-associated genes' (CRISPR-Cas), the potential of CRISPR-Cas to revolutionise molecular medicine, and the ethical and regulatory hurdles facing its application. PMID:27601107

  10. Engineered CRISPR-Cas9 nucleases with altered PAM specificities

    PubMed Central

    Kleinstiver, Benjamin P.; Prew, Michelle S.; Tsai, Shengdar Q.; Topkar, Ved; Nguyen, Nhu T.; Zheng, Zongli; Gonzales, Andrew P.W.; Li, Zhuyun; Peterson, Randall T.; Yeh, Jing-Ruey Joanna; Aryee, Martin J.; Joung, J. Keith

    2015-01-01

    Although CRISPR-Cas9 nucleases are widely used for genome editing1, 2, the range of sequences that Cas9 can recognize is constrained by the need for a specific protospacer adjacent motif (PAM)3–6. As a result, it can often be difficult to target double-stranded breaks (DSBs) with the precision that is necessary for various genome editing applications. The ability to engineer Cas9 derivatives with purposefully altered PAM specificities would address this limitation. Here we show that the commonly used Streptococcus pyogenes Cas9 (SpCas9) can be modified to recognize alternative PAM sequences using structural information, bacterial selection-based directed evolution, and combinatorial design. These altered PAM specificity variants enable robust editing of endogenous gene sites in zebrafish and human cells not currently targetable by wild-type SpCas9, and their genome-wide specificities are comparable to wild-type SpCas9 as judged by GUIDE-Seq analysis7. In addition, we identified and characterized another SpCas9 variant that exhibits improved specificity in human cells, possessing better discrimination against off-target sites with non-canonical NAG and NGA PAMs and/or mismatched spacers. We also found that two smaller-size Cas9 orthologues, Streptococcus thermophilus Cas9 (St1Cas9) and Staphylococcus aureus Cas9 (SaCas9), function efficiently in the bacterial selection systems and in human cells, suggesting that our engineering strategies could be extended to Cas9s from other species. Our findings provide broadly useful SpCas9 variants and, more importantly, establish the feasibility of engineering a wide range of Cas9s with altered and improved PAM specificities. PMID:26098369

  11. Engineered CRISPR-Cas9 nucleases with altered PAM specificities.

    PubMed

    Kleinstiver, Benjamin P; Prew, Michelle S; Tsai, Shengdar Q; Topkar, Ved V; Nguyen, Nhu T; Zheng, Zongli; Gonzales, Andrew P W; Li, Zhuyun; Peterson, Randall T; Yeh, Jing-Ruey Joanna; Aryee, Martin J; Joung, J Keith

    2015-07-23

    Although CRISPR-Cas9 nucleases are widely used for genome editing, the range of sequences that Cas9 can recognize is constrained by the need for a specific protospacer adjacent motif (PAM). As a result, it can often be difficult to target double-stranded breaks (DSBs) with the precision that is necessary for various genome-editing applications. The ability to engineer Cas9 derivatives with purposefully altered PAM specificities would address this limitation. Here we show that the commonly used Streptococcus pyogenes Cas9 (SpCas9) can be modified to recognize alternative PAM sequences using structural information, bacterial selection-based directed evolution, and combinatorial design. These altered PAM specificity variants enable robust editing of endogenous gene sites in zebrafish and human cells not currently targetable by wild-type SpCas9, and their genome-wide specificities are comparable to wild-type SpCas9 as judged by GUIDE-seq analysis. In addition, we identify and characterize another SpCas9 variant that exhibits improved specificity in human cells, possessing better discrimination against off-target sites with non-canonical NAG and NGA PAMs and/or mismatched spacers. We also find that two smaller-size Cas9 orthologues, Streptococcus thermophilus Cas9 (St1Cas9) and Staphylococcus aureus Cas9 (SaCas9), function efficiently in the bacterial selection systems and in human cells, suggesting that our engineering strategies could be extended to Cas9s from other species. Our findings provide broadly useful SpCas9 variants and, more importantly, establish the feasibility of engineering a wide range of Cas9s with altered and improved PAM specificities. PMID:26098369

  12. 48 CFR 30.201-1 - CAS applicability.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... CONTRACTING REQUIREMENTS COST ACCOUNTING STANDARDS ADMINISTRATION CAS Program Requirements 30.201-1 CAS applicability. See 48 CFR 9903.201-1 (FAR appendix). ... 48 Federal Acquisition Regulations System 1 2012-10-01 2012-10-01 false CAS applicability....

  13. 48 CFR 30.201-2 - Types of CAS coverage.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... CONTRACTING REQUIREMENTS COST ACCOUNTING STANDARDS ADMINISTRATION CAS Program Requirements 30.201-2 Types of CAS coverage. See 48 CFR 9903.201-2 (FAR appendix). ... 48 Federal Acquisition Regulations System 1 2014-10-01 2014-10-01 false Types of CAS coverage....

  14. 48 CFR 30.201-1 - CAS applicability.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... CONTRACTING REQUIREMENTS COST ACCOUNTING STANDARDS ADMINISTRATION CAS Program Requirements 30.201-1 CAS applicability. See 48 CFR 9903.201-1 (FAR appendix). ... 48 Federal Acquisition Regulations System 1 2013-10-01 2013-10-01 false CAS applicability....

  15. 48 CFR 30.201-2 - Types of CAS coverage.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... CONTRACTING REQUIREMENTS COST ACCOUNTING STANDARDS ADMINISTRATION CAS Program Requirements 30.201-2 Types of CAS coverage. See 48 CFR 9903.201-2 (FAR appendix). ... 48 Federal Acquisition Regulations System 1 2012-10-01 2012-10-01 false Types of CAS coverage....

  16. 48 CFR 9903.201-1 - CAS applicability.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 48 Federal Acquisition Regulations System 7 2012-10-01 2012-10-01 false CAS applicability. 9903... ACCOUNTING STANDARDS CONTRACT COVERAGE CAS Program Requirements 9903.201-1 CAS applicability. (a) This subsection describes the rules for determining whether a proposed contract or subcontract is exempt from...

  17. 48 CFR 30.201-2 - Types of CAS coverage.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... CONTRACTING REQUIREMENTS COST ACCOUNTING STANDARDS ADMINISTRATION CAS Program Requirements 30.201-2 Types of CAS coverage. See 48 CFR 9903.201-2 (FAR appendix). ... 48 Federal Acquisition Regulations System 1 2013-10-01 2013-10-01 false Types of CAS coverage....

  18. 48 CFR 9903.201-1 - CAS applicability.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 48 Federal Acquisition Regulations System 7 2010-10-01 2010-10-01 false CAS applicability. 9903... ACCOUNTING STANDARDS CONTRACT COVERAGE CAS Program Requirements 9903.201-1 CAS applicability. (a) This subsection describes the rules for determining whether a proposed contract or subcontract is exempt from...

  19. 48 CFR 30.201-2 - Types of CAS coverage.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... CONTRACTING REQUIREMENTS COST ACCOUNTING STANDARDS ADMINISTRATION CAS Program Requirements 30.201-2 Types of CAS coverage. See 48 CFR 9903.201-2 (FAR appendix). ... 48 Federal Acquisition Regulations System 1 2011-10-01 2011-10-01 false Types of CAS coverage....

  20. 48 CFR 30.201-1 - CAS applicability.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... CONTRACTING REQUIREMENTS COST ACCOUNTING STANDARDS ADMINISTRATION CAS Program Requirements 30.201-1 CAS applicability. See 48 CFR 9903.201-1 (FAR appendix). ... 48 Federal Acquisition Regulations System 1 2011-10-01 2011-10-01 false CAS applicability....

  1. 48 CFR 9903.201-1 - CAS applicability.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 48 Federal Acquisition Regulations System 7 2014-10-01 2014-10-01 false CAS applicability. 9903... ACCOUNTING STANDARDS CONTRACT COVERAGE CAS Program Requirements 9903.201-1 CAS applicability. (a) This subsection describes the rules for determining whether a proposed contract or subcontract is exempt from...

  2. 48 CFR 30.201-1 - CAS applicability.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... CONTRACTING REQUIREMENTS COST ACCOUNTING STANDARDS ADMINISTRATION CAS Program Requirements 30.201-1 CAS applicability. See 48 CFR 9903.201-1 (FAR appendix). ... 48 Federal Acquisition Regulations System 1 2014-10-01 2014-10-01 false CAS applicability....

  3. 48 CFR 9903.201-1 - CAS applicability.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 48 Federal Acquisition Regulations System 7 2013-10-01 2012-10-01 true CAS applicability. 9903.201... ACCOUNTING STANDARDS CONTRACT COVERAGE CAS Program Requirements 9903.201-1 CAS applicability. (a) This subsection describes the rules for determining whether a proposed contract or subcontract is exempt from...

  4. 48 CFR 30.201-2 - Types of CAS coverage.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... CONTRACTING REQUIREMENTS COST ACCOUNTING STANDARDS ADMINISTRATION CAS Program Requirements 30.201-2 Types of CAS coverage. See 48 CFR 9903.201-2 (FAR appendix). ... 48 Federal Acquisition Regulations System 1 2010-10-01 2010-10-01 false Types of CAS coverage....

  5. 48 CFR 30.201-1 - CAS applicability.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... CONTRACTING REQUIREMENTS COST ACCOUNTING STANDARDS ADMINISTRATION CAS Program Requirements 30.201-1 CAS applicability. See 48 CFR 9903.201-1 (FAR appendix). ... 48 Federal Acquisition Regulations System 1 2010-10-01 2010-10-01 false CAS applicability....

  6. DEVELOPMENTS AND APPLICATIONS OF CFD SIMULATIONS OF MICROMETEOROLOGY AND POLLUTION TRANSPORT IN SUPPORT OF AIR QUALITY MODELING

    EPA Science Inventory

    Development and application of computational fluid dynamics (CFD) simulations are being advanced through case studies for simulating air pollutant concentrations from sources within open fields and within complex urban building environments. CFD applications have been under deve...

  7. A Method for Estimating Urban Background Concentrations in Support of Hybrid Air Pollution Modeling for Environmental Health Studies

    EPA Science Inventory

    Exposure studies rely on detailed characterization of air quality, either from sparsely located routine ambient monitors or from central monitoring sites that may lack spatial representativeness. Alternatively, some studies use models of various complexities to characterize local...

  8. High-resolution air pollution modeling for urban environments in support of dense multi-platform networks

    NASA Astrophysics Data System (ADS)

    Berchet, Antoine; Zink, Katrin; Arfire, Adrian; Marjovi, Ali; Martinoli, Alcherio; Emmenegger, Lukas; Brunner, Dominik

    2015-04-01

    As the fraction of people living in urban areas is rapidly increasing worldwide, the impact of air quality on human health in cities is a growing concern not only in developing countries but also in Europe despite the achievements of European air quality legislation. One obstacle to the quantitative assessment of the connections between health and air quality is the very high temporal and spatial variability of air pollutant concentrations within cities. Yet, an important issue for obtaining accurate and spatially highly resolved air pollution data is the trade-off between the high costs of accurate air pollution sensors and the number of such devices required for succinctly monitoring a given geographical area. The OpenSense 2 project aims at establishing air quality data at very high temporal and spatial resolution in the cities of Lausanne and Zurich in Switzerland in order to provide reliable information for epidemiologic studies and for the design of air pollution controls and urban planning. Towards this goal, observations from both stationary reference monitoring stations and low-cost mobile sensors (including sensing platforms anchored on public transport vehicles) are combined with high-resolution air quality modeling throughout the two cities. As a first step, we simulate the 3-dimensional, high-resolution dispersion and distribution of key pollutants using the GRAMM/GRAL modeling system. The GRAMM meteorological meso-scale model calculates wind fields at 100 m resolution accounting for the complex topography and land use within and around the two cities. GRAMM outputs are then used to drive the building-resolving dispersion model GRAL at 5-10m resolution. Further key inputs for GRAL are high resolution emission inventories and the 3-D building structure which are available for both cities. Here, in order to evaluate the ability of the GRAMM/GRAL modeling system to reproduce air pollutant distributions within the two cities of Lausanne and Zurich, we

  9. Structural plasticity and in vivo activity of Cas1 from the type I-F CRISPR-Cas system.

    PubMed

    Wilkinson, Max E; Nakatani, Yoshio; Staals, Raymond H J; Kieper, Sebastian N; Opel-Reading, Helen K; McKenzie, Rebecca E; Fineran, Peter C; Krause, Kurt L

    2016-04-15

    CRISPR-Cas systems are adaptive immune systems in prokaryotes that provide protection against viruses and other foreign DNA. In the adaptation stage, foreign DNA is integrated into CRISPR (clustered regularly interspaced short palindromic repeat) arrays as new spacers. These spacers are used in the interference stage to guide effector CRISPR associated (Cas) protein(s) to target complementary foreign invading DNA. Cas1 is the integrase enzyme that is central to the catalysis of spacer integration. There are many diverse types of CRISPR-Cas systems, including type I-F systems, which are typified by a unique Cas1-Cas2-3 adaptation complex. In the present study we characterize the Cas1 protein of the potato phytopathogen Pectobacterium atrosepticum, an important model organism for understanding spacer acquisition in type I-F CRISPR-Cas systems. We demonstrate by mutagenesis that Cas1 is essential for adaptation in vivo and requires a conserved aspartic acid residue. By X-ray crystallography, we show that although P. atrosepticum Cas1 adopts a fold conserved among other Cas1 proteins, it possesses remarkable asymmetry as a result of structural plasticity. In particular, we resolve for the first time a flexible, asymmetric loop that may be unique to type I-F Cas1 proteins, and we discuss the implications of these structural features for DNA binding and enzymatic activity. PMID:26929403

  10. Historic light curve of V890 Cas

    NASA Astrophysics Data System (ADS)

    Nesci, R.

    2016-05-01

    The variability of V890 Cas is studied with 87 -band plates of the Asiago archive. The star shows variations of about 5 mag with an average magnitude =13 and a period of 486 days. An 5.0 color index is also derived near the maximum luminosity.

  11. Using the CAS Standards in Assessment Projects

    ERIC Educational Resources Information Center

    Dean, Laura A.

    2013-01-01

    This chapter provides an overview of the use of professional standards of practice in assessment and of the Council for the Advancement of Standards in Higher Education (CAS). It outlines a model for conducting program self-studies and discusses the importance of implementing change based on assessment results.

  12. Lessons Learned on Management of CAS Development.

    ERIC Educational Resources Information Center

    Boyadjieff, Kiril

    1995-01-01

    Computer-assisted studies (CAS) attract foreign language professionals' attention due to the reliability of personal computers, the decreasing cost of available technology, and the new generation of students for whom electronic media are a familiar habitat. This article focuses on a project of the Defense Language Institute that produced over…

  13. A novel air quality analysis and prediction system for São Paulo, Brazil to support decision-making

    NASA Astrophysics Data System (ADS)

    Hoshyaripour, Gholam Ali; Brasseur, Guy; Andrade, Maria Fatima; Gavidia-Calderón, Mario; Bouarar, Idir

    2016-04-01

    The extensive economic development and urbanization in southeastern Brazil (SEB) in recent decades have notably degraded the air quality with adverse impacts on human health. Since the Metropolitan Area of São Paulo (MASP) accommodates the majority of the economic growth in SEB, it overwhelmingly suffers from the air pollution. Consequently, there is a strong demand for developing ever-better assessment mechanisms to monitor the air quality and to assist the decision makers to mitigate the air pollution in MASP. Here we present the results of an air quality modeling system designed for SEB with focuses on MASP. The Weather Research and Forecast model with Chemistry (WRF-Chem) is used considering the anthropogenic, biomass-burning and biogenic emissions within a 1000×1500 km domain with resolution of 10 km. FINN and MEGAN are used for the biomass-burning and biogenic emissions, respectively. For the anthropogenic emissions we use a local bottom-up inventory for the transport sector and the HTAPv2 global inventory for all other sectors. The bottom-up inventory accounts for the traffic patterns, vehicle types and their emission factors in the area and thus could be used to evaluate the effect of changes in these parameters on air quality in MASP. The model outputs are compered to the satellite and ground-based observations for O3 and NOx. The results show that using the bottom-up or top-down inventories individually can result in a huge deviation between the predictions and observations. On the other hand, combining the inventories significantly enhances the forecast accuracy. It also provides a powerful tool to quantify the effects of traffic and vehicle emission policies on air quality in MASP.

  14. CasHRA (Cas9-facilitated Homologous Recombination Assembly) method of constructing megabase-sized DNA.

    PubMed

    Zhou, Jianting; Wu, Ronghai; Xue, Xiaoli; Qin, Zhongjun

    2016-08-19

    Current DNA assembly methods for preparing highly purified linear subassemblies require complex and time-consuming in vitro manipulations that hinder their ability to construct megabase-sized DNAs (e.g. synthetic genomes). We have developed a new method designated 'CasHRA (Cas9-facilitated Homologous Recombination Assembly)' that directly uses large circular DNAs in a one-step in vivo assembly process. The large circular DNAs are co-introduced into Saccharomyces cerevisiae by protoplast fusion, and they are cleaved by RNA-guided Cas9 nuclease to release the linear DNA segments for subsequent assembly by the endogenous homologous recombination system. The CasHRA method allows efficient assembly of multiple large DNA segments in vivo; thus, this approach should be useful in the last stage of genome construction. As a proof of concept, we combined CasHRA with an upstream assembly method (Gibson procedure of genome assembly) and successfully constructed a 1.03 Mb MGE-syn1.0 (Minimal Genome of Escherichia coli) that contained 449 essential genes and 267 important growth genes. We expect that CasHRA will be widely used in megabase-sized genome constructions. PMID:27220470

  15. United States Air Force 611th Air Support Group/Civil Engineering Squadron Elmendorf AFB, Alaska. Risk assessment Bullen Point Radar Installation, Alaska. Final report

    SciTech Connect

    Karmi, S.

    1996-03-18

    This document contains the baseline human health risk assessment and the ecological risk assessment (ERA) for the Bullen Point Distant Early Warning (DEW) Line radar installation. Five sites at the Bullen Point radar installation underwent remedial investigations (RIs) during the summer of 1993. The presence of chemical contamination in the soil, sediments, and surface water at the installation was evaluated and reported in the Bullen Point Remedial Investigation/Feasibility Study (RI/FS) (U.S. Air Force 1996). The analytical data reported in the RI/FS form the basis for the human health and ecological risk assessments. The primary chemicals of concern (COCs) at the five sites are diesel and gasoline from past spills and/or leaks.

  16. [Life support of the Mars exploration crew. Control of a zeolite system for carbon dioxide removal from space cabin air within a closed air regeneration cycle].

    PubMed

    Chekov, Iu F

    2009-01-01

    The author describes a zeolite system for carbon dioxide removal integrated into a closed air regeneration cycle aboard spacecraft. The continuous operation of a double-adsorbent regeneration system with pCO2-dependable productivity is maintained through programmable setting of adsorption (desorption) semicycle time. The optimal system regulation curve is presented within the space of statistical performance family obtained in quasi-steady operating modes with controlled parameters of the recurrent adsorption-desorption cycle. The automatically changing system productivity ensures continuous intake of concentrated CO2. Control of the adsorption-desorption process is based on calculation of the differential adsorption (desorption) heat from gradient of adsorbent and test inert substance temperatures. The adaptive algorithm of digital control is implemented through the standard spacecraft interface with the board computer system and programmable microprocessor-based controllers. PMID:19621802

  17. Programmable plasmid interference by the CRISPR-Cas system in Thermococcus kodakarensis

    PubMed Central

    Elmore, Joshua R.; Yokooji, Yuusuke; Sato, Takaaki; Olson, Sara; Glover, III, Claiborne V.C.; Graveley, Brenton R.; Atomi, Haruyuki; Terns, Rebecca M.; Terns, Michael P.

    2013-01-01

    CRISPR-Cas systems are RNA-guided immune systems that protect prokaryotes against viruses and other invaders. The CRISPR locus encodes crRNAs that recognize invading nucleic acid sequences and trigger silencing by the associated Cas proteins. There are multiple CRISPR-Cas systems with distinct compositions and mechanistic processes. Thermococcus kodakarensis (Tko) is a hyperthermophilic euryarchaeon that has both a Type I-A Csa and a Type I-B Cst CRISPR-Cas system. We have analyzed the expression and composition of crRNAs from the three CRISPRs in Tko by RNA deep sequencing and northern analysis. Our results indicate that crRNAs associated with these two CRISPR-Cas systems include an 8-nucleotide conserved sequence tag at the 5′ end. We challenged Tko with plasmid invaders containing sequences targeted by endogenous crRNAs and observed active CRISPR-Cas-mediated silencing. Plasmid silencing was dependent on complementarity with a crRNA as well as on a sequence element found immediately adjacent to the crRNA recognition site in the target termed the PAM (protospacer adjacent motif). Silencing occurred independently of the orientation of the target sequence in the plasmid, and appears to occur at the DNA level, presumably via DNA degradation. In addition, we have directed silencing of an invader plasmid by genetically engineering the chromosomal CRISPR locus to express customized crRNAs directed against the plasmid. Our results support CRISPR engineering as a feasible approach to develop prokaryotic strains that are resistant to infection for use in industry. PMID:23535213

  18. In vivo genome editing using Staphylococcus aureus Cas9

    PubMed Central

    Ran, F. Ann; Cong, Le; Yan, Winston X.; Scott, David A.; Gootenberg, Jonathan S.; Kriz, Andrea J.; Zetsche, Bernd; Shalem, Ophir; Wu, Xuebing; Makarova, Kira S.; Koonin, Eugene; Sharp, Phillip A.; Zhang, Feng

    2015-01-01

    The RNA-guided endonuclease Cas9 has emerged as a versatile genome-editing platform. However, the size of the commonly used Cas9 from Streptococcus pyogenes (SpCas9) limits its utility for basic research and therapeutic applications that employ the highly versatile adeno-associated virus (AAV) delivery vehicle. Here, we characterize six smaller Cas9 orthologs and show that Cas9 from Staphylococcus aureus (SaCas9) can edit the genome with efficiencies similar to those of SpCas9, while being >1kb shorter. We packaged SaCas9 and its sgRNA expression cassette into a single AAV vector and targeted the cholesterol regulatory gene Pcsk9 in the mouse liver. Within one week of injection, we observed >40% gene modification, accompanied by significant reductions in serum Pcsk9 and total cholesterol levels. We further demonstrate the power of using BLESS to assess the genome-wide targeting specificity of SaCas9 and SpCas9, and show that SaCas9 can mediate genome editing in vivo with high specificity. PMID:25830891

  19. Structure and Engineering of Francisella novicida Cas9

    PubMed Central

    Hirano, Hisato; Gootenberg, Jonathan S.; Horii, Takuro; Abudayyeh, Omar O.; Kimura, Mika; Hsu, Patrick D.; Nakane, Takanori; Ishitani, Ryuichiro; Hatada, Izuho; Zhang, Feng; Nishimasu, Hiroshi; Nureki, Osamu

    2016-01-01

    Summary The RNA-guided endonuclease Cas9 cleaves double-stranded DNA targets complementary to the guide RNA, and has been applied to programmable genome editing. Cas9-mediated cleavage requires a protospacer adjacent motif (PAM) juxtaposed with the DNA target sequence, thus constricting the range of targetable sites. Here, we report the 1.7 Å resolution crystal structures of Cas9 from Francisella novicida (FnCas9), one of the largest Cas9 orthologs, in complex with a guide RNA and its PAM-containing DNA targets. A structural comparison of FnCas9 with other Cas9 orthologs revealed striking conserved and divergent features among distantly related CRISPR-Cas9 systems. We found that FnCas9 recognizes the 5′-NGG-3′ PAM, and used the structural information to create a variant that can recognize the more relaxed 5′-YG-3′ PAM. Furthermore, we demonstrated that pre-assembled FnCas9 ribonucleoprotein complexes can be microinjected into mouse zygotes to edit endogenous sites with the 5′-YG-3′ PAMs, thus expanding the target space of the CRISPR-Cas9 toolbox. PMID:26875867

  20. Structure and Engineering of Francisella novicida Cas9.

    PubMed

    Hirano, Hisato; Gootenberg, Jonathan S; Horii, Takuro; Abudayyeh, Omar O; Kimura, Mika; Hsu, Patrick D; Nakane, Takanori; Ishitani, Ryuichiro; Hatada, Izuho; Zhang, Feng; Nishimasu, Hiroshi; Nureki, Osamu

    2016-02-25

    The RNA-guided endonuclease Cas9 cleaves double-stranded DNA targets complementary to the guide RNA and has been applied to programmable genome editing. Cas9-mediated cleavage requires a protospacer adjacent motif (PAM) juxtaposed with the DNA target sequence, thus constricting the range of targetable sites. Here, we report the 1.7 Å resolution crystal structures of Cas9 from Francisella novicida (FnCas9), one of the largest Cas9 orthologs, in complex with a guide RNA and its PAM-containing DNA targets. A structural comparison of FnCas9 with other Cas9 orthologs revealed striking conserved and divergent features among distantly related CRISPR-Cas9 systems. We found that FnCas9 recognizes the 5'-NGG-3' PAM, and used the structural information to create a variant that can recognize the more relaxed 5'-YG-3' PAM. Furthermore, we demonstrated that the FnCas9-ribonucleoprotein complex can be microinjected into mouse zygotes to edit endogenous sites with the 5'-YG-3' PAM, thus expanding the target space of the CRISPR-Cas9 toolbox. PMID:26875867

  1. Photoactivatable CRISPR-Cas9 for optogenetic genome editing.

    PubMed

    Nihongaki, Yuta; Kawano, Fuun; Nakajima, Takahiro; Sato, Moritoshi

    2015-07-01

    We describe an engineered photoactivatable Cas9 (paCas9) that enables optogenetic control of CRISPR-Cas9 genome editing in human cells. paCas9 consists of split Cas9 fragments and photoinducible dimerization domains named Magnets. In response to blue light irradiation, paCas9 expressed in human embryonic kidney 293T cells induces targeted genome sequence modifications through both nonhomologous end joining and homology-directed repair pathways. Genome editing activity can be switched off simply by extinguishing the light. We also demonstrate activation of paCas9 in spatial patterns determined by the sites of irradiation. Optogenetic control of targeted genome editing should facilitate improved understanding of complex gene networks and could prove useful in biomedical applications. PMID:26076431

  2. Information Requirements for Supervisory Air Traffic Controllers in Support of a Mid-Term Wake Vortex Departure System

    NASA Technical Reports Server (NTRS)

    Lohr, Gary W.; Williams, Daniel M.; Trujillo, Anna C.; Johnson, Edward J.; Domino, David A.

    2008-01-01

    A concept focusing on wind dependent departure operations has been developed the current version of this concept is called the Wake Turbulence Mitigation for Departures (WTMD). This concept takes advantage the fact that cross winds of sufficient velocity blow wakes generated by "heavy" and B757 category aircraft on the downwind runway away from the upwind runway. Supervisory Air Traffic Controllers would be responsible for authorization of the Procedure. An investigation of the information requirements necessary to for Supervisors to approve monitor and terminate the Procedure was conducted. Results clearly indicated that the requisite information is currently available in air traffic control towers and that additional information was not required.

  3. Unification of Cas protein families and a simple scenario for the origin and evolution of CRISPR-Cas systems

    PubMed Central

    2011-01-01

    Background The CRISPR-Cas adaptive immunity systems that are present in most Archaea and many Bacteria function by incorporating fragments of alien genomes into specific genomic loci, transcribing the inserts and using the transcripts as guide RNAs to destroy the genome of the cognate virus or plasmid. This RNA interference-like immune response is mediated by numerous, diverse and rapidly evolving Cas (CRISPR-associated) proteins, several of which form the Cascade complex involved in the processing of CRISPR transcripts and cleavage of the target DNA. Comparative analysis of the Cas protein sequences and structures led to the classification of the CRISPR-Cas systems into three Types (I, II and III). Results A detailed comparison of the available sequences and structures of Cas proteins revealed several unnoticed homologous relationships. The Repeat-Associated Mysterious Proteins (RAMPs) containing a distinct form of the RNA Recognition Motif (RRM) domain, which are major components of the CRISPR-Cas systems, were classified into three large groups, Cas5, Cas6 and Cas7. Each of these groups includes many previously uncharacterized proteins now shown to adopt the RAMP structure. Evidence is presented that large subunits contained in most of the CRISPR-Cas systems could be homologous to Cas10 proteins which contain a polymerase-like Palm domain and are predicted to be enzymatically active in Type III CRISPR-Cas systems but inactivated in Type I systems. These findings, the fact that the CRISPR polymerases, RAMPs and Cas2 all contain core RRM domains, and distinct gene arrangements in the three types of CRISPR-Cas systems together provide for a simple scenario for origin and evolution of the CRISPR-Cas machinery. Under this scenario, the CRISPR-Cas system originated in thermophilic Archaea and subsequently spread horizontally among prokaryotes. Conclusions Because of the extreme diversity of CRISPR-Cas systems, in-depth sequence and structure comparison continue to

  4. SITE CHARACTERIZATION OF AREA 6, DOVER AIR FORCE BASE, IN SUPPORT OF NATURAL ATTENUATION AND ENHANCED ANAEROBIC BIOREMEDIATION PROJECTS

    EPA Science Inventory

    A field program for site characterization of targeted study areas at the Dover Air Force Base was conducted between January 16, 1995, and March 9, 1995. The stated objectives of the investigation, "to characterize the stratigraphy, depth to groundwater, groundwater flow directio...

  5. Workshop in Support of the Secondary National Ambient Air Quality Standards for Nitrogen (NOx) and Sulfur Oxides (SOx)

    EPA Science Inventory

    EPA is announcing a workshop to discuss policy-relevant science to Inform EPA’s "Review of the Secondary National Ambient Air Quality Standards (NAAQS) for Oxides of Nitrogen and Sulfur" report. The workshop is being organized by EPA’s Office of Research and Development’s, Nation...

  6. “Estimating Regional Background Air Quality using Space/Time Ordinary Kriging to Support Exposure Studies”

    EPA Science Inventory

    Local-scale dispersion models are increasingly being used to perform exposure assessments. These types of models, while able to characterize local-scale air quality at increasing spatial scale, however, lack the ability to include background concentration in their overall estimat...

  7. Use of depuration compounds in passive air samplers: results from active sampling-supported field deployment, potential uses, and recommendations.

    PubMed

    Moeckel, Claudia; Harner, Tom; Nizzetto, Luca; Strandberg, Bo; Lindroth, Anders; Jones, Kevin C

    2009-05-01

    Depuration compounds (DCs) are added to passive air samplers (PAS) prior to deployment to account for the wind-dependency of the sampling rate for gas-phase compounds. This correction is particularly useful for providing comparable data for samplers that are deployed in different environments and subject to different meteorological conditions such as wind speeds. Two types of PAS--the polyurethane foam (PUF) disk sampler and semipermeable membrane devices (SPMDs)--were deployed at eight heights on a 100 m tower to test whether the DC approach could yield air concentrations profiles for PCBs and organochlorine pesticides and account for the wind speed gradient with height. Average wind speeds ranged from 0.3 to 4.5 m s(-1) over the 40 day deployment, increasing with height Two low volume active air samples (AAS), one collected at 25 m and one at 73 m over the 40 day deployment showed no significant concentration differences for target compounds. As expected, the target compounds taken up by PAS reflected the wind profile with height This wind-dependency of the PAS was also reflected in the results of the DCs. A correction based on the DC approach successfully accounted for the effect of wind on PAS sampling rates, yielding a profile consistent with the AAS. Interestingly, in terms of absolute air concentrations, there were differences between the AAS and PAS-derived values for some target compounds. These were attributed to different sampling characteristics of the two approaches that may have resulted in slightly different air masses being sampled. Based on the results of this study, guidelines are presented for the use of DCs and for the calibration of PAS using AAS. PMID:19534139

  8. Substrate generation for endonucleases of CRISPR/cas systems.

    PubMed

    Zoephel, Judith; Dwarakanath, Srivatsa; Richter, Hagen; Plagens, André; Randau, Lennart

    2012-01-01

    The interaction of viruses and their prokaryotic hosts shaped the evolution of bacterial and archaeal life. Prokaryotes developed several strategies to evade viral attacks that include restriction modification, abortive infection and CRISPR/Cas systems. These adaptive immune systems found in many Bacteria and most Archaea consist of clustered regularly interspaced short palindromic repeat (CRISPR) sequences and a number of CRISPR associated (Cas) genes (Fig. 1) (1-3). Different sets of Cas proteins and repeats define at least three major divergent types of CRISPR/Cas systems (4). The universal proteins Cas1 and Cas2 are proposed to be involved in the uptake of viral DNA that will generate a new spacer element between two repeats at the 5' terminus of an extending CRISPR cluster (5). The entire cluster is transcribed into a precursor-crRNA containing all spacer and repeat sequences and is subsequently processed by an enzyme of the diverse Cas6 family into smaller crRNAs (6-8). These crRNAs consist of the spacer sequence flanked by a 5' terminal (8 nucleotides) and a 3' terminal tag derived from the repeat sequence (9). A repeated infection of the virus can now be blocked as the new crRNA will be directed by a Cas protein complex (Cascade) to the viral DNA and identify it as such via base complementarity(10). Finally, for CRISPR/Cas type 1 systems, the nuclease Cas3 will destroy the detected invader DNA (11,12) . These processes define CRISPR/Cas as an adaptive immune system of prokaryotes and opened a fascinating research field for the study of the involved Cas proteins. The function of many Cas proteins is still elusive and the causes for the apparent diversity of the CRISPR/Cas systems remain to be illuminated. Potential activities of most Cas proteins were predicted via detailed computational analyses. A major fraction of Cas proteins are either shown or proposed to function as endonucleases (4). Here, we present methods to generate crRNAs and precursor-cRNAs for

  9. Protein engineering of Cas9 for enhanced function

    PubMed Central

    Oakes, Benjamin L.; Nadler, Dana C.; Savage, David F.

    2015-01-01

    CRISPR/Cas systems act to protect the cell from invading nucleic acids in many bacteria and archaea. The bacterial immune protein Cas9 is a component of one of these CRISPR/Cas systems and has recently been adapted as a tool for genome editing. Cas9 is easily targeted to bind and cleave a DNA sequence via a complimentary RNA; this straightforward programmability has gained Cas9 rapid acceptance in the field of genetic engineering. While this technology has developed quickly, a number of challenges regarding Cas9 specificity, efficiency, fusion protein function, and spatiotemporal control within the cell remain. In this work, we develop a platform for constructing novel proteins to address these open questions. We demonstrate methods to either screen or select active Cas9 mutants and use the screening technique to isolate functional Cas9 variants with a heterologous PDZ domain inserted directly into the protein. As a proof of concept, these methods lay the groundwork for the future construction of diverse Cas9 proteins. Straightforward and accessible techniques for genetic editing are helping to elucidate biology in new and exciting ways; a platform to engineer new functionalities into Cas9 will help forge the next generation of genome modifying tools. PMID:25398355

  10. CRISPR-Cas9-assisted recombineering in Lactobacillus reuteri.

    PubMed

    Oh, Jee-Hwan; van Pijkeren, Jan-Peter

    2014-01-01

    Clustered regularly interspaced palindromic repeats (CRISPRs) and the CRISPR-associated (Cas) nuclease protect bacteria and archeae from foreign DNA by site-specific cleavage of incoming DNA. Type-II CRISPR-Cas systems, such as the Streptococcus pyogenes CRISPR-Cas9 system, can be adapted such that Cas9 can be guided to a user-defined site in the chromosome to introduce double-stranded breaks. Here we have developed and optimized CRISPR-Cas9 function in the lactic acid bacterium Lactobacillus reuteri ATCC PTA 6475. We established proof-of-concept showing that CRISPR-Cas9 selection combined with single-stranded DNA (ssDNA) recombineering is a realistic approach to identify at high efficiencies edited cells in a lactic acid bacterium. We show for three independent targets that subtle changes in the bacterial genome can be recovered at efficiencies ranging from 90 to 100%. By combining CRISPR-Cas9 and recombineering, we successfully applied codon saturation mutagenesis in the L. reuteri chromosome. Also, CRISPR-Cas9 selection is critical to identify low-efficiency events such as oligonucleotide-mediated chromosome deletions. This also means that CRISPR-Cas9 selection will allow identification of recombinant cells in bacteria with low recombineering efficiencies, eliminating the need for ssDNA recombineering optimization procedures. We envision that CRISPR-Cas genome editing has the potential to change the landscape of genome editing in lactic acid bacteria, and other Gram-positive bacteria. PMID:25074379

  11. Costs of CRISPR-Cas-mediated resistance in Streptococcus thermophilus

    PubMed Central

    Vale, Pedro F.; Lafforgue, Guillaume; Gatchitch, Francois; Gardan, Rozenn; Moineau, Sylvain; Gandon, Sylvain

    2015-01-01

    CRISPR-Cas is a form of adaptive sequence-specific immunity in microbes. This system offers unique opportunities for the study of coevolution between bacteria and their viral pathogens, bacteriophages. A full understanding of the coevolutionary dynamics of CRISPR-Cas requires knowing the magnitude of the cost of resisting infection. Here, using the gram-positive bacterium Streptococcus thermophilus and its associated virulent phage 2972, a well-established model system harbouring at least two type II functional CRISPR-Cas systems, we obtained different fitness measures based on growth assays in isolation or in pairwise competition. We measured the fitness cost associated with different components of this adaptive immune system: the cost of Cas protein expression, the constitutive cost of increasing immune memory through additional spacers, and the conditional costs of immunity during phage exposure. We found that Cas protein expression is particularly costly, as Cas-deficient mutants achieved higher competitive abilities than the wild-type strain with functional Cas proteins. Increasing immune memory by acquiring up to four phage-derived spacers was not associated with fitness costs. In addition, the activation of the CRISPR-Cas system during phage exposure induces significant but small fitness costs. Together these results suggest that the costs of the CRISPR-Cas system arise mainly due to the maintenance of the defence system. We discuss the implications of these results for the evolution of CRISPR-Cas-mediated immunity. PMID:26224708

  12. Expanding the catalog of cas genes with metagenomes.

    PubMed

    Zhang, Quan; Doak, Thomas G; Ye, Yuzhen

    2014-02-01

    The CRISPR (clusters of regularly interspaced short palindromic repeats)-Cas adaptive immune system is an important defense system in bacteria, providing targeted defense against invasions of foreign nucleic acids. CRISPR-Cas systems consist of CRISPR loci and cas (CRISPR-associated) genes: sequence segments of invaders are incorporated into host genomes at CRISPR loci to generate specificity, while adjacent cas genes encode proteins that mediate the defense process. We pursued an integrated approach to identifying putative cas genes from genomes and metagenomes, combining similarity searches with genomic neighborhood analysis. Application of our approach to bacterial genomes and human microbiome datasets allowed us to significantly expand the collection of cas genes: the sequence space of the Cas9 family, the key player in the recently engineered RNA-guided platforms for genome editing in eukaryotes, is expanded by at least two-fold with metagenomic datasets. We found genes in cas loci encoding other functions, for example, toxins and antitoxins, confirming the recently discovered potential of coupling between adaptive immunity and the dormancy/suicide systems. We further identified 24 novel Cas families; one novel family contains 20 proteins, all identified from the human microbiome datasets, illustrating the importance of metagenomics projects in expanding the diversity of cas genes. PMID:24319142

  13. Precision Targeted Mutagenesis via Cas9 Paired Nickases in Rice

    PubMed Central

    Mikami, Masafumi; Toki, Seiichi; Endo, Masaki

    2016-01-01

    Recent reports of CRISPR- (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR-associated protein 9) mediated heritable mutagenesis in plants highlight the need for accuracy of the mutagenesis directed by this system. Off-target mutations are an important issue when considering functional gene analysis, as well as the molecular breeding of crop plants with large genome size, i.e. with many duplicated genes, and where the whole-genome sequence is still lacking. In mammals, off-target mutations can be suppressed by using Cas9 paired nickases together with paired guide RNAs (gRNAs). However, the performance of Cas9 paired nickases has not yet been fully assessed in plants. Here, we analyzed on- and off-target mutation frequency in rice calli and regenerated plants using Cas9 nuclease or Cas9 nickase with paired gRNAs. When Cas9 paired nickases were used, off-target mutations were fully suppressed in rice calli and regenerated plants. However, on-target mutation frequency also decreased compared with that induced by the Cas9 paired nucleases system. Since the gRNA sequence determines specific binding of Cas9 protein–gRNA ribonucleoproteins at the targeted sequence, the on-target mutation frequency of Cas9 paired nickases depends on the design of paired gRNAs. Our results suggest that a combination of gRNAs that can induce mutations at high efficiency with Cas9 nuclease should be used together with Cas9 nickase. Furthermore, we confirmed that a combination of gRNAs containing a one nucleotide (1 nt) mismatch toward the target sequence could not induce mutations when expressed with Cas9 nickase. Our results clearly show the effectiveness of Cas9 paired nickases in delivering on-target specific mutations. PMID:26936792

  14. Efficient Mitochondrial Genome Editing by CRISPR/Cas9

    PubMed Central

    Jo, Areum; Ham, Sangwoo; Lee, Gum Hwa; Lee, Yun-Il; Kim, SangSeong; Lee, Yun-Song; Shin, Joo-Ho; Lee, Yunjong

    2015-01-01

    The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 system has been widely used for nuclear DNA editing to generate mutations or correct specific disease alleles. Despite its flexible application, it has not been determined if CRISPR/Cas9, originally identified as a bacterial defense system against virus, can be targeted to mitochondria for mtDNA editing. Here, we show that regular FLAG-Cas9 can localize to mitochondria to edit mitochondrial DNA with sgRNAs targeting specific loci of the mitochondrial genome. Expression of FLAG-Cas9 together with gRNA targeting Cox1 and Cox3 leads to cleavage of the specific mtDNA loci. In addition, we observed disruption of mitochondrial protein homeostasis following mtDNA truncation or cleavage by CRISPR/Cas9. To overcome nonspecific distribution of FLAG-Cas9, we also created a mitochondria-targeted Cas9 (mitoCas9). This new version of Cas9 localizes only to mitochondria; together with expression of gRNA targeting mtDNA, there is specific cleavage of mtDNA. MitoCas9-induced reduction of mtDNA and its transcription leads to mitochondrial membrane potential disruption and cell growth inhibition. This mitoCas9 could be applied to edit mtDNA together with gRNA expression vectors without affecting genomic DNA. In this brief study, we demonstrate that mtDNA editing is possible using CRISPR/Cas9. Moreover, our development of mitoCas9 with specific localization to the mitochondria should facilitate its application for mitochondrial genome editing. PMID:26448933

  15. Breaking-Cas-interactive design of guide RNAs for CRISPR-Cas experiments for ENSEMBL genomes.

    PubMed

    Oliveros, Juan C; Franch, Mònica; Tabas-Madrid, Daniel; San-León, David; Montoliu, Lluis; Cubas, Pilar; Pazos, Florencio

    2016-07-01

    The CRISPR/Cas technology is enabling targeted genome editing in multiple organisms with unprecedented accuracy and specificity by using RNA-guided nucleases. A critical point when planning a CRISPR/Cas experiment is the design of the guide RNA (gRNA), which directs the nuclease and associated machinery to the desired genomic location. This gRNA has to fulfil the requirements of the nuclease and lack homology with other genome sites that could lead to off-target effects. Here we introduce the Breaking-Cas system for the design of gRNAs for CRISPR/Cas experiments, including those based in the Cas9 nuclease as well as others recently introduced. The server has unique features not available in other tools, including the possibility of using all eukaryotic genomes available in ENSEMBL (currently around 700), placing variable PAM sequences at 5' or 3' and setting the guide RNA length and the scores per nucleotides. It can be freely accessed at: http://bioinfogp.cnb.csic.es/tools/breakingcas, and the code is available upon request. PMID:27166368

  16. SU-E-T-552: Monte Carlo Calculation of Correction Factors for a Free-Air Ionization Chamber in Support of a National Air-Kerma Standard for Electronic Brachytherapy

    SciTech Connect

    Mille, M; Bergstrom, P

    2015-06-15

    Purpose: To use Monte Carlo radiation transport methods to calculate correction factors for a free-air ionization chamber in support of a national air-kerma standard for low-energy, miniature x-ray sources used for electronic brachytherapy (eBx). Methods: The NIST is establishing a calibration service for well-type ionization chambers used to characterize the strength of eBx sources prior to clinical use. The calibration approach involves establishing the well-chamber’s response to an eBx source whose air-kerma rate at a 50 cm distance is determined through a primary measurement performed using the Lamperti free-air ionization chamber. However, the free-air chamber measurements of charge or current can only be related to the reference air-kerma standard after applying several corrections, some of which are best determined via Monte Carlo simulation. To this end, a detailed geometric model of the Lamperti chamber was developed in the EGSnrc code based on the engineering drawings of the instrument. The egs-fac user code in EGSnrc was then used to calculate energy-dependent correction factors which account for missing or undesired ionization arising from effects such as: (1) attenuation and scatter of the x-rays in air; (2) primary electrons escaping the charge collection region; (3) lack of charged particle equilibrium; (4) atomic fluorescence and bremsstrahlung radiation. Results: Energy-dependent correction factors were calculated assuming a monoenergetic point source with the photon energy ranging from 2 keV to 60 keV in 2 keV increments. Sufficient photon histories were simulated so that the Monte Carlo statistical uncertainty of the correction factors was less than 0.01%. The correction factors for a specific eBx source will be determined by integrating these tabulated results over its measured x-ray spectrum. Conclusion: The correction factors calculated in this work are important for establishing a national standard for eBx which will help ensure that dose

  17. Spatio-temporal Spectral Variability in Cas A

    NASA Astrophysics Data System (ADS)

    Nambiar, Yamini; Kashyap, V.; Patnaude, D.

