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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. 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.

  3. 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.

  4. 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.

  5. 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.

  6. 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…

  7. 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.

  8. 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.

  9. 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).

  10. 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.

  11. Photocopy of drawing. OPERATIONS SUPPORT BUILDING. NASA, Cape Canaveral Air ...

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

    Photocopy of drawing. OPERATIONS SUPPORT BUILDING. NASA, Cape Canaveral Air Force Station, Florida. Drawing 86K01547, Maurice H. Connell & Associates, February, 1961. OPERATIONS SUPPORT BUILDING SITE PLAN. Sheet 2 of 34 - Cape Canaveral Air Force Station, Launch Complex 34, Operations Support Building, Freedom Road, Southwest of Launch Stand CX-34, Cape Canaveral, Brevard County, FL

  12. Photocopy of drawing. OPERATIONS SUPPORT BUILDING. NASA, Cape Canaveral Air ...

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

    Photocopy of drawing. OPERATIONS SUPPORT BUILDING. NASA, Cape Canaveral Air Force Station, Florida. Drawing 86K01547, Maurice H. Connell & Associates, February, 1961. OPERATIONS SUPPORT BUILDING ELEVATIONS AND SECTION. Sheet 5 of 34 - Cape Canaveral Air Force Station, Launch Complex 34, Operations Support Building, Freedom Road, Southwest of Launch Stand CX-34, Cape Canaveral, Brevard County, FL

  13. Photocopy of drawing. OPERATIONS SUPPORT BUILDING. NASA, Cape Canaveral Air ...

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

    Photocopy of drawing. OPERATIONS SUPPORT BUILDING. NASA, Cape Canaveral Air Force Station, Florida. Drawing 86K01547, Maurice H. Connell & Associates, February, 1961. OPERATIONS SUPPORT BUILDING ROOF PLAN, REFLECTED CEILING PLAN, AND DETAILS. Sheet 7 of 34 - Cape Canaveral Air Force Station, Launch Complex 34, Operations Support Building, Freedom Road, Southwest of Launch Stand CX-34, Cape Canaveral, Brevard County, FL

  14. Photocopy of drawing. OPERATIONS SUPPORT BUILDING. NASA, Cape Canaveral Air ...

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

    Photocopy of drawing. OPERATIONS SUPPORT BUILDING. NASA, Cape Canaveral Air Force Station, Florida. Drawing 86K01547, Maurice H. Connell & Associates, February, 1961. OPERATIONS SUPPORT BUILDING FLOOR PLAN AND SCHEDULES. Sheet 4 of 34 - Cape Canaveral Air Force Station, Launch Complex 34, Operations Support Building, Freedom Road, Southwest of Launch Stand CX-34, Cape Canaveral, Brevard County, FL

  15. 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.

  16. 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.

  17. 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.

  18. Multi-Storey Air-Supported Building Construction

    ERIC Educational Resources Information Center

    Pohl, J. G.; Cowan, H. J.

    1972-01-01

    Multistory buildings, supported by internal air pressure and surrounded by a thin, flexible or rigid membrane acting both as structural container and external cladding, are feasible and highly economical for a number of building applications. (Author)

  19. 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.

  20. Development of self-supporting air electrode SOFC

    SciTech Connect

    Nagata, M.; Iwasawa, C.; Yamaoka, S.; Seino, Y.; Ono, M.

    1995-12-31

    The authors are studying a self-supporting SOFC using the spray coating method, etc. A high-performance self-supporting air electrode has successfully been produced by the extrusion-sintering method, and a cell with its electrolyte and fuel electrode manufactured by the plasma spray coating method on the air electrode proved to have good performance. The maximum output density of a single cell is 0.31W/cm{sup 2}. Furthermore, the authors are developing a FGM (Functionally Gradient Material) film as the fuel electrode produced by the plasma spray coating method.

  1. 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

  2. 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.

  3. 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.

  4. Air Traffic Control Decision Support Tools for Noise Mitigation

    NASA Technical Reports Server (NTRS)

    Tobias, Leonard

    2001-01-01

    NASA has initiated a new five year program this year, the Quiet Aircraft Technology (QAT) Program, a program which will investigate airframe and engine system noise reduction. QAT will also address community noise impact. As part of this community noise impact component, NASA will investigate air traffic management (ATM) challenges in reducing noise. In particular, controller advisory automation aids will be developed to aid the air traffic controller in addressing noise concerns as he/she manages traffic in busy terminal areas. NASA has developed controller automation tools to address capacity concerns and the QAT strategy for ATM Low Noise Operations is to build upon this tool set to create added advisories for noise mitigation. The tools developed for capacity will be briefly reviewed, followed by the QAT plans to address ATM noise concerns. A major NASA goal in global civil aviation is to triple the aviation system throughput in all-weather conditions while maintaining safety. A centerpiece of this activity is the Center/TRACON Automation System (CTAS), an evolving suite of air traffic controller decision support tools (DSTs) to enhance capacity of arrivals and departures in both the enroute center and the TRACON. Two of these DSTs, the Traffic Management Advisor (TMA) and the passive Final approach Spacing Tool (pFAST), are in daily use at the Fort Worth Center and the Dallas/Fort Worth (DFW) TRACON, respectively, where capacity gains of 5-13% have been reported in recent NASA evaluations. Under the Federal Aviation Administration's (FAA) Free Flight Phase One Program, TMA and pFAST are each being implemented at six to eight additional sites. In addition, other DSTs are being developed by NASA under the umbrella of CTAS. This means that new software will be built upon CTAS, and the paradigm of real-time simulation evaluation followed by field site development and evaluation will be the pathway for the new tools. Additional information is included in the

  5. The CAS Classroom

    ERIC Educational Resources Information Center

    Garner, Sue

    2004-01-01

    The Victorian Curriculum and Assessment Authority (VCAA) Computer Algebra System (CAS)Pilot study (2001-2005) is monitoring the use of CAS in senior secondary mathematics. This article explores the author's experiences in the CAS classroom and delineates changes in teaching style, as a result of the introduction of CAS into the senior mathematics…

  6. 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...