    2014-01-01

    We have analyzed Chandra archival data of Cas A Supernova Remnant to identify regions with large spectral abnormalities and variability over the last decade. We use 8 ACIS-S observations spanning the years 2000 to 2012. We compute spectral hardness ratios in the soft/medium and medium/hard CSC bands over spatial scales corresponding to binning by 4, 8, 16, 32, and 64. We reduce the data and apply the latest calibration using the CIAO tool chandra_repro. We account for exposure variations using exposure maps and compute photon fluxes using the CIAO tool fluximage. We then renormalize the color light curves at each pixel and flag large departures from the norm by comparing with the observed spread in the renormalized color light curves. This allows regions with different intrinsic spectral properties to be compared. We flag deviations of >3σ from the renormalized mean at each epoch, and combine all such pixels to form a map of interesting regions in the remnant. We also identify pixels which have intrinsically abnormal hardness ratios at each epoch. We show that there exist many sites on Cas A where abnormal variations in the spectrum exist. Specifically, we find that many of the identified regions coincide with prominent features of the SNR, such as the edge of the remnant, the central compact object, and numerous knots. In addition, we find various other locations 1000) where there is indication of an atypical spectral signature. The full region lists, along with analysis scripts and the figures and tables shown in this poster, are stored on the Harvard Dataverse Network, at http://dx.doi.org/10.7910/DVN1/22634 YN thanks ABRHS and Young Einsteins Science Club for support and guidance. VK and DP acknowledge support during this project from the Chandra X-Ray Center.

  18. United States Air Force 611th air support group 611th Civil Engineer Squadron, Elmendorf AFB, Alaska. Eareckson Air Station, Alaska Remedial Investigation/Feasibility Study Report, final. Volume 2. Report for July 1994-September 1996

    SciTech Connect

    1995-08-30

    The Volume II has been prepared to present the findings of the environmental investigation activities conducted at Eareckson Air Station. The RI/FS report has been divided into four primary volumes. This volume, Volume II, discusses 13 source areas that are recommended to be proposed for no further action.

  19. Topography Analysis and Visualization Software Supports a Guided Comparative Planetology Education Exhibit at the Smithsonian's Air and Space Museum

    NASA Technical Reports Server (NTRS)

    Roark, J. H.; Masuoka, C. M.; Frey, H. V.; Keller, J.; Williams, S.

    2005-01-01

    The Planetary Geodynamics Laboratory (http://geodynamics.gsfc.nasa.gov) of NASA s Goddard Space Flight Center designed, produced and recently delivered a "museum-friendly" version of GRIDVIEW, a grid visualization and analysis application, to the Smithsonian's National Air and Space Museum where it will be used in a guided comparative planetology education exhibit. The software was designed to enable museum visitors to interact with the same Earth and Mars topographic data and tools typically used by planetary scientists, and experience the thrill of discovery while learning about the geologic differences between Earth and Mars.

  20. Advances in therapeutic CRISPR/Cas9 genome editing.

    PubMed

    Savić, Nataša; Schwank, Gerald

    2016-02-01

    Targeted nucleases are widely used as tools for genome editing. Two years ago the clustered regularly interspaced short palindromic repeat (CRISPR)-associated Cas9 nuclease was used for the first time, and since then has largely revolutionized the field. The tremendous success of the CRISPR/Cas9 genome editing tool is powered by the ease design principle of the guide RNA that targets Cas9 to the desired DNA locus, and by the high specificity and efficiency of CRISPR/Cas9-generated DNA breaks. Several studies recently used CRISPR/Cas9 to successfully modulate disease-causing alleles in vivo in animal models and ex vivo in somatic and induced pluripotent stem cells, raising hope for therapeutic genome editing in the clinics. In this review, we will summarize and discuss such preclinical CRISPR/Cas9 gene therapy reports. PMID:26470680

  1. Inducible in vivo genome editing with CRISPR/Cas9

    PubMed Central

    O'Rourke, Kevin P; Muley, Ashlesha; Kastenhuber, Edward R; Livshits, Geulah; Tschaharganeh, Darjus F; Socci, Nicholas D; Lowe, Scott W

    2015-01-01

    CRISPR/Cas9-based genome editing enables the rapid genetic manipulation of any genomic locus without the need for gene targeting by homologous recombination. Here we describe a conditional transgenic approach that allows temporal control of CRISPR/Cas9 activity for inducible genome editing in adult mice. We show that doxycycline-regulated Cas9 induction enables widespread gene disruption in multiple tissues and that limiting the duration of Cas9 expression or using a Cas9D10A (Cas9n) variant, can regulate the frequency and size of target gene modifications, respectively. Further, we show that the inducible CRISPR (iCRISPR) system can be used effectively to create biallelic mutation in multiple target loci and thus, provides a flexible and fast platform to study loss of function phenotypes in vivo. PMID:25690852

  2. Science Education Supporting Weather Broadcasters On-Air and in the Classroom with NASA "Mini-Education Supplements"

    NASA Technical Reports Server (NTRS)

    Shepherd, J. Marshall; Starr, David OC. (Technical Monitor)

    2001-01-01

    NASA-Goddard Space Flight Center has initiated a new project designed to expand on existing news services and add value to classrooms through the development and distribution of two-minute 'mini-supplements' which give context and teach about current weather and Earth research phenomena. The innovative mini-supplements provide raw materials for weather forecasters to build news stories around NASA related missions without having to edit the more traditional and cumbersome long-form video format. The supplements cover different weather and climate topics and include NASA data, animations, video footage, and interviews with scientists. The supplements also include a curriculum package with educational lessons, educator guide, and hand-on activities. One goal is to give on-air broadcasters who are the primary science educators for the general public what they need to 'teach' about the science related to NASA research behind weather and climate news. This goal achieves increasing public literacy and assures higher accuracy and quality science reporting by the media. The other goal is to enable on-air broadcasters to serve as distributors of high quality, standards-based educational curricula and supplemental material when they visit 8-12 grade classrooms. The focus of 'pilot effort' centers around the success of NASA's Tropical Rainfall Measuring Mission (TRMM) but is likely expandable to other NASA earth or space science missions.

  3. Evaporated CaS thin films for AC electroluminescence devices

    NASA Astrophysics Data System (ADS)

    Kobayashi, H.; Tanaka, S.; Shanker, V.; Shiiki, M.; Deguchi, H.

    1985-08-01

    The growth behavior of evaporated CaS thin films has been investigated to achieve bright electroluminescence. The crystallinity of CaS films is improved with substrate temperature and for temperatures higher than 300°C, the films orient to the (200) plane. Sulfur coevaporation further helps to form a more perfect film even at lower temperatures. A CaS: Ce,Cl electroluminescent thin film device has been fabricated with a brightness of 650 cd/m 2.

  4. Cellular apoptosis susceptibility (CAS) is linked to integrin β1 and required for tumor cell migration and invasion in hepatocellular carcinoma (HCC)

    PubMed Central

    Winkler, Juliane; Roessler, Stephanie; Sticht, Carsten; DiGuilio, Amanda L.; Drucker, Elisabeth; Holzer, Kerstin; Eiteneuer, Eva; Herpel, Esther; Breuhahn, Kai; Gretz, Norbert; Schirmacher, Peter; Ori, Alessandro; Singer, Stephan

    2016-01-01

    Importins and exportins represent an integral part of the nucleocytoplasmic transport machinery with fundamental importance for eukaryotic cell function. A variety of malignancies including hepatocellular carcinoma (HCC) show de-regulation of nuclear transport factors such as overexpression of the exportin Cellular Apoptosis Susceptibility (CAS). The functional implications of CAS in hepatocarcinogenesis remain, however, poorly understood. Here we integrated proteomics, transcriptomics and functional assays with patient data to further characterize the role of CAS in HCC. By analyzing ∼ 1700 proteins using quantitative mass spectrometry in HCC cells we found that CAS depletion by RNAi leads to de-regulation of integrins, particularly down-regulation of integrin β1. Consistent with this finding, CAS knockdown resulted in substantially reduced migration and invasion of HCC cell lines as analyzed by 2D ‘scratch’ and invasion chamber assays, respectively. Supporting the potential in vivo relevance, high expression levels of CAS in HCC tissue samples were associated with macroangioinvasion and poorer patient outcome. Our data suggest a previously unanticipated link between CAS and integrin signaling which correlates with an aggressive HCC phenotype. PMID:27015362

  5. Foreign DNA capture during CRISPR–Cas adaptive immunity

    PubMed Central

    Nuñez, James K.; Harrington, Lucas B.; Kranzusch, Philip J.; Engelman, Alan N.; Doudna, Jennifer A.

    2015-01-01

    Bacteria and archaea generate adaptive immunity against phages and plasmids by integrating foreign DNA of specific 30–40 base pair (bp) lengths into clustered regularly interspaced short palindromic repeats (CRISPR) loci as spacer segments1–6. The universally conserved Cas1–Cas2 integrase complex catalyzes spacer acquisition using a direct nucleophilic integration mechanism similar to retroviral integrases and transposases7–13. How the Cas1–Cas2 complex selects foreign DNA substrates for integration remains unknown. Here we present X-ray crystal structures of the Escherichia coli Cas1–Cas2 complex bound to cognate 33 nucleotide (nt) protospacer DNA substrates. The protein complex creates a curved binding surface spanning the length of the DNA and splays the ends of the protospacer to allow each terminal nucleophilic 3′–OH to enter a channel leading into the Cas1 active sites. Phosphodiester backbone interactions between the protospacer and the proteins explain the sequence-nonspecific substrate selection observed in vivo2–4. Our results uncover the structural basis for foreign DNA capture and the mechanism by which Cas1–Cas2 functions as a molecular ruler to dictate the sequence architecture of CRISPR loci. PMID:26503043

  6. CRISPR-Cas9-guided Genome Engineering in C. elegans

    PubMed Central

    Kim, Hyun-Min; Colaiácovo, Monica P.

    2016-01-01

    The CRISPR (clustered regularly interspaced short palindromic repeats)-Cas (CRISPR-associated) system is successfully being used for efficient and targeted genome editing in various organisms including the nematode C. elegans. Recent studies developed various CRISPR-Cas9 approaches to enhance genome engineering via two major DNA double-strand break repair pathways: non-homologous end joining and homologous recombination. Here we describe a protocol for Cas9-mediated C. elegans genome editing together with single guide RNA (sgRNA) and repair template cloning and injection methods required for delivering Cas9, sgRNAs and repair template DNA into the C. elegans germline. PMID:27366893

  7. Foreign DNA capture during CRISPR-Cas adaptive immunity.

    PubMed

    Nuñez, James K; Harrington, Lucas B; Kranzusch, Philip J; Engelman, Alan N; Doudna, Jennifer A

    2015-11-26

    Bacteria and archaea generate adaptive immunity against phages and plasmids by integrating foreign DNA of specific 30-40-base-pair lengths into clustered regularly interspaced short palindromic repeat (CRISPR) loci as spacer segments. The universally conserved Cas1-Cas2 integrase complex catalyses spacer acquisition using a direct nucleophilic integration mechanism similar to retroviral integrases and transposases. How the Cas1-Cas2 complex selects foreign DNA substrates for integration remains unknown. Here we present X-ray crystal structures of the Escherichia coli Cas1-Cas2 complex bound to cognate 33-nucleotide protospacer DNA substrates. The protein complex creates a curved binding surface spanning the length of the DNA and splays the ends of the protospacer to allow each terminal nucleophilic 3'-OH to enter a channel leading into the Cas1 active sites. Phosphodiester backbone interactions between the protospacer and the proteins explain the sequence-nonspecific substrate selection observed in vivo. Our results uncover the structural basis for foreign DNA capture and the mechanism by which Cas1-Cas2 functions as a molecular ruler to dictate the sequence architecture of CRISPR loci. PMID:26503043

  8. Cas9 as a versatile tool for engineering biology

    PubMed Central

    Mali, Prashant; Esvelt, Kevin M; Church, George M

    2014-01-01

    RNA-guided Cas9 nucleases derived from clustered regularly interspaced short palindromic repeats (CRISPR)-Cas systems have dramatically transformed our ability to edit the genomes of diverse organisms. We believe tools and techniques based on Cas9, a single unifying factor capable of colocalizing RNA, DNA and protein, will grant unprecedented control over cellular organization, regulation and behavior. Here we describe the Cas9 targeting methodology, detail current and prospective engineering advances and suggest potential applications ranging from basic science to the clinic. PMID:24076990

  9. Optical Control of CRISPR/Cas9 Gene Editing

    PubMed Central

    Hemphill, James; Borchardt, Erin K.; Brown, Kalyn; Asokan, Aravind; Deiters, Alexander

    2016-01-01

    The CRISPR/Cas9 system has emerged as an important tool in biomedical research for a wide range of applications, with significant potential for genome engineering and gene therapy. In order to achieve conditional control of the CRISPR/Cas9 system, a genetically encoded light-activated Cas9 was engineered through the site-specific installation of a caged lysine amino acid. Several potential lysine residues were identified as viable caging sites that can be modified to optically control Cas9 function, as demonstrated through optical activation and deactivation of both exogenous and endogenous gene function. PMID:25905628

  10. United States Air Force 611th air support group 611th Civil Engineer Squadron, Elmendorf AFB, Alaska. Eareckson Air Station, Alaska. Remedial investigation/feasibility study report, final. Volume 1. Report for July 1994-September 1996

    SciTech Connect

    1995-08-25

    The Report, has been prepared to present the findings of the environmental investigation activities conducted at Eareckson Air Station. The RI/FS report has been divided into four primary volumes. This volume, Volume I, provides general information about environmental activities conducted at Eareckson AS to date (Section 1.0), summarizes 1993 and 1994 investigation activities (Section 2.0), outlines the decision process (Section 3.0), and presents basewide findings (Section 4.0).

  11. A Fourth-Generation Approach to Decision Support in a Private University. AIR Annual 1984 Forum Paper.

    ERIC Educational Resources Information Center

    Glover, Robert H.

    The demand for decision-support systems (DSSs) at the University of Hartford and the use of current software systems based on an information center approach are discussed. It is noted that declining enrollment demand and financial stress in tuition and enrollment-dependent universities requires a flexible and dynamic DSS for effective planning and…

  12. Microoxic Niches within the Thylakoid Stroma of Air-Grown Chlamydomonas reinhardtii Protect [FeFe]-Hydrogenase and Support Hydrogen Production under Fully Aerobic Environment.

    PubMed

    Liran, Oded; Semyatich, Rinat; Milrad, Yuval; Eilenberg, Haviva; Weiner, Iddo; Yacoby, Iftach

    2016-09-01

    Photosynthetic hydrogen production in the microalga Chlamydomonas reinhardtii is catalyzed by two [FeFe]-hydrogenase isoforms, HydA1 and HydA2, both irreversibly inactivated upon a few seconds exposure to atmospheric oxygen. Until recently, it was thought that hydrogenase is not active in air-grown microalgal cells. In contrast, we show that the entire pool of cellular [FeFe]-hydrogenase remains active in air-grown cells due to efficient scavenging of oxygen. Using membrane inlet mass spectrometry, (18)O2 isotope, and various inhibitors, we were able to dissect the various oxygen uptake mechanisms. We found that both chlororespiration, catalyzed by plastid terminal oxidase, and Mehler reactions, catalyzed by photosystem I and Flavodiiron proteins, significantly contribute to oxygen uptake rate. This rate is considerably enhanced with increasing light, thus forming local anaerobic niches at the proximity of the stromal face of the thylakoid membrane. Furthermore, we found that in transition to high light, the hydrogen production rate is significantly enhanced for a short duration (100 s), thus indicating that [FeFe]-hydrogenase functions as an immediate sink for surplus electrons in aerobic as well as in anaerobic environments. In summary, we show that an anaerobic locality in the chloroplast preserves [FeFe]-hydrogenase activity and supports continuous hydrogen production in air-grown microalgal cells. PMID:27443604

  13. Tenth anniversary of CAS ONLINE service : What CAS services should be in the new era of chemical information

    NASA Astrophysics Data System (ADS)

    Kostakos, Charles N.

    Chemical Abstracts Service celebrated 10th anniversary of CAS online information service in 1990. A speech given on the occasion reviewed history of the CAS ONLINE, in relation to its most important benefits for scientists and engineers. The development of STN international, the network through which CAS ONLINE is accessible around the world, was also discussed in the speech. The CAS ONLINE now contains a wide variety of files relating to chemical field including CA file, Registry file. CA previews,. CASREACT, CIN. MARPAT, etc for supplying chemical information worldwide.

  14. P130Cas Src-Binding and Substrate Domains Have Distinct Roles in Sustaining Focal Adhesion Disassembly and Promoting Cell Migration

    PubMed Central

    Meenderink, Leslie M.; Ryzhova, Larisa M.; Donato, Dominique M.; Gochberg, Daniel F.; Kaverina, Irina; Hanks, Steven K.

    2010-01-01

    The docking protein p130Cas is a prominent Src substrate found in focal adhesions (FAs) and is implicated in regulating critical aspects of cell motility including FA disassembly and protrusion of the leading edge plasma membrane. To better understand how p130Cas acts to promote these events we examined requirements for established p130Cas signaling motifs including the SH3-binding site of the Src binding domain (SBD) and the tyrosine phosphorylation sites within the substrate domain (SD). Expression of wild type p130Cas in Cas −/− mouse embryo fibroblasts resulted in enhanced cell migration associated with increased leading-edge actin flux, increased rates of FA assembly/disassembly, and uninterrupted FA turnover. Variants lacking either the SD phosphorylation sites or the SBD SH3-binding motif were able to partially restore the migration response, while only a variant lacking both signaling functions was fully defective. Notably, the migration defects associated with p130Cas signaling-deficient variants correlated with longer FA lifetimes resulting from aborted FA disassembly attempts. However the SD mutational variant was fully defective in increasing actin assembly at the protruding leading edge and FA assembly/disassembly rates, indicating that SD phosphorylation is the sole p130Cas signaling function in regulating these processes. Our results provide the first quantitative evidence supporting roles for p130Cas SD tyrosine phosphorylation in promoting both leading edge actin flux and FA turnover during cell migration, while further revealing that the p130Cas SBD has a function in cell migration and sustained FA disassembly that is distinct from its known role of promoting SD tyrosine phosphorylation. PMID:20976150

  15. Recent Advances in Genome Editing Using CRISPR/Cas9

    PubMed Central

    Ding, Yuduan; Li, Hong; Chen, Ling-Ling; Xie, Kabin

    2016-01-01

    The CRISPR (clustered regularly interspaced short palindromic repeat)-Cas9 (CRISPR-associated nuclease 9) system is a versatile tool for genome engineering that uses a guide RNA (gRNA) to target Cas9 to a specific sequence. This simple RNA-guided genome-editing technology has become a revolutionary tool in biology and has many innovative applications in different fields. In this review, we briefly introduce the Cas9-mediated genome-editing method, summarize the recent advances in CRISPR/Cas9 technology, and discuss their implications for plant research. To date, targeted gene knockout using the Cas9/gRNA system has been established in many plant species, and the targeting efficiency and capacity of Cas9 has been improved by optimizing its expression and that of its gRNA. The CRISPR/Cas9 system can also be used for sequence-specific mutagenesis/integration and transcriptional control of target genes. We also discuss off-target effects and the constraint that the protospacer-adjacent motif (PAM) puts on CRISPR/Cas9 genome engineering. To address these problems, a number of bioinformatic tools are available to help design specific gRNAs, and new Cas9 variants and orthologs with high fidelity and alternative PAM specificities have been engineered. Owing to these recent efforts, the CRISPR/Cas9 system is becoming a revolutionary and flexible tool for genome engineering. Adoption of the CRISPR/Cas9 technology in plant research would enable the investigation of plant biology at an unprecedented depth and create innovative applications in precise crop breeding. PMID:27252719

  16. Cas9 Variants Expand the Target Repertoire in Caenorhabditis elegans.

    PubMed

    Bell, Ryan T; Fu, Becky X H; Fire, Andrew Z

    2016-02-01

    The proliferation of CRISPR/Cas9-based methods in Caenorhabditis elegans has enabled efficient genome editing and precise genomic tethering of Cas9 fusion proteins. Experimental designs using CRISPR/Cas9 are currently limited by the need for a protospacer adjacent motif (PAM) in the target with the sequence NGG. Here we report the characterization of two modified Cas9 proteins in C. elegans that recognize NGA and NGCG PAMs. We found that each variant could stimulate homologous recombination with a donor template at multiple loci and that PAM specificity was comparable to that of wild-type Cas9. To directly compare effectiveness, we used CRISPR/Cas9 genome editing to generate a set of assay strains with a common single-guide RNA (sgRNA) target sequence, but that differ in the juxtaposed PAM (NGG, NGA, or NGCG). In this controlled setting, we determined that the NGA PAM Cas9 variant can be as effective as wild-type Cas9. We similarly edited a genomic target to study the influence of the base following the NGA PAM. Using four strains with four NGAN PAMs differing only at the fourth position and adjacent to the same sgRNA target, we observed that efficient homologous replacement was attainable with any base in the fourth position, with an NGAG PAM being the most effective. In addition to demonstrating the utility of two Cas9 mutants in C. elegans and providing reagents that permit CRISPR/Cas9 experiments with fewer restrictions on potential targets, we established a means to benchmark the efficiency of different Cas9::PAM combinations that avoids variations owing to differences in the sgRNA sequence. PMID:26680661

  17. 40 CFR Appendix: Table 1 to... - List of Hazardous Air Pollutants (HAP) for Subpart HHH

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 40 Protection of Environment 11 2010-07-01 2010-07-01 true List of Hazardous Air Pollutants (HAP... AIR POLLUTANTS FOR SOURCE CATEGORIES National Emission Standards for Hazardous Air Pollutants From... HHH of Part 63—List of Hazardous Air Pollutants (HAP) for Subpart HHH CAS Number a Chemical name...

  18. Multiplexed pancreatic genome engineering and cancer induction by transfection-based CRISPR/Cas9 delivery in mice

    PubMed Central

    Maresch, Roman; Mueller, Sebastian; Veltkamp, Christian; Öllinger, Rupert; Friedrich, Mathias; Heid, Irina; Steiger, Katja; Weber, Julia; Engleitner, Thomas; Barenboim, Maxim; Klein, Sabine; Louzada, Sandra; Banerjee, Ruby; Strong, Alexander; Stauber, Teresa; Gross, Nina; Geumann, Ulf; Lange, Sebastian; Ringelhan, Marc; Varela, Ignacio; Unger, Kristian; Yang, Fengtang; Schmid, Roland M.; Vassiliou, George S.; Braren, Rickmer; Schneider, Günter; Heikenwalder, Mathias; Bradley, Allan; Saur, Dieter; Rad, Roland

    2016-01-01

    Mouse transgenesis has provided fundamental insights into pancreatic cancer, but is limited by the long duration of allele/model generation. Here we show transfection-based multiplexed delivery of CRISPR/Cas9 to the pancreas of adult mice, allowing simultaneous editing of multiple gene sets in individual cells. We use the method to induce pancreatic cancer and exploit CRISPR/Cas9 mutational signatures for phylogenetic tracking of metastatic disease. Our results demonstrate that CRISPR/Cas9-multiplexing enables key applications, such as combinatorial gene-network analysis, in vivo synthetic lethality screening and chromosome engineering. Negative-selection screening in the pancreas using multiplexed-CRISPR/Cas9 confirms the vulnerability of pancreatic cells to Brca2-inactivation in a Kras-mutant context. We also demonstrate modelling of chromosomal deletions and targeted somatic engineering of inter-chromosomal translocations, offering multifaceted opportunities to study complex structural variation, a hallmark of pancreatic cancer. The low-frequency mosaic pattern of transfection-based CRISPR/Cas9 delivery faithfully recapitulates the stochastic nature of human tumorigenesis, supporting wide applicability for biological/preclinical research. PMID:26916719

  19. Multiplexed pancreatic genome engineering and cancer induction by transfection-based CRISPR/Cas9 delivery in mice.

    PubMed

    Maresch, Roman; Mueller, Sebastian; Veltkamp, Christian; Öllinger, Rupert; Friedrich, Mathias; Heid, Irina; Steiger, Katja; Weber, Julia; Engleitner, Thomas; Barenboim, Maxim; Klein, Sabine; Louzada, Sandra; Banerjee, Ruby; Strong, Alexander; Stauber, Teresa; Gross, Nina; Geumann, Ulf; Lange, Sebastian; Ringelhan, Marc; Varela, Ignacio; Unger, Kristian; Yang, Fengtang; Schmid, Roland M; Vassiliou, George S; Braren, Rickmer; Schneider, Günter; Heikenwalder, Mathias; Bradley, Allan; Saur, Dieter; Rad, Roland

    2016-01-01

    Mouse transgenesis has provided fundamental insights into pancreatic cancer, but is limited by the long duration of allele/model generation. Here we show transfection-based multiplexed delivery of CRISPR/Cas9 to the pancreas of adult mice, allowing simultaneous editing of multiple gene sets in individual cells. We use the method to induce pancreatic cancer and exploit CRISPR/Cas9 mutational signatures for phylogenetic tracking of metastatic disease. Our results demonstrate that CRISPR/Cas9-multiplexing enables key applications, such as combinatorial gene-network analysis, in vivo synthetic lethality screening and chromosome engineering. Negative-selection screening in the pancreas using multiplexed-CRISPR/Cas9 confirms the vulnerability of pancreatic cells to Brca2-inactivation in a Kras-mutant context. We also demonstrate modelling of chromosomal deletions and targeted somatic engineering of inter-chromosomal translocations, offering multifaceted opportunities to study complex structural variation, a hallmark of pancreatic cancer. The low-frequency mosaic pattern of transfection-based CRISPR/Cas9 delivery faithfully recapitulates the stochastic nature of human tumorigenesis, supporting wide applicability for biological/preclinical research. PMID:26916719

  20. Tracking air-dropped drogues and dyes from aircraft in support of ERTS-1 circulation studies. [Delaware Bay

    NASA Technical Reports Server (NTRS)

    Klemas, V. (Principal Investigator); Davis, G.; Wang, H.

    1974-01-01

    The author has identified the following significant results. For two years ERTS-1 has been employed to investigate current circulation patterns in Delaware Bay under different tidal, flow, and wind conditions. Since sufficient numbers of current meters and boats are not available, air-droppable drogues and dye packs have been developed and tested. The drogues consist of a styrofoam float and a line to which is attached a stainless steel biplane. The length of the line determines at what depth currents will be monitored. The floats are color coded to distinguish their movement and mark the depth of the biplanes. Simultaneously floating and anchored dye packs of fluorescein dye have been deployed from aircraft. The movement of the dye and drogues is tracked by sequential aerial photography, using fixed markers on shore or on buoys as reference points to calibrate the scale and direction of drogue movement. The current data obtained by this technique is then used to annotate current circulation maps derived from ERTS-1 imagery.

  1. An Assessment of Hickam Air Force Base's Capability to Support Strategic Airlift Throughput when Operating under an Avian Flu Pandemic

    SciTech Connect

    Brigantic, Robert T.; Campbell, James R.; Doctor, Pamela G.; Johnson, Alan; Coomber, P.

    2006-03-10

    Hickam Air Force Base (AFB), Hawaii provides an ideal waypoint for U.S. strategic airlift aircraft to refuel and receive other services on their way to Northeast and Southeast Asia from the continental United States. Hickam AFB also serves as a critical aerial port of debarkation (APOD) for deploying U.S. forces and equipment to more distant lands as needed. Making use of the United States Transportation Command’s Aerial Port of Debarkation Plus model, this paper examines the ability of Hickam AFB to serve in its important role as an APOD when operating under the effects of a major avian flu pandemic. In this regard, the major influence on Hickam AFB will be a serious degradation to the number of available personnel to service aircraft and operate Hickam AFB’s aerial port. It is noted that the results presented herein are based on simplistic attrition rate assumptions. Nonetheless, it is envisioned that this work is applicable to more realistic input attrition rates as avian flu epidemiological models are refined, as well as attrition associated with other types of contagious pandemic disease or willful biological warfare attack.

  2. Performance Evaluation of Activated Carbon Nanofiber as Carbon Supports to Improve the Cyclability of Li-Air Batteries.

    PubMed

    Park, Inyeong; Kim, Heeyun; Shim, Sang Eun; Baeck, Sung-Hyeon

    2015-11-01

    This study addresses the effects of the pore structures of carbon materials used as cathodes for non-aqueous lithium-air batteries on cycle life. Carbon Nanofibers (CNFs) were synthesized by electrospinning polyacrylonitrile (PAN) and carbonization. The synthesized CNF was then converted to activated carbon nanofibers (ACNFs) under flowing CO2. The specific surface areas CNFs were increased on activation. ACNFs were arranged randomly to form a web-like structure providing both oxygen pathways and a means of discharging products. To examine the electrochemical properties of ACNF, charge-discharge tests were conducted using a Swagelok-type cell at a constant current density of 0.2 mA/cm2; impedance tests were also conducted. ACNF sheet electrodes had cycle lives of up to 50 cycles, which was attributed to high surface area and porosity, although overpotentials for both charge and discharge were high. This cycling performance showed that the pore structure of sheet ACNF is more suitable for the transport of oxygen and for the storage of discharge products than carbon powders. PMID:26726643

  3. Multi-Level Wild Land Fire Fighting Management Support System for an Optimized Guidance of Ground and Air Forces

    NASA Astrophysics Data System (ADS)

    Almer, Alexander; Schnabel, Thomas; Perko, Roland; Raggam, Johann; Köfler, Armin; Feischl, Richard

    2016-04-01

    Climate change will lead to a dramatic increase in damage from forest fires in Europe by the end of this century. In the Mediterranean region, the average annual area affected by forest fires has quadrupled since the 1960s (WWF, 2012). The number of forest fires is also on the increase in Central and Northern Europe. The Austrian forest fire database shows a total of 584 fires for the period 2012 to 2014, while even large areas of Sweden were hit by forest fires in August 2014, which were brought under control only after two weeks of intense fire-fighting efforts supported by European civil protection modules. Based on these facts, the improvements in forest fire control are a major international issue in the quest to protect human lives and resources as well as to reduce the negative environmental impact of these fires to a minimum. Within this paper the development of a multi-functional airborne management support system within the frame of the Austrian national safety and security research programme (KIRAS) is described. The main goal of the developments is to assist crisis management tasks of civil emergency teams and armed forces in disaster management by providing multi spectral, near real-time airborne image data products. As time, flexibility and reliability as well as objective information are crucial aspects in emergency management, the used components are tailored to meet these requirements. An airborne multi-functional management support system was developed as part of the national funded project AIRWATCH, which enables real-time monitoring of natural disasters based on optical and thermal images. Airborne image acquisition, a broadband line of sight downlink and near real-time processing solutions allow the generation of an up-to-date geo-referenced situation map. Furthermore, this paper presents ongoing developments for innovative extensions and research activities designed to optimize command operations in national and international fire

  4. CRISPR-Cas9 Genome Engineering in Saccharomyces cerevisiae Cells.

    PubMed

    Ryan, Owen W; Poddar, Snigdha; Cate, Jamie H D

    2016-01-01

    This protocol describes a method for CRISPR-Cas9-mediated genome editing that results in scarless and marker-free integrations of DNA into Saccharomyces cerevisiae genomes. DNA integration results from cotransforming (1) a single plasmid (pCAS) that coexpresses the Cas9 endonuclease and a uniquely engineered single guide RNA (sgRNA) expression cassette and (2) a linear DNA molecule that is used to repair the chromosomal DNA damage by homology-directed repair. For target specificity, the pCAS plasmid requires only a single cloning modification: replacing the 20-bp guide RNA sequence within the sgRNA cassette. This CRISPR-Cas9 protocol includes methods for (1) cloning the unique target sequence into pCAS, (2) assembly of the double-stranded DNA repair oligonucleotides, and (3) cotransformation of pCAS and linear repair DNA into yeast cells. The protocol is technically facile and requires no special equipment. It can be used in any S. cerevisiae strain, including industrial polyploid isolates. Therefore, this CRISPR-Cas9-based DNA integration protocol is achievable by virtually any yeast genetics and molecular biology laboratory. PMID:27250940

  5. Interacting Parallel Constructions of Knowledge in a CAS Context

    ERIC Educational Resources Information Center

    Kidron, Ivy; Dreyfus, Tommy

    2010-01-01

    We consider the influence of a CAS context on a learner's process of constructing a justification for the bifurcations in a logistic dynamical process. We describe how instrumentation led to cognitive constructions and how the roles of the learner and the CAS intertwine, especially close to the branching and combining of constructing actions. The…

  6. Semiotic and Discursive Variables in CAS-Based Didactical Engineering.

    ERIC Educational Resources Information Center

    Winslow, Carl

    2003-01-01

    Presents a semiotic analysis of the potential of computer algebra systems (CAS) for enabling mathematical activity on a conceptual level higher than usual. Illustrates examples of theoretical points from a development project in the context of a first year university course in calculus. Discusses how CAS may be used in a didactical analysis and in…

  7. 48 CFR 970.3002-1 - CAS applicability.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ....3002-1 CAS applicability. The provisions of 48 CFR part 30 and 48 CFR chapter 99 (FAR Appendix) shall... 48 Federal Acquisition Regulations System 5 2010-10-01 2010-10-01 false CAS applicability. 970.3002-1 Section 970.3002-1 Federal Acquisition Regulations System DEPARTMENT OF ENERGY...

  8. 48 CFR 970.3002-1 - CAS applicability.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ....3002-1 CAS applicability. The provisions of 48 CFR part 30 and 48 CFR chapter 99 (FAR Appendix) shall... 48 Federal Acquisition Regulations System 5 2011-10-01 2011-10-01 false CAS applicability. 970.3002-1 Section 970.3002-1 Federal Acquisition Regulations System DEPARTMENT OF ENERGY...

  9. 48 CFR 970.3002-1 - CAS applicability.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ....3002-1 CAS applicability. The provisions of 48 CFR part 30 and 48 CFR chapter 99 (FAR Appendix) shall... 48 Federal Acquisition Regulations System 5 2014-10-01 2014-10-01 false CAS applicability. 970.3002-1 Section 970.3002-1 Federal Acquisition Regulations System DEPARTMENT OF ENERGY...

  10. Teaching Undergraduate Mathematics Using CAS Technology: Issues and Prospects

    ERIC Educational Resources Information Center

    Tobin, Patrick C.; Weiss, Vida

    2016-01-01

    The use of handheld CAS technology in undergraduate mathematics courses in Australia is paradoxically shrinking under sustained disapproval or disdain from the professional mathematics community. Mathematics education specialists argue with their mathematics colleagues over a range of issues in course development and this use of CAS or even…

  11. 48 CFR 970.3002-1 - CAS applicability.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ....3002-1 CAS applicability. The provisions of 48 CFR part 30 and 48 CFR chapter 99 (FAR Appendix) shall... 48 Federal Acquisition Regulations System 5 2013-10-01 2013-10-01 false CAS applicability. 970.3002-1 Section 970.3002-1 Federal Acquisition Regulations System DEPARTMENT OF ENERGY...

  12. 48 CFR 970.3002-1 - CAS applicability.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ....3002-1 CAS applicability. The provisions of 48 CFR part 30 and 48 CFR chapter 99 (FAR Appendix) shall... 48 Federal Acquisition Regulations System 5 2012-10-01 2012-10-01 false CAS applicability. 970.3002-1 Section 970.3002-1 Federal Acquisition Regulations System DEPARTMENT OF ENERGY...

  13. From Calculus to Dynamical Systems through DGS and CAS

    ERIC Educational Resources Information Center

    García, Jeanett López; Zamudio, Jorge Javier Jiménez

    2015-01-01

    Several factors have motivated the use of CAS or DGS in the teaching-learning process, such as: the development of new technologies, the availability of computers, and the widespread use of the Internet, among others. Even more, the trend to include CAS and DGS in the curricula of some undergraduate studies has resulted in the instruction of the…

  14. Optimization of genome editing through CRISPR-Cas9 engineering.

    PubMed

    Zhang, Jian-Hua; Adikaram, Poorni; Pandey, Mritunjay; Genis, Allison; Simonds, William F

    2016-04-01

    CRISPR (Clustered Regularly-Interspaced Short Palindromic Repeats)-Cas9 (CRISPR associated protein 9) has rapidly become the most promising genome editing tool with great potential to revolutionize medicine. Through guidance of a 20 nucleotide RNA (gRNA), CRISPR-Cas9 finds and cuts target protospacer DNA precisely 3 base pairs upstream of a PAM (Protospacer Adjacent Motif). The broken DNA ends are repaired by either NHEJ (Non-Homologous End Joining) resulting in small indels, or by HDR (Homology Directed Repair) for precise gene or nucleotide replacement. Theoretically, CRISPR-Cas9 could be used to modify any genomic sequences, thereby providing a simple, easy, and cost effective means of genome wide gene editing. However, the off-target activity of CRISPR-Cas9 that cuts DNA sites with imperfect matches with gRNA have been of significant concern because clinical applications require 100% accuracy. Additionally, CRISPR-Cas9 has unpredictable efficiency among different DNA target sites and the PAM requirements greatly restrict its genome editing frequency. A large number of efforts have been made to address these impeding issues, but much more is needed to fully realize the medical potential of CRISPR-Cas9. In this article, we summarize the existing problems and current advances of the CRISPR-Cas9 technology and provide perspectives for the ultimate perfection of Cas9-mediated genome editing. PMID:27340770

  15. Simplified CRISPR-Cas genome editing for Saccharomyces cerevisiae.

    PubMed

    Generoso, Wesley Cardoso; Gottardi, Manuela; Oreb, Mislav; Boles, Eckhard

    2016-08-01

    CRISPR-Cas has become a powerful technique for genetic engineering of yeast. Here, we present an improved version by using only one single plasmid expressing Cas9 and one or two guide-RNAs. A high gene deletion efficiency was achieved even with simultaneous recombination cloning of the plasmid and deletion in industrial strains. PMID:27327211

  16. Recent Progress in CRISPR/Cas9 Technology.

    PubMed

    Mei, Yue; Wang, Yan; Chen, Huiqian; Sun, Zhong Sheng; Ju, Xing-Da

    2016-02-20

    The clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 system, a simple and efficient tool for genome editing, has experienced rapid progress in its technology and applicability in the past two years. Here, we review the recent advances in CRISPR/Cas9 technology and the ways that have been adopted to expand our capacity for precise genome manipulation. First, we introduce the mechanism of CRISPR/Cas9, including its biochemical and structural implications. Second, we highlight the latest improvements in the CRISPR/Cas9 system, especially Cas9 protein modifications for customization. Third, we review its current applications, in which the versatile CRISPR/Cas9 system was employed to edit the genome, epigenome, or RNA of various organisms. Although CRISPR/Cas9 allows convenient genome editing accompanied by many benefits, we should not ignore the significant ethical and biosafety concerns that it raises. Finally, we discuss the prospective applications and challenges of several promising techniques adapted from CRISPR/Cas9. PMID:26924689

  17. Comparison of Highly Resolved Model-Based Exposure Metrics for Traffic-Related Air Pollutants to Support Environmental Health Studies

    PubMed Central

    Chang, Shih Ying; Vizuete, William; Breen, Michael; Isakov, Vlad; Arunachalam, Saravanan

    2015-01-01

    Human exposure to air pollution in many studies is represented by ambient concentrations from space-time kriging of observed values. Space-time kriging techniques based on a limited number of ambient monitors may fail to capture the concentration from local sources. Further, because people spend more time indoors, using ambient concentration to represent exposure may cause error. To quantify the associated exposure error, we computed a series of six different hourly-based exposure metrics at 16,095 Census blocks of three Counties in North Carolina for CO, NOx, PM2.5, and elemental carbon (EC) during 2012. These metrics include ambient background concentration from space-time ordinary kriging (STOK), ambient on-road concentration from the Research LINE source dispersion model (R-LINE), a hybrid concentration combining STOK and R-LINE, and their associated indoor concentrations from an indoor infiltration mass balance model. Using a hybrid-based indoor concentration as the standard, the comparison showed that outdoor STOK metrics yielded large error at both population (67% to 93%) and individual level (average bias between −10% to 95%). For pollutants with significant contribution from on-road emission (EC and NOx), the on-road based indoor metric performs the best at the population level (error less than 52%). At the individual level, however, the STOK-based indoor concentration performs the best (average bias below 30%). For PM2.5, due to the relatively low contribution from on-road emission (7%), STOK-based indoor metric performs the best at both population (error below 40%) and individual level (error below 25%). The results of the study will help future epidemiology studies to select appropriate exposure metric and reduce potential bias in exposure characterization. PMID:26670242

  18. Comparison of Highly Resolved Model-Based Exposure Metrics for Traffic-Related Air Pollutants to Support Environmental Health Studies.

    PubMed

    Chang, Shih Ying; Vizuete, William; Breen, Michael; Isakov, Vlad; Arunachalam, Saravanan

    2015-12-01

    Human exposure to air pollution in many studies is represented by ambient concentrations from space-time kriging of observed values. Space-time kriging techniques based on a limited number of ambient monitors may fail to capture the concentration from local sources. Further, because people spend more time indoors, using ambient concentration to represent exposure may cause error. To quantify the associated exposure error, we computed a series of six different hourly-based exposure metrics at 16,095 Census blocks of three Counties in North Carolina for CO, NO(x), PM(2.5), and elemental carbon (EC) during 2012. These metrics include ambient background concentration from space-time ordinary kriging (STOK), ambient on-road concentration from the Research LINE source dispersion model (R-LINE), a hybrid concentration combining STOK and R-LINE, and their associated indoor concentrations from an indoor infiltration mass balance model. Using a hybrid-based indoor concentration as the standard, the comparison showed that outdoor STOK metrics yielded large error at both population (67% to 93%) and individual level (average bias between -10% to 95%). For pollutants with significant contribution from on-road emission (EC and NO(x)), the on-road based indoor metric performs the best at the population level (error less than 52%). At the individual level, however, the STOK-based indoor concentration performs the best (average bias below 30%). For PM(2.5), due to the relatively low contribution from on-road emission (7%), STOK-based indoor metric performs the best at both population (error below 40%) and individual level (error below 25%). The results of the study will help future epidemiology studies to select appropriate exposure metric and reduce potential bias in exposure characterization. PMID:26670242

  19. Cas9-mediated targeting of viral RNA in eukaryotic cells

    PubMed Central

    Price, Aryn A.; Sampson, Timothy R.; Ratner, Hannah K.; Grakoui, Arash; Weiss, David S.

    2015-01-01

    Clustered, regularly interspaced, short palindromic repeats–CRISPR associated (CRISPR-Cas) systems are prokaryotic RNA-directed endonuclease machineries that act as an adaptive immune system against foreign genetic elements. Using small CRISPR RNAs that provide specificity, Cas proteins recognize and degrade nucleic acids. Our previous work demonstrated that the Cas9 endonuclease from Francisella novicida (FnCas9) is capable of targeting endogenous bacterial RNA. Here, we show that FnCas9 can be directed by an engineered RNA-targeting guide RNA to target and inhibit a human +ssRNA virus, hepatitis C virus, within eukaryotic cells. This work reveals a versatile and portable RNA-targeting system that can effectively function in eukaryotic cells and be programmed as an antiviral defense. PMID:25918406

  20. Conditional Control of CRISPR/Cas9 Function.

    PubMed

    Zhou, Wenyuan; Deiters, Alexander

    2016-04-25

    The recently discovered CRISPR/Cas9 endonuclease system, comprised of a guide RNA for the recognition of a DNA target and the Cas9 nuclease protein for binding and processing the target, has been extensively studied and has been widely applied in genome editing, synthetic biology, and transcriptional modulation in cells and animals. Toward more precise genomic modification and further expansion of the CRISPR/Cas9 system as a spatiotemporally controlled gene regulatory system, several approaches of conditional activation of Cas9 function using small molecules and light have recently been developed. These methods have led to improvements in the genome editing specificity of the CRISPR/Cas9 system and enabled its activation with temporal and spatial precision. PMID:26996256

  1. Comprehensive Protocols for CRISPR/Cas9-based Gene Editing in Human Pluripotent Stem Cells.

    PubMed

    Santos, David P; Kiskinis, Evangelos; Eggan, Kevin; Merkle, Florian T

    2016-01-01

    Genome editing of human pluripotent stem cells (hPSCs) with the CRISPR/Cas9 system has the potential to revolutionize hPSC-based disease modeling, drug screening, and transplantation therapy. Here, we aim to provide a single resource to enable groups, even those with limited experience with hPSC culture or the CRISPR/Cas9 system, to successfully perform genome editing. The methods are presented in detail and are supported by a theoretical framework to allow for the incorporation of inevitable improvements in the rapidly evolving gene-editing field. We describe protocols to generate hPSC lines with gene-specific knock-outs, small targeted mutations, or knock-in reporters. © 2016 by John Wiley & Sons, Inc. PMID:27532820

  2. Implementing the CAS Standards: The Implementation of the CAS Standards in Student Affairs as a Comprehensive Assessment Approach

    ERIC Educational Resources Information Center

    Dorman, Jesse A.

    2012-01-01

    The increasing use of the CAS standards as a comprehensive assessment approach in divisions of student affairs necessitates a more in-depth understanding of how the CAS standards are being implemented in these settings. In response to increasing calls for improvement, accountability and professionalism in student affairs (Bresciani, 2006; Cooper…

  3. Clean Air Slots Amid Dense Atmospheric Pollution in Southern Africa

    NASA Technical Reports Server (NTRS)

    Hobbs, Peter V.

    2003-01-01

    During the flights of the University of Washington's Convair-580 in the Southern African Regional Science Initiative (SAFARI 2000) in southern Africa, a phenomenon was observed that has not been reported previously. This was the occurrence of thin layers of remarkably clean air, sandwiched between heavily polluted air, which persisted for many hours during the day. Photographs are shown of these clean air slots (CAS), and particle concentrations and light scattering coefficients in and around such slot are presented. An explanation is proposed for the propensity of CAS to form in southern Africa during the dry season.

  4. Surveillance system for air pollutants by combination of the decision support system COMPAS and optical remote sensing systems

    NASA Astrophysics Data System (ADS)

    Flassak, Thomas; de Witt, Helmut; Hahnfeld, Peter; Knaup, Andreas; Kramer, Lothar

    1995-09-01

    COMPAS is a decision support system designed to assist in the assessment of the consequences of accidental releases of toxic and flammable substances. One of the key elements of COMPAS is a feedback algorithm which allows us to calculate the source term with the aid of concentration measurements. Up to now the feedback technique is applied to concentration measurements done with test tubes or conventional point sensors. In this paper the extension of the actual method is presented which is the combination of COMPAS and an optical remote sensing system like the KAYSER-THREDE K300 FTIR system. Active remote sensing methods based on FTIR are, among other applications, ideal for the so-called fence line monitoring of the diffuse emissions and accidental releases from industrial facilities, since from the FTIR spectra averaged concentration levels along the measurement path can be achieved. The line-averaged concentrations are ideally suited as on-line input for COMPAS' feedback technique. Uncertainties in the assessment of the source term related with both shortcomings of the dispersion model itself and also problems of a feedback strategy based on point measurements are reduced.

  5. Preliminary assessment report for Army Aviation Support Facility 2, Installation 25075, Westover Air Force Base, Chicopee, Massachusetts. Installation Restoration Program

    SciTech Connect

    Haffenden, R.; Flaim, S.

    1993-08-01

    This report presents the results of the preliminary assessment (PA) conducted by Argonne National Laboratory at the Massachusetts Army National Guard (MAARNG) property known as the Army Aviation Support Facility 2 (AASF 2) near Chicopee, Massachusetts. Preliminary assessments of federal facilities are being conducted to compile the information necessary for completing preremedial activities and to provide a basis for establishing corrective actions in response to releases of hazardous substances. The principal objective of the PA is to characterize the site accurately and determine the need for further action by examining site activities, quantities of hazardous substances present, and potential pathways by which contamination could affect public health and the environment. The AASF 2 is a 10-acre site located in the western portion of Massachusetts, in the town of Chicopee, in the county of Hampden. The facilities included in this PA are Building 7400, adjacent paved areas, grassy areas, and the hazardous waste drum storage buildings. The environmentally significant operations (ESOS) associated with the property are (1) the waste drum storage area, (2) abandoned underground storage tanks (USTs), and (3) refueling activities.

  6. Algorithm and simulation development in support of response strategies for contamination events in air and water systems.

    SciTech Connect

    Waanders, Bart Van Bloemen

    2006-01-01

    Chemical/Biological/Radiological (CBR) contamination events pose a considerable threat to our nation's infrastructure, especially in large internal facilities, external flows, and water distribution systems. Because physical security can only be enforced to a limited degree, deployment of early warning systems is being considered. However to achieve reliable and efficient functionality, several complex questions must be answered: (1) where should sensors be placed, (2) how can sparse sensor information be efficiently used to determine the location of the original intrusion, (3) what are the model and data uncertainties, (4) how should these uncertainties be handled, and (5) how can our algorithms and forward simulations be sufficiently improved to achieve real time performance? This report presents the results of a three year algorithmic and application development to support the identification, mitigation, and risk assessment of CBR contamination events. The main thrust of this investigation was to develop (1) computationally efficient algorithms for strategically placing sensors, (2) identification process of contamination events by using sparse observations, (3) characterization of uncertainty through developing accurate demands forecasts and through investigating uncertain simulation model parameters, (4) risk assessment capabilities, and (5) reduced order modeling methods. The development effort was focused on water distribution systems, large internal facilities, and outdoor areas.

  7. Interference activity of a minimal Type I CRISPR–Cas system from Shewanella putrefaciens

    PubMed Central

    Dwarakanath, Srivatsa; Brenzinger, Susanne; Gleditzsch, Daniel; Plagens, André; Klingl, Andreas; Thormann, Kai; Randau, Lennart

    2015-01-01

    Type I CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)–Cas (CRISPR-associated) systems exist in bacterial and archaeal organisms and provide immunity against foreign DNA. The Cas protein content of the DNA interference complexes (termed Cascade) varies between different CRISPR-Cas subtypes. A minimal variant of the Type I-F system was identified in proteobacterial species including Shewanella putrefaciens CN-32. This variant lacks a large subunit (Csy1), Csy2 and Csy3 and contains two unclassified cas genes. The genome of S. putrefaciens CN-32 contains only five Cas proteins (Cas1, Cas3, Cas6f, Cas1821 and Cas1822) and a single CRISPR array with 81 spacers. RNA-Seq analyses revealed the transcription of this array and the maturation of crRNAs (CRISPR RNAs). Interference assays based on plasmid conjugation demonstrated that this CRISPR-Cas system is active in vivo and that activity is dependent on the recognition of the dinucleotide GG PAM (Protospacer Adjacent Motif) sequence and crRNA abundance. The deletion of cas1821 and cas1822 reduced the cellular crRNA pool. Recombinant Cas1821 was shown to form helical filaments bound to RNA molecules, which suggests its role as the Cascade backbone protein. A Cascade complex was isolated which contained multiple Cas1821 copies, Cas1822, Cas6f and mature crRNAs. PMID:26350210

  8. Long-term High-Resolution Spectroscopy of γ Cas, ζ Tau, and π Aqr

    NASA Astrophysics Data System (ADS)

    Bjorkman, K. S.; Miroshnichenko, A. S.; Krugov, V. D.

    2000-05-01

    High-resolution spectroscopic data (λ λ 5285-6595 Angstroms) for three bright classical Be stars with unusual Hα profiles (γ Cas, ζ Tau, and π Aqr) have been obtained during the time period 1993-2000 at the Ritter Observatory of the University of Toledo. The data for γ Cas are supplemented by medium-resolution spectroscopy taken at the Terskol station of the Main Astronomical Observatory of the Ukranian Academy of Sciences. The stars show the presence of an additional variable (central) emission peak in the Hα line, which has a double-peaked profile in most stars of this type. Long-term radial velocity (RV) variations of H I, He I, Si II, and Fe II lines are detected in γ Cas and ζ Tau. The RV of the central peak of Hα in ζ Tau seem to follow the binary orbital motion (period 132.9 days) on top of the long-term variations (period 1420 days). In γ Cas this peak shows a constant trend towards negative velocities since 1993, which is opposite to the behavior of other emission lines. Short-term RV variations of the Hα emission peak with a period 84 days are found in π Aqr during its weak-disk phase (since 1996). This work has been supported in part by NASA grant NAG5-8054 to the Univ. of Toledo, and by a Cottrell Scholars Award to KSB from the Research Corporation. Support for Ritter Observatory has been provided in part by NSF grant AST-9024802, and in part by a grant from the Fund for Astrophysical Research.

  9. Installation-Restoration Program. Phase 2. Confirmation/quantification. Stage 1. Problem confirmation study: Otis Air National Guard Base, Massachusetts, Air National Guard Support Center, Andrews Air Force Base, Maryland. Final technical report, November 1983-July 1985

    SciTech Connect

    Kraybill, R.L.; Smart, G.R.; Bopp, F.

    1985-09-04

    A Problem Confirmation Study was performed at seven sites on Otis Air National Guard Base: the Current and Former Training Areas, the Base Landfill, the Nondestructive Inspection Laboratory, the Fuel Test Dump Site, the Railyard Fuel Pumping Station, and the Petrol Fuel Storage Area. The field investigation was conducted in two stages, in November 1983 through January 1984, and in October through December 1984. Resampling was performed at selected locations in April and July 1985. A total of 11 monitor wells were installed and sampled and test-pit investigations were conducted at six sites. In addition, the contents of a sump tank, and two header pipes for fuel-transmission lines were sampled. Analytes included TOC, TOX, cyanide, phenols, Safe Drinking Water metals, pesticides and herbicides, and in the second round, priority-pollutant volatile organic compounds and a GC fingerprint scan for fuel products. On the basis of the field-work findings, it is concluded that, to date, water-quality impacts on ground water from past activities have been minimal.

  10. Pilomatricome: étude de 22 cas

    PubMed Central

    Nasreddine, Fatima Zahra; Hali, Fouzia; Chiheb, Soumiya

    2016-01-01

    Le pilomatricome est une tumeur cutanée fréquente et bénigne du follicule pileux chez l'enfant. C'est une tumeur annexielle souvent méconnue et confondue avec d'autres lésions cutanées. Les localisations habituelles sont la tête et le cou. Le but de ce travail est de rapporter une série de 22 cas comportant des formes inhabituelles colligées au service de dermatologie sur une période allant de Janvier 2006 jusqu'au Mai 2015. L’étude a concerné 16 femmes et 6 hommes. La moyenne d’âge était de 23,3 ans (4-80 ans). La localisation cervico faciale a été observée dans 12 cas, 2 patients avaient des localisations multiples, un garçon de 4ans avait une localisation au niveau fronto-temporal et une fillette de 14 ans avait une localisation au niveau du visage et de l'avant-bras, et un patient de 48 ans avait une localisation sous unguéale. L'aspect clinique était typique dans tous les cas avec des nodules sous cutanés de consistance pierreuse. Tous les patients ont bénéficié d'une exérèse des nodules sous anesthésie locale. L’étude histologique était en faveur d'un épithélioma momifié de Malherbe d'exérèse complète sans signes de malignité. Aucun patient n'a présenté de rechute. L'originalité de notre étude réside dans la présence de localisations exceptionnelles au niveau latéro-vertébral, des membres et sous-unguéale, l’âge de survenue inhabituel à 80 ans et la présence de localisations multiples signalées chez 2 enfants. PMID:27516819

  11. Air Pollution Training Programs.

    ERIC Educational Resources Information Center

    Public Health Service (DHEW), Rockville, MD.

    This catalog lists the universities, both supported and not supported by the Division of Air Pollution, which offer graduate programs in the field of air pollution. The catalog briefly describes the programs and their entrance requirements, the requirements, qualifications and terms of special fellowships offered by the Division of Air Pollution.…

  12. Nucleosome breathing and remodeling constrain CRISPR-Cas9 function

    PubMed Central

    Isaac, R Stefan; Jiang, Fuguo; Doudna, Jennifer A; Lim, Wendell A; Narlikar, Geeta J; Almeida, Ricardo

    2016-01-01

    The CRISPR-Cas9 bacterial surveillance system has become a versatile tool for genome editing and gene regulation in eukaryotic cells, yet how CRISPR-Cas9 contends with the barriers presented by eukaryotic chromatin is poorly understood. Here we investigate how the smallest unit of chromatin, a nucleosome, constrains the activity of the CRISPR-Cas9 system. We find that nucleosomes assembled on native DNA sequences are permissive to Cas9 action. However, the accessibility of nucleosomal DNA to Cas9 is variable over several orders of magnitude depending on dynamic properties of the DNA sequence and the distance of the PAM site from the nucleosome dyad. We further find that chromatin remodeling enzymes stimulate Cas9 activity on nucleosomal templates. Our findings imply that the spontaneous breathing of nucleosomal DNA together with the action of chromatin remodelers allow Cas9 to effectively act on chromatin in vivo. DOI: http://dx.doi.org/10.7554/eLife.13450.001 PMID:27130520

  13. CRISPR/Cas9 Based Genome Editing of Penicillium chrysogenum.

    PubMed

    Pohl, C; Kiel, J A K W; Driessen, A J M; Bovenberg, R A L; Nygård, Y

    2016-07-15

    CRISPR/Cas9 based systems have emerged as versatile platforms for precision genome editing in a wide range of organisms. Here we have developed powerful CRISPR/Cas9 tools for marker-based and marker-free genome modifications in Penicillium chrysogenum, a model filamentous fungus and industrially relevant cell factory. The developed CRISPR/Cas9 toolbox is highly flexible and allows editing of new targets with minimal cloning efforts. The Cas9 protein and the sgRNA can be either delivered during transformation, as preassembled CRISPR-Cas9 ribonucleoproteins (RNPs) or expressed from an AMA1 based plasmid within the cell. The direct delivery of the Cas9 protein with in vitro synthesized sgRNA to the cells allows for a transient method for genome engineering that may rapidly be applicable for other filamentous fungi. The expression of Cas9 from an AMA1 based vector was shown to be highly efficient for marker-free gene deletions. PMID:27072635

  14. HPCCP/CAS Workshop Proceedings 1998

    NASA Technical Reports Server (NTRS)

    Schulbach, Catherine; Mata, Ellen (Editor); Schulbach, Catherine (Editor)

    1999-01-01

    This publication is a collection of extended abstracts of presentations given at the HPCCP/CAS (High Performance Computing and Communications Program/Computational Aerosciences Project) Workshop held on August 24-26, 1998, at NASA Ames Research Center, Moffett Field, California. The objective of the Workshop was to bring together the aerospace high performance computing community, consisting of airframe and propulsion companies, independent software vendors, university researchers, and government scientists and engineers. The Workshop was sponsored by the HPCCP Office at NASA Ames Research Center. The Workshop consisted of over 40 presentations, including an overview of NASA's High Performance Computing and Communications Program and the Computational Aerosciences Project; ten sessions of papers representative of the high performance computing research conducted within the Program by the aerospace industry, academia, NASA, and other government laboratories; two panel sessions; and a special presentation by Mr. James Bailey.

  15. Adaptation in CRISPR-Cas Systems.

    PubMed

    Sternberg, Samuel H; Richter, Hagen; Charpentier, Emmanuelle; Qimron, Udi

    2016-03-17

    Clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) proteins constitute an adaptive immune system in prokaryotes. The system preserves memories of prior infections by integrating short segments of foreign DNA, termed spacers, into the CRISPR array in a process termed adaptation. During the past 3 years, significant progress has been made on the genetic requirements and molecular mechanisms of adaptation. Here we review these recent advances, with a focus on the experimental approaches that have been developed, the insights they generated, and a proposed mechanism for self- versus non-self-discrimination during the process of spacer selection. We further describe the regulation of adaptation and the protein players involved in this fascinating process that allows bacteria and archaea to harbor adaptive immunity. PMID:26949040

  16. Characterization and evolution of Salmonella CRISPR-Cas systems.

    PubMed

    Shariat, Nikki; Timme, Ruth E; Pettengill, James B; Barrangou, Rodolphe; Dudley, Edward G

    2015-02-01

    Prokaryotic CRISPR-Cas (clustered regularly interspaced short palindromic repeats and CRISPR-associated genes) systems provide adaptive immunity from invasive genetic elements and encompass three essential features: (i) cas genes, (ii) a CRISPR array composed of spacers and direct repeats and (iii) an AT-rich leader sequence upstream of the array. We performed in-depth sequence analysis of the CRISPR-Cas systems in >600 Salmonella, representing four clinically prevalent serovars. Each CRISPR-Cas feature is extremely conserved in the Salmonella, and the CRISPR1 locus is more highly conserved than CRISPR2. Array composition is serovar-specific, although no convincing evidence of recent spacer acquisition against exogenous nucleic acids exists. Only 12% of spacers match phage and plasmid sequences and self-targeting spacers are associated with direct repeat variants. High nucleotide identity (>99.9%) exists across the cas operon among isolates of a single serovar and in some cases this conservation extends across divergent serovars. These observations reflect historical CRISPR-Cas immune activity, showing that this locus has ceased undergoing adaptive events. Intriguingly, the high level of conservation across divergent serovars shows that the genetic integrity of these inactive loci is maintained over time, contrasting with the canonical view that inactive CRISPR loci degenerate over time. This thorough characterization of Salmonella CRISPR-Cas systems presents new insights into Salmonella CRISPR evolution, particularly with respect to cas gene conservation, leader sequences, organization of direct repeats and protospacer matches. Collectively, our data suggest that Salmonella CRISPR-Cas systems are no longer immunogenic; rather, their impressive conservation indicates they may have an alternative function in Salmonella. PMID:25479838

  17. CRISPR-Cas9-Mediated Genome Editing in Leishmania donovani

    PubMed Central

    Zhang, Wen-Wei

    2015-01-01

    ABSTRACT The prokaryotic CRISPR (clustered regularly interspaced short palindromic repeat)-Cas9, an RNA-guided endonuclease, has been shown to mediate efficient genome editing in a wide variety of organisms. In the present study, the CRISPR-Cas9 system has been adapted to Leishmania donovani, a protozoan parasite that causes fatal human visceral leishmaniasis. We introduced the Cas9 nuclease into L. donovani and generated guide RNA (gRNA) expression vectors by using the L. donovani rRNA promoter and the hepatitis delta virus (HDV) ribozyme. It is demonstrated within that L. donovani mainly used homology-directed repair (HDR) and microhomology-mediated end joining (MMEJ) to repair the Cas9 nuclease-created double-strand DNA break (DSB). The nonhomologous end-joining (NHEJ) pathway appears to be absent in L. donovani. With this CRISPR-Cas9 system, it was possible to generate knockouts without selection by insertion of an oligonucleotide donor with stop codons and 25-nucleotide homology arms into the Cas9 cleavage site. Likewise, we disrupted and precisely tagged endogenous genes by inserting a bleomycin drug selection marker and GFP gene into the Cas9 cleavage site. With the use of Hammerhead and HDV ribozymes, a double-gRNA expression vector that further improved gene-targeting efficiency was developed, and it was used to make precise deletion of the 3-kb miltefosine transporter gene (LdMT). In addition, this study identified a novel single point mutation caused by CRISPR-Cas9 in LdMT (M381T) that led to miltefosine resistance, a concern for the only available oral antileishmanial drug. Together, these results demonstrate that the CRISPR-Cas9 system represents an effective genome engineering tool for L. donovani. PMID:26199327

  18. 41 CFR 102-33.435 - What CAS cost and utilization data must we report?

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 41 Public Contracts and Property Management 3 2012-01-01 2012-01-01 false What CAS cost and... Services (cas) Cost and Utilization Data § 102-33.435 What CAS cost and utilization data must we report? You must report the costs and flying hours for each CAS aircraft you hire. You must also report...

  19. 41 CFR 102-33.130 - If we hire CAS, what are our management responsibilities?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 41 Public Contracts and Property Management 3 2010-07-01 2010-07-01 false If we hire CAS, what are... § 102-33.130 If we hire CAS, what are our management responsibilities? If you hire CAS, you are... agreements; (b) Accounting for the cost of your aircraft and services hired as CAS; (c) Accounting for use...

  20. 41 CFR 102-33.440 - Who must report CAS cost and utilization data?

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 41 Public Contracts and Property Management 3 2014-01-01 2014-01-01 false Who must report CAS cost... (cas) Cost and Utilization Data § 102-33.440 Who must report CAS cost and utilization data? Executive agencies, except the Armed Forces and U.S. intelligence agencies, must report CAS cost and utilization...

  1. 41 CFR 102-33.130 - If we hire CAS, what are our management responsibilities?

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 41 Public Contracts and Property Management 3 2011-01-01 2011-01-01 false If we hire CAS, what are... § 102-33.130 If we hire CAS, what are our management responsibilities? If you hire CAS, you are... agreements; (b) Accounting for the cost of your aircraft and services hired as CAS; (c) Accounting for use...

  2. 41 CFR 102-33.435 - What CAS cost and utilization data must we report?

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 41 Public Contracts and Property Management 3 2014-01-01 2014-01-01 false What CAS cost and... Services (cas) Cost and Utilization Data § 102-33.435 What CAS cost and utilization data must we report? You must report the costs and flying hours for each CAS aircraft you hire. You must also report...

  3. 41 CFR 102-33.440 - Who must report CAS cost and utilization data?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 41 Public Contracts and Property Management 3 2013-07-01 2013-07-01 false Who must report CAS cost... (cas) Cost and Utilization Data § 102-33.440 Who must report CAS cost and utilization data? Executive agencies, except the Armed Forces and U.S. intelligence agencies, must report CAS cost and utilization...

  4. 41 CFR 102-33.435 - What CAS cost and utilization data must we report?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 41 Public Contracts and Property Management 3 2010-07-01 2010-07-01 false What CAS cost and... Services (cas) Cost and Utilization Data § 102-33.435 What CAS cost and utilization data must we report? You must report the costs and flying hours for each CAS aircraft you hire. You must also report...

  5. 41 CFR 102-33.130 - If we hire CAS, what are our management responsibilities?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 41 Public Contracts and Property Management 3 2013-07-01 2013-07-01 false If we hire CAS, what are... § 102-33.130 If we hire CAS, what are our management responsibilities? If you hire CAS, you are... agreements; (b) Accounting for the cost of your aircraft and services hired as CAS; (c) Accounting for use...

  6. 41 CFR 102-33.440 - Who must report CAS cost and utilization data?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 41 Public Contracts and Property Management 3 2010-07-01 2010-07-01 false Who must report CAS cost... (cas) Cost and Utilization Data § 102-33.440 Who must report CAS cost and utilization data? Executive agencies, except the Armed Forces and U.S. intelligence agencies, must report CAS cost and utilization...

  7. 41 CFR 102-33.435 - What CAS cost and utilization data must we report?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 41 Public Contracts and Property Management 3 2013-07-01 2013-07-01 false What CAS cost and... Services (cas) Cost and Utilization Data § 102-33.435 What CAS cost and utilization data must we report? You must report the costs and flying hours for each CAS aircraft you hire. You must also report...

  8. 41 CFR 102-33.440 - Who must report CAS cost and utilization data?

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 41 Public Contracts and Property Management 3 2011-01-01 2011-01-01 false Who must report CAS cost... (cas) Cost and Utilization Data § 102-33.440 Who must report CAS cost and utilization data? Executive agencies, except the Armed Forces and U.S. intelligence agencies, must report CAS cost and utilization...

  9. 41 CFR 102-33.130 - If we hire CAS, what are our management responsibilities?

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 41 Public Contracts and Property Management 3 2012-01-01 2012-01-01 false If we hire CAS, what are... § 102-33.130 If we hire CAS, what are our management responsibilities? If you hire CAS, you are... agreements; (b) Accounting for the cost of your aircraft and services hired as CAS; (c) Accounting for use...

  10. 41 CFR 102-33.435 - What CAS cost and utilization data must we report?

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 41 Public Contracts and Property Management 3 2011-01-01 2011-01-01 false What CAS cost and... Services (cas) Cost and Utilization Data § 102-33.435 What CAS cost and utilization data must we report? You must report the costs and flying hours for each CAS aircraft you hire. You must also report...

  11. 41 CFR 102-33.440 - Who must report CAS cost and utilization data?

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 41 Public Contracts and Property Management 3 2012-01-01 2012-01-01 false Who must report CAS cost... (cas) Cost and Utilization Data § 102-33.440 Who must report CAS cost and utilization data? Executive agencies, except the Armed Forces and U.S. intelligence agencies, must report CAS cost and utilization...

  12. 41 CFR 102-33.130 - If we hire CAS, what are our management responsibilities?

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 41 Public Contracts and Property Management 3 2014-01-01 2014-01-01 false If we hire CAS, what are... § 102-33.130 If we hire CAS, what are our management responsibilities? If you hire CAS, you are... agreements; (b) Accounting for the cost of your aircraft and services hired as CAS; (c) Accounting for use...

  13. Determination of the Physical Dimensions of μ Cas

    NASA Astrophysics Data System (ADS)

    Bach, K.; Kang, W.

    2015-07-01

    We investigate the physical properties of the nearby astrometric binary μ Cas based on a spectroscopic analysis and evolutionary calculations. The chemical composition of the μ Cas system has been determined based on high resolution spectroscopy from BOES. Combining our spectroscopic analysis with observations from Hipparcos and CHARA, we determine the evolutionary status of μ Cas. With well-constrained stellar parameters, the internal structure and frequency modes have also been calculated. The ultimate goal of this study is to describe the physical processes inside stars through a comprehensive modeling.

  14. Grossesse intra murale à propos d'un cas

    PubMed Central

    de Tové, Kofi-Mensa Savi; Salifou, Kabibou; Imorou, Rachidi Sidi; Biaou, Olivier; Boco, Vicentia

    2015-01-01

    La grossesse intra-murale est la variété la plus rare de grossesse extra-utérine. Il s'agit de la localisation de l’œuf dans l’épaisseur du myomètre. En cas de retard diagnostic, l’évolution peut être catastrophique avec rupture utérine et hémorragie cataclysmique. L’échographie permet dans certains cas un diagnostic pré opératoire. Les auteurs rapportent un cas survenu chez une patiente aux antécédents de curetage. PMID:26448812

  15. Introducing the TI-Nspire CAS Learning Technology

    NASA Astrophysics Data System (ADS)

    Osbourne, Michael

    2008-03-01

    Engage your students! In this hands-on workshop, we will demonstrate how TI's newest graphing technology can be integrated into the physics classroom. Come experience both the TI-NspireT CAS Handheld and the TI- NspireT CAS Computer Software. The new TI-Nspire learning technology has been developed to allow students to explore physics and to better understand concepts through the manipulation of complex formulas,graphing, spreadsheets, data analysis, and simulations. During the workshop you will receive an overview of the technology, sample activities, along with the opportunity to experience how TI-Nspire CAS can be effectively integrated into the physics classroom. Limited to 20 participants.

  16. Functional Analysis of Bacteriophage Immunity through a Type I-E CRISPR-Cas System in Vibrio cholerae and Its Application in Bacteriophage Genome Engineering

    PubMed Central

    Box, Allison M.; McGuffie, Matthew J.; O'Hara, Brendan J.

    2015-01-01

    ABSTRACT The classical and El Tor biotypes of Vibrio cholerae serogroup O1, the etiological agent of cholera, are responsible for the sixth and seventh (current) pandemics, respectively. A genomic island (GI), GI-24, previously identified in a classical biotype strain of V. cholerae, is predicted to encode clustered regularly interspaced short palindromic repeat (CRISPR)-associated proteins (Cas proteins); however, experimental evidence in support of CRISPR activity in V. cholerae has not been documented. Here, we show that CRISPR-Cas is ubiquitous in strains of the classical biotype but excluded from strains of the El Tor biotype. We also provide in silico evidence to suggest that CRISPR-Cas actively contributes to phage resistance in classical strains. We demonstrate that transfer of GI-24 to V. cholerae El Tor via natural transformation enables CRISPR-Cas-mediated resistance to bacteriophage CP-T1 under laboratory conditions. To elucidate the sequence requirements of this type I-E CRISPR-Cas system, we engineered a plasmid-based system allowing the directed targeting of a region of interest. Through screening for phage mutants that escape CRISPR-Cas-mediated resistance, we show that CRISPR targets must be accompanied by a 3′ TT protospacer-adjacent motif (PAM) for efficient interference. Finally, we demonstrate that efficient editing of V. cholerae lytic phage genomes can be performed by simultaneously introducing an editing template that allows homologous recombination and escape from CRISPR-Cas targeting. IMPORTANCE Cholera, caused by the facultative pathogen Vibrio cholerae, remains a serious public health threat. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) provide prokaryotes with sequence-specific protection from invading nucleic acids, including bacteriophages. In this work, we show that one genomic feature differentiating sixth pandemic (classical biotype) strains from seventh pandemic (El Tor

  17. On Cas A, Cassini, Comets, and King Charles

    NASA Astrophysics Data System (ADS)

    Soria, Roberto; Balestrieri, Riccardo; Ohtsuka, Yasuyo

    2013-03-01

    We re-examine the long-standing problem of the date of the Cassiopeia A supernova (SN), in view of recent claims that it might be the 1630 'noon-star' seen at the birth of King Charles II. We do not support this identification, based on the expected brightness of a Type-IIb SN (too faint to be seen in daylight), the extrapolated motion of the ejecta (inconsistent with a date earlier than 1650), the lack of any scientific follow-up observations, the lack of any mention of it in Asian archives. The origin of the 1630 noon-star event (if real) remains a mystery; there was a bright comet in 1630 June but no evidence to determine whether or not it was visible in daylight. Instead, we present French reports about a fourth-magnitude star discovered by Cassini in Cassiopeia in or shortly before 1671, which was not seen before or since. The brightness is consistent with what we expect for the Cas A SN; the date is consistent with the extrapolated motion of the ejecta. We argue that this source could be the long-sought SN.

  18. The Structural Biology of CRISPR-Cas Systems

    PubMed Central

    Jiang, Fuguo; Doudna, Jennifer A.