  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. 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...

  9. 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...

  10. 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…

  11. 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

  12. 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.

  13. 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

  14. 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...

  15. Data Communications Requirements for CAS Applications

    NASA Technical Reports Server (NTRS)

    McCabe, James D.; Blaylock, Bruce T. (Technical Monitor)

    1994-01-01

    This paper discusses the long-haul data communications requirements needed to support visualization of Computational Aeroscience (CAS) simulations. A scientific computing model is presented, from which current communications requirements are derived. This model is then expanded to describe the CAS Collaborative Visualization Environment, in which scientists from multiple NASA centers will collaborate on multidisciplinary, time-accurate simulations. An analysis of this model, along with data from the TFCC Workload Model, shows that CAS data communications requirements will increase to 10-100 megabytes/second by 1996.

  16. 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.

  17. 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.

  18. 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 ...

  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. Catalytic wet air oxidation of chlorophenols over supported ruthenium catalysts.

    PubMed

    Li, Ning; Descorme, Claude; Besson, Michèle

    2007-07-31

    A series of noble metal (Pt, Pd, Ru) loaded zirconia catalysts were evaluated in the catalytic wet air oxidation (CWAO) of mono-chlorophenols (2-CP, 3-CP, 4-CP) under relatively mild reaction conditions. Among the investigated noble metals, Ru appeared to be the best to promote the CWAO of CPs as far as incipient-wetness impregnation was used to prepare all the catalysts. The position of the chlorine substitution on the aromatic ring was also shown to have a significant effect on the CP reactivity in the CWAO over 3wt.% Ru/ZrO(2). 2-CP was relatively easier to degradate compared to 3-CP and 4-CP. One reason could be the higher adsorption of 2-CP on the catalyst surface. Further investigations suggested that 3wt.% Ru/ZrO(2) is a very efficient catalyst in the CWAO of 2-CP as far as high 2-CP conversion and TOC abatement could still be reached at even lower temperature (393K) and lower total pressure (3MPa). Additionally, the conversion of 2-CP was demonstrated to increase with the initial pH of the 2-CP solution. The dechlorination reaction is promoted at higher pH. In all cases, the adsorption of the reactants and the reaction intermediates was shown to play a major role. All parameters that would control the molecule speciation in solution or the catalyst surface properties would have a key effect. PMID:17513043

  2. 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.

  3. 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.

  4. 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.

  5. 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.

  6. 75 FR 30159 - Automatic Dependent Surveillance-Broadcast (ADS-B) Out Performance Requirements To Support Air...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-05-28

    ...) Out Performance Requirements To Support Air Traffic Control (ATC) Service; Final Rule #0;#0;Federal...--Broadcast (ADS-B) Out Performance Requirements To Support Air Traffic Control (ATC) Service AGENCY: Federal... air traffic control from a radar-based system to a satellite-derived aircraft location system....

  7. 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 ...

  8. Design and implementation of an air monitoring program in support of a brownfields redevelopment program

    SciTech Connect

    Maisel, B.E.; Hunt, G.T.; Devaney, R.J. Jr.; Gauvin, M.R.

    1998-12-31

    EPA`s Brownfields Economic Redevelopment Initiative has sparked renewal of industrial and commercial parcels otherwise idled or under-utilized because of real or perceived environmental contamination. In certain cases, restoring such parcels to productive economic use requires a redevelopment effort protective of human health and welfare through minimizing offsite migration of environmental contaminants during cleanup, demolition and remediation activities. To support these objectives, an air monitoring program is often required as an integral element of a comprehensive brownfields redevelopment effort. This paper presents a strategic framework for design and execution of an ambient air monitoring program in support of a brownfields remediation effort ongoing in Lawrence, MA. Based on site characterization, the program included sample collection and laboratory analysis of ambient air samples for polychlorinated biphenyls (PCBs), polychlorinated dibenzodioxins and polychlorinated dibenzofurans (PCDDs/PCDFs), total suspended particulate (TSP), inhalable particulate (PM10), and lead. The program included four monitoring phases, identified as background, wintertime, demolition/remediation and post-demolition. Air sampling occurred over a 16 month period during 1996--97, during which time nine sampling locations were utilized to produce approximately 1,500 ambient air samples. Following strict data review and validation procedures, ambient air data interpretation focused on the following: evaluation of upwind/downwind sample pairs, comparison of ambient levels to existing regulatory standards, relation of ambient levels to data reported in the open literature, and, determination of normal seasonal variations in existing background burden, comparison of ambient levels measured during site activity to background levels.

  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. 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.

  11. 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.

  12. 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.

  13. 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...

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

    PubMed

    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

  17. 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

  18. 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."

  19. 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".

  20. 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

  1. 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.

  2. Fluidic and air-stable supported lipid bilayer and cell-mimicking microarrays.

    PubMed

    Deng, Yang; Wang, Yini; Holtz, Bryan; Li, Jingyi; Traaseth, Nathan; Veglia, Gianluigi; Stottrup, Benjamin J; Elde, Robert; Pei, Duanqing; Guo, Athena; Zhu, X-Y

    2008-05-14

    As drug delivery, therapy, and medical imaging are becoming increasingly cell-specific, there is a critical need for high fidelity and high-throughput screening methods for cell surface interactions. Cell membrane-mimicking surfaces, i.e., supported lipid bilayers (SLBs), are currently not sufficiently robust to meet this need. Here we describe a method of forming fluidic and air-stable SLBs through tethered and dispersed cholesterol groups incorporated into the bottom leaflet. Achieving air stability allows us to easily fabricate SLB microarrays from direct robotic spotting of vesicle solutions. We demonstrate their application as cell membrane-mimicking microarrays by reconstituting peripheral as well as integral membrane components that can be recognized by their respective targets. These demonstrations establish the viability of the fluidic and air-stable SLB platform for generating content microarrays in high throughput studies, e.g., the screening of drugs and nanomedicine targeting cell surface receptors.

  3. 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.

  4. 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.