    2015-01-01

    Prokaryotic CRISPR-Cas genomic loci encode RNA-mediated adaptive immune systems that bear some functional similarities with eukaryotic RNA interference. Acquired and heritable immunity against bacteriophage and plasmids begins with integration of ~30 base pair foreign DNA sequences into the host genome. CRISPR-derived transcripts assemble with CRISPR-associated (Cas) proteins to target complementary nucleic acids for degradation. Here we review recent advances in the structural biology of these targeting complexes, with a focus on structural studies of the multisubunit Type I CRISPR RNA-guided surveillance and the Cas9 DNA endonuclease found in Type II CRISPR-Cas systems. These complexes have distinct structures that are each capable of site-specific double-stranded DNA binding and local helix unwinding. PMID:25723899

  19. CAS Standards and Guidelines for Student Services/Development Programs.

    ERIC Educational Resources Information Center

    NACADA Journal, 1986

    1986-01-01

    The Council for the Advancement of Standards for Student Services/Development Programs (CAS) developed and adopted standards and interpretive guidelines for specific functional areas of student services/development programs within postsecondary educational institutions. These guidelines are presented. (MLW)

  20. CRISPR-Cas9-Guided Genome Engineering in C. elegans.

    PubMed

    Kim, Hyun-Min; Colaiácovo, Monica P

    2016-01-01

    The CRISPR (clustered regularly interspaced short palindromic repeats)-Cas (CRISPR-associated) system is successfully being used for efficient and targeted genome editing in various organisms, including the nematode C. elegans. Recent studies have developed various CRISPR-Cas9 approaches to enhance genome engineering via two major DNA double-strand break repair pathways: non-homologous end joining and homologous recombination. Here we describe a protocol for Cas9-mediated C. elegans genome editing together with single guide RNA (sgRNA) and repair template cloning, as well as injection methods required for delivering Cas9, sgRNAs, and repair template DNA into the C. elegans germline. © 2016 by John Wiley & Sons, Inc. PMID:27366893

  1. Methods for Optimizing CRISPR-Cas9 Genome Editing Specificity.

    PubMed

    Tycko, Josh; Myer, Vic E; Hsu, Patrick D

    2016-08-01

    Advances in the development of delivery, repair, and specificity strategies for the CRISPR-Cas9 genome engineering toolbox are helping researchers understand gene function with unprecedented precision and sensitivity. CRISPR-Cas9 also holds enormous therapeutic potential for the treatment of genetic disorders by directly correcting disease-causing mutations. Although the Cas9 protein has been shown to bind and cleave DNA at off-target sites, the field of Cas9 specificity is rapidly progressing, with marked improvements in guide RNA selection, protein and guide engineering, novel enzymes, and off-target detection methods. We review important challenges and breakthroughs in the field as a comprehensive practical guide to interested users of genome editing technologies, highlighting key tools and strategies for optimizing specificity. The genome editing community should now strive to standardize such methods for measuring and reporting off-target activity, while keeping in mind that the goal for specificity should be continued improvement and vigilance. PMID:27494557

  2. Engineering the Caenorhabditis elegans genome with CRISPR/Cas9.

    PubMed

    Waaijers, Selma; Boxem, Mike

    2014-08-01

    The development in early 2013 of CRISPR/Cas9-based genome engineering promises to dramatically advance our ability to alter the genomes of model systems at will. A single, easily produced targeting RNA guides the Cas9 endonuclease to a specific DNA sequence where it creates a double strand break. Imprecise repair of the break can yield mutations, while homologous recombination with a repair template can be used to effect specific changes to the genome. The tremendous potential of this system led several groups to independently adapt it for use in Caenorhabditiselegans, where it was successfully used to generate mutations and to create tailored genome changes through homologous recombination. Here, we review the different approaches taken to adapt CRISPR/Cas9 for C. elegans, and provide practical guidelines for CRISPR/Cas9-based genome engineering. PMID:24685391

  3. CRISPR-Cas systems: Prokaryotes upgrade to adaptive immunity.

    PubMed

    Barrangou, Rodolphe; Marraffini, Luciano A

    2014-04-24

    Clustered regularly interspaced short palindromic repeats (CRISPR), and associated proteins (Cas) comprise the CRISPR-Cas system, which confers adaptive immunity against exogenic elements in many bacteria and most archaea. CRISPR-mediated immunization occurs through the uptake of DNA from invasive genetic elements such as plasmids and viruses, followed by its integration into CRISPR loci. These loci are subsequently transcribed and processed into small interfering RNAs that guide nucleases for specific cleavage of complementary sequences. Conceptually, CRISPR-Cas shares functional features with the mammalian adaptive immune system, while also exhibiting characteristics of Lamarckian evolution. Because immune markers spliced from exogenous agents are integrated iteratively in CRISPR loci, they constitute a genetic record of vaccination events and reflect environmental conditions and changes over time. Cas endonucleases, which can be reprogrammed by small guide RNAs have shown unprecedented potential and flexibility for genome editing and can be repurposed for numerous DNA targeting applications including transcriptional control. PMID:24766887

  4. CRISPR-Cas systems: prokaryotes upgrade to adaptive immunity

    PubMed Central

    Barrangou, Rodolphe; Marraffini, Luciano A.

    2014-01-01

    Summary Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR), and associated proteins (Cas) comprise the CRISPR-Cas system, which confers adaptive immunity against exogenic elements in many bacteria and most archaea. CRISPR-mediated immunization occurs through the uptake of DNA from invasive genetic elements such as plasmids and viruses, followed by its integration into CRISPR loci. These loci are subsequently transcribed and processed into small interfering RNAs that guide nucleases for specific cleavage of complementary sequences. Conceptually, CRISPR-Cas shares functional features with the mammalian adaptive immune system, while also exhibiting characteristics of Lamarckian evolution. Because immune markers spliced from exogenous agents are integrated iteratively in CRISPR loci, they constitute a genetic record of vaccination events and reflect environmental conditions and changes over time. Cas endonucleases, which can be reprogrammed by small guide RNAs have shown unprecedented potential and flexibility for genome editing, and can be repurposed for numerous DNA targeting applications including transcriptional control. PMID:24766887

  5. An updated evolutionary classification of CRISPR-Cas systems.

    PubMed

    Makarova, Kira S; Wolf, Yuri I; Alkhnbashi, Omer S; Costa, Fabrizio; Shah, Shiraz A; Saunders, Sita J; Barrangou, Rodolphe; Brouns, Stan J J; Charpentier, Emmanuelle; Haft, Daniel H; Horvath, Philippe; Moineau, Sylvain; Mojica, Francisco J M; Terns, Rebecca M; Terns, Michael P; White, Malcolm F; Yakunin, Alexander F; Garrett, Roger A; van der Oost, John; Backofen, Rolf; Koonin, Eugene V

    2015-11-01

    The evolution of CRISPR-cas loci, which encode adaptive immune systems in archaea and bacteria, involves rapid changes, in particular numerous rearrangements of the locus architecture and horizontal transfer of complete loci or individual modules. These dynamics complicate straightforward phylogenetic classification, but here we present an approach combining the analysis of signature protein families and features of the architecture of cas loci that unambiguously partitions most CRISPR-cas loci into distinct classes, types and subtypes. The new classification retains the overall structure of the previous version but is expanded to now encompass two classes, five types and 16 subtypes. The relative stability of the classification suggests that the most prevalent variants of CRISPR-Cas systems are already known. However, the existence of rare, currently unclassifiable variants implies that additional types and subtypes remain to be characterized. PMID:26411297

  6. Comparison of Cas9 activators in multiple species.

    PubMed

    Chavez, Alejandro; Tuttle, Marcelle; Pruitt, Benjamin W; Ewen-Campen, Ben; Chari, Raj; Ter-Ovanesyan, Dmitry; Haque, Sabina J; Cecchi, Ryan J; Kowal, Emma J K; Buchthal, Joanna; Housden, Benjamin E; Perrimon, Norbert; Collins, James J; Church, George

    2016-07-01

    Several programmable transcription factors exist based on the versatile Cas9 protein, yet their relative potency and effectiveness across various cell types and species remain unexplored. Here, we compare Cas9 activator systems and examine their ability to induce robust gene expression in several human, mouse, and fly cell lines. We also explore the potential for improved activation through the combination of the most potent activator systems, and we assess the role of cooperativity in maximizing gene expression. PMID:27214048

  7. Potential pitfalls of CRISPR/Cas9-mediated genome editing.

    PubMed

    Peng, Rongxue; Lin, Guigao; Li, Jinming

    2016-04-01

    Recently, a novel technique named the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein (Cas)9 system has been rapidly developed. This genome editing tool has improved our ability tremendously with respect to exploring the pathogenesis of diseases and correcting disease mutations, as well as phenotypes. With a short guide RNA, Cas9 can be precisely directed to target sites, and functions as an endonuclease to efficiently produce breaks in DNA double strands. Over the past 30 years, CRISPR has evolved from the 'curious sequences of unknown biological function' into a promising genome editing tool. As a result of the incessant development in the CRISPR/Cas9 system, Cas9 co-expressed with custom guide RNAs has been successfully used in a variety of cells and organisms. This genome editing technology can also be applied to synthetic biology, functional genomic screening, transcriptional modulation and gene therapy. However, although CRISPR/Cas9 has a broad range of action in science, there are several aspects that affect its efficiency and specificity, including Cas9 activity, target site selection and short guide RNA design, delivery methods, off-target effects and the incidence of homology-directed repair. In the present review, we highlight the factors that affect the utilization of CRISPR/Cas9, as well as possible strategies for handling any problems. Addressing these issues will allow us to take better advantage of this technique. In addition, we also review the history and rapid development of the CRISPR/Cas system from the time of its initial discovery in 2012. PMID:26535798

  8. Target specificity of the CRISPR-Cas9 system

    PubMed Central

    Wu, Xuebing; Kriz, Andrea J.; Sharp, Phillip A.

    2015-01-01

    The CRISPR-Cas9 system, naturally a defense mechanism in prokaryotes, has been repurposed as an RNA-guided DNA targeting platform. It has been widely used for genome editing and transcriptome modulation, and has shown great promise in correcting mutations in human genetic diseases. Off-target effects are a critical issue for all of these applications. Here we review the current status on the target specificity of the CRISPR-Cas9 system. PMID:25722925

  9. Involvement of the CasK/R two-component system in optimal unsaturation of the Bacillus cereus fatty acids during low-temperature growth.

    PubMed

    Diomandé, Sara Esther; Nguyen-the, Christophe; Abee, Tjakko; Tempelaars, Marcel H; Broussolle, Véronique; Brillard, Julien

    2015-11-20

    Bacillus cereus sensu lato is composed of a set of ubiquitous strains including human pathogens that can survive a range of food processing conditions, grow in refrigerated food, and sometimes cause food poisoning. We previously identified the two-component system CasK/R that plays a key role in cold adaptation. To better understand the CasK/R-controlled mechanisms that support low-temperature adaptation, we performed a transcriptomic analysis on the ATCC 14579 strain and its isogenic ∆casK/R mutant grown at 12°C. Several genes involved in fatty acid (FA) metabolism were downregulated in the mutant, including desA and desB encoding FA acyl-lipid desaturases that catalyze the formation of a double-bond on the FA chain in positions ∆5 and ∆10, respectively. A lower proportion of FAs presumably unsaturated by DesA was observed in the ΔcasK/R strain compared to the parental strain while no difference was found for FAs presumably unsaturated by DesB. Addition of phospholipids from egg yolk lecithin rich in unsaturated FAs, to growth medium, abolished the cold-growth impairment of ΔcasK/R suggesting that exogenous unsaturated FAs can support membrane-level modifications and thus compensate for the decreased production of these FAs in the B. cereus ∆casK/R mutant during growth at low temperature. Our findings indicate that CasK/R is involved in the regulation of FA metabolism, and is necessary for cold adaptation of B. cereus unless an exogenous source of unsaturated FAs is available. PMID:25987542

  10. Rational design of a split-Cas9 enzyme complex

    DOE PAGESBeta

    Wright, Addison V.; Sternberg, Samuel H.; Taylor, David W.; Staahl, Brett T.; Bardales, Jorge A.; Kornfeld, Jack E.; Doudna, Jennifer A.

    2015-02-23

    Cas9, an RNA-guided DNA endonuclease found in clustered regularly interspaced short palindromic repeats (CRISPR) bacterial immune systems, is a versatile tool for genome editing, transcriptional regulation, and cellular imaging applications. Structures of Streptococcus pyogenes Cas9 alone or bound to single-guide RNA (sgRNA) and target DNA revealed a bilobed protein architecture that undergoes major conformational changes upon guide RNA and DNA binding. To investigate the molecular determinants and relevance of the interlobe rearrangement for target recognition and cleavage, we designed a split-Cas9 enzyme in which the nuclease lobe and α-helical lobe are expressed as separate polypeptides. The lobes do not interactmore » on their own, the sgRNA recruits them into a ternary complex that recapitulates the activity of full-length Cas9 and catalyzes site-specific DNA cleavage. The use of a modified sgRNA abrogates split-Cas9 activity by preventing dimerization, allowing for the development of an inducible dimerization system. We propose that split-Cas9 can act as a highly regulatable platform for genome-engineering applications.« less

  11. Internal guide RNA interactions interfere with Cas9-mediated cleavage.

    PubMed

    Thyme, Summer B; Akhmetova, Laila; Montague, Tessa G; Valen, Eivind; Schier, Alexander F

    2016-01-01

    The CRISPR/Cas system uses guide RNAs (gRNAs) to direct sequence-specific DNA cleavage. Not every gRNA elicits cleavage and the mechanisms that govern gRNA activity have not been resolved. Low activity could result from either failure to form a functional Cas9-gRNA complex or inability to recognize targets in vivo. Here we show that both phenomena influence Cas9 activity by comparing mutagenesis rates in zebrafish embryos with in vitro cleavage assays. In vivo, our results suggest that genomic factors such as CTCF inhibit mutagenesis. Comparing near-identical gRNA sequences with different in vitro activities reveals that internal gRNA interactions reduce cleavage. Even though gRNAs containing these structures do not yield cleavage-competent complexes, they can compete with active gRNAs for binding to Cas9. These results reveal that both genomic context and internal gRNA interactions can interfere with Cas9-mediated cleavage and illuminate previously uncharacterized features of Cas9-gRNA complex formation. PMID:27282953

  12. CRISPR-Cas9 Based Engineering of Actinomycetal Genomes.

    PubMed

    Tong, Yaojun; Charusanti, Pep; Zhang, Lixin; Weber, Tilmann; Lee, Sang Yup

    2015-09-18

    Bacteria of the order Actinomycetales are one of the most important sources of pharmacologically active and industrially relevant secondary metabolites. Unfortunately, many of them are still recalcitrant to genetic manipulation, which is a bottleneck for systematic metabolic engineering. To facilitate the genetic manipulation of actinomycetes, we developed a highly efficient CRISPR-Cas9 system to delete gene(s) or gene cluster(s), implement precise gene replacements, and reversibly control gene expression in actinomycetes. We demonstrate our system by targeting two genes, actIORF1 (SCO5087) and actVB (SCO5092), from the actinorhodin biosynthetic gene cluster in Streptomyces coelicolor A3(2). Our CRISPR-Cas9 system successfully inactivated the targeted genes. When no templates for homology-directed repair (HDR) were present, the site-specific DNA double-strand breaks (DSBs) introduced by Cas9 were repaired through the error-prone nonhomologous end joining (NHEJ) pathway, resulting in a library of deletions with variable sizes around the targeted sequence. If templates for HDR were provided at the same time, precise deletions of the targeted gene were observed with near 100% frequency. Moreover, we developed a system to efficiently and reversibly control expression of target genes, deemed CRISPRi, based on a catalytically dead variant of Cas9 (dCas9). The CRISPR-Cas9 based system described here comprises a powerful and broadly applicable set of tools to manipulate actinomycetal genomes. PMID:25806970

  13. Rational design of a split-Cas9 enzyme complex

    PubMed Central

    Wright, Addison V.; Sternberg, Samuel H.; Taylor, David W.; Staahl, Brett T.; Bardales, Jorge A.; Kornfeld, Jack E.; Doudna, Jennifer A.

    2015-01-01

    Cas9, an RNA-guided DNA endonuclease found in clustered regularly interspaced short palindromic repeats (CRISPR) bacterial immune systems, is a versatile tool for genome editing, transcriptional regulation, and cellular imaging applications. Structures of Streptococcus pyogenes Cas9 alone or bound to single-guide RNA (sgRNA) and target DNA revealed a bilobed protein architecture that undergoes major conformational changes upon guide RNA and DNA binding. To investigate the molecular determinants and relevance of the interlobe rearrangement for target recognition and cleavage, we designed a split-Cas9 enzyme in which the nuclease lobe and α-helical lobe are expressed as separate polypeptides. Although the lobes do not interact on their own, the sgRNA recruits them into a ternary complex that recapitulates the activity of full-length Cas9 and catalyzes site-specific DNA cleavage. The use of a modified sgRNA abrogates split-Cas9 activity by preventing dimerization, allowing for the development of an inducible dimerization system. We propose that split-Cas9 can act as a highly regulatable platform for genome-engineering applications. PMID:25713377

  14. Internal guide RNA interactions interfere with Cas9-mediated cleavage

    PubMed Central

    Thyme, Summer B.; Akhmetova, Laila; Montague, Tessa G.; Valen, Eivind; Schier, Alexander F.

    2016-01-01

    The CRISPR/Cas system uses guide RNAs (gRNAs) to direct sequence-specific DNA cleavage. Not every gRNA elicits cleavage and the mechanisms that govern gRNA activity have not been resolved. Low activity could result from either failure to form a functional Cas9–gRNA complex or inability to recognize targets in vivo. Here we show that both phenomena influence Cas9 activity by comparing mutagenesis rates in zebrafish embryos with in vitro cleavage assays. In vivo, our results suggest that genomic factors such as CTCF inhibit mutagenesis. Comparing near-identical gRNA sequences with different in vitro activities reveals that internal gRNA interactions reduce cleavage. Even though gRNAs containing these structures do not yield cleavage-competent complexes, they can compete with active gRNAs for binding to Cas9. These results reveal that both genomic context and internal gRNA interactions can interfere with Cas9-mediated cleavage and illuminate previously uncharacterized features of Cas9–gRNA complex formation. PMID:27282953

  15. Rational design of a split-Cas9 enzyme complex

    SciTech Connect

    Wright, Addison V.; Sternberg, Samuel H.; Taylor, David W.; Staahl, Brett T.; Bardales, Jorge A.; Kornfeld, Jack E.; Doudna, Jennifer A.

    2015-02-23

    Cas9, an RNA-guided DNA endonuclease found in clustered regularly interspaced short palindromic repeats (CRISPR) bacterial immune systems, is a versatile tool for genome editing, transcriptional regulation, and cellular imaging applications. Structures of Streptococcus pyogenes Cas9 alone or bound to single-guide RNA (sgRNA) and target DNA revealed a bilobed protein architecture that undergoes major conformational changes upon guide RNA and DNA binding. To investigate the molecular determinants and relevance of the interlobe rearrangement for target recognition and cleavage, we designed a split-Cas9 enzyme in which the nuclease lobe and α-helical lobe are expressed as separate polypeptides. The lobes do not interact on their own, the sgRNA recruits them into a ternary complex that recapitulates the activity of full-length Cas9 and catalyzes site-specific DNA cleavage. The use of a modified sgRNA abrogates split-Cas9 activity by preventing dimerization, allowing for the development of an inducible dimerization system. We propose that split-Cas9 can act as a highly regulatable platform for genome-engineering applications.

  16. Monodisperse CoFe2O4 nanoparticles supported on Vulcan XC-72: High performance electrode materials for lithium-air and lithium-ion batteries

    NASA Astrophysics Data System (ADS)

    Şener, Tansel; Kayhan, Emine; Sevim, Melike; Metin, Önder

    2015-08-01

    Addressed herein is the preparation and the electrode performance of monodisperse CoFe2O4 nanoparticles (NPs) supported on Vulcan XC-72 for the Lithium-air battery (LAB) and Lithium-ion battery (LIB). Monodisperse CoFe2O4 NPs were synthesized by the thermal decomposition of cobalt(II) acetylacetonate and iron(III) acetylacetonate in oleylamine and oleic acid in the presence of 1,2-tetradecanediol and benzyl ether. As-prepared CoFe2O4 NPs with a particle size of 11 nm were then supported on Vulcan XC-72 (Vulcan-CoFe2O4) at different theoretical loadings (20, 40 and 60 wt % CoFe2O4 NPs) by using the simple liquid phase self assembly method. CoFe2O4 NPs dispersed on Vulcan-CoFe2O4 composites were characterized by transmission electron microscopy (TEM), powder X-ray diffraction (PXRD) and atomic absorption spectroscopy (AAS). The AAS analyses indicated that the Vulcan-CoFe2O4 composites with different loadings were included 3.7, 8.1 and 16.4 wt % CoFe2O4 on the metal basis. The electrode performance of Vulcan-CoFe2O4 composites were evaluated as the anode active material for LIB and cathode active material for LABs by performing the galvanostatic charge-discharge tests. The highest discharge capacity for both LAB (7510 mAh g(Vulcan+CoFe2O4)-1; 13380 mAh gCoFe2O4-1 @ 0.1C) and LIB (863 mAh g(Vulcan+CoFe2O4)-1; 9330 mAh gCoFe2O4-1@ 0.1C) was investigated with 16.4 wt % CoFe2O4.

  17. Highly specific targeted mutagenesis in plants using Staphylococcus aureus Cas9

    PubMed Central

    Kaya, Hidetaka; Mikami, Masafumi; Endo, Akira; Endo, Masaki; Toki, Seiichi

    2016-01-01

    The CRISPR/Cas9 system is an efficient and convenient tool for genome editing in plants. Cas9 nuclease derived from Streptococcus pyogenes (Sp) is commonly used in this system. Recently, Staphylococcus aureus Cas9 (SaCas9)-mediated genome editing was reported in human cells and Arabidopsis. Because SaCas9 (1053 a.a.) is smaller than SpCas9 (1368 a.a.), SaCas9 could have substantial advantages for delivering and expressing Cas9 protein, especially using virus vectors. Since the protospacer adjacent motif (PAM) sequence of SaCas9 (5′-NNGRRT-3′) differs from that of SpCas9 (5′-NGG-3′), the use of this alternative Cas9 nuclease could expand the selectivity at potential cleavage target sites of the CRISPR/Cas9 system. Here we show that SaCas9 can mutagenize target sequences in tobacco and rice with efficiencies similar to those of SpCas9. We also analyzed the base preference for ‘T’ at the 6th position of the SaCas9 PAM. Targeted mutagenesis efficiencies in target sequences with non-canonical PAMs (5′-NNGRRV-3′) were much lower than those with a canonical PAM (5′-NNGRRT-3′). The length of target sequence recognized by SaCas9 is one or two nucleotides longer than that recognized by SpCas9. Taken together, our results demonstrate that SaCas9 has higher sequence recognition capacity than SpCas9 and is useful for reducing off-target mutations in crop. PMID:27226350

  18. Highly specific targeted mutagenesis in plants using Staphylococcus aureus Cas9.

    PubMed

    Kaya, Hidetaka; Mikami, Masafumi; Endo, Akira; Endo, Masaki; Toki, Seiichi

    2016-01-01

    The CRISPR/Cas9 system is an efficient and convenient tool for genome editing in plants. Cas9 nuclease derived from Streptococcus pyogenes (Sp) is commonly used in this system. Recently, Staphylococcus aureus Cas9 (SaCas9)-mediated genome editing was reported in human cells and Arabidopsis. Because SaCas9 (1053 a.a.) is smaller than SpCas9 (1368 a.a.), SaCas9 could have substantial advantages for delivering and expressing Cas9 protein, especially using virus vectors. Since the protospacer adjacent motif (PAM) sequence of SaCas9 (5'-NNGRRT-3') differs from that of SpCas9 (5'-NGG-3'), the use of this alternative Cas9 nuclease could expand the selectivity at potential cleavage target sites of the CRISPR/Cas9 system. Here we show that SaCas9 can mutagenize target sequences in tobacco and rice with efficiencies similar to those of SpCas9. We also analyzed the base preference for 'T' at the 6th position of the SaCas9 PAM. Targeted mutagenesis efficiencies in target sequences with non-canonical PAMs (5'-NNGRRV-3') were much lower than those with a canonical PAM (5'-NNGRRT-3'). The length of target sequence recognized by SaCas9 is one or two nucleotides longer than that recognized by SpCas9. Taken together, our results demonstrate that SaCas9 has higher sequence recognition capacity than SpCas9 and is useful for reducing off-target mutations in crop. PMID:27226350

  19. Tolerance of chufa (Cyperus esculentus L.) plants, representing the higher plant compartment in bioregenerative life support systems, to super-optimal air temperatures

    NASA Astrophysics Data System (ADS)

    Shklavtsova, E. S.; Ushakova, S. A.; Shikhov, V. N.; Anishchenko, O. V.

    2013-01-01

    Plants intended to be included in the photosynthesizing compartment of the bioregenerative life support system (BLSS) need to be studied in terms of both their production parameters under optimal conditions and their tolerance to stress factors that might be caused by emergency situations. The purpose of this study was to investigate tolerance of chufa (Cyperus esculentus L.) plants to the super-optimal air temperature of 45 ± 1 °C as dependent upon PAR (photosynthetically active radiation) intensity and the duration of the exposure to the stress factor. Chufa plants were grown hydroponically, on expanded clay, under artificial light. The nutrient solution was Knop's mineral medium. Until the plants were 30 days old, they had been grown at 690 μmol m-2 s-1 PAR and air temperature 25 °C. Thirty-day-old plants were exposed to the temperature 45 °C for 6 h, 20 h, and 44 h at PAR intensities 690 μmol m-2 s-1 and 1150 μmol m-2 s-1. The exposure to the damaging air temperature for 44 h at 690 μmol m-2 s-1 PAR caused irreversible damage to PSA, resulting in leaf mortality. In chufa plants exposed to heat shock treatment at 690 μmol m-2 s-1 PAR for 6 h and 20 h, respiration exceeded photosynthesis, and CO2 release in the light was recorded. Functional activity of photosynthetic apparatus, estimated from parameters of pulse-modulated chlorophyll fluorescence in Photosystem 2 (PS 2), decreased 40% to 50%. After the exposure to the stress factor was finished, functional activity of PSA recovered its initial values, and apparent photosynthesis (Papparent) rate after a 20-h exposure to the stress factor was 2.6 times lower than before the elevation of the temperature. During the first hours of plant exposure to the temperature 45 °C at 1150 μmol m-2 s-1 PAR, respiration rate was higher than photosynthesis rate, but after 3-4 h of the exposure, photosynthetic processes exceeded oxidative ones and CO2 absorption in the light was recorded. At the end of the 6-h exposure

  20. Multiplex gene editing via CRISPR/Cas9 exhibits desirable muscle hypertrophy without detectable off-target effects in sheep

    PubMed Central

    Wang, Xiaolong; Niu, Yiyuan; Zhou, Jiankui; Yu, Honghao; Kou, Qifang; Lei, Anmin; Zhao, Xiaoe; Yan, Hailong; Cai, Bei; Shen, Qiaoyan; Zhou, Shiwei; Zhu, Haijing; Zhou, Guangxian; Niu, Wenzhi; Hua, Jinlian; Jiang, Yu; Huang, Xingxu; Ma, Baohua; Chen, Yulin

    2016-01-01

    The CRISPR/Cas9 system provides a flexible approach for genome engineering of genetic loci. Here, we successfully achieved precise gene targeting in sheep by co-injecting one-cell-stage embryos with Cas9 mRNA and RNA guides targeting three genes (MSTN, ASIP, and BCO2). We carefully examined the sgRNAs:Cas9-mediated targeting effects in injected embryos, somatic tissues, as well as gonads via cloning and sequencing. The targeting efficiencies in these three genes were within the range of 27–33% in generated lambs, and that of simultaneously targeting the three genes was 5.6%, which demonstrated that micro-injection of zygotes is an efficient approach for generating gene-modified sheep. Interestingly, we observed that disruption of the MSTN gene resulted in the desired muscle hypertrophy that is characterized by enlarged myofibers, thereby providing the first detailed evidence supporting that gene modifications had occurred at both the genetic and morphological levels. In addition, prescreening for the off-target effect of sgRNAs was performed on fibroblasts before microinjection, to ensure that no detectable off-target mutations from founder animals existed. Our findings suggested that the CRISPR/Cas9 method can be exploited as a powerful tool for livestock improvement by simultaneously targeting multiple genes that are responsible for economically significant traits. PMID:27562433

  1. Multiplex gene editing via CRISPR/Cas9 exhibits desirable muscle hypertrophy without detectable off-target effects in sheep.

    PubMed

    Wang, Xiaolong; Niu, Yiyuan; Zhou, Jiankui; Yu, Honghao; Kou, Qifang; Lei, Anmin; Zhao, Xiaoe; Yan, Hailong; Cai, Bei; Shen, Qiaoyan; Zhou, Shiwei; Zhu, Haijing; Zhou, Guangxian; Niu, Wenzhi; Hua, Jinlian; Jiang, Yu; Huang, Xingxu; Ma, Baohua; Chen, Yulin

    2016-01-01

    The CRISPR/Cas9 system provides a flexible approach for genome engineering of genetic loci. Here, we successfully achieved precise gene targeting in sheep by co-injecting one-cell-stage embryos with Cas9 mRNA and RNA guides targeting three genes (MSTN, ASIP, and BCO2). We carefully examined the sgRNAs:Cas9-mediated targeting effects in injected embryos, somatic tissues, as well as gonads via cloning and sequencing. The targeting efficiencies in these three genes were within the range of 27-33% in generated lambs, and that of simultaneously targeting the three genes was 5.6%, which demonstrated that micro-injection of zygotes is an efficient approach for generating gene-modified sheep. Interestingly, we observed that disruption of the MSTN gene resulted in the desired muscle hypertrophy that is characterized by enlarged myofibers, thereby providing the first detailed evidence supporting that gene modifications had occurred at both the genetic and morphological levels. In addition, prescreening for the off-target effect of sgRNAs was performed on fibroblasts before microinjection, to ensure that no detectable off-target mutations from founder animals existed. Our findings suggested that the CRISPR/Cas9 method can be exploited as a powerful tool for livestock improvement by simultaneously targeting multiple genes that are responsible for economically significant traits. PMID:27562433

  2. CRISPR/Cas9 somatic multiplex-mutagenesis for high-throughput functional cancer genomics in mice.

    PubMed

    Weber, Julia; Öllinger, Rupert; Friedrich, Mathias; Ehmer, Ursula; Barenboim, Maxim; Steiger, Katja; Heid, Irina; Mueller, Sebastian; Maresch, Roman; Engleitner, Thomas; Gross, Nina; Geumann, Ulf; Fu, Beiyuan; Segler, Angela; Yuan, Detian; Lange, Sebastian; Strong, Alexander; de la Rosa, Jorge; Esposito, Irene; Liu, Pentao; Cadiñanos, Juan; Vassiliou, George S; Schmid, Roland M; Schneider, Günter; Unger, Kristian; Yang, Fengtang; Braren, Rickmer; Heikenwälder, Mathias; Varela, Ignacio; Saur, Dieter; Bradley, Allan; Rad, Roland

    2015-11-10

    Here, we show CRISPR/Cas9-based targeted somatic multiplex-mutagenesis and its application for high-throughput analysis of gene function in mice. Using hepatic single guide RNA (sgRNA) delivery, we targeted large gene sets to induce hepatocellular carcinoma (HCC) and intrahepatic cholangiocarcinoma (ICC). We observed Darwinian selection of target genes, which suppress tumorigenesis in the respective cellular/tissue context, such as Pten or Cdkn2a, and conversely found low frequency of Brca1/2 alterations, explaining mutational spectra in human ICC/HCC. Our studies show that multiplexed CRISPR/Cas9 can be used for recessive genetic screening or high-throughput cancer gene validation in mice. The analysis of CRISPR/Cas9-induced tumors provided support for a major role of chromatin modifiers in hepatobiliary tumorigenesis, including that of ARID family proteins, which have recently been reported to be mutated in ICC/HCC. We have also comprehensively characterized the frequency and size of chromosomal alterations induced by combinatorial sgRNA delivery and describe related limitations of CRISPR/Cas9 multiplexing, as well as opportunities for chromosome engineering in the context of hepatobiliary tumorigenesis. Our study describes novel approaches to model and study cancer in a high-throughput multiplexed format that will facilitate the functional annotation of cancer genomes. PMID:26508638

  3. Correction of a genetic disease by CRISPR-Cas9-mediated gene editing in mouse spermatogonial stem cells

    PubMed Central

    Wu, Yuxuan; Zhou, Hai; Fan, Xiaoying; Zhang, Ying; Zhang, Man; Wang, Yinghua; Xie, Zhenfei; Bai, Meizhu; Yin, Qi; Liang, Dan; Tang, Wei; Liao, Jiaoyang; Zhou, Chikai; Liu, Wujuan; Zhu, Ping; Guo, Hongshan; Pan, Hong; Wu, Chunlian; Shi, Huijuan; Wu, Ligang; Tang, Fuchou; Li, Jinsong

    2015-01-01

    Spermatogonial stem cells (SSCs) can produce numerous male gametes after transplantation into recipient testes, presenting a valuable approach for gene therapy and continuous production of gene-modified animals. However, successful genetic manipulation of SSCs has been limited, partially due to complexity and low efficiency of currently available genetic editing techniques. Here, we show that efficient genetic modifications can be introduced into SSCs using the CRISPR-Cas9 system. We used the CRISPR-Cas9 system to mutate an EGFP transgene or the endogenous Crygc gene in SCCs. The mutated SSCs underwent spermatogenesis after transplantation into the seminiferous tubules of infertile mouse testes. Round spermatids were generated and, after injection into mature oocytes, supported the production of heterozygous offspring displaying the corresponding mutant phenotypes. Furthermore, a disease-causing mutation in Crygc (Crygc−/−) that pre-existed in SSCs could be readily repaired by CRISPR-Cas9-induced nonhomologous end joining (NHEJ) or homology-directed repair (HDR), resulting in SSC lines carrying the corrected gene with no evidence of off-target modifications as shown by whole-genome sequencing. Fertilization using round spermatids generated from these lines gave rise to offspring with the corrected phenotype at an efficiency of 100%. Our results demonstrate efficient gene editing in mouse SSCs by the CRISPR-Cas9 system, and provide the proof of principle of curing a genetic disease via gene correction in SSCs. PMID:25475058

  4. Treatability study in support of monitored natural attenuation at Site S-1, Zone 5, Kelly Air Force Base, Texas. Final report January--December 1998

    SciTech Connect

    Herrington, T.; Downey, D.

    1999-02-28

    This report presents the results of a treatability study (TS) to evaluate the potential effectiveness of monitored natural attenuation (MNA) as a remedial option for groundwater contaminated with chlorinated benzene compounds at Site S-1, located at Kelly Air Force Base (AFB), Texas. Although other contaminants were found at Site S-1 at relatively low concentrations, this TS will focus on the chlorinated benzene compounds present in the groundwater plume. Hydrogeologic and groundwater chemical data collected for this report can be used to evaluate the effectiveness of various engineered remedial options; however, the results of this TS will be used in support of MNA with long term monitoring (LTM) for restoration of groundwater contaminated with chlorinated benzene compounds. The work performed as part of the TS is not intended to fulfill the requirements of a contamination assessment report, a remedial action plan (RAP), or any other document specified in federal or state regulations; rather, it is provided for the use by the Base, its prime environmental contractors, and regulators to present information on the viability of the MNA alternative for chlorobenzene residuals at Site S-1.

  5. Creating Locally-Resolved Mobile-Source Emissions Inputs for Air Quality Modeling in Support of an Exposure Study in Detroit, Michigan, USA

    PubMed Central

    Snyder, Michelle; Arunachalam, Saravanan; Isakov, Vlad; Talgo, Kevin; Naess, Brian; Valencia, Alejandro; Omary, Mohammad; Davis, Neil; Cook, Rich; Hanna, Adel

    2014-01-01

    This work describes a methodology for modeling the impact of traffic-generated air pollutants in an urban area. This methodology presented here utilizes road network geometry, traffic volume, temporal allocation factors, fleet mixes, and emission factors to provide critical modeling inputs. These inputs, assembled from a variety of sources, are combined with meteorological inputs to generate link-based emissions for use in dispersion modeling to estimate pollutant concentration levels due to traffic. A case study implementing this methodology for a large health study is presented, including a sensitivity analysis of the modeling results reinforcing the importance of model inputs and identify those having greater relative impact, such as fleet mix. In addition, an example use of local measurements of fleet activity to supplement model inputs is described, and its impacts to the model outputs are discussed. We conclude that with detailed model inputs supported by local traffic measurements and meteorology, it is possible to capture the spatial and temporal patterns needed to accurately estimate exposure from traffic-related pollutants. PMID:25501000

  6. Comparison of Predicted and Experimental Heat-Transfer and Pressure-Drop Results for an Air-Cooled Plug Nozzle and Supporting Struts

    NASA Technical Reports Server (NTRS)

    Graber, E. J., Jr.; Clark, J. S.

    1972-01-01

    A calculational procedure is presented to analyze the heat-transfer and fluid-flow characteristics of a convectively air -cooled plug-nozzle operating on an afterburning turbojet engine. Anderson's method was used to predict hot-gas static pressures in the supersonic stream with fully expanded flow (high nozzle-pressure ratios); the results were excellent. For low nozzle-pressure ratios, the flow was assumed to expand one-dimensionally and isentropically to the plug back pressure. Wall temperatures predicted using this latter pressure distribution agreed well with the wall temperatures predicted using the measured hot-gas pressures (maximum deviation was about 30 K (54 deg R)). Either an in tegral boundary-layer technique or a simple pipe-flow equation may be used to calculate convective heat transfer from the hot gas to the wall. The simple pipeflow equation results in the prediction of slightly higher wall temperatures than does the integral technique. Experimental wall temperatures were generally in good agreement with the two predicted wall temperature distributions. Excellent agreement was noted b etween measured and predicted coolant static-pressure distributions. The plug-coolant temperature rise was generally overpredicted by about 22.2 K (40 deg R); possible explanations are offered. Although an an alysis of the struts, which support the plug, was purposely kept simple, reasonable results were obtained. Potential flow over an ellipse was used to calculate hot-gas static pressure; the results were satisfactory.

  7. Creating locally-resolved mobile-source emissions inputs for air quality modeling in support of an exposure study in Detroit, Michigan, USA.

    PubMed

    Snyder, Michelle; Arunachalam, Saravanan; Isakov, Vlad; Talgo, Kevin; Naess, Brian; Valencia, Alejandro; Omary, Mohammad; Davis, Neil; Cook, Rich; Hanna, Adel

    2014-12-01

    This work describes a methodology for modeling the impact of traffic-generated air pollutants in an urban area. This methodology presented here utilizes road network geometry, traffic volume, temporal allocation factors, fleet mixes, and emission factors to provide critical modeling inputs. These inputs, assembled from a variety of sources,are combined with meteorological inputs to generate link-based emissions for use in dispersion modeling to estimate pollutant concentration levels due to traffic. A case study implementing this methodology for a large health study is presented, including a sensitivity analysis of the modeling results reinforcing the importance of model inputs and identify those having greater relative impact, such as fleet mix. In addition, an example use of local measurements of fleet activity to supplement model inputs is described, and its impacts to the model outputs are discussed. We conclude that with detailed model inputs supported by local traffic measurements and meteorology, it is possible to capture the spatial and temporal patterns needed to accurately estimate exposure from traffic-related pollutants. PMID:25587603

  8. Creating locally-resolved mobile-source emissions inputs for air quality modeling in support of an exposure study in Detroit, Michigan, USA.

    PubMed

    Snyder, Michelle; Arunachalam, Saravanan; Isakov, Vlad; Talgo, Kevin; Naess, Brian; Valencia, Alejandro; Omary, Mohammad; Davis, Neil; Cook, Rich; Hanna, Adel

    2014-01-01

    This work describes a methodology for modeling the impact of traffic-generated air pollutants in an urban area. This methodology presented here utilizes road network geometry, traffic volume, temporal allocation factors, fleet mixes, and emission factors to provide critical modeling inputs. These inputs, assembled from a variety of sources, are combined with meteorological inputs to generate link-based emissions for use in dispersion modeling to estimate pollutant concentration levels due to traffic. A case study implementing this methodology for a large health study is presented, including a sensitivity analysis of the modeling results reinforcing the importance of model inputs and identify those having greater relative impact, such as fleet mix. In addition, an example use of local measurements of fleet activity to supplement model inputs is described, and its impacts to the model outputs are discussed. We conclude that with detailed model inputs supported by local traffic measurements and meteorology, it is possible to capture the spatial and temporal patterns needed to accurately estimate exposure from traffic-related pollutants. PMID:25501000

  9. A Sounding-based Severe Weather Tool to Support Daily Operations at Kennedy Space Center and Cape Canaveral Air Force Station

    NASA Technical Reports Server (NTRS)

    Bauman, William H.; Roeder, William P.

    2014-01-01

    People and property at Kennedy Space Center (KSC) and Cape Canaveral Air Force Station (CCAFS) are at risk when severe weather occurs. Strong winds, hail and tornadoes can injure individuals and cause costly damage to structures if not properly protected. NASA's Launch Services Program and Ground Systems Development and Operations Program and other KSC programs use the daily and weekly severe weather forecasts issued by the 45th Weather Squadron (45 WS) to determine if they need to limit an activity such as working on gantries, or protect property such as a vehicle on a pad. The 45 WS requested the Applied Meteorology Unit (AMU) develop a warm season (May-September) severe weather tool for use in the Meteorological Interactive Data Display System (MIDDS) based on the late morning, 1500 UTC (1100 local time), CCAFS (XMR) sounding. The 45 WS frequently makes decisions to issue a severe weather watch and other severe weather warning support products to NASA and the 45th Space Wing in the late morning, after the 1500 UTC sounding. The results of this work indicate that certain stability indices based on the late morning XMR soundings can depict differences between days with reported severe weather and days with no reported severe weather. The AMU determined a frequency of reported severe weather for the stability indices and implemented an operational tool in MIDDS.