  5. Fire hazards analysis of the Radioactive Waste Management Complex Air Support Buildings

    SciTech Connect

    Davis, M.L.; Satterwhite, D.G.

    1989-09-01

    This report describes the methods, analyses, results, and conclusions of a fire hazards risk analysis performed for the RWMC Air Support Buildings. An evaluation of the impact for adding a sprinkler system is also presented. Event and fault trees were used to model and analyze the waste storage process. Tables are presented indicating the fire initiators providing the highest potential for release of radioactive materials into the environment. Engineering insights drawn form the data are also provided.

  6. [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

  7. 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).

  8. Understanding and Supporting Web Developers: Design and Evaluation of a Web Accessibility Information Resource (WebAIR).

    PubMed

    Swallow, David; Petrie, Helen; Power, Christopher

    2016-01-01

    This paper describes the design and evaluation of a Web Accessibility Information Resource (WebAIR) for supporting web developers to create and evaluate accessible websites. WebAIR was designed with web developers in mind, recognising their current working practices and acknowledging their existing understanding of web accessibility. We conducted an evaluation with 32 professional web developers in which they used either WebAIR or an existing accessibility information resource, the Web Content Accessibility Guidelines, to identify accessibility problems. The findings indicate that several design decisions made in relation to the language, organisation, and volume of WebAIR were effective in supporting web developers to undertake web accessibility evaluations.

  9. 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...

  10. Spectra of Cas A's Highest Velocity Ejecta

    NASA Astrophysics Data System (ADS)

    Fesen, Robert A.; Milisavljevic, Dan

    2010-08-01

    The young age and close distance of the Galactic supernova remnant Cassiopeia A (Cas A) make it perhaps our best case study and clearest look at the explosion dynamics of a core-collapse supernova (CCSN). Interestingly, Cas A exhibits two nearly opposing streams of high velocity ejecta or `jets' in its NE and SW regions racing outward at speeds more than twice that of the main shell. The nature of these jets, however, and their possible association with an aspherical supernova explosion mechanism is controversial. A handful of existing low-resolution spectra of outer knots in the NE jet display chemical abundances hinting at an origin from the S-Si-Ca- Ar rich layer deep inside the progenitor. If these abundances could be firmly established in both the NE and SW jets, it would be very strong evidence in support of a highly asymmetrical explosion engine for Cas A's progenitor and, in turn, for CCSNe in general. We request KPNO 4m telescope + MARS time to obtain high quality multi-object spectroscopy of Cas A's highest velocity ejecta to measure their nitrogen, sulfur, oxygen, calcium, and argon abundances. These spectra will be analyzed with the metal-rich shock models of J. Raymond and then compared to current sets of CCSN models paying particular attention to knot composition vs. ejection velocity and ejecta mixing.

  11. 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

  12. [Elevated air temperatures tolerance of chufa (Cyperus esculentus L.), a phototroph component of life support systems].

    PubMed

    Shklavtsova, E S; Ushakova, S A; Shikhov, V N

    2011-01-01

    Resistance of biotechnical life support systems (BTLSS) to stress-factors depends, in addition to some other conditions, on tolerance of higher plants as part of the photosynthesizing component. Purpose of the investigations with chufa Cyperus esculentus L. cultivation on mineralized solid and liquid human wastes (according to Yu. Kudenko) was to test plant tolerance of air temperature rise to 45 degrees C. Tolerance was assessed as a function of nitrogen form in nutrient solutions and PAR intensity during thermal shock. PAR intensity was controlled at 150 W/m2 and air temperature--at 25 degrees C. Thermal shock was induced in 30-day plants with PAR = 150 or 250 W/m2. Twenty hours at 45 degrees C did not cause irreversible damage of the plant photosynthetic apparatus. Higher PAR intensity (250 W/m2) and nitrates in nutrient solution mitigates substantially the damaging effect of the stress factor PMID:21848217

  13. 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.

  14. 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.

  15. 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.

  16. 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…

  17. Multi-objective optimization to support rapid air operations mission planning

    NASA Astrophysics Data System (ADS)

    Gonsalves, Paul G.; Burge, Janet E.

    2005-05-01

    Within the context of military air operations, Time-sensitive targets (TSTs) are targets where modifiers such, "emerging, perishable, high-payoff, short dwell, or highly mobile" can be used. Time-critical targets (TCTs) further the criticality of TSTs with respect to achievement of mission objectives and a limited window of opportunity for attack. The importance of TST/TCTs within military air operations has been met with a significant investment in advanced technologies and platforms to meet these challenges. Developments in ISR systems, manned and unmanned air platforms, precision guided munitions, and network-centric warfare have made significant strides for ensuring timely prosecution of TSTs/TCTs. However, additional investments are needed to further decrease the targeting decision cycle. Given the operational needs for decision support systems to enable time-sensitive/time-critical targeting, we present a tool for the rapid generation and analysis of mission plan solutions to address TSTs/TCTs. Our system employs a genetic algorithm-based multi-objective optimization scheme that is well suited to the rapid generation of approximate solutions in a dynamic environment. Genetic Algorithms (GAs) allow for the effective exploration of the search space for potentially novel solutions, while addressing the multiple conflicting objectives that characterize the prosecution of TSTs/TCTs (e.g. probability of target destruction, time to accomplish task, level of disruption to other mission priorities, level of risk to friendly assets, etc.).

  18. 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.

  19. Development of an Aura Chemical Reanalysis in support Air Quality Applications

    NASA Astrophysics Data System (ADS)

    Pierce, R. B.; Lenzen, A.; Schaack, T.