  10. United States Air Force 611th Air Support Group/Civil Engineering Squadron, Elmendorf AFB, Alaska. Decision document for no further response action planned Oliktok Point Radar Installation, Alaska. Final report

    SciTech Connect

    Karmi, S.

    1996-06-03

    This Decision Document discusses the selection of no further action as the recommended action for four sites located at the Oliktok Point radar installation. The United States Air Force (Air Force) completed a Remedial Investigation/Feasibility Study and a Risk Assessment for the eight sites located at the Oliktok Point installation (U.S. Air Force 1996a,b). Based on the findings of these activities, four sites are recommended for no further action.

  11. United States Air Force 611th Air Support Group/Civil Engineering Squadron Elmendorf AFB, Alaska. Decision document for no further response action planned: Barter Island Radar Installation, Alaska. Final report, December 1995-May 1996

    SciTech Connect

    Karmi, S.; Madden, J.; Borsetti, R.

    1996-05-03

    This Decision Document discusses the selection of no further action as the recommended action for nine sites located at the Barter Island radar installation. The United States Air Force (Air Force) completed a Remedial Investigation/Feasibility Study and a Risk Assessment for the 14 sites located at the Barter Island installation (U.S. Air Force 1996a,b). Based on the findings of these activities, nine sites are recommended for no further action.

  12. A non-inheritable maternal Cas9-based multiple-gene editing system in mice

    PubMed Central

    Sakurai, Takayuki; Kamiyoshi, Akiko; Kawate, Hisaka; Mori, Chie; Watanabe, Satoshi; Tanaka, Megumu; Uetake, Ryuichi; Sato, Masahiro; Shindo, Takayuki

    2016-01-01

    The CRISPR/Cas9 system is capable of editing multiple genes through one-step zygote injection. The preexisting method is largely based on the co-injection of Cas9 DNA (or mRNA) and guide RNAs (gRNAs); however, it is unclear how many genes can be simultaneously edited by this method, and a reliable means to generate transgenic (Tg) animals with multiple gene editing has yet to be developed. Here, we employed non-inheritable maternal Cas9 (maCas9) protein derived from Tg mice with systemic Cas9 overexpression (Cas9 mice). The maCas9 protein in zygotes derived from mating or in vitro fertilization of Tg/+ oocytes and +/+ sperm could successfully edit the target genome. The efficiency of such maCas9-based genome editing was comparable to that of zygote microinjection–based genome editing widely used at present. Furthermore, we demonstrated a novel approach to create “Cas9 transgene-free” gene-modified mice using non-Tg (+/+) zygotes carrying maCas9. The maCas9 protein in mouse zygotes edited nine target loci simultaneously after injection with nine different gRNAs alone. Cas9 mouse-derived zygotes have the potential to facilitate the creation of genetically modified animals carrying the Cas9 transgene, enabling repeatable genome engineering and the production of Cas9 transgene-free mice. PMID:26817415

  13. A non-inheritable maternal Cas9-based multiple-gene editing system in mice.

    PubMed

    Sakurai, Takayuki; Kamiyoshi, Akiko; Kawate, Hisaka; Mori, Chie; Watanabe, Satoshi; Tanaka, Megumu; Uetake, Ryuichi; Sato, Masahiro; Shindo, Takayuki

    2016-01-01

    The CRISPR/Cas9 system is capable of editing multiple genes through one-step zygote injection. The preexisting method is largely based on the co-injection of Cas9 DNA (or mRNA) and guide RNAs (gRNAs); however, it is unclear how many genes can be simultaneously edited by this method, and a reliable means to generate transgenic (Tg) animals with multiple gene editing has yet to be developed. Here, we employed non-inheritable maternal Cas9 (maCas9) protein derived from Tg mice with systemic Cas9 overexpression (Cas9 mice). The maCas9 protein in zygotes derived from mating or in vitro fertilization of Tg/+ oocytes and +/+ sperm could successfully edit the target genome. The efficiency of such maCas9-based genome editing was comparable to that of zygote microinjection-based genome editing widely used at present. Furthermore, we demonstrated a novel approach to create "Cas9 transgene-free" gene-modified mice using non-Tg (+/+) zygotes carrying maCas9. The maCas9 protein in mouse zygotes edited nine target loci simultaneously after injection with nine different gRNAs alone. Cas9 mouse-derived zygotes have the potential to facilitate the creation of genetically modified animals carrying the Cas9 transgene, enabling repeatable genome engineering and the production of Cas9 transgene-free mice. PMID:26817415

  14. Zinc-air battery: understanding the structure and morphology changes of graphene-supported CoMn(2)O(4) bifunctional catalysts under practical rechargeable conditions.

    PubMed

    Prabu, Moni; Ramakrishnan, Prakash; Nara, Hiroki; Momma, Toshiyuki; Osaka, Tetsuya; Shanmugam, Sangaraju

    2014-10-01

    Nitrogen-doped/undoped thermally reduced graphene oxide (N-rGO) decorated with CoMn2O4 (CMO) nanoparticles were synthesized using a simple one-step hydrothermal method. The activity and stability of this hybrid catalyst were evaluated by preparing air electrodes with both primary and rechargeable zinc-air batteries that consume ambient air. Further, we investigated the relationship between the physical properties and the electrochemical results for hybrid electrodes at various cycles using X-ray diffraction, scanning electron microscopy, galvanodynamic charge-discharging and electrochemical impedance spectroscopy. The structural, morphological and electrocatalytic performances confirm that CMO/N-rGO is a promising material for safe, reliable, and long-lasting air cathodes for both primary and rechargeable zinc-air batteries that consume air under ambient condition. PMID:25192132

  15. The Climate Analysis Seoul (CAS) simulation for thermal characteristics in different urban locations in Seoul, Korea

    NASA Astrophysics Data System (ADS)

    Lee, Young-Gon; Lee, Jisu; Kim, Baek-Jo

    2016-04-01

    The Climate Analysis Seoul (CAS) being tailored for simulating detailed climate was applied to investigate the temperature distribution around the Seonjeongneung Royal Tombs that includes forest and grassland and in the surrounding high-density buildings in Seoul, Korea for a hot summer days of August 6-7, 2015. In this study, we used the digitalized topographical and land-use data of the 5 m resolution and wind and air temperature fields of 100 m resolution produced by the numerical model MetPhoMod (MPM), part of the CAS for the area of interest with an area of 25,000,000 m2. The difference of the temperatures compared with the representative meteorological station of Seoul was analyzed by considering the presence or absence of the Seonjeongneung Royal Tombs. For the case of heat wave issued in Seoul from August 6th, 06 LST to 7th, 18 LST, 2015, the simulation results presented that averaged temperatures in the green area were 2 °C lower than the one of the standard weather station, whereas they showed 1.5 °C higher when the green area was replaced with the building area. From this simulation result, we can suggest that the green area has a temperature reduction effect of 1-3 °C and its influence can be extended up to 150 m from the boundary of the green area. This fact reconfirms that the urban green area generates a micro-scale cool-island effect and this local phenomenon can reduce heat stress produced by surplus thermal sources of the surrounding buildings and paved roads during the hot summer days. Key words: Climate Analysis Seoul (CAS), Urban Green Area, Cool-Island Effect

  16. Design and verification by nonlinear simulation of a Mach/CAS control law for the NASA TCV B737 aircraft

    NASA Technical Reports Server (NTRS)

    Bruce, Kevin R.

    1986-01-01

    A Mach/CAS control system using an elevator was designed and developed for use on the NASA TCV B737 aircraft to support research in profile descent procedures and approach energy management. The system was designed using linear analysis techniques primarily. The results were confirmed and the system validated at additional flight conditions using a nonlinear 737 aircraft simulation. All design requirements were satisfied.

  17. Tuning CAS Application using AIMS: An Automated Instrumentation and Monitoring System

    NASA Technical Reports Server (NTRS)

    Mehra, P.; Sarukkai, S.; Schmidt, M.; Schulbach, C.; VanVoorst, B.; Yan, Jerry; Woodrow, Thomas (Technical Monitor)

    1994-01-01

    To bring together NASA's scientists and engineers and their counterparts in industry, other government agencies, and academia working in the Computational AeroSciences (CAS) field. This workshop is part of the technology transfer plan of the High Performance Computing and Communications Program (HPCCP). Specific objectives of this Workshop are to: (1) communicate the goals and objectives of HPCCP in the area of CAS; (2) promote and disseminate CAS technology within the appropriate technical communities, including NASA, industry, academia, and other government labs; (3) help promote synergy among CAS scientists; and (4) permit feedback from peer researchers in issues pacing the CAS field in general and the HPCCP CAS program in particular.

  18. Spectroscopic studies of three Cepheids with high positive pulsation period increments: SZ Cas, BY Cas, and RU Sct

    NASA Astrophysics Data System (ADS)

    Usenko, I. A.; Klochkova, V. G.

    2015-07-01

    Three high-resolution spectra have been taken at different times with the 6-m SAO RAS telescope (LYNX and PFES spectrographs) for three Cepheids exhibiting high positive period increments: the small-amplitude (DCEPS) SZ Cas and BY Cas and the classical (DCEP) RU Sct. SZ Cas and RU Sct are members of the Galactic open clusters χ and h Per and Trump 35, respectively. Analysis of the spectra has shown that the interstellar Na I D1 and D2 lines in all objects are considerably stronger than the atmospheric ones and are redshifted in SZ Cas and BY Cas and blushifted in RU Sct. The core of the H α absorption line in BY Cas has an asymmetric knifelike shape, while RU Sct exhibits an intense emission in the blue wing of this line. Such phenomena are observed in long-period Cepheids and bright hypergiants with an extended envelope. In this case, the strong Mg Ib 5183.62 Å and Ba II 5853.67, 6141.713, and 6496.90 Å lines with low χlow in SZ Cas and RU Sct also show characteristic knifelike profiles with an asymmetry in the red region, while the Ba II 4934.095 Å line shows similar profiles in the blue one. The absorption lines of neutral atoms and singly ionized metals with different lowerlevel excitation potentials exhibit different degrees of asymmetry: from a pronounced one with secondary components in BY Cas (similar to those in the small-amplitude Cepheid BG Cru pulsating in the first overtone and having an envelope) to its insignificance or virtual absence in SZ Cas and RU Sct. Analysis of the secular changes in mean T eff determined from photometric color indices and spectra over the last 55 years for these stars has revealed periodic fluctuations of 200 K for SZ Cas and BY Cas and 500 K for RU Sct. For SZ Cas and RU Sct, T eff determined in some years from some color indices show much lower values, which together with the temperature fluctuations can be associated with mass loss and dust formation. Based on these facts, we hypothesize the existence of

  19. The Use of Small-Particle Sized TiO2 Supported on Clays as Photocatalytic Materials: A Low- Cost Alternative Technology for the Degradation of Air Pollutants

    NASA Astrophysics Data System (ADS)

    Kibanova, D.; Trejo, M.; Destaillats, H.; Cervini-Silva, J.

    2007-05-01

    Assisted photocatalysis by TiO2 is an advanced oxidation process that has been employed for air and water remediation. Clays are natural porous materials bearing high surface areas and interlayer spacing that allows entrapment of small-sized particles. Pillared clays exchanged with small-sized TiO2 can constitute materials with interesting photocatalytic properties because high surface area values and large contents of mesospores, which enables analyte trapping. Furthermore, intercalation at the clay interlayer enables TiO2 to become more resistant to aggregation when in solution. Just recently it has been reported that clays can lead to increases in the photocatalytic activity of TiO2 when the mesopores size is adequate to host organic solutes and ensure their effective interaction with the TiO2 particles. In this paper we study the photocatalytic properties of small-sized TiO2 supported on the following clay samples: Montmorillonite [SWy-2, Na0.2Ca0.1Al2Si4O10(OH)2(H2O)10 ] from Crook Country, Wyoming, USA; Hectorite [SHCa-1, Na0.4Mg2.7Li0.3Si4O10(OH)2 ] from San Bernardino. Country, California, USA; Kaolinite [KGa-1b, Al2Si2O5(OH)4 ] from Washington Country, Georgia, USA. Deposition of TiO2 on the clay surface was conducted by using a sol-gel synthetic method. Anatase TiO2 particles transformation at the clay interlayer was achieved by thermic treatment at 180 °C. Material characterization was conducted using FTIR microspectroscopy, Scanning Electron Microscopy (SEM), and XRD analysis. The organic compound used as probe was ethanol

  20. Workstation-Based Real-Time Mesoscale Modeling Designed for Weather Support to Operations at the Kennedy Space Center and Cape Canaveral Air Station

    NASA Technical Reports Server (NTRS)

    Manobianco, John; Zack, John W.; Taylor, Gregory E.

    1996-01-01

    This paper describes the capabilities and operational utility of a version of the Mesoscale Atmospheric Simulation System (MASS) that has been developed to support operational weather forecasting at the Kennedy Space Center (KSC) and Cape Canaveral Air Station (CCAS). The implementation of local, mesoscale modeling systems at KSC/CCAS is designed to provide detailed short-range (less than 24 h) forecasts of winds, clouds, and hazardous weather such as thunderstorms. Short-range forecasting is a challenge for daily operations, and manned and unmanned launches since KSC/CCAS is located in central Florida where the weather during the warm season is dominated by mesoscale circulations like the sea breeze. For this application, MASS has been modified to run on a Stardent 3000 workstation. Workstation-based, real-time numerical modeling requires a compromise between the requirement to run the system fast enough so that the output can be used before expiration balanced against the desire to improve the simulations by increasing resolution and using more detailed physical parameterizations. It is now feasible to run high-resolution mesoscale models such as MASS on local workstations to provide timely forecasts at a fraction of the cost required to run these models on mainframe supercomputers. MASS has been running in the Applied Meteorology Unit (AMU) at KSC/CCAS since January 1994 for the purpose of system evaluation. In March 1995, the AMU began sending real-time MASS output to the forecasters and meteorologists at CCAS, Spaceflight Meteorology Group (Johnson Space Center, Houston, Texas), and the National Weather Service (Melbourne, Florida). However, MASS is not yet an operational system. The final decision whether to transition MASS for operational use will depend on a combination of forecaster feedback, the AMU's final evaluation results, and the life-cycle costs of the operational system.

  1. Pyo-pneumothorax tuberculeux: à propos de 18 cas

    PubMed Central

    Hicham, Souhi; Hanane, El Ouazzani; Hicham, Janah; Ismaïl, Rhorfi; Ahmed, Abid

    2016-01-01

    Le pyo-pneumothorax tuberculeux est une complication rare mais grave de la tuberculose pulmonaire évolutive. Nous rapportons une série de 18 cas de pyo-pneumothorax tuberculeux colligés au service de Pneumo-Phtisiologie de l'Hôpital Militaire d'Instruction Mohammed V de Rabat entre janvier 2005 et décembre 2009. Il s'agit de 15 hommes et 3 femmes d’âge moyen de 35 ans ±7 ans. 4 patients étaient diabétiques. Le tabagisme était retrouvé chez 9 cas. Le pyo-pneumothorax était du coté droit dans 13 cas. La radiographie thoracique avait montré des lésions cavitaires chez 15 patients et des lésions étendues et bilatérales chez 8 cas. La recherche de BK dans le liquide de tubage gastrique était positive chez 16 cas. Un drainage thoracique associé à un traitement antituberculeux selon le régime 2SRHZ/7RH et une kinésithérapie respiratoire ont été instaurés chez tous les cas. La durée moyenne de drainage pleural était de 4 semaines. Chez 3 cas on avait noté la persistance de la suppuration pleurale ayant nécessité une toilette pleurale sous thoracoscopie avec pleurectomie et exérèse pulmonaire limitée emportant la lésion parenchymateuse tuberculeuse et la persistance d'une volumineuse poche pleurale avec trouble ventilatoire restrictif ayant nécessité une décortication pleurale chirurgicale chez deux cas. L’évolution était favorable avec pachypleurite séquellaire minime chez le reste des cas. Le pyo-pneumothorax tuberculeux est une forme grave, qui est souvent en rapport avec une tuberculose cavitaire active. L’évolution est généralement trainante malgré le traitement antituberculeux et le drainage thoracique, d'où la nécessité d'un diagnostic et un traitement précoce de toute forme de tuberculose. PMID:27583090

  2. Repurposing CRISPR/Cas9 for in situ functional assays

    PubMed Central

    Malina, Abba; Mills, John R.; Cencic, Regina; Yan, Yifei; Fraser, James; Schippers, Laura M.; Paquet, Marilène; Dostie, Josée; Pelletier, Jerry

    2013-01-01

    RNAi combined with next-generation sequencing has proven to be a powerful and cost-effective genetic screening platform in mammalian cells. Still, this technology has its limitations and is incompatible with in situ mutagenesis screens on a genome-wide scale. Using p53 as a proof-of-principle target, we readapted the CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR associated 9) genome-editing system to demonstrate the feasibility of this methodology for targeted gene disruption positive selection assays. By using novel “all-in-one” lentiviral and retroviral delivery vectors heterologously expressing both a codon-optimized Cas9 and its synthetic guide RNA (sgRNA), we show robust selection for the CRISPR-modified Trp53 locus following drug treatment. Furthermore, by linking Cas9 expression to GFP fluorescence, we use an “all-in-one” system to track disrupted Trp53 in chemoresistant lymphomas in the Eμ-myc mouse model. Deep sequencing analysis of the tumor-derived endogenous Cas9-modified Trp53 locus revealed a wide spectrum of mutants that were enriched with seemingly limited off-target effects. Taken together, these results establish Cas9 genome editing as a powerful and practical approach for positive in situ genetic screens. PMID:24298059

  3. SD-CAS: Spin Dynamics by Computer Algebra System.

    PubMed

    Filip, Xenia; Filip, Claudiu

    2010-11-01

    A computer algebra tool for describing the Liouville-space quantum evolution of nuclear 1/2-spins is introduced and implemented within a computational framework named Spin Dynamics by Computer Algebra System (SD-CAS). A distinctive feature compared with numerical and previous computer algebra approaches to solving spin dynamics problems results from the fact that no matrix representation for spin operators is used in SD-CAS, which determines a full symbolic character to the performed computations. Spin correlations are stored in SD-CAS as four-entry nested lists of which size increases linearly with the number of spins into the system and are easily mapped into analytical expressions in terms of spin operator products. For the so defined SD-CAS spin correlations a set of specialized functions and procedures is introduced that are essential for implementing basic spin algebra operations, such as the spin operator products, commutators, and scalar products. They provide results in an abstract algebraic form: specific procedures to quantitatively evaluate such symbolic expressions with respect to the involved spin interaction parameters and experimental conditions are also discussed. Although the main focus in the present work is on laying the foundation for spin dynamics symbolic computation in NMR based on a non-matrix formalism, practical aspects are also considered throughout the theoretical development process. In particular, specific SD-CAS routines have been implemented using the YACAS computer algebra package (http://yacas.sourceforge.net), and their functionality was demonstrated on a few illustrative examples. PMID:20843716

  4. Measurement of Flux Density of Cas A at Low Frequencies

    NASA Astrophysics Data System (ADS)

    Patil, Ajinkya; Fisher, R.

    2012-01-01

    Cas A is used as a flux calibrator throughout the radio spectrum. Therefore it is important to know the spectral and secular variations in its flux density. Earlier observations by Scott et. al. (1969) and Baars et. al. (1972) suggested a secular decrease in flux density of Cas A at a rate of about 1% per year at all frequencies. However later observations by Erickson & Perley (1975) and Read (1977) indicated anomalously high flux from Cas A at 38 MHz. Also, these observations suggested that the original idea of faster decay of the flux density rate at low frequencies may be in error or that something more complex than simple decay is affecting the flux density at low frequencies. The source changes at 38 MHz still remains a mystery. We intend to present the results of follow up observations made from 1995 to 1998 with a three element interferometer in Green Bank operating in frequency range 30 to 120 MHz. We will discuss the problems at such low frequencies due to large beamwidth and unstable ionosphere. We will also discuss the strategies we have used so far to to find the flux density of Cas A by calculating the ratio of flux density of Cas A to that of Cyg A, assuming flux density of Cyg A to be constant. Above mentioned work was performed in summer student program sponsored by National Radio Astronomy Observatory.

  5. United States Air Force 611th Air Support Group/Civil Engineering Squadron Elmendorf AFB, Alaska. Decision document for no further response action planned: Bullen Point Radar Installation, Alaska. Final report

    SciTech Connect

    Karmi, S.

    1996-05-24

    This Decision Document discusses the selection of no further action as the recommended action for two sites located at the Bullen Point radar installation. The United States Air Force (Air Force) completed a Remedial Investigation/Feasibility Study and a Risk Assessment for the five sites located at the Bullen Point installation (U.S. Air Force 1996a,b). Based on the findings of these activities, two sites are recommended for no further action. Sites at the Bullen Point radar installation recommended for no further action are: Old Landfill/Dump Site East (LF06) and Drum Storage Area (SS10).

  6. Displacement of p130Cas from focal adhesions links actomyosin contraction to cell migration.

    PubMed

    Machiyama, Hiroaki; Hirata, Hiroaki; Loh, Xia Kun; Kanchi, Madhu Mathi; Fujita, Hideaki; Tan, Song Hui; Kawauchi, Keiko; Sawada, Yasuhiro

    2014-08-15

    Cell adhesion complexes provide platforms where cell-generated forces are transmitted to the extracellular matrix (ECM). Tyrosine phosphorylation of focal adhesion proteins is crucial for cells to communicate with the extracellular environment. However, the mechanisms that transmit actin cytoskeletal motion to the extracellular environment to drive cell migration are poorly understood. We find that the movement of p130Cas (Cas, also known as BCAR1), a mechanosensor at focal adhesions, correlates with actin retrograde flow and depends upon actomyosin contraction and phosphorylation of the Cas substrate domain (CasSD). This indicates that CasSD phosphorylation underpins the physical link between Cas and the actin cytoskeleton. Fluorescence recovery after photobleaching (FRAP) experiments reveal that CasSD phosphorylation, as opposed to the association of Cas with Src, facilitates Cas displacement from adhesion complexes in migrating cells. Furthermore, the stabilization of Src-Cas binding and inhibition of myosin II, both of which sustain CasSD phosphorylation but mitigate Cas displacement from adhesion sites, retard cell migration. These results indicate that Cas promotes cell migration by linking actomyosin contractions to the adhesion complexes through a dynamic interaction with Src as well as through the phosphorylation-dependent association with the actin cytoskeleton. PMID:24928898

  7. Support for and reported compliance among smokers with smoke-free policies in air-conditioned hospitality venues in Malaysia and Thailand: findings from the International Tobacco Control Southeast Asia Survey.

    PubMed

    Yong, Hua-Hie; Foong, Kin; Borland, Ron; Omar, Maizurah; Hamann, Stephen; Sirirassamee, Buppha; Fong, Geoffrey T; Fotuhi, Omid; Hyland, Andrew

    2010-01-01

    This study examined support for and reported compliance with smoke-free policy in air-conditioned restaurants and other similar places among adult smokers in Malaysia and Thailand. Baseline data (early 2005) from the International Tobacco Control Southeast Asia Survey (ITC-SEA), conducted face-to-face in Malaysia and Thailand (n = 4005), were used. Among those attending venues, reported total smoking bans in indoor air-conditioned places such as restaurants, coffee shops, and karaoke lounges were 40% and 57% in Malaysia and Thailand, respectively. Support for a total ban in air-conditioned venues was high and similar for both countries (82% Malaysian and 90% Thai smokers who believed there was a total ban), but self-reported compliance with bans in such venues was significantly higher in Thailand than in Malaysia (95% vs 51%, P < .001). As expected, reporting a ban in air-conditioned venues was associated with a greater support for a ban in such venues in both countries. PMID:20032039

  8. U.S. ENVIRONMENTAL PROTECTION AGENCY'S PM SUPERSITES PROGRAM - A MAJOR SUCCESSFUL COLLABORATIVE AIR QUALITY PROGRAM SUPPORTING STATES AND REGIONAL ORGANIZATIONS IN THEIR APPROACHES TO REDUCE PM LEVELS IN AIR ON URBAN AND REGIONAL SCALES

    EPA Science Inventory

    The U.S. Environmental Protection Agency's Particulate Matter (PM) Supersites Program (Program) is a nationwide air quality methods, measurement, modeling, and data analysis program initiated through cooperative agreements with leading universities in the United States. The Progr...

  9. GENOME EDITING IN HUMAN CELLS USING CRISPR/CAS NUCLEASES

    PubMed Central

    Wyvekens, Nicolas; Tsai, Shengdar; Joung, J. Keith

    2016-01-01

    The clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas) system has been broadly adopted for highly efficient genome editing in a variety of model organisms and human cell types. Unlike previous genome editing technologies such as Zinc Finger Nucleases (ZFNs) and Transcription Activator-Like Effector Nucleases (TALENs), the CRISPR/Cas technology does not require complex protein engineering and can be utilized by any researcher proficient in basic molecular biology and cell culture techniques. Here we describe protocols for design and cloning of vectors expressing single or multiplex gRNAs, for transient transfection of human cell lines, and for quantitation of mutation frequencies by T7 Endonuclease I assay. These protocols also include guidance for using two improvements that increase the specificity of CRISPR/Cas nucleases: truncated gRNAs and dimeric RNA-guided FokI nucleases. PMID:26423589

  10. CRISPR-Cas9: A Revolutionary Tool for Cancer Modelling

    PubMed Central

    Torres-Ruiz, Raul; Rodriguez-Perales, Sandra

    2015-01-01

    The cancer-modelling field is now experiencing a conversion with the recent emergence of the RNA-programmable CRISPR-Cas9 system, a flexible methodology to produce essentially any desired modification in the genome. Cancer is a multistep process that involves many genetic mutations and other genome rearrangements. Despite their importance, it is difficult to recapitulate the degree of genetic complexity found in patient tumors. The CRISPR-Cas9 system for genome editing has been proven as a robust technology that makes it possible to generate cellular and animal models that recapitulate those cooperative alterations rapidly and at low cost. In this review, we will discuss the innovative applications of the CRISPR-Cas9 system to generate new models, providing a new way to interrogate the development and progression of cancers. PMID:26389881

  11. DNA fragment editing of genomes by CRISPR/Cas9.

    PubMed

    Jinhuan, Li; Jia, Shou; Qiang, Wu

    2015-10-01

    The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated nuclease 9 (Cas9) system from bacteria and archaea emerged recently as a new powerful technology of genome editing in virtually any organism. Due to its simplicity and cost effectiveness, a revolutionary change of genetics has occurred. Here, we summarize the recent development of DNA fragment editing methods by CRISPR/Cas9 and describe targeted DNA fragment deletions, inversions, duplications, insertions, and translocations. The efficient method of DNA fragment editing provides a powerful tool for studying gene function, regulatory elements, tissue development, and disease progression. Finally, we discuss the prospects of CRISPR/Cas9 system and the potential applications of other types of CRISPR system. PMID:26496751

  12. CRISPR-Cas9: A Revolutionary Tool for Cancer Modelling.

    PubMed

    Torres-Ruiz, Raul; Rodriguez-Perales, Sandra

    2015-01-01

    The cancer-modelling field is now experiencing a conversion with the recent emergence of the RNA-programmable CRISPR-Cas9 system, a flexible methodology to produce essentially any desired modification in the genome. Cancer is a multistep process that involves many genetic mutations and other genome rearrangements. Despite their importance, it is difficult to recapitulate the degree of genetic complexity found in patient tumors. The CRISPR-Cas9 system for genome editing has been proven as a robust technology that makes it possible to generate cellular and animal models that recapitulate those cooperative alterations rapidly and at low cost. In this review, we will discuss the innovative applications of the CRISPR-Cas9 system to generate new models, providing a new way to interrogate the development and progression of cancers. PMID:26389881

  13. Applications of CRISPR-Cas systems in neuroscience

    PubMed Central

    Heidenreich, Matthias; Zhang, Feng

    2016-01-01

    Genome editing tools, and in particular those based on CRISPR-Cas systems, are accelerating the pace of biological research and enabling targeted genetic interrogation in virtually any organism and cell type. These tools have opened the door to the development of new model systems for studying the complexity of the nervous system, including animal and stem cell-derived in vitro models. Precise and efficient gene editing using CRISPR-Cas systems has the potential to advance both basic and translational neuroscience research. PMID:26656253

  14. High-throughput functional genomics using CRISPR-Cas9

    PubMed Central

    Shalem, Ophir; Sanjana, Neville E.; Zhang, Feng

    2015-01-01

    Forward genetic screens are powerful tools for the discovery and functional annotation of genetic elements. Recently, the RNA-guided CRISPR (clustered regularly interspaced short palindromic repeat)-associated Cas9 nuclease has been combined with genome-scale guide RNA libraries for unbiased, phenotypic screening. In this Review, we describe recent advances using Cas9 for genome-scale screens, including knockout approaches that inactivate genomic loci and strategies that modulate transcriptional activity. We discuss practical aspects of screen design, provide comparisons with RNA interference (RNAi) screening, and outline future applications and challenges. PMID:25854182

  15. Mouse Genome Editing using CRISPR/Cas System

    PubMed Central

    Harms, Donald W; Quadros, Rolen M; Seruggia, Davide; Ohtsuka, Masato; Takahashi, Gou

    2015-01-01

    The availability of techniques to create desired genetic mutations has enabled the laboratory mouse as an extensively used model organism in biomedical research including human genetics. A new addition to this existing technical repertoire is the CRISPR/Cas system. Specifically, this system allows editing of the mouse genome much faster than the previously used techniques and more importantly multiple mutations can be created in a single experiment. Here we provide protocols for preparation of CRISPR/Cas reagents and microinjection into one cell mouse embryos to create knockout or knock-in mouse models. PMID:25271839

  16. Les sarcomes des tissus mous: à propos de 33 cas

    PubMed Central

    Abdou, Jiddou; Elkabous, Mustapha; M'rabti, Hind; Errihani, Hassan

    2015-01-01

    L'objectif de cette étude est de rapporter les particularités épidémiologiques, cliniques, histologiques, thérapeutiques et évolutives des sarcomes des tissus mous à l'Institut National d'Oncologie et de définir les facteurs influençant la survie des patients. C'est une étude rétrospective de 33 cas de sarcome des tissus mous, colligés entre janvier 2008 et décembre 2010. Les critères d’éligibilité étaient un âge supérieur à 16 ans, une épreuve histologique d'un sarcome des tissus mous à l'exclusion des tumeurs stromales gastro-intestinales (GIST). Les items recueillis étaient: épidémiologiques, cliniques, histologiques, Radiologiques, et thérapeutiques. Des analyses univariées puis multivariées ont été réalisées à la recherche de facteurs influençant la survie à 2 ans. Il s'agit de 33 cas, 17 Hommes et 16 Femmes, l’âge moyen était de 43,21 ans (Extrêmes= 18-76 ans). La tumeur était localisée aux extrémités dans 24 cas (72,72%). Le type histologique prédominant était le Liposarcome dans 9 cas (27,27%). Le stade tumoral était localisé dans 25 cas (75,8%), métastatique dans 8 cas (24,2%). Vingt-cinq tumeurs ont été traitées chirurgicalement dont 21 cas (84%) de chirurgie conservatrice et 4 cas (16%) de chirurgie radicale. La radiothérapie a été réalisée chez 10 patients (30,3%). La chimiothérapie a été faite chez 20 patients. En analyse univariée les facteurs pronostiques étaient l’âge (p=0,03) et le stade tumoral (p=0,09). L’âge et le stade tumoral sont des facteurs pronostiques influençant la survie des sarcomes des tissus mous. PMID:27022434

  17. V845 Cas - ein RRab-Stern mit Blazhko-Effekt

    NASA Astrophysics Data System (ADS)

    Maintz, Gisela

    2015-02-01

    CCD observations of V845 Cas (RA= 23 26 15.02, DE= +57 23 55.5 (2000)) were obtained at my private observatory. For V845 Cas 13 maxima were gained, showing a light curve, that varies from epoch to epoch. The irregularity of the lightcurves of V845 Cas is due to the Blazhko effect. Revised elements are given as: V845 Cas Max = 2456221.3450 + 0.570845 * E.

  18. Functional annotation of native enhancers with a Cas9-histone demethylase fusion.

    PubMed

    Kearns, Nicola A; Pham, Hannah; Tabak, Barbara; Genga, Ryan M; Silverstein, Noah J; Garber, Manuel; Maehr, René

    2015-05-01

    Understanding of mammalian enhancers is limited by the lack of a technology to rapidly and thoroughly test the cell type-specific function. Here, we use a nuclease-deficient Cas9 (dCas9)-histone demethylase fusion to functionally characterize previously described and new enhancer elements for their roles in the embryonic stem cell state. Further, we distinguish the mechanism of action of dCas9-LSD1 at enhancers from previous dCas9-effectors. PMID:25775043

  19. Cas9 gRNA engineering for genome editing, activation and repression

    PubMed Central

    Kiani, Samira; Chavez, Alejandro; Tuttle, Marcelle; Hall, Richard N; Chari, Raj; Ter-Ovanesyan, Dmitry; Qian, Jason; Pruitt, Benjamin W; Beal, Jacob; Vora, Suhani; Buchthal, Joanna; Kowal, Emma J K; Ebrahimkhani, Mohammad R; Collins, James J; Weiss, Ron; Church, George

    2015-01-01

    We demonstrate that by altering the length of Cas9-associated guide RNA(gRNA) we were able to control Cas9 nuclease activity and simultaneously perform genome editing and transcriptional regulation with a single Cas9 protein. We exploited these principles to engineer mammalian synthetic circuits with combined transcriptional regulation and kill functions governed by a single multifunctional Cas9 protein. PMID:26344044

  20. Cas9 gRNA engineering for genome editing, activation and repression.

    PubMed

    Kiani, Samira; Chavez, Alejandro; Tuttle, Marcelle; Hall, Richard N; Chari, Raj; Ter-Ovanesyan, Dmitry; Qian, Jason; Pruitt, Benjamin W; Beal, Jacob; Vora, Suhani; Buchthal, Joanna; Kowal, Emma J K; Ebrahimkhani, Mohammad R; Collins, James J; Weiss, Ron; Church, George

    2015-11-01

    We demonstrate that by altering the length of Cas9-associated guide RNA (gRNA) we were able to control Cas9 nuclease activity and simultaneously perform genome editing and transcriptional regulation with a single Cas9 protein. We exploited these principles to engineer mammalian synthetic circuits with combined transcriptional regulation and kill functions governed by a single multifunctional Cas9 protein. PMID:26344044

  1. The Cas6e ribonuclease is not required for interference and adaptation by the E. coli type I-E CRISPR-Cas system

    PubMed Central

    Semenova, Ekaterina; Kuznedelov, Konstantin; Datsenko, Kirill A.; Boudry, Pierre M.; Savitskaya, Ekaterina E.; Medvedeva, Sofia; Beloglazova, Natalia; Logacheva, Maria; Yakunin, Alexander F.; Severinov, Konstantin

    2015-01-01

    CRISPR-Cas are small RNA-based adaptive prokaryotic immunity systems protecting cells from foreign DNA or RNA. Type I CRISPR-Cas systems are composed of a multiprotein complex (Cascade) that, when bound to CRISPR RNA (crRNA), can recognize double-stranded DNA targets and recruit the Cas3 nuclease to destroy target-containing DNA. In the Escherichia coli type I-E CRISPR-Cas system, crRNAs are generated upon transcription of CRISPR arrays consisting of multiple palindromic repeats and intervening spacers through the function of Cas6e endoribonuclease, which cleaves at specific positions of repeat sequences of the CRISPR array transcript. Cas6e is also a component of Cascade. Here, we show that when mature unit-sized crRNAs are provided in a Cas6e-independent manner by transcription termination, the CRISPR-Cas system can function without Cas6e. The results should allow facile interrogation of various targets by type I-E CRISPR-Cas system in E. coli using unit-sized crRNAs generated by transcription. PMID:26013814

  2. CRISPR/Cas-Mediated Site-Specific Mutagenesis in Arabidopsis thaliana Using Cas9 Nucleases and Paired Nickases.

    PubMed

    Schiml, Simon; Fauser, Friedrich; Puchta, Holger

    2016-01-01

    The CRISPR/Cas system has recently become the most important tool for genome engineering due to its simple architecture that allows for rapidly changing the target sequence and its applicability to organisms throughout all kingdoms of life. The need for an easy-to-use and reliable nuclease is especially high in plant research, as precise genome modifications are almost impossible to achieve by Agrobacterium-mediated transformation and the regeneration of plants from protoplast cultures is very labor intensive. Here, we describe the application of the Cas9 nuclease to Arabidopsis thaliana for the induction of heritable targeted mutations, which may also be used for other plant species. To cover the concern for off-target activity, we also describe the generation of stable mutants using paired Cas9 nickases. PMID:27557689

  3. Efficient in vivo deletion of a large imprinted lncRNA by CRISPR/Cas9.

    PubMed

    Han, Jinxiong; Zhang, Jun; Chen, Li; Shen, Bin; Zhou, Jiankui; Hu, Bian; Du, Yinan; Tate, Peri H; Huang, Xingxu; Zhang, Wensheng

    2014-01-01

    Recent genome-wide studies have revealed that the majority of the mouse genome is transcribed as non-coding RNAs (ncRNAs) and growing evidence supports the importance of ncRNAs in regulating gene expression and epigenetic processes. However, the low efficiency of conventional gene targeting strategies has hindered the functional study of ncRNAs in vivo, particularly in generating large fragment deletions of long non-coding RNAs (lncRNAs) with multiple expression variants. The bacterial clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) system has recently been applied as an efficient tool for engineering site-specific mutations of protein-coding genes in the genome. In this study, we explored the potential of using the CRISPR/Cas9 system to generate large genomic deletions of lncRNAs in mice. We developed an efficient one-step strategy to target the maternally expressed lncRNA, Rian, on chromosome 12 in mice. We showed that paired sgRNAs can precisely generate large deletions up to 23kb and the deletion efficiency can be further improved up to 33% by combining multiple sgRNAs. The deletion successfully abolished the expression of Rian from the maternally inherited allele, validating the biological relevance of the mutations in studying an imprinted locus. Mutation of Rian has differential effects on expression of nearby genes in different somatic tissues. Taken together, we have established a robust one-step method to engineer large deletions to knockout lncRNA genes with the CRISPR/Cas9 system. Our work will facilitate future functional studies of other lncRNAs in vivo. PMID:25137067

  4. Cerebral protection vs no cerebral protection: timing of stroke with CAS.

    PubMed

    Tietke, M; Jansen, O

    2009-12-01

    Since its development in the late 1980s the discussion of risk and benefit of carotid artery stenting (CAS) compared to carotid endarterectomy (CEA) is still ongoing. In this discussion the increasing technical inventions in CAS like PDs, different stent designs and materials and their effect upon the complication rates are playing increasingly important roles. We present a review of mono- or oligocentre studies with greater collectives, one review study and multi centre studies SPACE I, EVA-3S and preliminary data from the ICSS-trial, in which subgroups of patients were treated with or without protection devices. Despite the results of several retrospective studies mostly with historic comparator cohorts rather than concurrent the results of the most recent multi centre prospective randomized trials seem to show a benefit for unprotected stenting. The complication rates for protected vs. unprotected groups in SPACE I showed 8.3% vs. 6.5% and the pooled data from SPACE I and EVA-3S 8.1% vs. 7.3%. A subgroup analysis of SPACE I aiming at the impact of the stent design on peri-interventional complication rate shows that the pOE rate was significantly lower in patients treated with a closed cell stent (5.7%, 95% CI: 3.7-8.3%) than in those treated with an open cell stent (11.0%, 95% CI: 6.2-17.8%) (P=0.047). The most recent data from prospective multi centre trials support the presumption that PDs do not reduce, but may increase the periinterventional complication rate. Whereas many older single centre studies promoting PDs do show conceptual problems. The stent design appears to have a more important impact on the resulting complications in CAS, than the PDs. PMID:19935606

  5. The abundances of major elements in Cas A and Tycho supernova remnants

    NASA Technical Reports Server (NTRS)

    Gorenstein, P.