    2015-12-01

    We present results of chemical data assimilation experiments utilizing the NOAA National Environmental Satellite, Data, and Information Service (NESDIS), University of Wisconsin Space Science and Engineering (SSEC) Real-time Air Quality Modeling System (RAQMS) in conjunction with the NOAA National Centers for Environmental Prediction (NCEP) Operational Gridpoint Statistical Interpolation (GSI) 3-dimensional variational data assimilation system. The impact of assimilating NASA Ozone Monitoring Instrument (OMI) total column ozone, OMI tropospheric nitrogen dioxide columns, and Microwave Limb Sounder (MLS) stratospheric ozone profiles on background ozone is assessed using measurements from the 2010 NSF High-performance Instrumented Airborne Platform for Environmental Research (HIAPER) Pole-to-Pole Observation (HIPPO) and NOAA California Nexus (CalNex) campaigns. Results show that the RAQMS/GSI Chemical Reanalysis is able to provide very good estimates of background ozone and large-scale ozone variability and is suitable for use in constraining regional air quality modeling activities. These experiments are being used to guide the development of a multi-year global chemical and aerosol reanalysis using NASA Aura and A-Train measurements to support air quality applications.

  20. 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

  1. 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...

  2. Hygienic support of the ISS air quality (main achievements and prospects)

    NASA Astrophysics Data System (ADS)

    Moukhamedieva, Lana; Tsarkov, Dmitriy; Pakhomova, Anna

    Hygienic preventive measures during pre-flight processing of manned spaceships, selection of polymeric materials, sanitary-hygienic evaluation of cargo and scientific hardware to be used on the ISS and life support systems allow to maintain air quality in limits of regulatory requirements. However, graduate increase of total air contamination by harmful chemicals is observed as service life of the ISS gets longer. It is caused by polymeric materials used on the station overall quantity rise, by additional contamination brought by cargo spacecrafts and modules docking to the ISS and by the cargo. At the same time the range of contaminants that are typical for off-gassing from polymeric materials where modern stabilizers, plasticizers, flame retarders and other additives are used gets wider. In resolving the matters of the ISS service life extension the main question of hygienic researches is to determine real safe operation life of the polymeric material used in structures and hardware of the station, including: begin{itemize} research of polymers degradation (ageing) and its effect on intensity of off gassing and its toxicity; begin{itemize} introduction of polymers with minimal volatile organic compounds off gassing under conditions of space flight and thermal-oxidative degradation. In order to ensure human safety during long-term flight it is important to develop: begin{itemize} real-time air quality monitoring systems, including on-line analysis of highly toxic contaminants evolving during thermo-oxidative degradation of polymer materials and during blowouts of toxic contaminants; begin{itemize} hygienic standards of contaminants level for extended duration of flight up to 3 years. It is essential to develop an automated control system for on-line monitoring of toxicological status and to develop hygienic and engineer measures of its management in order to ensure crew members safety during off-nominal situation.

  3. Support vector data description for detecting the air-ground interface in ground penetrating radar signals

    NASA Astrophysics Data System (ADS)

    Wood, Joshua; Wilson, Joseph

    2011-06-01

    In using GPR images for landmine detection it is often useful to identify the air-ground interface in the GRP signal for alignment purposes. A common simple technique for doing this is to assume that the highest return in an A-scan is from the reflection due to the ground and to use that as the location of the interface. However there are many situations, such as the presence of nose clutter or shallow sub-surface objects, that can cause the global maximum estimate to be incorrect. A Support Vector Data Description (SVDD) is a one-class classifier related to the SVM which encloses the class in a hyper-sphere as opposed to using a hyper-plane as a decision boundary. We apply SVDD to the problem of detection of the air-ground interface by treating each sample in an A-scan, with some number of leading and trailing samples, as a feature vector. Training is done using a set of feature vectors based on known interfaces and detection is done by creating feature vectors from each of the samples in an A-scan, applying the trained SVDD to them and selecting the one with the least distance from the center of the hyper-sphere. We compare this approach with the global maximum approach, examining both the performance on human truthed data and how each method affects false alarm and true positive rates when used as the alignment method in mine detection algorithms.

  4. Stability of rigid rotors supported by air foil bearings: Comparison of two fundamental approaches

    NASA Astrophysics Data System (ADS)

    Larsen, Jon S.; Santos, Ilmar F.; von Osmanski, Sebastian

    2016-10-01

    High speed direct drive motors enable the use of Air Foil Bearings (AFB) in a wide range of applications due to the elimination of gear forces. Unfortunately, AFB supported rotors are lightly damped, and an accurate prediction of their Onset Speed of Instability (OSI) is therefore important. This paper compares two fundamental methods for predicting the OSI. One is based on a nonlinear time domain simulation and another is based on a linearised frequency domain method and a perturbation of the Reynolds equation. Both methods are based on equivalent models and should predict similar results. Significant discrepancies are observed leading to the question, is the classical frequency domain method sufficiently accurate? The discrepancies and possible explanations are discussed in detail.

  5. 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

  6. 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.

  7. 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.

  8. Development and case study of a science-based software platform to support policy making on air quality.

    PubMed

    Zhu, Yun; Lao, Yanwen; Jang, Carey; Lin, Chen-Jen; Xing, Jia; Wang, Shuxiao; Fu, Joshua S; Deng, Shuang; Xie, Junping; Long, Shicheng

    2015-01-01

    This article describes the development and implementations of a novel software platform that supports real-time, science-based policy making on air quality through a user-friendly interface. The software, RSM-VAT, uses a response surface modeling (RSM) methodology and serves as a visualization and analysis tool (VAT) for three-dimensional air quality data obtained by atmospheric models. The software features a number of powerful and intuitive data visualization functions for illustrating the complex nonlinear relationship between emission reductions and air quality benefits. The case study of contiguous U.S. demonstrates that the enhanced RSM-VAT is capable of reproducing the air quality model results with Normalized Mean Bias <2% and assisting in air quality policy making in near real time.

  9. Characterization of organic air emissions from the Certification and Segregation Building and Air Support Weather Shield II at the Radioactive Waste Management Complex

    SciTech Connect

    Shoop, D.S.; Jackson, J.M.; Jolley, J.G.; Izbicki, K.J.