    1995-01-01

    The objective of this program was to map the abundances of major elements such as O, Si, S, and Fe in the supernova remnants, Tycho and Cas A. The approach was based upon using archival cosmic X-ray data from several space missions, notably, the Einstein Observatory, EXOSAT, ROSAT, BBSRT, and ASCA. Two of the missions, Einstein and ROSAT, had high resolution telescopes that provided excellent images, but no spectral information. Two missions with much poorer resolution telescopes, BBXRT and ASCA, gave good spectral information through pulse height of signals in their cooled solid state detector, but rather crude spatial information. Our goal was to extract spectral information from the combined analysis of the Einstein and ROSAT images of Cas A and Tycho and to verify or refine the spectral map by checking its agreement with the BBSRT or ASCA spectra results for larger regions. In particular, we note that the Einstein and ROSAT telescopes have different spectral responses. The Einstein bandwidth includes the 2-4 keV region which is absent from ROSAT. Hence, by forming linear combinations of the Einstein and ROSAT images, we are able to resolve the contributions of the 0.5-2 keV band from the 2-4 keV band. The former contains lines of O and Fe while the latter is dominated by Si and S. We correct for the expansion that has taken place in the remnants during the ten-year interval between the Einstein and ROSAT measurements, but we must assume that no significant spectral changes have occurred during that time. The analysis of the Tycho SNR was completed and the results have been published. A copy of the paper is included. The analysis of Cas A has proved to be more complicated. It is continuing with support from another program. Part of the problem may be due to difficulties in the aspect information which is needed to precisely register the ROSAT and Einstein images.

  6. Creation of targeted genomic deletions using TALEN or CRISPR/Cas nuclease pairs in one-cell mouse embryos

    PubMed Central

    Brandl, Christina; Ortiz, Oskar; Röttig, Bernhard; Wefers, Benedikt; Wurst, Wolfgang; Kühn, Ralf

    2014-01-01

    The use of TALEN and CRISPR/CAS nucleases is becoming increasingly popular as a means to edit single target sites in one-cell mouse embryos. Nevertheless, an area that has received less attention concerns the engineering of structural genome variants and the necessary religation of two distant double-strand breaks. Herein, we applied pairs of TALEN or sgRNAs and Cas9 to create deletions in the Rab38 gene. We found that the deletion of 3.2 or 9.3 kb, but not of 30 kb, occurs at a frequency of 6–37%. This is sufficient for the direct production of mutants by embryo microinjection. Therefore, deletions up to ∼10 kb can be readily achieved for modeling human disease alleles. This work represents an important step towards the establishment of new protocols that support the ligation of remote DSB ends to achieve even larger rearrangements. PMID:25685662

  7. On the Integration of Computer Algebra Systems (CAS) by Canadian Mathematicians: Results of a National Survey

    ERIC Educational Resources Information Center

    Buteau, Chantal; Jarvis, Daniel H.; Lavicza, Zsolt

    2014-01-01

    In this article, we outline the findings of a Canadian survey study (N = 302) that focused on the extent of computer algebra systems (CAS)-based technology use in postsecondary mathematics instruction. Results suggest that a considerable number of Canadian mathematicians use CAS in research and teaching. CAS use in research was found to be the…

  8. After 16 Years of Publishing Standards, Do CAS Standards Make a Difference?

    ERIC Educational Resources Information Center

    Arminio, Jan; Gochenauer, Patty

    2004-01-01

    Using members of professional associations who are a part of the Council for the Advancement of Standards in Higher Education (CAS) consortia as a sample, this study investigated who uses CAS Standards, how and why they are used, and whether CAS Standards are associated with enhanced student learning. Using a quantitative analysis, this study…

  9. Creating Genome Modifications in C. elegans Using the CRISPR/Cas9 System.

    PubMed

    Calarco, John A; Friedland, Ari E

    2015-01-01

    The clustered, regularly interspaced, short, palindromic repeat (CRISPR)-associated (CAS) nuclease Cas9 has been used in many organisms to generate specific mutations and transgene insertions. Here we describe a method using the S. pyogenes Cas9 in C. elegans that provides a convenient and effective approach for making heritable changes to the worm genome. PMID:26423968

  10. 48 CFR 9904.412-64.1 - Transition Method for the CAS Pension Harmonization Rule.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... CAS Pension Harmonization Rule. 9904.412-64.1 Section 9904.412-64.1 Federal Acquisition Regulations... Transition Method for the CAS Pension Harmonization Rule. Contractors or subcontractors that become subject... through 7 Notes (Note 1) Actuarial Accrued Liability $2,470,500 $14,225,000 2 CAS Actuarial Value...

  11. 48 CFR 9904.412-64.1 - Transition Method for the CAS Pension Harmonization Rule.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... CAS Pension Harmonization Rule. 9904.412-64.1 Section 9904.412-64.1 Federal Acquisition Regulations... Transition Method for the CAS Pension Harmonization Rule. Contractors or subcontractors that become subject... through 7 Notes (Note 1) Actuarial Accrued Liability $2,470,500 $14,225,000 2 CAS Actuarial Value...

  12. 48 CFR 9904.412-60.1 - Illustrations-CAS Pension Harmonization Rule.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 48 Federal Acquisition Regulations System 7 2013-10-01 2012-10-01 true Illustrations-CAS Pension... AND COST ACCOUNTING STANDARDS COST ACCOUNTING STANDARDS 9904.412-60.1 Illustrations—CAS Pension... cost on or after the Applicability Date of the CAS Harmonization Rule. The illustrations present...

  13. 48 CFR 9904.412-60.1 - Illustrations-CAS Pension Harmonization Rule.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 48 Federal Acquisition Regulations System 7 2014-10-01 2014-10-01 false Illustrations-CAS Pension... AND COST ACCOUNTING STANDARDS COST ACCOUNTING STANDARDS 9904.412-60.1 Illustrations—CAS Pension... cost on or after the Applicability Date of the CAS Harmonization Rule. The illustrations present...

  14. 48 CFR 9904.412-60.1 - Illustrations-CAS Pension Harmonization Rule.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 48 Federal Acquisition Regulations System 7 2012-10-01 2012-10-01 false Illustrations-CAS Pension... AND COST ACCOUNTING STANDARDS COST ACCOUNTING STANDARDS 9904.412-60.1 Illustrations—CAS Pension... cost on or after the Applicability Date of the CAS Harmonization Rule. The illustrations present...

  15. 48 CFR 9904.413-64.1 - Transition Method for the CAS Pension Harmonization Rule.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... CAS Pension Harmonization Rule. 9904.413-64.1 Section 9904.413-64.1 Federal Acquisition Regulations... Transition Method for the CAS Pension Harmonization Rule. The transition method for the CAS Pension Harmonization Rule under this Standard shall be in accordance with 9904.412.64.1 Transition Method for...

  16. Mismatch Negativity Responses in Children with a Diagnosis of Childhood Apraxia of Speech (CAS)

    ERIC Educational Resources Information Center

    Froud, Karen; Khamis-Dakwar, Reem

    2012-01-01

    Purpose: To evaluate whether a hypothesis suggesting that apraxia of speech results from phonological overspecification could be relevant for childhood apraxia of speech (CAS). Method: High-density EEG was recorded from 5 children with CAS and 5 matched controls, ages 5-8 years, with and without CAS, as they listened to randomized sequences of CV…

  17. 48 CFR 9904.413-64.1 - Transition Method for the CAS Pension Harmonization Rule.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... CAS Pension Harmonization Rule. 9904.413-64.1 Section 9904.413-64.1 Federal Acquisition Regulations... Transition Method for the CAS Pension Harmonization Rule. The transition method for the CAS Pension Harmonization Rule under this Standard shall be in accordance with 9904.412.64.1 Transition Method for...

  18. 48 CFR 9904.413-64.1 - Transition Method for the CAS Pension Harmonization Rule.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... CAS Pension Harmonization Rule. 9904.413-64.1 Section 9904.413-64.1 Federal Acquisition Regulations... Transition Method for the CAS Pension Harmonization Rule. The transition method for the CAS Pension Harmonization Rule under this Standard shall be in accordance with 9904.412.64.1 Transition Method for...

  19. The Impact on Student Achievement of When CAS Technology Is Introduced

    ERIC Educational Resources Information Center

    Driver, David

    2012-01-01

    When a Computer Algebra System (CAS) is used as a pedagogical and functional tool in class and as a functional tool in exams, its effect on student achievement can be quite profound. The timing of when students are first introduced to a CAS has an impact on gains in student achievement. In this action research project, the CAS calculator was…

  20. 48 CFR 9904.412-64.1 - Transition Method for the CAS Pension Harmonization Rule.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... CAS Pension Harmonization Rule. 9904.412-64.1 Section 9904.412-64.1 Federal Acquisition Regulations... Transition Method for the CAS Pension Harmonization Rule. Contractors or subcontractors that become subject... through 7 Notes (Note 1) Actuarial Accrued Liability $2,470,500 $14,225,000 2 CAS Actuarial Value...

  1. Guide RNAs: A Glimpse at the Sequences that Drive CRISPR-Cas Systems.

    PubMed

    Briner, Alexandra E; Barrangou, Rodolphe

    2016-01-01

    CRISPR-Cas systems provide adaptive immunity in bacteria and archaea. Although there are two main classes of CRISPR-Cas systems defined by gene content, interfering RNA biogenesis, and effector proteins, Type II systems have recently been exploited on a broad scale to develop next-generation genetic engineering and genome-editing tools. Conveniently, Type II systems are streamlined and rely on a single protein, Cas9, and a guide RNA molecule, comprised of a CRISPR RNA (crRNA) and trans-acting CRISPR RNA (tracrRNA), to achieve effective and programmable nucleic acid targeting and cleavage. Currently, most commercially available Cas9-based genome-editing tools use the CRISPR-Cas system from Streptococcus pyogenes (SpyCas9), although many orthogonal Type II systems are available for diverse and multiplexable genome engineering applications. Here, we discuss the biological significance of Type II CRISPR-Cas elements, including the tracrRNA, crRNA, Cas9, and protospacer-adjacent motif (PAM), and look at the native function of these elements to understand how they can be engineered, enhanced, and optimized for genome editing applications. Additionally, we discuss the basis for orthogonal Cas9 and guide RNA systems that would allow researchers to concurrently use multiple Cas9-based systems for different purposes. Understanding the native function of endogenous Type II CRISPR-Cas systems can lead to new Cas9 tool development to expand the genetic manipulation toolbox. PMID:27371605

  2. The role of Cas8 in type I CRISPR interference

    PubMed Central

    Cass, Simon D.B.; Haas, Karina A.; Stoll, Britta; Alkhnbashi, Omer S.; Sharma, Kundan; Urlaub, Henning; Backofen, Rolf; Marchfelder, Anita; Bolt, Edward L.

    2015-01-01

    CRISPR (clustered regularly interspaced short palindromic repeat) systems provide bacteria and archaea with adaptive immunity to repel invasive genetic elements. Type I systems use ‘cascade’ [CRISPR-associated (Cas) complex for antiviral defence] ribonucleoprotein complexes to target invader DNA, by base pairing CRISPR RNA (crRNA) to protospacers. Cascade identifies PAMs (protospacer adjacent motifs) on invader DNA, triggering R-loop formation and subsequent DNA degradation by Cas3. Cas8 is a candidate PAM recognition factor in some cascades. We analysed Cas8 homologues from type IB CRISPR systems in archaea Haloferax volcanii (Hvo) and Methanothermobacter thermautotrophicus (Mth). Cas8 was essential for CRISPR interference in Hvo and purified Mth Cas8 protein responded to PAM sequence when binding to nucleic acids. Cas8 interacted physically with Cas5–Cas7–crRNA complex, stimulating binding to PAM containing substrates. Mutation of conserved Cas8 amino acid residues abolished interference in vivo and altered catalytic activity of Cas8 protein in vitro. This is experimental evidence that Cas8 is important for targeting Cascade to invader DNA. PMID:26182359

  3. The role of Cas8 in type I CRISPR interference.

    PubMed

    Cass, Simon D B; Haas, Karina A; Stoll, Britta; Alkhnbashi, Omer S; Sharma, Kundan; Urlaub, Henning; Backofen, Rolf; Marchfelder, Anita; Bolt, Edward L

    2015-01-01

    CRISPR (clustered regularly interspaced short palindromic repeat) systems provide bacteria and archaea with adaptive immunity to repel invasive genetic elements. Type I systems use 'cascade' [CRISPR-associated (Cas) complex for antiviral defence] ribonucleoprotein complexes to target invader DNA, by base pairing CRISPR RNA (crRNA) to protospacers. Cascade identifies PAMs (protospacer adjacent motifs) on invader DNA, triggering R-loop formation and subsequent DNA degradation by Cas3. Cas8 is a candidate PAM recognition factor in some cascades. We analysed Cas8 homologues from type IB CRISPR systems in archaea Haloferax volcanii (Hvo) and Methanothermobacter thermautotrophicus (Mth). Cas8 was essential for CRISPR interference in Hvo and purified Mth Cas8 protein responded to PAM sequence when binding to nucleic acids. Cas8 interacted physically with Cas5-Cas7-crRNA complex, stimulating binding to PAM containing substrates. Mutation of conserved Cas8 amino acid residues abolished interference in vivo and altered catalytic activity of Cas8 protein in vitro. This is experimental evidence that Cas8 is important for targeting Cascade to invader DNA. PMID:26182359

  4. Statistical Analysis of Model Data for Operational Space Launch Weather Support at Kennedy Space Center and Cape Canaveral Air Force Station

    NASA Technical Reports Server (NTRS)

    Bauman, William H., III

    2010-01-01

    The 12-km resolution North American Mesoscale (NAM) model (MesoNAM) is used by the 45th Weather Squadron (45 WS) Launch Weather Officers at Kennedy Space Center (KSC) and Cape Canaveral Air Force Station (CCAFS) to support space launch weather operations. The 45 WS tasked the Applied Meteorology Unit to conduct an objective statistics-based analysis of MesoNAM output compared to wind tower mesonet observations and then develop a an operational tool to display the results. The National Centers for Environmental Prediction began running the current version of the MesoNAM in mid-August 2006. The period of record for the dataset was 1 September 2006 - 31 January 2010. The AMU evaluated MesoNAM hourly forecasts from 0 to 84 hours based on model initialization times of 00, 06, 12 and 18 UTC. The MesoNAM forecast winds, temperature and dew point were compared to the observed values of these parameters from the sensors in the KSC/CCAFS wind tower network. The data sets were stratified by model initialization time, month and onshore/offshore flow for each wind tower. Statistics computed included bias (mean difference), standard deviation of the bias, root mean square error (RMSE) and a hypothesis test for bias = O. Twelve wind towers located in close proximity to key launch complexes were used for the statistical analysis with the sensors on the towers positioned at varying heights to include 6 ft, 30 ft, 54 ft, 60 ft, 90 ft, 162 ft, 204 ft and 230 ft depending on the launch vehicle and associated weather launch commit criteria being evaluated. These twelve wind towers support activities for the Space Shuttle (launch and landing), Delta IV, Atlas V and Falcon 9 launch vehicles. For all twelve towers, the results indicate a diurnal signal in the bias of temperature (T) and weaker but discernable diurnal signal in the bias of dewpoint temperature (T(sub d)) in the MesoNAM forecasts. Also, the standard deviation of the bias and RMSE of T, T(sub d), wind speed and wind

  5. Apport de l'imagerie dans le diagnostic des sacroiliites infectieuses : à propos de 19 cas

    PubMed Central

    Abid, Hanen; Chaabouni, Salim; Frikha, Faten; Toumi, Nozha; Souissi, Basma; Lahiani, Dorra; Bahloul, Zouhir; Ben Mahfoudh, Khaireddine

    2014-01-01

    Les sacro-iliites infectieuses méritent d’être mieux connues. Leur diagnostic est souvent retardé en raison d'une symptomatologie trompeuse et des diffcultés d'exploration de l'articulation sacro-iliaque. Notre travail est basé sur une étude rétrospective portant sur les cas de SII, recueillis sur une période comprise entre 1997 et 2008 dans notre centre universitaire Sfax-Tunisie. Le diagnostic de sacro-iliite était retenu en présence d'arguments cliniques et radiologiques d'atteinte sacroiliaque. Nous rapportons dix neuf cas de sacroiliites infectieuses (10 hommes et 9 femmes), avec un âge moyen de 32 ans. L'atteinte était unilatérale dans tous les cas. Les radiographies standard faites dans tous les cas ont été suggestives dans 14 cas et normales dans les autres cas. La TDM faite dans 13 cas a montré, un abcès des parties molles dans 8 cas et un séquestre osseux dans 2 cas. L'IRM réalisée dans 8 cas, a objectivé une infiltration des parties molles dans tous les cas et un abcès dans 3 cas. Le germe a été identifié dans 9 cas (3 cas de tuberculose, 3 cas de brucellose, 2 sacro-iliites à pyogène et un cas de candidose). Cette identification était faite par biopsie dans 3 cas, hémocultures dans 2 cas, prélèvement au niveau de la porte d'entrée dans 1 cas et sérodiagnostic dans 3 cas. Pour les autres cas, l'origine pyogène a été retenue sur des arguments cliniques et biologiques. L'imagerie joue un rôle primordial dans le diagnostic précoce et l'orientation étiologique d'une sacroiliite infectieuse. PMID:25120884

  6. Equivalent Expressions Using CAS and Paper-and-Pencil Techniques

    ERIC Educational Resources Information Center

    Fonger, Nicole L.

    2014-01-01

    How can the key concept of equivalent expressions be addressed so that students strengthen their representational fluency with symbols, graphs, and numbers? How can research inform the synergistic use of both paper-and-pencil analysis and computer algebra systems (CAS) in a classroom learning environment? These and other related questions have…

  7. Preparing Students to Take SOA/CAS Exam FM/2

    ERIC Educational Resources Information Center

    Marchand, Richard J.

    2014-01-01

    This paper provides suggestions for preparing students to take the actuarial examination on financial mathematics, SOA/CAS Exam FM/2. It is based on current practices employed at Slippery Rock University, a small public liberal arts university. Detailed descriptions of our Theory of Interest course and subsequent Exam FM/2 prep course are provided…

  8. Research Needed on the Use of CAS Standards and Guidelines.

    ERIC Educational Resources Information Center

    Creamer, Don G.

    2003-01-01

    This article suggests research projects that would extend the knowledge base about the use of Council for the Advancement of Standards in Higher Education (CAS) standards and guidelines in useful ways. Included are five research questions and specific research methodologies to guide researchers. (Contains 20 references.) (Author)

  9. Prospective Mathematics Teachers' Interactions with CAS-Based Textbook Elements

    ERIC Educational Resources Information Center

    Davis, Jon D.

    2015-01-01

    This study investigated how a group of 10 prospective secondary mathematics teachers (PST) read, evaluated, and adapted a textbook lesson involving the symbolic manipulation capabilities of computer algebra systems (CASS). PST read the entire lesson and tended to focus on the organizing question at the beginning of the student lesson and the CAS-S…

  10. Does CAS Use Disadvantage Girls in VCE Mathematics?

    ERIC Educational Resources Information Center

    Forgasz, Helen; Tan, Hazel

    2010-01-01

    In 2009, four mathematics subjects were offered at the year 12 level in the Victorian Certificate of Education (VCE). The two subjects at the intermediate level--Mathematical Methods and Mathematical Methods CAS--run in parallel, that is, a student can be enrolled in only one or the other, the choice being made at the school level. The curricular…

  11. Targeted mutagenesis in chicken using CRISPR/Cas9 system.

    PubMed

    Oishi, Isao; Yoshii, Kyoko; Miyahara, Daichi; Kagami, Hiroshi; Tagami, Takahiro

    2016-01-01

    The CRISPR/Cas9 system is a simple and powerful tool for genome editing in various organisms including livestock animals. However, the system has not been applied to poultry because of the difficulty in accessing their zygotes. Here we report the implementation of CRISPR/Cas9-mediated gene targeting in chickens. Two egg white genes, ovalbumin and ovomucoid, were efficiently (>90%) mutagenized in cultured chicken primordial germ cells (PGCs) by transfection of circular plasmids encoding Cas9, a single guide RNA, and a gene encoding drug resistance, followed by transient antibiotic selection. We transplanted CRISPR-induced mutant-ovomucoid PGCs into recipient chicken embryos and established three germline chimeric roosters (G0). All of the roosters had donor-derived mutant-ovomucoid spermatozoa, and the two with a high transmission rate of donor-derived gametes produced heterozygous mutant ovomucoid chickens as about half of their donor-derived offspring in the next generation (G1). Furthermore, we generated ovomucoid homozygous mutant offspring (G2) by crossing the G1 mutant chickens. Taken together, these results demonstrate that the CRISPR/Cas9 system is a simple and effective gene-targeting method in chickens. PMID:27050479

  12. Problem Solving in Calculus with Symbolic Geometry and CAS

    ERIC Educational Resources Information Center

    Todd, Philip; Wiechmann, James

    2008-01-01

    Computer algebra systems (CAS) have been around for a number of years, as has dynamic geometry. Symbolic geometry software is new. It bears a superficial similarity to dynamic geometry software, but differs in that problems may be set up involving symbolic variables and constants, and measurements are given as symbolic expressions. Mathematical…

  13. CRISPR/Cas9-targeted mutagenesis in Caenorhabditis elegans.

    PubMed

    Waaijers, Selma; Portegijs, Vincent; Kerver, Jana; Lemmens, Bennie B L G; Tijsterman, Marcel; van den Heuvel, Sander; Boxem, Mike

    2013-11-01

    The generation of genetic mutants in Caenorhabditis elegans has long relied on the selection of mutations in large-scale screens. Directed mutagenesis of specific loci in the genome would greatly speed up analysis of gene function. Here, we adapt the CRISPR/Cas9 system to generate mutations at specific sites in the C. elegans genome. PMID:23979586

  14. CRISPR/Cas9 advances engineering of microbial cell factories.

    PubMed

    Jakočiūnas, Tadas; Jensen, Michael K; Keasling, Jay D

    2016-03-01

    One of the key drivers for successful metabolic engineering in microbes is the efficacy by which genomes can be edited. As such there are many methods to choose from when aiming to modify genomes, especially those of model organisms like yeast and bacteria. In recent years, clustered regularly interspaced palindromic repeats (CRISPR) and its associated proteins (Cas) have become the method of choice for precision genome engineering in many organisms due to their orthogonality, versatility and efficacy. Here we review the strategies adopted for implementation of RNA-guided CRISPR/Cas9 genome editing with special emphasis on their application for metabolic engineering of yeast and bacteria. Also, examples of how nuclease-deficient Cas9 has been applied for RNA-guided transcriptional regulation of target genes will be reviewed, as well as tools available for computer-aided design of guide-RNAs will be highlighted. Finally, this review will provide a perspective on the immediate challenges and opportunities foreseen by the use of CRISPR/Cas9 genome engineering and regulation in the context of metabolic engineering. PMID:26707540

  15. CRISPR-Cas9 genome editing in Drosophila

    PubMed Central

    Gratz, Scott J.; Rubinstein, C. Dustin; Harrison, Melissa M.; Wildonger, Jill; O’Connor-Giles, Kate M.

    2015-01-01

    The CRISPR-Cas9 system has transformed genome engineering of model organisms from possible to practical. CRISPR-Cas9 can be readily programmed to generate sequence-specific double-strand breaks that disrupt targeted loci when repaired by error-prone non-homologous end joining or to catalyze precise genome modification through homology-directed repair (HDR). Here we describe a streamlined approach for rapid and highly efficient engineering of the Drosophila genome via CRISPR-Cas9-mediated HDR. In this approach, transgenic flies expressing Cas9 are injected with plasmids to express guide RNAs (gRNAs) and positively marked donor templates. We detail target site selection; gRNA plasmid generation; donor template design and construction; and the generation, identification and molecular confirmation of engineered lines. We also present alternative approaches and highlight key considerations for experimental design. The approach outlined here can be used to rapidly and reliably generate a variety of engineered modifications, including genomic deletions and replacements, precise sequence edits, and incorporation of protein tags. PMID:26131852

  16. Indispensable Manual Calculation Skills in a CAS Environment.

    ERIC Educational Resources Information Center

    Herget, Wilfried; Heugl, Helmut; Kutzler, Bernhard; Lehmann, Eberhard

    Which manual calculation skills are still needed when students use graphic/symbolic calculators or computers with computer algebra systems (CAS)? What should students be able to do manually, i.e. just using paper and pencil? This text is the outcome of a two-day discussion on these questions, held by the four authors. Our answers and proposals are…

  17. Some Reflections on CAS Assisted Proofs of Theorems

    ERIC Educational Resources Information Center

    Dana-Picard, Thierry

    2005-01-01

    A mathematician's work consists of proving theorems, calculating, and making mathematics understandable. An assistant for all three components is a Computer Algebra System. We describe and discuss various CAS-assisted processes for proving theorems, and discuss the constraints which can appear regarding efficiency, confidence in the result and…

  18. Technological Discourse on CAS-Based Operative Knowledge

    ERIC Educational Resources Information Center

    Mann, Giora; Dana-Picard, Thierry; Zehavi, Nurit

    2007-01-01

    This article begins with a comparison of two groups of teachers, working on the same tasks in Analytic Geometry. One group has only basic experience in CAS-assisted problem solving, and the other group has extensive experience. The comparison is discussed in terms of the interplay between reflection, operative knowledge and execution. The findings…

  19. Duchenne muscular dystrophy: CRISPR/Cas9 treatment.

    PubMed

    Mendell, Jerry R; Rodino-Klapac, Louise R

    2016-05-01

    A novel approach to gene correction by genome editing shows great promise as a treatment for Duchenne muscular dystrophy (DMD). CRISPR/Cas9 delivered by adeno-associated virus to a mouse model for DMD demonstrated improvement in function and histology. PMID:26926391

  20. New Insight in Mathematics by Live CAS Documents.

    ERIC Educational Resources Information Center

    Cnop, Ivan

    Traditional education in mathematics is mostly a matter of hindsight, and many mathematics texts offer little opportunity for students and learners to gain insight. This paper shows how experiments in CAS (computer algebra systems) can lead to new ways of handling problems, new conjectures, new visualizations, new proofs, new correspondences…

  1. Targeted mutagenesis in chicken using CRISPR/Cas9 system

    PubMed Central

    Oishi, Isao; Yoshii, Kyoko; Miyahara, Daichi; Kagami, Hiroshi; Tagami, Takahiro

    2016-01-01

    The CRISPR/Cas9 system is a simple and powerful tool for genome editing in various organisms including livestock animals. However, the system has not been applied to poultry because of the difficulty in accessing their zygotes. Here we report the implementation of CRISPR/Cas9-mediated gene targeting in chickens. Two egg white genes, ovalbumin and ovomucoid, were efficiently (>90%) mutagenized in cultured chicken primordial germ cells (PGCs) by transfection of circular plasmids encoding Cas9, a single guide RNA, and a gene encoding drug resistance, followed by transient antibiotic selection. We transplanted CRISPR-induced mutant-ovomucoid PGCs into recipient chicken embryos and established three germline chimeric roosters (G0). All of the roosters had donor-derived mutant-ovomucoid spermatozoa, and the two with a high transmission rate of donor-derived gametes produced heterozygous mutant ovomucoid chickens as about half of their donor-derived offspring in the next generation (G1). Furthermore, we generated ovomucoid homozygous mutant offspring (G2) by crossing the G1 mutant chickens. Taken together, these results demonstrate that the CRISPR/Cas9 system is a simple and effective gene-targeting method in chickens. PMID:27050479

  2. Maximizing mutagenesis with solubilized CRISPR-Cas9 ribonucleoprotein complexes.

    PubMed

    Burger, Alexa; Lindsay, Helen; Felker, Anastasia; Hess, Christopher; Anders, Carolin; Chiavacci, Elena; Zaugg, Jonas; Weber, Lukas M; Catena, Raul; Jinek, Martin; Robinson, Mark D; Mosimann, Christian

    2016-06-01

    CRISPR-Cas9 enables efficient sequence-specific mutagenesis for creating somatic or germline mutants of model organisms. Key constraints in vivo remain the expression and delivery of active Cas9-sgRNA ribonucleoprotein complexes (RNPs) with minimal toxicity, variable mutagenesis efficiencies depending on targeting sequence, and high mutation mosaicism. Here, we apply in vitro assembled, fluorescent Cas9-sgRNA RNPs in solubilizing salt solution to achieve maximal mutagenesis efficiency in zebrafish embryos. MiSeq-based sequence analysis of targeted loci in individual embryos using CrispRVariants, a customized software tool for mutagenesis quantification and visualization, reveals efficient bi-allelic mutagenesis that reaches saturation at several tested gene loci. Such virtually complete mutagenesis exposes loss-of-function phenotypes for candidate genes in somatic mutant embryos for subsequent generation of stable germline mutants. We further show that targeting of non-coding elements in gene regulatory regions using saturating mutagenesis uncovers functional control elements in transgenic reporters and endogenous genes in injected embryos. Our results establish that optimally solubilized, in vitro assembled fluorescent Cas9-sgRNA RNPs provide a reproducible reagent for direct and scalable loss-of-function studies and applications beyond zebrafish experiments that require maximal DNA cutting efficiency in vivo. PMID:27130213

  3. Genome Editing with CRISPR-Cas9: Can It Get Any Better?

    PubMed

    Haeussler, Maximilian; Concordet, Jean-Paul

    2016-05-20

    The CRISPR-Cas revolution is taking place in virtually all fields of life sciences. Harnessing DNA cleavage with the CRISPR-Cas9 system of Streptococcus pyogenes has proven to be extraordinarily simple and efficient, relying only on the design of a synthetic single guide RNA (sgRNA) and its co-expression with Cas9. Here, we review the progress in the design of sgRNA from the original dual RNA guide for S. pyogenes and Staphylococcus aureus Cas9 (SpCas9 and SaCas9). New assays for genome-wide identification of off-targets have provided important insights into the issue of cleavage specificity in vivo. At the same time, the on-target activity of thousands of guides has been determined. These data have led to numerous online tools that facilitate the selection of guide RNAs in target sequences. It appears that for most basic research applications, cleavage activity can be maximized and off-targets minimized by carefully choosing guide RNAs based on computational predictions. Moreover, recent studies of Cas proteins have further improved the flexibility and precision of the CRISPR-Cas toolkit for genome editing. Inspired by the crystal structure of the complex of sgRNA-SpCas9 bound to target DNA, several variants of SpCas9 have recently been engineered, either with novel protospacer adjacent motifs (PAMs) or with drastically reduced off-targets. Novel Cas9 and Cas9-like proteins called Cpf1 have also been characterized from other bacteria and will benefit from the insights obtained from SpCas9. Genome editing with CRISPR-Cas9 may also progress with better understanding and control of cellular DNA repair pathways activated after Cas9-induced DNA cleavage. PMID:27210042

  4. U.S.-MEXICO BORDER PROGRAM ARIZONA BORDER STUDY--VOCS IN AIR ANALYTICAL RESULTS

    EPA Science Inventory

    The VOCs in Air data set contains analytical results for measurements of up to 45 volatile organic compounds (VOCs) in 183 air samples over 86 households. Results include samples taken using active and passive techniques. The primary VOCs of interest include benzene (CAS# 71-43...

  5. 40 CFR Table 1 to Subpart Xx of... - Hazardous Air Pollutants

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 11 2014-07-01 2014-07-01 false Hazardous Air Pollutants 1 Table 1 to Subpart XX of Part 63 Protection of Environment ENVIRONMENTAL PROTECTION AGENCY (CONTINUED) AIR PROGRAMS... pollutant CAS No. Benzene 71432 1,3-Butadiene 106990 Cumene 98828 Ethyl benzene 100414 Hexane...

  6. Production of genome-edited pluripotent stem cells and mice by CRISPR/Cas [Review].

    PubMed

    Horii, Takuro; Hatada, Izuho

    2016-03-31

    Clustered regularly at interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) nucleases, so-called CRISPR/Cas, was recently developed as an epoch-making genome engineering technology. This system only requires Cas9 nuclease and single-guide RNA complementary to a target locus. CRISPR/Cas enables the generation of knockout cells and animals in a single step. This system can also be used to generate multiple mutations and knockin in a single step, which is not possible using other methods. In this review, we provide an overview of genome editing by CRISPR/Cas in pluripotent stem cells and mice. PMID:26743444

  7. Mobile CRISPR/Cas-Mediated Bacteriophage Resistance in Lactococcus lactis

    PubMed Central

    Millen, Anne M.; Horvath, Philippe; Boyaval, Patrick; Romero, Dennis A.

    2012-01-01

    Lactococcus lactis is a biotechnological workhorse for food fermentations and potentially therapeutic products and is therefore widely consumed by humans. It is predominantly used as a starter microbe for fermented dairy products, and specialized strains have adapted from a plant environment through reductive evolution and horizontal gene transfer as evidenced by the association of adventitious traits with mobile elements. Specifically, L. lactis has armed itself with a myriad of plasmid-encoded bacteriophage defensive systems to protect against viral predation. This known arsenal had not included CRISPR/Cas (clustered regularly interspaced short palindromic repeats/CRISPR-associated proteins), which forms a remarkable microbial immunity system against invading DNA. Although CRISPR/Cas systems are common in the genomes of closely related lactic acid bacteria (LAB), none was identified within the eight published lactococcal genomes. Furthermore, a PCR-based search of the common LAB CRISPR/Cas systems (Types I and II) in 383 industrial L. lactis strains proved unsuccessful. Here we describe a novel, Type III, self-transmissible, plasmid-encoded, phage-interfering CRISPR/Cas discovered in L. lactis. The native CRISPR spacers confer resistance based on sequence identity to corresponding lactococcal phage. The interference is directed at phages problematic to the dairy industry, indicative of a responsive system. Moreover, targeting could be modified by engineering the spacer content. The 62.8-kb plasmid was shown to be conjugally transferrable to various strains. Its mobility should facilitate dissemination within microbial communities and provide a readily applicable system to naturally introduce CRISPR/Cas to industrially relevant strains for enhanced phage resistance and prevention against acquisition of undesirable genes. PMID:23240053

  8. Adenovirus-Mediated Somatic Genome Editing of Pten by CRISPR/Cas9 in Mouse Liver in Spite of Cas9-Specific Immune Responses.

    PubMed

    Wang, Dan; Mou, Haiwei; Li, Shaoyong; Li, Yingxiang; Hough, Soren; Tran, Karen; Li, Jia; Yin, Hao; Anderson, Daniel G; Sontheimer, Erik J; Weng, Zhiping; Gao, Guangping; Xue, Wen

    2015-07-01

    CRISPR/Cas9 derived from the bacterial adaptive immunity pathway is a powerful tool for genome editing, but the safety profiles of in vivo delivered Cas9 (including host immune responses to the bacterial Cas9 protein) have not been comprehensively investigated in model organisms. Nonalcoholic steatohepatitis (NASH) is a prevalent human liver disease characterized by excessive fat accumulation in the liver. In this study, we used adenovirus (Ad) vector to deliver a Streptococcus pyogenes-derived Cas9 system (SpCas9) targeting Pten, a gene involved in NASH and a negative regulator of the PI3K-AKT pathway, in mouse liver. We found that the Ad vector mediated efficient Pten gene editing even in the presence of typical Ad vector-associated immunotoxicity in the liver. Four months after vector infusion, mice receiving the Pten gene-editing Ad vector showed massive hepatomegaly and features of NASH, consistent with the phenotypes following Cre-loxP-induced Pten deficiency in mouse liver. We also detected induction of humoral immunity against SpCas9 and the potential presence of an SpCas9-specific cellular immune response. Our findings provide a strategy to model human liver diseases in mice and highlight the importance considering Cas9-specific immune responses in future translational studies involving in vivo delivery of CRISPR/Cas9. PMID:26086867

  9. Anti-cas spacers in orphan CRISPR4 arrays prevent uptake of active CRISPR-Cas I-F systems.

    PubMed

    Almendros, Cristóbal; Guzmán, Noemí M; García-Martínez, Jesús; Mojica, Francisco J M

    2016-01-01

    Archaea and bacteria harbour clustered regularly interspaced short palindromic repeats (CRISPR) loci. These arrays encode RNA molecules (crRNA), each containing a sequence of a single repeat-intervening spacer. The crRNAs guide CRISPR-associated (Cas) proteins to cleave nucleic acids complementary to the crRNA spacer, thus interfering with targeted foreign elements. Notably, pre-existing spacers may trigger the acquisition of new spacers from the target molecule by means of a primed adaptation mechanism. Here, we show that naturally occurring orphan CRISPR arrays that contain spacers matching sequences of the cognate (absent) cas genes are able to elicit both primed adaptation and direct interference against genetic elements carrying those genes. Our findings show the existence of an anti-cas mechanism that prevents the transfer of a fully equipped CRISPR-Cas system. Hence, they suggest that CRISPR immunity may be undesired by particular prokaryotes, potentially because they could limit possibilities for gaining favourable sequences by lateral transfer. PMID:27573106

  10. Observation d'un minimum plat pour RZ Cas [Observation of a flat minimum of RZ Cas

    NASA Astrophysics Data System (ADS)

    Dumont, M.

    1995-07-01

    We observed a minimum of RZ Cas during the night 15/16 august 1991 with the 76 cm telescope of the Fungfraujoch Observatory. We found: Min = HJD 2,448,484.50062 and observed a flat minimum during 10 minutes.

  11. Structural Basis for the Altered PAM Specificities of Engineered CRISPR-Cas9.

    PubMed

    Hirano, Seiichi; Nishimasu, Hiroshi; Ishitani, Ryuichiro; Nureki, Osamu

    2016-03-17

    The RNA-guided endonuclease Cas9 cleaves double-stranded DNA targets bearing a PAM (protospacer adjacent motif) and complementarity to the guide RNA. A recent study showed that, whereas wild-type Streptococcus pyogenes Cas9 (SpCas9) recognizes the 5'-NGG-3' PAM, the engineered VQR, EQR, and VRER SpCas9 variants recognize the 5'-NGA-3', 5'-NGAG-3', and 5'-NGCG-3' PAMs, respectively, thus expanding the targetable sequences in Cas9-mediated genome editing applications. Here, we present the high-resolution crystal structures of the three SpCas9 variants in complexes with a single-guide RNA and its altered PAM-containing, partially double-stranded DNA targets. A structural comparison of the three SpCas9 variants with wild-type SpCas9 revealed that the multiple mutations synergistically induce an unexpected displacement in the phosphodiester backbone of the PAM duplex, thereby allowing the SpCas9 variants to directly recognize the altered PAM nucleotides. Our findings explain the altered PAM specificities of the SpCas9 variants and establish a framework for further rational engineering of CRISPR-Cas9. PMID:26990991

  12. Chemical and Biophysical Modulation of Cas9 for Tunable Genome Engineering.

    PubMed

    Nuñez, James K; Harrington, Lucas B; Doudna, Jennifer A

    2016-03-18

    The application of the CRISPR-Cas9 system for genome engineering has revolutionized the ability to interrogate genomes of mammalian cells. Programming the Cas9 endonuclease to induce DNA breaks at specified sites is achieved by simply modifying the sequence of its cognate guide RNA. Although Cas9-mediated genome editing has been shown to be highly specific, cleavage events at off-target sites have also been reported. Minimizing, and eventually abolishing, unwanted off-target cleavage remains a major goal of the CRISPR-Cas9 technology before its implementation for therapeutic use. Recent efforts have turned to chemical biology and biophysical approaches to engineer inducible genome editing systems for controlling Cas9 activity at the transcriptional and protein levels. Here, we review recent advancements to modulate Cas9-mediated genome editing by engineering split-Cas9 constructs, inteins, small molecules, protein-based dimerizing domains, and light-inducible systems. PMID:26857072

  13. Comparison of NASA OMI and MLS Ozone Products with US Forest Service Ground-based Ozone Monitoring Data for US Forest Service Air Quality / Forest Management Decision Support

    NASA Astrophysics Data System (ADS)

    Barrett, S.; Brooks, A.; Moussa, Y.; Spencer, T.; Thompson, J.

    2013-12-01

    Tropospheric ozone, formed when nitrogen oxides (NOx) and volatile organic compounds (VOCs) react with sunlight, is a significant threat to the health of US National Forests. Approximately one third of ozone is absorbed by plants during the uptake of carbon dioxide. This increases the vegetation's susceptibility to drought, beetle infestation, and wildfire. Currently the US Forest Service has ground monitoring stations sparsely located across the country. This project looks specifically at the area surrounding several Class I Wilderness Areas in the Appalachian region. These areas are the highest priority for protection from air pollutants. The Forest Service must interpolate ozone concentrations for areas between these monitoring stations. Class I Wilderness Areas are designated by the Forest Service and are defined as a total 5000 acres or greater when the Clean Air Act was passed in 1977. This Act mandated that the EPA create national ambient air quality standards (NAAQS) for six major air pollutants including ground-level ozone. This project assessed the feasibility of incorporating NASA ozone data into Forest Service ozone monitoring in an effort to enhance the accuracy and precision of ozone exposure measurements in Class I Wilderness Areas and other federally managed lands in order to aid in complying with the Clean Air Act of 1977. This was accomplished by establishing a method of comparison between a preliminary data product produced at the Goddard Space Flight Center that uses OMI/MLS data to derive global tropospheric ozone measurements and Forest Service ozone monitoring station measurements. Once a methodology for comparison was established, statistical comparisons of these data were performed to assess the quantitative differences.

  14. Nuclease Activity of Legionella pneumophila Cas2 Promotes Intracellular Infection of Amoebal Host Cells

    PubMed Central

    Gunderson, Felizza F.; Mallama, Celeste A.; Fairbairn, Stephanie G.

    2014-01-01

    Legionella pneumophila, the primary agent of Legionnaires' disease, flourishes in both natural and man-made environments by growing in a wide variety of aquatic amoebae. Recently, we determined that the Cas2 protein of L. pneumophila promotes intracellular infection of Acanthamoeba castellanii and Hartmannella vermiformis, the two amoebae most commonly linked to cases of disease. The Cas2 family of proteins is best known for its role in the bacterial and archeal clustered regularly interspaced short palindromic repeat (CRISPR)–CRISPR-associated protein (Cas) system that constitutes a form of adaptive immunity against phage and plasmid. However, the infection event mediated by L. pneumophila Cas2 appeared to be distinct from this function, because cas2 mutants exhibited infectivity defects in the absence of added phage or plasmid and since mutants lacking the CRISPR array or any one of the other cas genes were not impaired in infection ability. We now report that the Cas2 protein of L. pneumophila has both RNase and DNase activities, with the RNase activity being more pronounced. By characterizing a catalytically deficient version of Cas2, we determined that nuclease activity is critical for promoting infection of amoebae. Also, introduction of Cas2, but not its catalytic mutant form, into a strain of L. pneumophila that naturally lacks a CRISPR-Cas locus caused that strain to be 40- to 80-fold more infective for amoebae, unequivocally demonstrating that Cas2 facilitates the infection process independently of any other component encoded within the CRISPR-Cas locus. Finally, a cas2 mutant was impaired for infection of Willaertia magna but not Naegleria lovaniensis, suggesting that Cas2 promotes infection of most but not all amoebal hosts. PMID:25547789

  15. Air Pollution

    MedlinePlus

    Air pollution is a mixture of solid particles and gases in the air. Car emissions, chemicals from factories, dust, ... a gas, is a major part of air pollution in cities. When ozone forms air pollution, it's ...

  16. Air Pollution

    MedlinePlus

    Air pollution is a mixture of solid particles and gases in the air. Car emissions, chemicals from factories, ... Ozone, a gas, is a major part of air pollution in cities. When ozone forms air pollution, it's ...

  17. A CRISPR-Cas9 Gene Drive System Targeting Female Reproduction in the Malaria Mosquito vector Anopheles gambiae

    PubMed Central

    Hammond, Andrew; Galizi, Roberto; Kyrou, Kyros; Simoni, Alekos; Siniscalchi, Carla; Katsanos, Dimitris; Gribble, Matthew; Baker, Dean; Marois, Eric; Russell, Steven; Burt, Austin; Windbichler, Nikolai; Crisanti, Andrea; Nolan, Tony

    2016-01-01

    Gene-drive systems that enable super-Mendelian inheritance of a transgene have the potential to modify insect populations over a timeframe of a few years [AU please provide a real estimate, this seems vague]. We describe CRISPR-Cas9 endonuclease constructs that function as gene-drive systems in Anopheles gambiae, the main vector for malaria [AU:OK?]. We identified three genes (AGAP005958, AGAP011377 and AGAP007280) that confer a recessive female sterility phenotype upon disruption, and inserted into each locus CRISPR-Cas9 gene-drive constructs designed to target and edit each gene [AU:OK?]. For each locus targeted we observed strong gene drive at the molecular level, with transmission rates to progeny of 91 to 99.6%. Population modelling and cage experiments indicate that a CRISPR-Cas9 construct targeting one of these loci, AGAP007280, meets the minimum requirement for a gene drive targeting female reproduction in an insect population. These findings could expedite the development of gene drives to control suppress mosquito populations to levels that do not support malaria transmission. PMID:26641531

  18. A CRISPR-Cas9 gene drive system targeting female reproduction in the malaria mosquito vector Anopheles gambiae.

    PubMed

    Hammond, Andrew; Galizi, Roberto; Kyrou, Kyros; Simoni, Alekos; Siniscalchi, Carla; Katsanos, Dimitris; Gribble, Matthew; Baker, Dean; Marois, Eric; Russell, Steven; Burt, Austin; Windbichler, Nikolai; Crisanti, Andrea; Nolan, Tony

    2016-01-01

    Gene drive systems that enable super-Mendelian inheritance of a transgene have the potential to modify insect populations over a timeframe of a few years. We describe CRISPR-Cas9 endonuclease constructs that function as gene drive systems in Anopheles gambiae, the main vector for malaria. We identified three genes (AGAP005958, AGAP011377 and AGAP007280) that confer a recessive female-sterility phenotype upon disruption, and inserted into each locus CRISPR-Cas9 gene drive constructs designed to target and edit each gene. For each targeted locus we observed a strong gene drive at the molecular level, with transmission rates to progeny of 91.4 to 99.6%. Population modeling and cage experiments indicate that a CRISPR-Cas9 construct targeting one of these loci, AGAP007280, meets the minimum requirement for a gene drive targeting female reproduction in an insect population. These findings could expedite the development of gene drives to suppress mosquito populations to levels that do not support malaria transmission. PMID:26641531

  19. BCAR3/AND-34 can signal independent of complex formation with CAS family members or the presence of p130Cas

    PubMed Central

    Borre, Pierre Vanden; Near, Richard I.; Makkinje, Anthony; Mostoslavsky, Gustavo; Lerner, Adam

    2011-01-01

    BCAR3 binds to the carboxy-terminus of p130Cas, a focal adhesion adapter protein. Both BCAR3 and p130Cas have been linked to resistance to anti-estrogens in breast cancer, Rac activation and cell motility. Using R743A BCAR3, a point mutant that has lost the ability to bind p130Cas, we find that BCAR3-p130Cas complex formation is not required for BCAR3-mediated anti-estrogen resistance, Rac activation or discohesion of epithelial breast cancer cells. Complex formation was also not required for BCAR3-induced lamellipodia formation in BALB/c-3T3 fibroblasts but was required for optimal BCAR3-induced motility. Although both wildtype and R743A BCAR3 induced phosphorylation of p130Cas and the related adapter protein HEF1/NEDD9, chimeric NSP3:BCAR3 experiments demonstrate that such phosphorylation does not correlate with BCAR3-induced anti-estrogen resistance or lamellipodia formation. Wildtype but not R743A BCAR3 induced lamellipodia formation and augmented cell motility in p130Cas-/- murine embryonic fibroblasts (MEFs), suggesting that while p130Cas itself is not strictly required for these endpoints, complex formation with other CAS family members is, at least in cells lacking p130Cas. Overall, our work suggests that many, but not all, BCAR3-mediated signaling events in epithelial and mesenchymal cells are independent of p130Cas association. These studies also indicate that disruption of the BCAR3-p130Cas complex is unlikely to reverse BCAR3-mediated anti-estrogen resistance. PMID:21262352

  20. Parasitose intestinale simulant une appendicite à l’échographie: à propos de deux cas

    PubMed Central

    Gabrielle, Andrianah Emmylou Prisca; Hasina, Rajaonarison NyOny Narindra Lova; Ravelonarivo, RandrianantenainaFarlahy; Ahmad, Ahmad

    2015-01-01

    L'objectif de ce travail est d'apporter un diagnostic différentiel d'appendicite à l’échographie sur la région de la fosse iliaque droite, destiné notamment aux échographistes mal entrainés ou débutant. Nous rapportons deux cas de parasitoses intestinales découvertes au niveau de la fosse iliaque droite simulant une appendicite. Le premier cas concernait une femme de 45 ans, sans antécédent particulier, référée pour échographie abdominale suite à une douleur de la fosse iliaque droite, apyrétique, sans trouble de transit. L'examen échographique par voie sus-pubienne de la fosse iliaque droite (FID) objectivait une structure arrondie à paroi épaisse, de 8 mm de diamètre en coupe transversale et tubulée à extrémité borgne en coupe longitudinale, à contenu hétérogène hypoéchogène en coupe longitudinale et une sensibilité de cette zone au passage de la sonde. La biologie présentait une hyperéosinophilie et à l'examen des selles par recherche de kyste, amibe et oeufs parasitaires, on décelait des oeufs parasitaires. Le deuxième cas était un garçon de 15 ans, sans antécédent particulier. Son échographie était indiquée suite à une douleur abdominale aigue fébrile. Les images échographiques sur la FID montraient une structure arrondie de 7 mm de diamètre et à paroi épaisse hyperéchogène en coupe transversale et en coupe longitudinale, elle était tubulée borgne, immobile et sensible au passage de la sonde. La numération formule sanguine rapportait une hyperleucocytose à prédominance polynucléaire. Au total, la parasitose intestinale peut mimer un aspect échographique d'une appendicite, que tout échographiste notamment les débutants et peu expérimentésdoit éliminer. PMID:26587169

  1. An Effective Strategy for Reliably Isolating Heritable and Cas9-Free Arabidopsis Mutants Generated by CRISPR/Cas9-Mediated Genome Editing.

    PubMed

    Gao, Xiuhua; Chen, Jilin; Dai, Xinhua; Zhang, Da; Zhao, Yunde

    2016-07-01

    Mutations generated by CRISPR/Cas9 in Arabidopsis (Arabidopsis thaliana) are often somatic and are rarely heritable. Isolation of mutations in Cas9-free Arabidopsis plants can ensure the stable transmission of the identified mutations to next generations, but the process is laborious and inefficient. Here, we present a simple visual screen for Cas9-free T2 seeds, allowing us to quickly obtain Cas9-free Arabidopsis mutants in the T2 generation. To demonstrate this in principle, we targeted two sites in the AUXIN-BINDING PROTEIN1 (ABP1) gene, whose function as a membrane-associated auxin receptor has been challenged recently. We obtained many T1 plants with detectable mutations near the target sites, but only a small fraction of T1 plants yielded Cas9-free abp1 mutations in the T2 generation. Moreover, the mutations did not segregate in Mendelian fashion in the T2 generation. However, mutations identified in the Cas9-free T2 plants were stably transmitted to the T3 generation following Mendelian genetics. To further simplify the screening procedure, we simultaneously targeted two sites in ABP1 to generate large deletions, which can be easily identified by PCR. We successfully generated two abp1 alleles that contained 1,141- and 711-bp deletions in the ABP1 gene. All of the Cas9-free abp1 alleles we generated were stable and heritable. The method described here allows for effectively isolating Cas9-free heritable CRISPR mutants in Arabidopsis. PMID:27208253

  2. An Effective Strategy for Reliably Isolating Heritable and Cas9-Free Arabidopsis Mutants Generated by CRISPR/Cas9-Mediated Genome Editing1[OPEN

    PubMed Central

    Gao, Xiuhua; Chen, Jilin; Dai, Xinhua; Zhang, Da

    2016-01-01

    Mutations generated by CRISPR/Cas9 in Arabidopsis (Arabidopsis thaliana) are often somatic and are rarely heritable. Isolation of mutations in Cas9-free Arabidopsis plants can ensure the stable transmission of the identified mutations to next generations, but the process is laborious and inefficient. Here, we present a simple visual screen for Cas9-free T2 seeds, allowing us to quickly obtain Cas9-free Arabidopsis mutants in the T2 generation. To demonstrate this in principle, we targeted two sites in the AUXIN-BINDING PROTEIN1 (ABP1) gene, whose function as a membrane-associated auxin receptor has been challenged recently. We obtained many T1 plants with detectable mutations near the target sites, but only a small fraction of T1 plants yielded Cas9-free abp1 mutations in the T2 generation. Moreover, the mutations did not segregate in Mendelian fashion in the T2 generation. However, mutations identified in the Cas9-free T2 plants were stably transmitted to the T3 generation following Mendelian genetics. To further simplify the screening procedure, we simultaneously targeted two sites in ABP1 to generate large deletions, which can be easily identified by PCR. We successfully generated two abp1 alleles that contained 1,141- and 711-bp deletions in the ABP1 gene. All of the Cas9-free abp1 alleles we generated were stable and heritable. The method described here allows for effectively isolating Cas9-free heritable CRISPR mutants in Arabidopsis. PMID:27208253

  3. Broadening Staphylococcus aureus Cas9 Targeting Range by Modifying PAM Recognition

    PubMed Central

    Kleinstiver, Benjamin P.; Prew, Michelle S.; Tsai, Shengdar Q.; Nguyen, Nhu T.; Topkar, Ved V.; Zheng, Zongli; Joung, J. Keith

    2015-01-01

    CRISPR-Cas9 nucleases are primarily guided by RNA-DNA interactions but also require Cas9-mediated recognition of a protospacer adjacent motif (PAM). While potentially advantageous for specificity, extended PAM sequences limit the targeting range of Cas9 orthologues for genome editing. One possible strategy to relieve this restriction is to relax specificities for certain positions within the PAM. Here we used molecular evolution to modify the NNGRRT PAM specificity of Staphylococcus aureus Cas9 (SaCas9). One variant we identified, referred to as KKH SaCas9, shows robust genome editing activities at endogenous human target sites with NNNRRT PAMs. Importantly, using GUIDE-seq, we show that both wild-type and KKH SaCas9 induce comparable numbers of off-target effects in human cells. KKH SaCas9 increased the targeting range of SaCas9 by nearly two- to four-fold. Our molecular evolution strategy does not require structural information and therefore should be applicable to a wide range of Cas9 orthologues. PMID:26524662

  4. Nucleosomes impede Cas9 access to DNA in vivo and in vitro.

    PubMed

    Horlbeck, Max A; Witkowsky, Lea B; Guglielmi, Benjamin; Replogle, Joseph M; Gilbert, Luke A; Villalta, Jacqueline E; Torigoe, Sharon E; Tjian, Robert; Weissman, Jonathan S

    2016-01-01

    The prokaryotic CRISPR (clustered regularly interspaced palindromic repeats)-associated protein, Cas9, has been widely adopted as a tool for editing, imaging, and regulating eukaryotic genomes. However, our understanding of how to select single-guide RNAs (sgRNAs) that mediate efficient Cas9 activity is incomplete, as we lack insight into how chromatin impacts Cas9 targeting. To address this gap, we analyzed large-scale genetic screens performed in human cell lines using either nuclease-active or nuclease-dead Cas9 (dCas9). We observed that highly active sgRNAs for Cas9 and dCas9 were found almost exclusively in regions of low nucleosome occupancy. In vitro experiments demonstrated that nucleosomes in fact directly impede Cas9 binding and cleavage, while chromatin remodeling can restore Cas9 access. Our results reveal a critical role of eukaryotic chromatin in dictating the targeting specificity of this transplanted bacterial enzyme, and provide rules for selecting Cas9 target sites distinct from and complementary to those based on sequence properties. PMID:26987018

  5. Nucleosomes impede Cas9 access to DNA in vivo and in vitro

    PubMed Central

    Horlbeck, Max A; Witkowsky, Lea B; Guglielmi, Benjamin; Replogle, Joseph M; Gilbert, Luke A; Villalta, Jacqueline E; Torigoe, Sharon E; Tjian, Robert; Weissman, Jonathan S

    2016-01-01

    The prokaryotic CRISPR (clustered regularly interspaced palindromic repeats)-associated protein, Cas9, has been widely adopted as a tool for editing, imaging, and regulating eukaryotic genomes. However, our understanding of how to select single-guide RNAs (sgRNAs) that mediate efficient Cas9 activity is incomplete, as we lack insight into how chromatin impacts Cas9 targeting. To address this gap, we analyzed large-scale genetic screens performed in human cell lines using either nuclease-active or nuclease-dead Cas9 (dCas9). We observed that highly active sgRNAs for Cas9 and dCas9 were found almost exclusively in regions of low nucleosome occupancy. In vitro experiments demonstrated that nucleosomes in fact directly impede Cas9 binding and cleavage, while chromatin remodeling can restore Cas9 access. Our results reveal a critical role of eukaryotic chromatin in dictating the targeting specificity of this transplanted bacterial enzyme, and provide rules for selecting Cas9 target sites distinct from and complementary to those based on sequence properties. DOI: http://dx.doi.org/10.7554/eLife.12677.001 PMID:26987018

  6. CRISPR/Cas9-induced knockout and knock-in mutations in Chlamydomonas reinhardtii

    PubMed Central

    Shin, Sung-Eun; Lim, Jong-Min; Koh, Hyun Gi; Kim, Eun Kyung; Kang, Nam Kyu; Jeon, Seungjib; Kwon, Sohee; Shin, Won-Sub; Lee, Bongsoo; Hwangbo, Kwon; Kim, Jungeun; Ye, Sung Hyeok; Yun, Jae-Young; Seo, Hogyun; Oh, Hee-Mock; Kim, Kyung-Jin; Kim, Jin-Soo; Jeong, Won-Joong; Chang, Yong Keun; Jeong, Byeong-ryool

    2016-01-01

    Genome editing is crucial for genetic engineering of organisms for improved traits, particularly in microalgae due to the urgent necessity for the next generation biofuel production. The most advanced CRISPR/Cas9 system is simple, efficient and accurate in some organisms; however, it has proven extremely difficult in microalgae including the model alga Chlamydomonas. We solved this problem by delivering Cas9 ribonucleoproteins (RNPs) comprising the Cas9 protein and sgRNAs to avoid cytotoxicity and off-targeting associated with vector-driven expression of Cas9. We obtained CRISPR/Cas9-induced mutations at three loci including MAA7, CpSRP43 and ChlM, and targeted mutagenic efficiency was improved up to 100 fold compared to the first report of transgenic Cas9-induced mutagenesis. Interestingly, we found that unrelated vectors used for the selection purpose were predominantly integrated at the Cas9 cut site, indicative of NHEJ-mediated knock-in events. As expected with Cas9 RNPs, no off-targeting was found in one of the mutagenic screens. In conclusion, we improved the knockout efficiency by using Cas9 RNPs, which opens great opportunities not only for biological research but also industrial applications in Chlamydomonas and other microalgae. Findings of the NHEJ-mediated knock-in events will allow applications of the CRISPR/Cas9 system in microalgae, including “safe harboring” techniques shown in other organisms. PMID:27291619

  7. CRISPR/Cas9-induced knockout and knock-in mutations in Chlamydomonas reinhardtii.

    PubMed

    Shin, Sung-Eun; Lim, Jong-Min; Koh, Hyun Gi; Kim, Eun Kyung; Kang, Nam Kyu; Jeon, Seungjib; Kwon, Sohee; Shin, Won-Sub; Lee, Bongsoo; Hwangbo, Kwon; Kim, Jungeun; Ye, Sung Hyeok; Yun, Jae-Young; Seo, Hogyun; Oh, Hee-Mock; Kim, Kyung-Jin; Kim, Jin-Soo; Jeong, Won-Joong; Chang, Yong Keun; Jeong, Byeong-Ryool

    2016-01-01

    Genome editing is crucial for genetic engineering of organisms for improved traits, particularly in microalgae due to the urgent necessity for the next generation biofuel production. The most advanced CRISPR/Cas9 system is simple, efficient and accurate in some organisms; however, it has proven extremely difficult in microalgae including the model alga Chlamydomonas. We solved this problem by delivering Cas9 ribonucleoproteins (RNPs) comprising the Cas9 protein and sgRNAs to avoid cytotoxicity and off-targeting associated with vector-driven expression of Cas9. We obtained CRISPR/Cas9-induced mutations at three loci including MAA7, CpSRP43 and ChlM, and targeted mutagenic efficiency was improved up to 100 fold compared to the first report of transgenic Cas9-induced mutagenesis. Interestingly, we found that unrelated vectors used for the selection purpose were predominantly integrated at the Cas9 cut site, indicative of NHEJ-mediated knock-in events. As expected with Cas9 RNPs, no off-targeting was found in one of the mutagenic screens. In conclusion, we improved the knockout efficiency by using Cas9 RNPs, which opens great opportunities not only for biological research but also industrial applications in Chlamydomonas and other microalgae. Findings of the NHEJ-mediated knock-in events will allow applications of the CRISPR/Cas9 system in microalgae, including "safe harboring" techniques shown in other organisms. PMID:27291619

  8. Force Sensing by Extension of the Src Family Kinase Substrate, p130Cas

    PubMed Central

    Sawada, Yasuhiro; Tamada, Masako; Dubin-Thaler, Benjamin J.; Cherniavskaya, Oksana; Sakai, Ryuichi; Tanaka, Sakae; Sheetz, Michael P.

    2009-01-01

    SUMMARY Physical force is implicated in many cell functions. However, the molecular mechanisms of force sensing are poorly understood. Here, we show that mechanical extension of p130Cas (Cas) in vitro and in vivo causes phosphorylation by Src family kinases with no apparent change in Src kinase activity and that Cas phosphorylation is involved in stretch-dependent activation of the small GTPase, Rap1. In vitro, we mechanically extended bacterially expressed Cas substrate domain (CasSD) and found a remarkable enhancement of phosphorylation by specific kinases. Using an antibody that recognized extended CasSD in vitro, Cas extension in intact cells was observed in the peripheral regions of spreading cells where higher traction forces are expected and phosphorylated Cas was detected, suggesting that the in vitro extension and phosphorylation of CasSD is relevant to physiological force transduction. Thus, Cas acts as a primary force-sensor through extension of the substrate domain, which primes it for phosphorylation. PMID:17129785

  9. Highly efficient targeted chromosome deletions using CRISPR/Cas9.

    PubMed

    He, Zuyong; Proudfoot, Chris; Mileham, Alan J; McLaren, David G; Whitelaw, C Bruce A; Lillico, Simon G

    2015-05-01

    The CRISPR/Cas9 system has emerged as an intriguing new technology for genome engineering. It utilizes the bacterial endonuclease Cas9 which, when delivered to eukaryotic cells in conjunction with a user-specified small guide RNA (gRNA), cleaves the chromosomal DNA at the target site. Here we show that concurrent delivery of gRNAs designed to target two different sites in a human chromosome introduce DNA double-strand breaks in the chromosome and give rise to targeted deletions of the intervening genomic segment. Predetermined genomic DNA segments ranging from several-hundred base pairs to 1 Mbp can be precisely deleted at frequencies of 1-10%, with no apparent correlation between the size of the deleted fragment and the deletion frequency. The high efficiency of this technique holds promise for large genomic deletions that could be useful in generation of cell and animal models with engineered chromosomes. PMID:25362885

  10. Energy biotechnology in the CRISPR-Cas9 era.

    PubMed

    Estrela, Raissa; Cate, Jamie Harrison Doudna

    2016-04-01

    The production of bioenergy from plant biomass previously relied on using microorganisms that rapidly and efficiently convert simple sugars into fuels and chemicals. However, to exploit the far more abundant carbon fixed in plant cell walls, future industrial production hosts will need to be engineered to leverage the most efficient biochemical pathways and most robust traits that can be found in nature. The CRISPR-Cas9 genome editing technology now enables writing the genome at will, which will allow biotechnology to become an 'information science.' This review covers recent advances in using CRISPR-Cas9 to engineer the genomes of a wide variety of organisms that could be use in the industrial production of biofuels and renewable chemicals. PMID:26874259

  11. Application of CRISPR/Cas9 Technology to HBV

    PubMed Central

    Lin, Guigao; Zhang, Kuo; Li, Jinming

    2015-01-01

    More than 240 million people around the world are chronically infected with hepatitis B virus (HBV). Nucleos(t)ide analogs and interferon are the only two families of drugs to treat HBV currently. However, none of these anti-virals directly target the stable nuclear covalently closed circular DNA (cccDNA), which acts as a transcription template for viral mRNA and pre-genomic RNA synthesis and secures virus persistence. Thus, the fact that only a small number of patients treated achieve sustained viral response (SVR) or cure, highlights the need for new therapies against HBV. The clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 gene editing system can specifically target the conserved regions of the HBV genome. This results in robust viral suppression and provides a promising tool for eradicating the virus. In this review, we discuss the function and application of the CRISPR/Cas9 system as a novel therapy for HBV. PMID:26540039

  12. Chromosome engineering in zygotes with CRISPR/Cas9

    PubMed Central

    Boroviak, Katharina; Doe, Brendan; Banerjee, Ruby; Yang, Fengtang

    2016-01-01

    SUMMARY Deletions, duplications, and inversions of large genomic regions covering several genes are an important class of disease causing variants in humans. Modeling these structural variants in mice requires multistep processes in ES cells, which has limited their availability. Mutant mice containing small insertions, deletions, and single nucleotide polymorphisms can be reliably generated using CRISPR/Cas9 directly in mouse zygotes. Large structural variants can be generated using CRISPR/Cas9 in ES cells, but it has not been possible to generate these directly in zygotes. We now demonstrate the direct generation of deletions, duplications and inversions of up to one million base pairs by zygote injection. genesis 54:78–85, 2016. © 2016 The Authors. genesis Published by Wiley Periodicals, Inc. PMID:26742453

  13. Next Generation Prokaryotic Engineering: The CRISPR-Cas Toolkit.

    PubMed

    Mougiakos, Ioannis; Bosma, Elleke F; de Vos, Willem M; van Kranenburg, Richard; van der Oost, John

    2016-07-01

    The increasing demand for environmentally friendly production processes of green chemicals and fuels has stimulated research in microbial metabolic engineering. CRISPR-Cas-based tools for genome editing and expression control have enabled fast, easy, and accurate strain development for established production platform organisms, such as Escherichia coli and Saccharomyces cerevisiae. However, the growing interest in alternative production hosts, for which genome editing options are generally limited, requires further developing such engineering tools. In this review, we discuss established and emerging CRISPR-Cas-based tools for genome editing and transcription control of model and non-model prokaryotes, and we analyse the possibilities for further improvement and expansion of these tools for next generation prokaryotic engineering. PMID:26944793

  14. Photometry of stars in the Cas OB5 Associations

    NASA Astrophysics Data System (ADS)

    Tanriver, Mehmet; Keskin, Ahmet

    2016-07-01

    OB associations are a grouping of very young associations, contain 10-100 very hot massive stars, spectral types O and B. Also, the OB associations contain low and intermediate mass stars, too. Association members are believed to form within the same small volume inside a giant molecular cloud. Once the surrounding dust and gas is blown away, the remaining stars become not tied up and begin to drift separately. It is believed that the majority of all stars in the Milky Way were formed in OB associations. O type stars are short-lived, and will be at an end as supernovae after roundly a million years. OB associations are generally only a few million years in age or less. In this study, the photometry of UU Cas and field star which been Cas OB5 association member was carried out. Light curves and color diagrams are given in the study.

  15. Analysis of estimation algorithms for CDTI and CAS applications

    NASA Technical Reports Server (NTRS)

    Goka, T.

    1985-01-01

    Estimation algorithms for Cockpit Display of Traffic Information (CDTI) and Collision Avoidance System (CAS) applications were analyzed and/or developed. The algorithms are based on actual or projected operational and performance characteristics of an Enhanced TCAS II traffic sensor developed by Bendix and the Federal Aviation Administration. Three algorithm areas are examined and discussed. These are horizontal x and y, range and altitude estimation algorithms. Raw estimation errors are quantified using Monte Carlo simulations developed for each application; the raw errors are then used to infer impacts on the CDTI and CAS applications. Applications of smoothing algorithms to CDTI problems are also discussed briefly. Technical conclusions are summarized based on the analysis of simulation results.

  16. Dissociation of CAS phase in the uppermost lower mantle

    NASA Astrophysics Data System (ADS)

    Ishibashi, Kazufusa; Hirose, Kei; Sata, Nagayoshi; Ohishi, Yasuo

    2008-05-01

    The high-pressure stability limit of calcium aluminosilicate (CAS) phase has been examined in its end-member CaAl4Si2O11 composition at 18 39 GPa and 1,670 2,300 K in a laser-heated diamond-anvil cell (LHDAC). The in-situ synchrotron X-ray diffraction measurements revealed that the CAS phase decomposes into three-phase assemblage of cubic Al-bearing CaSiO3 perovskite, Al2O3 corundum, and SiO2 stishovite above 30 GPa and 2,000 K with a positive pressure temperature slope. Present results have important implications for the subsolidus mineral assemblage of subducted sediment and the melting phase relation of basalt in the lower mantle.

  17. Plasmocytome costal solitaire: à propos d'un cas

    PubMed Central

    Razafimanjato, Narindra Njarasoa Mihaja; Ravoatrarilandy, Manjakaniaina; Rakotoarisoa, Andriamihaja Jean Claude; Hasiniatsy, Rodrigue; Hunald, Allen Francis; Rakototiana, Auberlin Felantsoa; Rafaramino, Florine; Rakotovao, Hanitrala Jean Louis

    2014-01-01

    Les auteurs rapportent un cas de plasmocytome solitaire particulière par leur localisation costale. Le diagnostic est basé sur la mise en évidence d'une tumeur localisée, constituée de cellules plasmocytaires monoclonales cytologiquement identiques à celles du myélome multiple, en l'absence d'autres signes en faveur d'une forme disséminée. Nous rapportons un cas de plasmocytome solitaire à localisation costale et nous discutons les aspects diagnostiques et thérapeutiques de cette affection potentiellement menacée dans son évolution par la transformation en myélome multiple. PMID:25419306

  18. Inactivation of Cancer Mutations Utilizing CRISPR/Cas9.

    PubMed

    Gebler, Christina; Lohoff, Tim; Paszkowski-Rogacz, Maciej; Mircetic, Jovan; Chakraborty, Debojyoti; Camgoz, Aylin; Hamann, Martin V; Theis, Mirko; Thiede, Christian; Buchholz, Frank

    2017-01-01

    Although whole-genome sequencing has uncovered a large number of mutations that drive tumorigenesis, functional ratification for most mutations remains sparse. Here, we present an approach to test functional relevance of tumor mutations employing CRISPR/Cas9. Combining comprehensive sgRNA design and an efficient reporter assay to nominate efficient and selective sgRNAs, we establish a pipeline to dissect roles of cancer mutations with potential applicability to personalized medicine and future therapeutic use. PMID:27576906

  19. Motions of alloying additions in the CAS steelmaking operations

    NASA Astrophysics Data System (ADS)

    Mazumdar, D.; Guthrie, R. I. L.

    1993-08-01

    Water model studies in a pilot scale ladle ( D = 1.12 m and L = 0.93 m) were carried out to investigate the subsurface motion of both buoyant and sinking additions during the CAS (com-position adjustment by sealed argon bubbling systems) alloy addition procedure in steelmaking. This technique involves placing a refractory baffle around a rising gas/liquid plume within a stirred ladle of steel. Alloy additions are then made by projecting them into the slag-free region of steel within the baffled region. It was found that such particles while moving through the upwelling two-phase plume region can experience a significant reduction in drag forces, causing buoyant particles to penetrate more deeply than anticipated for a homogeneous fluid. Therefore, considering reduced drag on particles penetrating the upwelling gas liquid plume region, predictions were made for the trajectories of spherical-shaped particles using Newton’s law of motion. Predictions were in very reasonable agreement with those measured. Incorporating the velocity fields in industrial size vessels already reported by the present authors, trajectories of spherical-shaped additions (diameter ˜ 80 mm) in a 150-ton ladle during CAS operations were then predicted. The industrial implications of such trajectories, together with the alloy’s dissolution and dispersion behavior, were also analyzed. Finally, advantages of the CAS alloy addition procedure over conventional methods, in terms of the recovery rates of buoyant additions, are discussed.

  20. Targeted mutagenesis using CRISPR/Cas system in medaka

    PubMed Central

    Ansai, Satoshi; Kinoshita, Masato

    2014-01-01

    ABSTRACT Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) system-based RNA-guided endonuclease (RGEN) has recently emerged as a simple and efficient tool for targeted genome editing. In this study, we showed successful targeted mutagenesis using RGENs in medaka, Oryzias latipes. Somatic and heritable mutations were induced with high efficiency at the targeted genomic sequence on the DJ-1 gene in embryos that had been injected with the single guide RNA (sgRNA) transcribed by a T7 promoter and capped RNA encoding a Cas9 nuclease. The sgRNAs that were designed for the target genomic sequences without the 5′ end of GG required by the T7 promoter induced the targeted mutations. This suggests that the RGEN can target any sequence adjacent to an NGG protospacer adjacent motif (PAM) sequence, which occurs once every 8 bp. The off-target alterations at 2 genomic loci harboring double mismatches in the 18-bp targeting sequences were induced in the RGEN-injected embryos. However, we also found that the off-target effects could be reduced by lower dosages of sgRNA. Taken together, our results suggest that CRISPR/Cas-mediated RGENs may be an efficient and flexible tool for genome editing in medaka. PMID:24728957

  1. Embolie de liquide amniotique: à propos de deux cas

    PubMed Central

    Elbahraoui, Houda; Bouziane, Hanane; Elghanmi, Adil; Lakhdar, Amina; Elhanchi, Zaki; Ferhati, Driss

    2012-01-01

    L’embolie de liquide amniotique (ELA) est une complication imprévisible de l’accouchement, souvent fatale, associant un collapsus cardiovasculaire sévère, un syndrome de détresse respiratoire aiguë et une hémorragie avec coagulation intra vasculaire disséminée (CIVD). Dès l’évocation du diagnostic, la prise en charge doit être multidisciplinaire et intensive. ELA est responsable d’une mortalité maternelle et néonatale importante, son incidence est extrêmement variable selon les études et le taux de mortalité maternelle varie entre 26 et 86 % selon les études. Ces dix dernières années, le pronostic materno-fœtal semble en amélioration grâce aux progrès de prise en charge standardisée multidisciplinaire sur les lieux d’accouchement. Nous rapportons deux cas d’embolie de liquide amniotique. Le premier cas s’est manifesté au cours du travail et le deuxième cas est survenu dans les suites immédiates de l’accouchement. PMID:22655108

  2. CRISPR-Cas systems for genome editing, regulation and targeting

    PubMed Central

    Sander, Jeffry D.; Joung, J. Keith

    2014-01-01

    Targeted genome editing using engineered nucleases has rapidly transformed from a niche technology to a mainstream method used by many biological researchers. This widespread adoption has been largely fueled by the emergence of the clustered regularly interspaced short palindromic repeat (CRISPR) technology, an important new platform for generating RNA-guided nucleases (RGNs), such as Cas9, with customizable specificities. RGN-mediated genome editing is facile, rapid and has enabled the efficient modification of endogenous genes in a wide variety of biomedically important cell types and novel organisms that have traditionally been challenging to manipulate genetically. Furthermore, a modified version of the CRISPR-Cas9 system has been developed to recruit heterologous domains that can regulate endogenous gene expression or label specific genomic loci in living cells. Although the genome-wide specificities of CRISPR-Cas9 systems remain to be fully defined, the capabilities of these systems to perform targeted, highly efficient alterations of genome sequence and gene expression will undoubtedly transform biological research and spur the development of novel molecular therapeutics for human disease. PMID:24584096

  3. Climatological classification of five sectors in the Iberian Peninsula using columnar (AOD, α) and surface (PM10, PM2.5) aerosol data supported by air mass apportioning

    NASA Astrophysics Data System (ADS)

    Cachorro, Victoria; Mateos, David; Toledano, Carlos; Burgos, Maria A.; Bennouna, Yasmine; Torres, Benjamín; Fuertes, David; González, Ramiro; Guirado, Carmen; Román, Roberto; Velasco-Merino, Cristian; Marcos, Alberto; Calle, Abel; de Frutos, Angel M.

    2015-04-01

    The study of atmospheric aerosol over the Iberian Peninsula (IP) under a climatologic perspective is an interesting and meaningful aim due to the wide variety of conditions (geographical position, air masses, topography, among others) which cause a complex role of the distribution of aerosol properties. In the deeply investigation on the annual cycle and time evolution of the particulate matter lower than 10 µm (PM10, surface) and aerosol optical depth (AOD, columnar) in a large number of sites covering the period 2000-2013, five sectors can be distinguished in the IP. Both set of data belong to EMEP and AERONET networks respectively, as representative of aerosol air quality and climate studies, are complementary elements for a global aerosol research. The prevalence of fine-coarse particles is also analyzed over each sector. Seasonal bimodality of the PM10 annual cycle with a strong North-South gradient is observed in most sites, but this is only reported in the AOD climatology for the southern IP. The northern coast is clearly governed by the Atlantic Ocean influence, while the northeastern area is modulated by the Mediterranean Sea. The southern area, very close to the African continent, presents a large influence of desert dust intrusions. However, the southern Atlantic and Mediterranean coast present discrepancies and two sectors have been defined in this area. Finally, the center of the Peninsula is a mix of conditions, with north-south and east-west gradients of different magnitude. Overall, there is a relationship between PM10 and AOD with a proportional factor varying from 20 to 90, depending on the sector. The particular characteristic of PM10-AOD annual cycle of each geographical sector can be understood by the different climatology of the air mass origins observed at 500 and 1500 m (a.s.l.) and its apportioning to PM10 and AOD, respectively.

  4. The OceanFlux Greenhouse Gases methodology for deriving a sea surface climatology of CO2 fugacity in support of air-sea gas flux studies

    NASA Astrophysics Data System (ADS)

    Goddijn-Murphy, L. M.; Woolf, D. K.; Land, P. E.; Shutler, J. D.; Donlon, C.

    2015-07-01

    Climatologies, or long-term averages, of essential climate variables are useful for evaluating models and providing a baseline for studying anomalies. The Surface Ocean CO2 Atlas (SOCAT) has made millions of global underway sea surface measurements of CO2 publicly available, all in a uniform format and presented as fugacity, fCO2. As fCO2 is highly sensitive to temperature, the measurements are only valid for the instantaneous sea surface temperature (SST) that is measured concurrently with the in-water CO2 measurement. To create a climatology of fCO2 data suitable for calculating air-sea CO2 fluxes, it is therefore desirable to calculate fCO2 valid for a more consistent and averaged SST. This paper presents the OceanFlux Greenhouse Gases methodology for creating such a climatology. We recomputed SOCAT's fCO2 values for their respective measurement month and year using monthly composite SST data on a 1° × 1° grid from satellite Earth observation and then extrapolated the resulting fCO2 values to reference year 2010. The data were then spatially interpolated onto a 1° × 1° grid of the global oceans to produce 12 monthly fCO2 distributions for 2010, including the prediction errors of fCO2 produced by the spatial interpolation technique. The partial pressure of CO2 (pCO2) is also provided for those who prefer to use pCO2. The CO2 concentration difference between ocean and atmosphere is the thermodynamic driving force of the air-sea CO2 flux, and hence the presented fCO2 distributions can be used in air-sea gas flux calculations together with climatologies of other climate variables.

  5. The effectiveness of circulating aeration in air and high purity oxygen systems for control of VOC emissions from aeration basins

    SciTech Connect

    Zhu, H.; Keener, T.C.; Bishop, P.L.; Orton, T.L.; Wang, M.; Siddiqui, K.F.

    1997-12-31

    A simple steady state circulating aeration system (CAS) model has been used to study the effects of volatility and degradability on the fate of VOCs in both air and high purity oxygen (HPO) systems. With increase of circulation ratio in an air CAS, air emissions by stripping can be significantly reduced for compounds of low degradabilities and high volatilities. Enhancement of biodegradation is more significant for compounds of high degradabilities and volatilities. A large portion of VOCs will remain in the wastewater when circulation ratio is high, especially for VOCs that are difficult to degrade. In HPO systems, emissions by stripping are much less than air systems. However, VOCs will remain in the wastewater if they have poor degradabilities. Volatilities of VOCs are not important in HPO systems. Due to their wide range and large uncertainties, degradation rate constants are a major factor determining the effectiveness of a CAS for VOC emission control

  6. Generation of Two Noradrenergic-Specific Dopamine-Beta-Hydroxylase-FLPo Knock-In Mice Using CRISPR/Cas9-Mediated Targeting in Embryonic Stem Cells

    PubMed Central

    Sun, Jenny J.