    1994-12-01

    During the latter part of Fiscal Year (FY-92), a task was initiated to characterize the organic air emissions from the Certification and Segregation (C and S) Building [Waste Management Facility (WMF) 612] and the Air Support Weather Shield II (ASWS II or ASB II) (WMF 711) at the Radioactive Waste Management Complex (RWMC). The purpose of this task, titled the RWMC Organic Air Emissions Evaluation Task, was to identify and quantify the volatile organic compounds (VOCS) present in the ambient air in these two facilities and to estimate the organic air emissions. The VOCs were identified and quantified by implementing a dual method approach using two dissimilar analytical methodologies, Open-Path Fourier Transform Infrared Spectroscopy (OP-FTIR) and SUMMA canister sampling, following the US Environmental Protection Agency (EPA) analytical method TO-14. The data gathered were used in conjunction with the building`s ventilation rate to calculate an estimated organic air emissions rate. This report presents the data gathered during the performance of this task and relates the data to the relevant regulatory requirements.

  10. 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

  11. 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.

  12. 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.

  13. 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

  14. 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.

  15. 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.

  16. 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.

  17. 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.

  18. Public Support for Smoke-Free Air Strategies Among Smokers and Nonsmokers, New York City, 2010–2012

    PubMed Central

    Waddell, Elizabeth Needham; Mandel-Ricci, Jenna; Kansagra, Susan M.

    2014-01-01

    Introduction From 2010 through 2012, the New York City Department of Health and Mental Hygiene engaged in multiple smoke-free-air activities in collaboration with community, institution, and government partners. These included implementing a law prohibiting smoking in all parks and beaches as well as working to increase compliance with existing Smoke-free Air Act provisions. Methods We investigated trends in awareness of existing smoke-free rules publicized with new signage and public support for new smoke-free air strategies by using 3 waves of survey data from population-based samples of smoking and nonsmoking adults in New York City (2010–2012). Analyses adjusted for the influence of sociodemographic characteristics. Results Among both smokers and nonsmokers, we observed increased awareness of smoke-free regulations in outdoor areas around hospital entrances and grounds and in lines in outdoor waiting areas for buses and taxis. Regardless of smoking status, women, racial/ethnic minorities, and adults aged 25 to 44 years were more likely than men, non-Hispanic whites, and adults aged 65 years or older to support smoke-free air strategies. Conclusion New signage was successful in increasing population-wide awareness of rules. Our analysis of the association between demographic characteristics and support for tobacco control over time provide important contextual information for community education efforts on secondhand smoke and smoke-free air strategies. PMID:24480633

  19. Pilot Study on Alternative Pictorial Representations and Supporting Text of Sound Standing Waves of Air Columns in a Pipe

    NASA Astrophysics Data System (ADS)

    Zeng, Liang; Smith, Chris; Rodriguez, Jennifer; Corpuz, Edgar

    2011-10-01

    Alternative pictorial representations of sound standing waves of air columns in a pipe were drawn for the first three harmonics in an open-open pipe as well as in an open-closed pipe. Supporting text describing air molecule motion over time was also provided. These pictorial representations and supporting text were designed to reveal the main characteristics of the physical mechanisms of sound standing waves of air columns in pipes. Through a pilot study utilizing surveys and interviews, we validated our design and investigated the differences in the effects on student learning of underlying physics concepts between the new design and the existing one in an introductory physics textbook. The implications of our results for teaching were discussed.

  20. 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

    ... avionics on aircraft operating in Classes A, B, and C airspace, as well as certain other specified classes... 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....

  1. 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...

  2. “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...

  3. 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.

  4. 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%).

  5. 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…

  6. 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.

  7. 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....

  8. 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...

  9. Editorial input for the right price: tobacco industry support for a sheet metal indoor air quality manual.

    PubMed

    Campbell, Richard; Balbach, Edith

    2013-01-01

    Following legal action in the 1990s, internal tobacco industry documents became public, allowing unprecedented insight into the industry's relationships with outside organizations. During the 1980s and 1990s, the National Energy Management Institute (NEMI), established by the Sheet Metal Workers International Association and the Sheet Metal and Air Conditioning Contractors' National Association, (SMACNA) received tobacco industry funding to establish an indoor air quality services program. But the arrangement also required NEMI to serve as an advocate for industry efforts to defeat indoor smoking bans by arguing that ventilation was a more appropriate solution to environmental tobacco smoke. Drawing on tobacco industry documents, this paper describes a striking example of the ethical compromises that accompanied NEMI's collaboration with the tobacco industry, highlighting the solicitation of tobacco industry financial support for a SMACNA indoor air quality manual in exchange for sanitizing references to the health impact of environmental tobacco smoke prior to publication.

  10. 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.

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

    PubMed

    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.

  12. 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.

  13. Decision Support: The Keys to Success. AIR 1986 Annual Forum Paper.

    ERIC Educational Resources Information Center

    Oerly, Diane

    Characteristics of a decision support system (DSS) and factors that influence system design are described, along with a decision support database at the University of Missouri-Columbia. Reasons that the institutional research office is in a unique position to support decision-making are identified. A review of the literature of DSS briefly covers…

  14. Computer-supported Classes in Mathematics for Engineering Students

    NASA Astrophysics Data System (ADS)

    Stoynov, Y. D.

    2008-10-01

    A combined method for teaching and learning selected topics of Mathematics for engineering students is presented. Some points in approaching these topics using CAS DERIVE are discussed and CAS-support of students' learning is shown.

  15. The functions of social support as protective factors for suicidal ideation in a sample of air force personnel.

    PubMed

    Bryan, Craig J; Hernandez, Ann Marie

    2013-10-01

    This study examined various functions of social support (i.e., tangible, esteem, belonging, and appraisal) were examined as protective factors for suicidal ideation in a sample of 273 active duty Air Force Security Forces personnel. Generalized linear regression analyses were conducted to determine if various social support functions were differentially associated with the presence and severity of suicidal ideation, both as main effects and as moderators of emotional distress. None of the four social support functions differentiated suicidal from nonsuicidal Airmen, but esteem support (i.e., feeling respected, encouraged, and valued by others) was associated with significantly less severe suicidal ideation (B = -.074, SE = .025, p = .003). A significant interaction of tangible support (i.e., access to material resources) with emotional distress indicated that emotional distress was associated with more severe suicidal ideation only among Airmen reporting low levels of tangible support (B = .006, SE = .003, p = .018). When considered concurrently, both tangible and self-esteem functions of social support are differentially associated with decreased suicidal ideation among Airmen, but belonging (i.e., having someone to do things with) and appraisal (i.e., having someone to talk to about problems) functions were not. Findings suggest that different aspects of social support affect suicidal ideation in different ways.

  16. 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.

  17. 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.

  18. 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.

  19. 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, 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.

  20. 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

  1. 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.

  2. Spectroscopic Observations of UU Cas

    NASA Astrophysics Data System (ADS)

    Markov, H.; Vince, I.; Markova, N.; Djurasevic, G.

    2010-09-01

    High dispersion (R = 30000) spectra of the eclipsing binary system UU Cas are presented for the first time. Spectra covering 15 different phases were taken in two spectral regions - the vicinity of H_α and another centered on 5800Å. Seven spectral lines were identified in common. Four of them belonging to the brighter component were used to construct radial velocity curve. Some constrains on the spectral classification of the components are presented.

  3. 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.

  4. 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

  5. Genome modification by CRISPR/Cas9.

    PubMed

    Ma, Yuanwu; Zhang, Lianfeng; Huang, Xingxu

    2014-12-01

    Clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein (Cas)9-mediated genome modification enables us to edit the genomes of a variety of organisms rapidly and efficiently. The advantages of the CRISPR-Cas9 system have made it an increasingly popular genetic engineering tool for biological and therapeutic applications. Moreover, CRISPR-Cas9 has been employed to recruit functional domains that repress/activate gene expression or label specific genomic loci in living cells or organisms, in order to explore developmental mechanisms, gene expression regulation, and animal behavior. One major concern about this system is its specificity; although CRISPR-Cas9-mediated off-target mutation has been broadly studied, more efforts are required to further improve the specificity of CRISPR-Cas9. We will also discuss the potential applications of CRISPR-Cas9.

  6. 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.

  7. Prediction of DC Corona Onset Voltage for Rod-Plane Air Gaps by a Support Vector Machine

    NASA Astrophysics Data System (ADS)

    Jin, Shuo; Ruan, Jiangjun; Du, Zhiye; Zhu, Lin; Shu, Shengwen

    2016-10-01

    This paper proposes a new method to predict the corona onset voltage for a rod-plane air gap, based on the support vector machine (SVM). Because the SVM is not limited by the size, dimension and nonlinearity of the samples, this method can realize accurate prediction with few training data. Only electric field features are chosen as the input; no geometric parameter is included. Therefore, the experiment data of one kind of electrode can be used to predict the corona onset voltages of other electrodes with different sizes. With the experimental data obtained by ozone detection technology, and experimental data provided by the reference, the efficiency of the proposed method is validated. Accurate predicted results with an average relative less than 3% are obtained with only 6 experimental data. supported by National Natural Science Foundation of China (No. 51477120)

  8. 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.

  9. 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

  10. 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,…

  11. GLIMPSE: A decision support tool for simultaneously achieving our air quality management and climate change mitigation goals

    NASA Astrophysics Data System (ADS)

    Pinder, R. W.; Akhtar, F.; Loughlin, D. H.; Henze, D. K.; Bowman, K. W.

    2012-12-01

    Poor air quality, ecosystem damages, and climate change all are caused by the combustion of fossil fuels, yet environmental management often addresses each of these challenges separately. This can lead to sub-optimal strategies and unintended consequences. Here we present GLIMPSE -- a decision support tool for simultaneously achieving our air quality and climate change mitigation goals. GLIMPSE comprises of two types of models, (i) the adjoint of the GEOS-Chem chemical transport model, to calculate the relationship between emissions and impacts at high spatial resolution, and (ii) the MARKAL energy system model, to calculate the relationship between energy technologies and emissions. This presentation will demonstrate how GLIMPSE can be used to explore energy scenarios to better achieve both improved air quality and mitigate climate change. Second, this presentation will discuss how space-based observations can be incorporated into GLIMPSE to improve decision-making. NASA satellite products, namely ozone radiative forcing from the Tropospheric Emission Spectrometer (TES), are used to extend GLIMPSE to include the impact of emissions on ozone radiative forcing. This provides a much needed observational constraint on ozone radiative forcing.

  12. 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.

  13. Catalytic wet air oxidation of carboxylic acids on TiO{sub 2}-supported ruthenium catalysts

    SciTech Connect

    Beziat, J.C.; Besson, M.; Gallezot, P.; Durecu, S.

    1999-02-15

    The total oxidation of aqueous solutions of carboxylic acids by air was studied in a slurry reactor over the temperature range 180--100 C and oxygen partial pressure of 0.3--1.8 MPa in the presence of a 2.8%Ru/TiO{sub 2} catalyst. The influence of various parameters is presented: the catalytic wet air oxidation of succinic acid is 0 order with respect to succinic acid; the order with respect to oxygen pressure is 0.4, and the activation energy is ca. 125 kJ/mol. It was found that acetic acid, which is one of the intermediates, and CO{sub 2} have no retarding effect on the total organic carbon abatement rate of succinic acid. Substitution of one hydrogen atom of the methyl group in acetic acid by Cl, OH, or NH{sub 2} gives an increase of the oxidation rate. However, it was proposed that the low activity of acetic acid oxidation is due not only to the difficulty to oxidize the methyl group, but also to the low adsorption coefficient of acetic acid on ruthenium surface. Inorganic salts, such as sodium chloride, only slightly decrease the oxidation rate of acetic acid. The absence of metal ions (Ru, Ti) in the effluents after reaction and the absence of particle sintering indicate also a high stability of the catalyst under the conditions employed. The catalyst can be recycled without loss of activity after the second run. The activity becomes stable after the attainment of a steady-state coverage of the Ru particles by oxygen. The study of the effect of reduction-oxidation treatments of the catalyst showed that the activity depends on the oxidation state of the surface.

  14. 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

  15. 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.

  16. Aeronautical System Center's environmental compliance assessment and management program's cost-saving initiatives support the Air Force's acquisition reform initiative

    SciTech Connect

    Meanor, T.