    2016-01-01

    CRISPR/Cas9 mediated DNA double strand cutting is emerging as a powerful approach to increase rates of homologous recombination of large targeting vectors, but the optimization of parameters, equipment and expertise required remain barriers to successful mouse generation by single-step zygote injection. Here, we sought to apply CRISPR/Cas9 methods to traditional embryonic stem (ES) cell targeting followed by blastocyst injection to overcome the common issues of difficult vector construction and low targeting efficiency. To facilitate the study of noradrenergic function, which is implicated in myriad behavioral and physiological processes, we generated two different mouse lines that express FLPo recombinase under control of the noradrenergic-specific Dopamine-Beta-Hydroxylase (DBH) gene. We found that by co-electroporating a circular vector expressing Cas9 and a locus-specific sgRNA, we could target FLPo to the DBH locus in ES cells with shortened 1 kb homology arms. Two different sites in the DBH gene were targeted; the translational start codon with 6–8% targeting efficiency, and the translational stop codon with 75% targeting efficiency. Using this approach, we established two mouse lines with DBH-specific expression of FLPo in brainstem catecholaminergic populations that are publically available on MMRRC (MMRRC_041575-UCD and MMRRC_041577-UCD). Altogether, this study supports simplified, high-efficiency Cas9/CRISPR-mediated targeting in embryonic stem cells for production of knock-in mouse lines in a wider variety of contexts than zygote injection alone. PMID:27441631

  7. Generation of Two Noradrenergic-Specific Dopamine-Beta-Hydroxylase-FLPo Knock-In Mice Using CRISPR/Cas9-Mediated Targeting in Embryonic Stem Cells.

    PubMed

    Sun, Jenny J; Ray, Russell

    2016-01-01

    CRISPR/Cas9 mediated DNA double strand cutting is emerging as a powerful approach to increase rates of homologous recombination of large targeting vectors, but the optimization of parameters, equipment and expertise required remain barriers to successful mouse generation by single-step zygote injection. Here, we sought to apply CRISPR/Cas9 methods to traditional embryonic stem (ES) cell targeting followed by blastocyst injection to overcome the common issues of difficult vector construction and low targeting efficiency. To facilitate the study of noradrenergic function, which is implicated in myriad behavioral and physiological processes, we generated two different mouse lines that express FLPo recombinase under control of the noradrenergic-specific Dopamine-Beta-Hydroxylase (DBH) gene. We found that by co-electroporating a circular vector expressing Cas9 and a locus-specific sgRNA, we could target FLPo to the DBH locus in ES cells with shortened 1 kb homology arms. Two different sites in the DBH gene were targeted; the translational start codon with 6-8% targeting efficiency, and the translational stop codon with 75% targeting efficiency. Using this approach, we established two mouse lines with DBH-specific expression of FLPo in brainstem catecholaminergic populations that are publically available on MMRRC (MMRRC_041575-UCD and MMRRC_041577-UCD). Altogether, this study supports simplified, high-efficiency Cas9/CRISPR-mediated targeting in embryonic stem cells for production of knock-in mouse lines in a wider variety of contexts than zygote injection alone. PMID:27441631

  8. Application of support vector machine method for the analysis of absorption spectra of exhaled air of patients with broncho-pulmonary diseases

    NASA Astrophysics Data System (ADS)

    Bukreeva, Ekaterina B.; Bulanova, Anna A.; Kistenev, Yury V.; Kuzmin, Dmitry A.; Tuzikov, Sergei A.; Yumov, Evgeny L.

    2014-11-01

    The results of the joint use of laser photoacoustic spectroscopy and chemometrics methods in gas analysis of exhaled air of patients with respiratory diseases (chronic obstructive pulmonary disease, pneumonia and lung cancer) are presented. The absorption spectra of exhaled breath of all volunteers were measured, the classification methods of the scans of the absorption spectra were applied, the sensitivity/specificity of the classification results were determined. It were obtained a result of nosological in pairs classification for all investigated volunteers, indices of sensitivity and specificity.

  9. Mobile phone tracking: in support of modelling traffic-related air pollution contribution to individual exposure and its implications for public health impact assessment.

    PubMed

    Liu, Hai-Ying; Skjetne, Erik; Kobernus, Mike

    2013-01-01

    We propose a new approach to assess the impact of traffic-related air pollution on public health by mapping personal trajectories using mobile phone tracking technology in an urban environment. Although this approach is not based on any empirical studies, we believe that this method has great potential and deserves serious attention. Mobile phone tracking technology makes it feasible to generate millions of personal trajectories and thereby cover a large fraction of an urban population. Through analysis, personal trajectories are not only associated to persons, but it can also be associated with vehicles, vehicle type, vehicle speed, vehicle emission rates, and sources of vehicle emissions. Pollution levels can be estimated by dispersion models from calculated traffic emissions. Traffic pollution exposure to individuals can be estimated based on the exposure along the individual human trajectories in the estimated pollution concentration fields by utilizing modelling tools. By data integration, one may identify trajectory patterns of particularly exposed human groups. The approach of personal trajectories may open a new paradigm in understanding urban dynamics and new perspectives in population-wide empirical public health research. This new approach can be further applied to individual commuter route planning, land use planning, urban traffic network planning, and used by authorities to formulate air pollution mitigation policies and regulations. PMID:24188173

  10. Mobile phone tracking: in support of modelling traffic-related air pollution contribution to individual exposure and its implications for public health impact assessment

    PubMed Central

    2013-01-01

    We propose a new approach to assess the impact of traffic-related air pollution on public health by mapping personal trajectories using mobile phone tracking technology in an urban environment. Although this approach is not based on any empirical studies, we believe that this method has great potential and deserves serious attention. Mobile phone tracking technology makes it feasible to generate millions of personal trajectories and thereby cover a large fraction of an urban population. Through analysis, personal trajectories are not only associated to persons, but it can also be associated with vehicles, vehicle type, vehicle speed, vehicle emission rates, and sources of vehicle emissions. Pollution levels can be estimated by dispersion models from calculated traffic emissions. Traffic pollution exposure to individuals can be estimated based on the exposure along the individual human trajectories in the estimated pollution concentration fields by utilizing modelling tools. By data integration, one may identify trajectory patterns of particularly exposed human groups. The approach of personal trajectories may open a new paradigm in understanding urban dynamics and new perspectives in population-wide empirical public health research. This new approach can be further applied to individual commuter route planning, land use planning, urban traffic network planning, and used by authorities to formulate air pollution mitigation policies and regulations. PMID:24188173

  11. How Do Pre-Service Teachers Cope with a Literacy Intervention Program in a Remote Indigenous Community? The Community Action Support Program in the Northern Territory, Australia

    ERIC Educational Resources Information Center

    Naidoo, Loshini

    2012-01-01

    This paper examines a new community education initiative, Community Action Support (CAS) that helps facilitate learning in Indigenous young people from Tennant Creek in the Northern Territory. CAS is an innovative partnership program between the Australian Literacy and Numeracy Foundation and the University of Western Sydney. The core aim of the…

  12. Direct CRISPR spacer acquisition from RNA by a natural reverse transcriptase-Cas1 fusion protein.

    PubMed

    Silas, Sukrit; Mohr, Georg; Sidote, David J; Markham, Laura M; Sanchez-Amat, Antonio; Bhaya, Devaki; Lambowitz, Alan M; Fire, Andrew Z

    2016-02-26

    CRISPR systems mediate adaptive immunity in diverse prokaryotes. CRISPR-associated Cas1 and Cas2 proteins have been shown to enable adaptation to new threats in type I and II CRISPR systems by the acquisition of short segments of DNA (spacers) from invasive elements. In several type III CRISPR systems, Cas1 is naturally fused to a reverse transcriptase (RT). In the marine bacterium Marinomonas mediterranea (MMB-1), we showed that a RT-Cas1 fusion protein enables the acquisition of RNA spacers in vivo in a RT-dependent manner. In vitro, the MMB-1 RT-Cas1 and Cas2 proteins catalyze the ligation of RNA segments into the CRISPR array, which is followed by reverse transcription. These observations outline a host-mediated mechanism for reverse information flow from RNA to DNA. PMID:26917774

  13. Evidence for the widespread distribution of CRISPR-Cas system in the Phylum Cyanobacteria

    PubMed Central

    Cai, Fei; Axen, Seth D.; Kerfeld, Cheryl A.

    2013-01-01

    Members of the phylum Cyanobacteria inhabit ecologically diverse environments. However, the CRISPR-Cas (clustered regularly interspaced short palindromic repeats, CRISPR associated genes), an extremely adaptable defense system, has not been surveyed in this phylum. We analyzed 126 cyanobacterial genomes and, surprisingly, found CRISPR-Cas in the majority except the marine subclade (Synechococcus and Prochlorococcus), in which cyanophages are a known force shaping their evolution. Multiple observations of CRISPR loci in the absence of cas1/cas2 genes may represent an early stage of losing a CRISPR-Cas locus. Our findings reveal the widespread distribution of their role in the phylum Cyanobacteria and provide a first step to systematically understanding CRISPR-Cas systems in cyanobacteria. PMID:23628889

  14. Transcriptional Regulation with CRISPR/Cas9 Effectors in Mammalian Cells.

    PubMed

    Pham, Hannah; Kearns, Nicola A; Maehr, René

    2016-01-01

    CRISPR/Cas9-based regulation of gene expression provides the scientific community with a new high-throughput tool to dissect the role of genes in molecular processes and cellular functions. Single-guide RNAs allow for recruitment of a nuclease-dead Cas9 protein and transcriptional Cas9-effector fusion proteins to specific genomic loci, thereby modulating gene expression. We describe the application of a CRISPR-Cas9 effector system from Streptococcus pyogenes for transcriptional regulation in mammalian cells resulting in activation or repression of transcription. We present methods for appropriate target site selection, sgRNA design, and delivery of dCas9 and dCas9-effector system components into cells through lentiviral transgenesis to modulate transcription. PMID:26463376

  15. Generation of an Oocyte-Specific Cas9 Transgenic Mouse for Genome Editing

    PubMed Central

    Zhang, Linlin; Zhou, Jiankui; Han, Jinxiong; Hu, Bian; Hou, Ningning; Shi, Yun; Huang, Xingxu

    2016-01-01

    The CRISPR/Cas9 system has been developed as an easy-handle and multiplexable approach for engineering eukaryotic genomes by zygote microinjection of Cas9 and sgRNA, while preparing Cas9 for microinjection is laborious and introducing inconsistency into the experiment. Here, we describe a modified strategy for gene targeting through using oocyte-specific Cas9 transgenic mouse. With this mouse line, we successfully achieve precise gene targeting by injection of sgRNAs only into one-cell-stage embryos. Through comprehensive analysis, we also show allele complexity and off-target mutagenesis induced by this strategy is obviously lower than Cas9 mRNA/sgRNA injection. Thus, injection of sgRNAs into oocyte-specific Cas9 transgenic mouse embryo provides a convenient, efficient and reliable approach for mouse genome editing. PMID:27119535

  16. Long-term monitoring of orbital modulation and secondary-star irradiation in Nova Cas 1995 (V723 Cas)

    NASA Astrophysics Data System (ADS)

    Ochner, P.; Moschini, F.; Munari, U.; Frigo, A.

    2015-11-01

    We present optical spectroscopy collected at seven epochs and BVRCIC photometry obtained at 1227 epochs of nova V723 Cas, covering the time interval between 2007 and 2015. The mean magnitude during this period, stable at ˜3 mag brighter than in quiescence, and the continuous presence of strong [Fe X] and other high-ionization emission lines, indicates that the nuclear burning at the surface of the white dwarf is continuing 20 years past the initial outburst. The light curve shows a large amplitude (2 mag) orbital modulation, which is governed by the visibility of the irradiated side of the secondary star. Our observations do not confirm the reported increase with time of the orbital period of V723 Cas, a period of P=16.638 383 ± 0.000 025 h satisfying equally well all available observations in all bands. Our observations also do not confirm the presence of an additional periodicity around P=15.2397 h from which V723 Cas was classified as an intermediate-polar system.

  17. Introduction of p130cas signaling complex formation upon integrin-mediated cell adhesion: a role for Src family kinases.

    PubMed Central

    Vuori, K; Hirai, H; Aizawa, S; Ruoslahti, E

    1996-01-01

    Integrin-mediated cell adhesion triggers intracellular signaling cascades, including tyrosine phosphorylation of intracellular proteins. Among these are the focal adhesion proteins p130cas (Cas) and focal adhesion kinase (FAK). Here we identify the kinase(s) mediating integrin-induced Cas phosphorylation and characterize protein-protein interactions mediated by phosphorylated Cas. We found that expression of a constitutively active FAK in fibroblasts results in a consecutive tyrosine phosphorylation of Cas. This effect required the autophosphorylation site of FAK, which is a binding site for Src family kinases. Integrin-mediated phosphorylation of Cas was not, however, compromised in fibroblasts lacking FAK. In contrast, adhesion-induced tyrosine phosphorylation of Cas was reduced in cells lacking Src, whereas enhanced phosphorylation of Cas was observed Csk- cells, in which Src kinases are activated. These results suggest that Src kinases are responsible for the integrin-mediated tyrosine phosphorylation of Cas. FAK seems not to be necessary for phosphorylation of Cas, but when autophosphorylated, FAK may recruit Src family kinases to phosphorylate Cas. Cas was found to form complexes with Src homology 2 (SH2) domain-containing signaling molecules, such as the SH2/SH3 adapter protein Crk, following integrin-induced tyrosine phosphorylation. Guanine nucleotide exchange factors C3G and Sos were found in the Cas-Crk complex upon integrin ligand binding. These observations suggest that Cas serves as a docking protein and may transduce signals to downstream signaling pathways following integrin-mediated cell adhesion. PMID:8649368

  18. 40 CFR 63.61 - Deletion of methyl ethyl ketone from the list of hazardous air pollutants.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... the list of hazardous air pollutants. 63.61 Section 63.61 Protection of Environment ENVIRONMENTAL... POLLUTANTS FOR SOURCE CATEGORIES List of Hazardous Air Pollutants, Petitions Process, Lesser Quantity... pollutants. The substance methyl ethyl ketone (MEK, 2-Butanone) (CAS Number 78-93-3) is deleted from the...

  19. Highly efficient heritable plant genome engineering using Cas9 orthologues from Streptococcus thermophilus and Staphylococcus aureus.

    PubMed

    Steinert, Jeannette; Schiml, Simon; Fauser, Friedrich; Puchta, Holger

    2015-12-01

    The application of the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas system of Streptococcus pyogenes (SpCas9) is currently revolutionizing genome engineering in plants. However, synthetic plant biology will require more complex manipulations of genomes and transcriptomes. The simultaneous addressing of different specific genomic sites with independent enzyme activities within the same cell is a key to this issue. Such approaches can be achieved by the adaptation of additional bacterial orthologues of the CRISPR/Cas system for use in plant cells. Here, we show that codon-optimised Cas9 orthologues from Streptococcus thermophilus (St1Cas9) and Staphylococcus aureus (SaCas9) can both be used to induce error-prone non-homologous end-joining-mediated targeted mutagenesis in the model plant Arabidopsis thaliana at frequencies at least comparable to those that have previously been reported for the S. pyogenes CRISPR/Cas system. Stable inheritance of the induced targeted mutations of the ADH1 gene was demonstrated for both St1Cas9- and SaCas9-based systems at high frequencies. We were also able to demonstrate that the SaCas9 and SpCas9 proteins enhance homologous recombination via the induction of double-strand breaks only in the presence of their species-specific single guide (sg) RNAs. These proteins are not prone to inter-species interference with heterologous sgRNA expression constructs. Thus, the CRISPR/Cas systems of S. pyogenes and S. aureus should be appropriate for simultaneously addressing different sequence motifs with different enzyme activities in the same plant cell. PMID:26576927

  20. Functional annotation of native enhancers with a Cas9 -histone demethylase fusion

    PubMed Central

    Tabak, Barbara; Genga, Ryan M; Silverstein, Noah J; Garber, Manuel; Maehr, René

    2015-01-01

    Understanding of mammalian enhancer function is limited by the lack of a technology to rapidly and thoroughly test their cell type-specific function. Here, we use a nuclease-deficient (d)Cas9 histone demethylase fusion to functionally characterize previously described and novel enhancer elements for their roles in the embryonic stem cell state. Further, we distinguish the mechanism of action of dCas9-LSD1 at enhancers from previous dCas9-effectors. PMID:25775043

  1. Efficient Production of Gene-Modified Mice using Staphylococcus aureus Cas9

    PubMed Central

    Zhang, Xiya; Liang, Puping; Ding, Chenhui; Zhang, Zhen; Zhou, Jianwen; Xie, Xiaowei; Huang, Rui; Sun, Ying; Sun, Hongwei; Zhang, Jinran; Xu, Yanwen; Songyang, Zhou; Huang, Junjiu

    2016-01-01

    The CRISPR/Cas system is an efficient genome-editing tool to modify genes in mouse zygotes. However, only the Streptococcus pyogenes Cas9 (SpCas9) has been systematically tested for generating gene-modified mice. The protospacer adjacent motif (PAM, 5′-NGG-3′) recognized by SpCas9 limits the number of potential target sites for this system. Staphylococcus aureus Cas9 (SaCas9), with its smaller size and unique PAM (5′-NNGRRT-3′) preferences, presents an alternative for genome editing in zygotes. Here, we showed that SaCas9 could efficiently and specifically edit the X-linked gene Slx2 and the autosomal gene Zp1 in mouse zygotes. SaCas9-mediated disruption of the tyrosinase (Tyr) gene led to C57BL/6J mice with mosaic coat color. Furthermore, multiplex targeting proved efficient multiple genes disruption when we co-injected gRNAs targeting Slx2, Zp1, and Tyr together with SaCas9 mRNA. We were also able to insert a Flag tag at the C-terminus of histone H1c, when a Flag-encoding single-stranded DNA oligo was co-introduced into mouse zygotes with SaCas9 mRNA and the gRNA. These results indicate that SaCas9 can specifically cleave the target gene locus, leading to successful gene knock-out and precise knock-in in mouse zygotes, and highlight the potential of using SaCas9 for genome editing in preimplantation embryos and producing gene-modified animal models. PMID:27586692

  2. Prediction and Validation of Native and Engineered Cas9 Guide Sequences.

    PubMed

    Briner, Alexandra E; Henriksen, Emily D; Barrangou, Rodolphe

    2016-01-01

    Cas9-based technologies rely on native elements of Type II CRISPR-Cas bacterial immune systems, including the trans-activating CRISPR RNA (tracrRNA), CRISPR RNA (crRNA), Cas9 protein, and protospacer-adjacent motif (PAM). The tracrRNA and crRNA form an RNA duplex that guides the Cas9 endonuclease to complementary nucleic acid sequences. Mechanistically, Cas9 initiates interactions by binding to the target PAM sequence and interrogating the target DNA in a 3'-to-5' manner. Complementarity between the guide RNA and the target DNA is key. In natural systems, precise cleavage occurs when the target DNA sequence contains a PAM flanking a sequence homologous to the crRNA spacer sequence. Currently, the majority of commercial Cas9-based genome-editing tools are derived from the Type II CRISPR-Cas system of Streptococcus pyogenes However, a diverse set of Type II CRISPR-Cas systems exist in nature that are potentially valuable for genome engineering applications. Exploitation of these systems requires prediction and validation of both native and engineered dual and single guide RNAs to drive Cas9 functionality. Here, we discuss how to identify the elements of these immune systems to develop next-generation Cas9-based genome-editing tools. We first discuss how to predict tracrRNA sequences and suggest a method for designing single guide RNAs containing only critical structural modules. We then outline how to predict the PAM sequence, which is crucial for determining potential targets for Cas9. Finally, validation of the system elements through transcriptome analysis and interference assays is essential for developing next-generation Cas9-based genome-editing tools. PMID:27371591

  3. Modified RNAs in CRISPR/Cas9: An Old Trick Works Again.

    PubMed

    Latorre, Alfonso; Latorre, Ana; Somoza, Álvaro

    2016-03-01

    Old tricks, new dog: CRISPR/Cas9 is a powerful tool for gene editing that requires an endonuclease (Cas9) and RNA strands. It has been shown that chemical modification of the RNA structures, an approach that has been used to improve the efficiency of RNA interference, can also be applied to enhance the activity of CRISPR/Cas9 and reduce its off-target effects. PMID:26880106

  4. Efficient Production of Gene-Modified Mice using Staphylococcus aureus Cas9.

    PubMed

    Zhang, Xiya; Liang, Puping; Ding, Chenhui; Zhang, Zhen; Zhou, Jianwen; Xie, Xiaowei; Huang, Rui; Sun, Ying; Sun, Hongwei; Zhang, Jinran; Xu, Yanwen; Songyang, Zhou; Huang, Junjiu

    2016-01-01

    The CRISPR/Cas system is an efficient genome-editing tool to modify genes in mouse zygotes. However, only the Streptococcus pyogenes Cas9 (SpCas9) has been systematically tested for generating gene-modified mice. The protospacer adjacent motif (PAM, 5'-NGG-3') recognized by SpCas9 limits the number of potential target sites for this system. Staphylococcus aureus Cas9 (SaCas9), with its smaller size and unique PAM (5'-NNGRRT-3') preferences, presents an alternative for genome editing in zygotes. Here, we showed that SaCas9 could efficiently and specifically edit the X-linked gene Slx2 and the autosomal gene Zp1 in mouse zygotes. SaCas9-mediated disruption of the tyrosinase (Tyr) gene led to C57BL/6J mice with mosaic coat color. Furthermore, multiplex targeting proved efficient multiple genes disruption when we co-injected gRNAs targeting Slx2, Zp1, and Tyr together with SaCas9 mRNA. We were also able to insert a Flag tag at the C-terminus of histone H1c, when a Flag-encoding single-stranded DNA oligo was co-introduced into mouse zygotes with SaCas9 mRNA and the gRNA. These results indicate that SaCas9 can specifically cleave the target gene locus, leading to successful gene knock-out and precise knock-in in mouse zygotes, and highlight the potential of using SaCas9 for genome editing in preimplantation embryos and producing gene-modified animal models. PMID:27586692

  5. Environmental Scanning in Support of Planning and Decision Making: Case Studies at Selected Institutions of Higher Education. AIR 1990 Annual Forum Paper.

    ERIC Educational Resources Information Center

    Pritchett, Mark S.

    This paper presents a comprehensive case analysis of formal environmental scanning processes in three different institutions of higher education. The study focuses on how environmental scanning activities are organized and used to support institutional planning and decision-making. Special emphasis is placed on describing how environmental…

  6. Air Abrasion

    MedlinePlus

    ... delivered directly to your desktop! more... What Is Air Abrasion? Article Chapters What Is Air Abrasion? What Happens? The Pros and Cons Will I Feel Anything? Is Air Abrasion for Everyone? print full article print this ...

  7. Development of standardized air-blown coal gasifier/gas turbine concepts for future electric power systems. Volume 5, Appendix D: Cost support information: Final report

    SciTech Connect

    Sadowski, R.S.; Brown, M.J.; Harriz, J.T.; Ostrowski, E.

    1991-01-01

    The cost estimate provided for the DOE sponsored study of Air Blown Coal Gasification was developed from vendor quotes obtained directly for the equipment needed in the 50 MW, 100 MW, and 200 MW sized plants and from quotes from other jobs that have been referenced to apply to the particular cycle. Quotes were generally obtained for the 100 MW cycle and a scale up/down factor was used to generate the cost estimates for the 200 MW and 50 MW cycles, respectively. Information from GTPro (property of Thermoflow, Inc.) was used to estimate the cost of the 200 MW and 50 MW gas turbine, HRSG, and steam turbines. To available the use of GTPro`s estimated values for this equipment, a comparison was made between the quotes obtained for the 100 MW cycle (ABB GT 11N combustion turbine and a HSRG) against the estimated values by GTPro.

  8. Generating Mouse Models Using CRISPR-Cas9-Mediated Genome Editing.

    PubMed

    Qin, Wenning; Kutny, Peter M; Maser, Richard S; Dion, Stephanie L; Lamont, Jeffrey D; Zhang, Yingfan; Perry, Greggory A; Wang, Haoyi

    2016-01-01

    The CRISPR-Cas9 system in bacteria and archaea has recently been exploited for genome editing in various model organisms, including mice. The CRISPR-Cas9 reagents can be delivered directly into the mouse zygote to derive a mutant animal carrying targeted genetic modifications. The major components of the system include the guide RNA, which provides target specificity, the Cas9 nuclease that creates the DNA double-strand break, and the donor oligonucleotide or plasmid carrying the intended mutation flanked by sequences homologous to the target site. Here we describe the general considerations and experimental protocols for creating genetically modified mice using the CRISPR-Cas9 system. PMID:26928663

  9. Modulating the Cascade architecture of a minimal Type I-F CRISPR-Cas system

    PubMed Central

    Gleditzsch, Daniel; Müller-Esparza, Hanna; Pausch, Patrick; Sharma, Kundan; Dwarakanath, Srivatsa; Urlaub, Henning; Bange, Gert; Randau, Lennart

    2016-01-01

    Shewanella putrefaciens CN-32 contains a single Type I-Fv CRISPR-Cas system which confers adaptive immunity against bacteriophage infection. Three Cas proteins (Cas6f, Cas7fv, Cas5fv) and mature CRISPR RNAs were shown to be required for the assembly of an interference complex termed Cascade. The Cas protein-CRISPR RNA interaction sites within this complex were identified via mass spectrometry. Additional Cas proteins, commonly described as large and small subunits, that are present in all other investigated Cascade structures, were not detected. We introduced this minimal Type I system in Escherichia coli and show that it provides heterologous protection against lambda phage. The absence of a large subunit suggests that the length of the crRNA might not be fixed and recombinant Cascade complexes with drastically shortened and elongated crRNAs were engineered. Size-exclusion chromatography and small-angle X-ray scattering analyses revealed that the number of Cas7fv backbone subunits is adjusted in these shortened and extended Cascade variants. Larger Cascade complexes can still confer immunity against lambda phage infection in E. coli. Minimized Type I CRISPR-Cas systems expand our understanding of the evolution of Cascade assembly and diversity. Their adjustable crRNA length opens the possibility for customizing target DNA specificity. PMID:27216815

  10. Multiple mechanisms for CRISPR-Cas inhibition by anti-CRISPR proteins.

    PubMed

    Bondy-Denomy, Joseph; Garcia, Bianca; Strum, Scott; Du, Mingjian; Rollins, MaryClare F; Hidalgo-Reyes, Yurima; Wiedenheft, Blake; Maxwell, Karen L; Davidson, Alan R

    2015-10-01

    The battle for survival between bacteria and the viruses that infect them (phages) has led to the evolution of many bacterial defence systems and phage-encoded antagonists of these systems. Clustered regularly interspaced short palindromic repeats (CRISPR) and the CRISPR-associated (cas) genes comprise an adaptive immune system that is one of the most widespread means by which bacteria defend themselves against phages. We identified the first examples of proteins produced by phages that inhibit a CRISPR-Cas system. Here we performed biochemical and in vivo investigations of three of these anti-CRISPR proteins, and show that each inhibits CRISPR-Cas activity through a distinct mechanism. Two block the DNA-binding activity of the CRISPR-Cas complex, yet do this by interacting with different protein subunits, and using steric or non-steric modes of inhibition. The third anti-CRISPR protein operates by binding to the Cas3 helicase-nuclease and preventing its recruitment to the DNA-bound CRISPR-Cas complex. In vivo, this anti-CRISPR can convert the CRISPR-Cas system into a transcriptional repressor, providing the first example-to our knowledge-of modulation of CRISPR-Cas activity by a protein interactor. The diverse sequences and mechanisms of action of these anti-CRISPR proteins imply an independent evolution, and foreshadow the existence of other means by which proteins may alter CRISPR-Cas function. PMID:26416740

  11. Modulating the Cascade architecture of a minimal Type I-F CRISPR-Cas system.

    PubMed

    Gleditzsch, Daniel; Müller-Esparza, Hanna; Pausch, Patrick; Sharma, Kundan; Dwarakanath, Srivatsa; Urlaub, Henning; Bange, Gert; Randau, Lennart

    2016-07-01

    Shewanella putrefaciens CN-32 contains a single Type I-Fv CRISPR-Cas system which confers adaptive immunity against bacteriophage infection. Three Cas proteins (Cas6f, Cas7fv, Cas5fv) and mature CRISPR RNAs were shown to be required for the assembly of an interference complex termed Cascade. The Cas protein-CRISPR RNA interaction sites within this complex were identified via mass spectrometry. Additional Cas proteins, commonly described as large and small subunits, that are present in all other investigated Cascade structures, were not detected. We introduced this minimal Type I system in Escherichia coli and show that it provides heterologous protection against lambda phage. The absence of a large subunit suggests that the length of the crRNA might not be fixed and recombinant Cascade complexes with drastically shortened and elongated crRNAs were engineered. Size-exclusion chromatography and small-angle X-ray scattering analyses revealed that the number of Cas7fv backbone subunits is adjusted in these shortened and extended Cascade variants. Larger Cascade complexes can still confer immunity against lambda phage infection in E. coli Minimized Type I CRISPR-Cas systems expand our understanding of the evolution of Cascade assembly and diversity. Their adjustable crRNA length opens the possibility for customizing target DNA specificity. PMID:27216815

  12. Development of an intein-mediated split–Cas9 system for gene therapy

    PubMed Central

    Truong, Dong-Jiunn Jeffery; Kühner, Karin; Kühn, Ralf; Werfel, Stanislas; Engelhardt, Stefan; Wurst, Wolfgang; Ortiz, Oskar

    2015-01-01

    Using CRISPR/Cas9, it is possible to target virtually any gene in any organism. A major limitation to its application in gene therapy is the size of Cas9 (>4 kb), impeding its efficient delivery via recombinant adeno-associated virus (rAAV). Therefore, we developed a split–Cas9 system, bypassing the packaging limit using split-inteins. Each Cas9 half was fused to the corresponding split-intein moiety and, only upon co-expression, the intein-mediated trans-splicing occurs and the full Cas9 protein is reconstituted. We demonstrated that the nuclease activity of our split-intein system is comparable to wild-type Cas9, shown by a genome-integrated surrogate reporter and by targeting three different endogenous genes. An analogously designed split-Cas9D10A nickase version showed similar activity as Cas9D10A. Moreover, we showed that the double nick strategy increased the homologous directed recombination (HDR). In addition, we explored the possibility of delivering the repair template accommodated on the same dual-plasmid system, by transient transfection, showing an efficient HDR. Most importantly, we revealed for the first time that intein-mediated split–Cas9 can be packaged, delivered and its nuclease activity reconstituted efficiently, in cells via rAAV. PMID:26082496

  13. CRISPR-Cas: New Tools for Genetic Manipulations from Bacterial Immunity Systems.

    PubMed

    Jiang, Wenyan; Marraffini, Luciano A

    2015-01-01

    Prokaryotic CRISPR-Cas loci encode proteins that function as an adaptive immune system against infectious viruses and plasmids. Immunity is mediated by Cas nucleases and small RNA guides, which specify a cleavage site within the genome of the invader. In type II CRISPR-Cas systems, the RNA-guided Cas9 nuclease cleaves the DNA. Cas9 can be reprogrammed to create double-strand DNA breaks in the genomes of a variety of organisms, from bacteria to human cells. Repair of Cas9 lesions by homologous recombination or nonhomologous end joining mechanisms can lead to the introduction of specific nucleotide substitutions or indel mutations, respectively. Furthermore, a nuclease-null Cas9 has been developed to regulate endogenous gene expression and to label genomic loci in living cells. Targeted genome editing and gene regulation mediated by Cas9 are easy to program, scale, and multiplex, allowing researchers to decipher the causal link between genetic and phenotypic variation. In this review, we describe the most notable applications of Cas9 in basic biology, translational medicine, synthetic biology, biotechnology, and other fields. PMID:26209264

  14. Development of an intein-mediated split-Cas9 system for gene therapy.

    PubMed

    Truong, Dong-Jiunn Jeffery; Kühner, Karin; Kühn, Ralf; Werfel, Stanislas; Engelhardt, Stefan; Wurst, Wolfgang; Ortiz, Oskar

    2015-07-27

    Using CRISPR/Cas9, it is possible to target virtually any gene in any organism. A major limitation to its application in gene therapy is the size of Cas9 (>4 kb), impeding its efficient delivery via recombinant adeno-associated virus (rAAV). Therefore, we developed a split-Cas9 system, bypassing the packaging limit using split-inteins. Each Cas9 half was fused to the corresponding split-intein moiety and, only upon co-expression, the intein-mediated trans-splicing occurs and the full Cas9 protein is reconstituted. We demonstrated that the nuclease activity of our split-intein system is comparable to wild-type Cas9, shown by a genome-integrated surrogate reporter and by targeting three different endogenous genes. An analogously designed split-Cas9D10A nickase version showed similar activity as Cas9D10A. Moreover, we showed that the double nick strategy increased the homologous directed recombination (HDR). In addition, we explored the possibility of delivering the repair template accommodated on the same dual-plasmid system, by transient transfection, showing an efficient HDR. Most importantly, we revealed for the first time that intein-mediated split-Cas9 can be packaged, delivered and its nuclease activity reconstituted efficiently, in cells via rAAV. PMID:26082496

  15. Targeted Gene Manipulation in Plants Using the CRISPR/Cas Technology.

    PubMed

    Zhang, Dandan; Li, Zhenxiang; Li, Jian-Feng

    2016-05-20

    The CRISPR/Cas technology is emerging as a revolutionary genome editing tool in diverse organisms including plants, and has quickly evolved into a suite of versatile tools for sequence-specific gene manipulations beyond genome editing. Here, we review the most recent applications of the CRISPR/Cas toolkit in plants and also discuss key factors for improving CRISPR/Cas performance and strategies for reducing the off-target effects. Novel technical breakthroughs in mammalian research regarding the CRISPR/Cas toolkit will also be incorporated into this review in hope to stimulate prospective users from the plant research community to fully explore the potential of these technologies. PMID:27165865

  16. Cas3 is a single-stranded DNA nuclease and ATP-dependent helicase in the CRISPR/Cas immune system.

    PubMed

    Sinkunas, Tomas; Gasiunas, Giedrius; Fremaux, Christophe; Barrangou, Rodolphe; Horvath, Philippe; Siksnys, Virginijus

    2011-04-01

    Clustered regularly interspaced short palindromic repeat (CRISPR) is a recently discovered adaptive prokaryotic immune system that provides acquired immunity against foreign nucleic acids by utilizing small guide crRNAs (CRISPR RNAs) to interfere with invading viruses and plasmids. In Escherichia coli, Cas3 is essential for crRNA-guided interference with virus proliferation. Cas3 contains N-terminal HD phosphohydrolase and C-terminal Superfamily 2 (SF2) helicase domains. Here, we provide the first report of the cloning, expression, purification and in vitro functional analysis of the Cas3 protein of the Streptococcus thermophilus CRISPR4 (Ecoli subtype) system. Cas3 possesses a single-stranded DNA (ssDNA)-stimulated ATPase activity, which is coupled to unwinding of DNA/DNA and RNA/DNA duplexes. Cas3 also shows ATP-independent nuclease activity located in the HD domain with a preference for ssDNA substrates. To dissect the contribution of individual domains, Cas3 separation-of-function mutants (ATPase(+)/nuclease(-) and ATPase(-)/nuclease(+)) were obtained by site-directed mutagenesis. We propose that the Cas3 ATPase/helicase domain acts as a motor protein, which assists delivery of the nuclease activity to Cascade-crRNA complex targeting foreign DNA. PMID:21343909

  17. Cas5d protein processes pre-crRNA and assembles into a Cascade-like interference complex in Subtype I-C/Dvulg CRISPR-Cas system

    PubMed Central

    Nam, Ki Hyun; Haitjema, Charles; Liu, Xueqi; Ding, Fran; Wang, Hongwei; DeLisa, Matthew P.; Ke, Ailong

    2012-01-01

    SUMMARY Clustered regularly interspaced short palindromic repeats (CRISPRs), together with an operon of CRISPR-associated (Cas) proteins, form an RNA-based prokaryotic immune system against exogenous genetic elements. Cas5 family proteins are found in several Type I CRISPR-Cas systems. Here we report the molecular function of Subtype I-C/Dvulg Cas5d from B. halodurans. We show that Cas5d cleaves pre-crRNA into unit length by recognizing both the hairpin structure and the 3′ single stranded sequence in the CRISPR repeat region. Cas5d structure reveals a ferredoxin domain-based architecture and a catalytic triad formed by Y46, K116 and H117 residues. We further show that after pre-crRNA processing, Cas5d assembles with crRNA, Csd1, and Csd2 proteins to form a multi-subunit interference complex similar to E. coli Cascade (CRISPR-associated complex for antiviral defense) in architecture. Our results suggest that formation of a crRNA-presenting Cascade-like complex is likely a common theme among Type I CRISPR subtypes. PMID:22841292

  18. CRISPR/Cas mutagenesis of soybean and Medicago truncatula using a new web-tool and a modified Cas9 enzyme.

    PubMed

    Michno, Jean-Michel; Wang, Xiaobo; Liu, Junqi; Curtin, Shaun J; Kono, Thomas Jy; Stupar, Robert M

    2015-10-01

    The CRISPR/Cas9 system is rapidly becoming the reagent of choice for targeted mutagenesis and gene editing in crop species. There are currently intense research efforts in the crop sciences to identify efficient CRISPR/Cas9 platforms to carry out targeted mutagenesis and gene editing projects. These efforts typically result in the incremental tweaking of various platform components including the identification of crop-specific promoters and terminators for optimal expression of the Cas9 enzyme and identification of promoters for expression of the CRISPR guide RNA. In this report, we demonstrate the development of an online web tool for fast identification of CRISPR/Cas9 target loci within soybean gene models, and generic DNA sequences. The web-tool described in this work can quickly identify a high number of potential CRISPR/Cas9 target sites, including restriction enzyme sites that can facilitate the detection of new mutations. In conjunction with the web tool, a soybean codon-optimized CRISPR/Cas9 platform was designed to direct double-stranded breaks to the targeted loci in hairy root transformed cells. The modified Cas9 enzyme was shown to successfully mutate target genes in somatic cells of 2 legume species, soybean and Medicago truncatula. These new tools may help facilitate targeted mutagenesis in legume and other plant species. PMID:26479970

  19. Cas5d Protein Processes Pre-crRNA and Assembles into a Cascade-like Interference Complex in Subtype I-C/Dvulg CRISPR-Cas System

    SciTech Connect

    Nam, Ki Hyun; Haitjema, Charles; Liu, Xueqi; Ding, Fran; Wang, Hongwei; DeLisa, Matthew P.; Ke, Ailong

    2012-10-10

    Clustered regularly interspaced short palindromic repeats (CRISPRs), together with an operon of CRISPR-associated (Cas) proteins, form an RNA-based prokaryotic immune system against exogenous genetic elements. Cas5 family proteins are found in several type I CRISPR-Cas systems. Here, we report the molecular function of subtype I-C/Dvulg Cas5d from Bacillus halodurans. We show that Cas5d cleaves pre-crRNA into unit length by recognizing both the hairpin structure and the 3 single stranded sequence in the CRISPR repeat region. Cas5d structure reveals a ferredoxin domain-based architecture and a catalytic triad formed by Y46, K116, and H117 residues. We further show that after pre-crRNA processing, Cas5d assembles with crRNA, Csd1, and Csd2 proteins to form a multi-sub-unit interference complex similar to Escherichia coli Cascade (CRISPR-associated complex for antiviral defense) in architecture. Our results suggest that formation of a crRNA-presenting Cascade-like complex is likely a common theme among type I CRISPR subtypes.

  20. Breathing zone air sampler

    DOEpatents

    Tobin, John

    1989-01-01

    A sampling apparatus is provided which comprises a sampler for sampling air in the breathing zone of a wearer of the apparatus and a support for the sampler preferably in the form of a pair of eyeglasses. The sampler comprises a sampling assembly supported on the frame of the eyeglasses and including a pair of sample transport tubes which are suspended, in use, centrally of the frame so as to be disposed on opposite sides of the nose of the wearer and which each include an inlet therein that, in use, is disposed adjacent to a respective nostril of the nose of the wearer. A filter holder connected to sample transport tubes supports a removable filter for filtering out particulate material in the air sampled by the apparatus. The sample apparatus is connected to a pump for drawing air into the apparatus through the tube inlets so that the air passes through the filter.