    1999-07-01

    The Environmental Management directorate of ASC (ASC/EM) has the responsibility of providing government oversight for the Government Owned Contractor Operated Aircraft and Missile plants (GOCOs). This oversight is manifested as a landlord role where Air Force provides the funding required to maintain the plant facilities including buildings and utilities as well as environmental systems. By agreement the companies operating the plants are required to operate them in accordance with environmental law. Presently the GOCOs include Air Force Plant (AFP) 6 in Marietta Ga., AFP 4 in Fort Worth, Tx., AFP 44 in Tucson, Az., AFP 42 in Palmdale, Ca., and AFP PJKS in Denver, Co. Lockheed Martin corporation operates AFPs 4,6, PJKS and a portion of AFP 42 while AFP 44 is operated by Raytheon Missile Systems Company. Other GOCOs at AFP 42 are Northrup-Grumman, Boeing, and Cabaco, the facilities engineer. Since 1992 the Environmental Management division has conducted its Environmental Compliance Assessment and Management Program assessments (ECAMP) annually at each of the plants. Using DOD's ECAMP Team Guide and teams comprised of both Air Force and consultant engineering personnel, each plant is assessed for its environmental compliance well being. In the face of rising operational costs and diminishing budgets ASC/EM performed a comprehensive review of its ECAMP. As a result, the basic ECAMP program was improved to reduce costs without compromising on quality of the effort. The program retained its emphasis in providing a snap-shot evaluation of each Air Force plant's environmental compliance health supported by complete but tailored protocol assessments.

  17. 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.

  18. 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)

  19. CasA mediates Cas3-catalyzed target degradation during CRISPR RNA-guided interference

    PubMed Central

    Hochstrasser, Megan L.; Taylor, David W.; Bhat, Prashant; Guegler, Chantal K.; Sternberg, Samuel H.; Nogales, Eva; Doudna, Jennifer A.

    2014-01-01

    In bacteria, the clustered regularly interspaced short palindromic repeats (CRISPR)–associated (Cas) DNA-targeting complex Cascade (CRISPR-associated complex for antiviral defense) uses CRISPR RNA (crRNA) guides to bind complementary DNA targets at sites adjacent to a trinucleotide signature sequence called the protospacer adjacent motif (PAM). The Cascade complex then recruits Cas3, a nuclease-helicase that catalyzes unwinding and cleavage of foreign double-stranded DNA (dsDNA) bearing a sequence matching that of the crRNA. Cascade comprises the CasA–E proteins and one crRNA, forming a structure that binds and unwinds dsDNA to form an R loop in which the target strand of the DNA base pairs with the 32-nt RNA guide sequence. Single-particle electron microscopy reconstructions of dsDNA-bound Cascade with and without Cas3 reveal that Cascade positions the PAM-proximal end of the DNA duplex at the CasA subunit and near the site of Cas3 association. The finding that the DNA target and Cas3 colocalize with CasA implicates this subunit in a key target-validation step during DNA interference. We show biochemically that base pairing of the PAM region is unnecessary for target binding but critical for Cas3-mediated degradation. In addition, the L1 loop of CasA, previously implicated in PAM recognition, is essential for Cas3 activation following target binding by Cascade. Together, these data show that the CasA subunit of Cascade functions as an essential partner of Cas3 by recognizing DNA target sites and positioning Cas3 adjacent to the PAM to ensure cleavage. PMID:24748111

  20. CasA mediates Cas3-catalyzed target degradation during CRISPR RNA-guided interference.

    PubMed

    Hochstrasser, Megan L; Taylor, David W; Bhat, Prashant; Guegler, Chantal K; Sternberg, Samuel H; Nogales, Eva; Doudna, Jennifer A

    2014-05-01

    In bacteria, the clustered regularly interspaced short palindromic repeats (CRISPR)-associated (Cas) DNA-targeting complex Cascade (CRISPR-associated complex for antiviral defense) uses CRISPR RNA (crRNA) guides to bind complementary DNA targets at sites adjacent to a trinucleotide signature sequence called the protospacer adjacent motif (PAM). The Cascade complex then recruits Cas3, a nuclease-helicase that catalyzes unwinding and cleavage of foreign double-stranded DNA (dsDNA) bearing a sequence matching that of the crRNA. Cascade comprises the CasA-E proteins and one crRNA, forming a structure that binds and unwinds dsDNA to form an R loop in which the target strand of the DNA base pairs with the 32-nt RNA guide sequence. Single-particle electron microscopy reconstructions of dsDNA-bound Cascade with and without Cas3 reveal that Cascade positions the PAM-proximal end of the DNA duplex at the CasA subunit and near the site of Cas3 association. The finding that the DNA target and Cas3 colocalize with CasA implicates this subunit in a key target-validation step during DNA interference. We show biochemically that base pairing of the PAM region is unnecessary for target binding but critical for Cas3-mediated degradation. In addition, the L1 loop of CasA, previously implicated in PAM recognition, is essential for Cas3 activation following target binding by Cascade. Together, these data show that the CasA subunit of Cascade functions as an essential partner of Cas3 by recognizing DNA target sites and positioning Cas3 adjacent to the PAM to ensure cleavage.

  1. Air-stable supported membranes for single-cell cytometry on PDMS microchips.

    PubMed

    Phillips, K Scott; Kang, Kyung Mo; Licata, Louise; Allbritton, Nancy L

    2010-04-01

    Protein-reinforced supported bilayer membranes (rSBMs) composed of phosphatidylcholine (PC), biotin-PE and Neutravidin were used to coat hybrid microchips composed of polydimethylsiloxane (PDMS) and glass. Since the coatings required a freshly oxidized, hydrophilic substrate, a novel method to rapidly connect reservoirs using plasma oxidation was first developed and found to support up to 5.2 N cm(-2) (1.5 N) pull-off force. rSBMs were then assembled in the oxidized hydrophilic channels. The electroosmotic mobility (mu(eo)) of rSBM-coated channels was measured over a 3 h time to evaluate the stability of the coatings for microchip electrophoresis. rSBM-coated microchips with a simple cross-design had excellent properties for microchip separations, yielding efficiencies of up to 700,000 plates m(-1) for fluorescent dyes and peptides. The separation performance of rSBM and PC-coated channels was evaluated after repeatedly drying and rehydrating the channels. The separation efficiency of fluorescein on PC-coated devices decreased by 40% after one dehydration cycle and nearly 75% after 3 cycles. In contrast for rSBM-coated devices there was no significant change in the fluorescein efficiency until the third cycle (10% decreased efficiency). rSBM-coated channels were also markedly more stable when placed in a dehydrated state during long-term storage compared to PC-coated channels, and showed reduced chip failure and no reduction in performance for up to one month of dehydrated storage. Finally, rSBM-coated devices were used to perform single-cell cytometry. Microchips that had been dehydrated, stored two weeks, and rehydrated prior to use demonstrated similar performance to newly coated devices for the separation of fluorescein and carboxyfluorescein from single cells. Thus rSBM-coated devices were rugged withstanding electric fields, prolonged storage under dehydrated conditions, and biofouling by cellular constituents while maintaining excellent separation

  2. 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.

  3. 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.

  4. Impact of urban air pollution on the allergenicity of Aspergillus fumigatus conidia: Outdoor exposure study supported by laboratory experiments.

    PubMed

    Lang-Yona, Naama; Shuster-Meiseles, Timor; Mazar, Yinon; Yarden, Oded; Rudich, Yinon

    2016-01-15

    Understanding the chemical interactions of common allergens in urban environments may help to decipher the general increase in susceptibility to allergies observed in recent decades. In this study, asexual conidia of the allergenic mold Aspergillus fumigatus were exposed to air pollution under natural (ambient) and controlled (laboratory) conditions. The allergenic activity was measured using two immunoassays and supported by a protein mass spectrometry analysis. The allergenicity of the conidia was found to increase by 2-5 fold compared to the control for short exposure times of up to 12h (accumulated exposure of about 50 ppb NO2 and 750 ppb O3), possibly due to nitration. At higher exposure times, the allergenicity increase lessened due to protein deamidation. These results indicate that during the first 12h of exposure, the allergenic potency of the fungal allergen A. fumigatus in polluted urban environments is expected to increase. Additional work is needed in order to determine if this behavior occurs for other allergens.

  5. 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.

  6. 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

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

    PubMed

    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

  8. 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

  9. 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…

  10. 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.

  11. Nucleosomes Inhibit Cas9 Endonuclease Activity in Vitro.

    PubMed

    Hinz, John M; Laughery, Marian F; Wyrick, John J

    2015-12-01

    During Cas9 genome editing in eukaryotic cells, the bacterial Cas9 enzyme cleaves DNA targets within chromatin. To understand how chromatin affects Cas9 targeting, we characterized Cas9 activity on nucleosome substrates in vitro. We find that Cas9 endonuclease activity is strongly inhibited when its target site is located within the nucleosome core. In contrast, the nucleosome structure does not affect Cas9 activity at a target site within the adjacent linker DNA. Analysis of target sites that partially overlap with the nucleosome edge indicates that the accessibility of the protospacer-adjacent motif (PAM) is the critical determinant of Cas9 activity on a nucleosome.

  12. 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...

  13. 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

  14. Cas9 specifies functional viral targets during CRISPR-Cas adaptation.

    PubMed

    Heler, Robert; Samai, Poulami; Modell, Joshua W; Weiner, Catherine; Goldberg, Gregory W; Bikard, David; Marraffini, Luciano A

    2015-03-12

    Clustered regularly interspaced short palindromic repeat (CRISPR) loci and their associated (Cas) proteins provide adaptive immunity against viral infection in prokaryotes. Upon infection, short phage sequences known as spacers integrate between CRISPR repeats and are transcribed into small RNA molecules that guide the Cas9 nuclease to the viral targets (protospacers). Streptococcus pyogenes Cas9 cleavage of the viral genome requires the presence of a 5'-NGG-3' protospacer adjacent motif (PAM) sequence immediately downstream of the viral target. It is not known whether and how viral sequences flanked by the correct PAM are chosen as new spacers. Here we show that Cas9 selects functional spacers by recognizing their PAM during spacer acquisition. The replacement of cas9 with alleles that lack the PAM recognition motif or recognize an NGGNG PAM eliminated or changed PAM specificity during spacer acquisition, respectively. Cas9 associates with other proteins of the acquisition machinery (Cas1, Cas2 and Csn2), presumably to provide PAM-specificity to this process. These results establish a new function for Cas9 in the genesis of prokaryotic immunological memory.

  15. Phylogeny of Cas9 determines functional exchangeability of dual-RNA and Cas9 among orthologous type II CRISPR-Cas systems

    PubMed Central

    Fonfara, Ines; Le Rhun, Anaïs; Chylinski, Krzysztof; Makarova, Kira S.; Lécrivain, Anne-Laure; Bzdrenga, Janek; Koonin, Eugene V.; Charpentier, Emmanuelle

    2014-01-01

    The CRISPR-Cas-derived RNA-guided Cas9 endonuclease is the key element of an emerging promising technology for genome engineering in a broad range of cells and organisms. The DNA-targeting mechanism of the type II CRISPR-Cas system involves maturation of tracrRNA:crRNA duplex (dual-RNA), which directs Cas9 to cleave invading DNA in a sequence-specific manner, dependent on the presence of a Protospacer Adjacent Motif (PAM) on the target. We show that evolution of dual-RNA and Cas9 in bacteria produced remarkable sequence diversity. We selected eight representatives of phylogenetically defined type II CRISPR-Cas groups to analyze possible coevolution of Cas9 and dual-RNA. We demonstrate that these two components are interchangeable only between closely related type II systems when the PAM sequence is adjusted to the investigated Cas9 protein. Comparison of the taxonomy of bacterial species that harbor type II CRISPR-Cas systems with the Cas9 phylogeny corroborates horizontal transfer of the CRISPR-Cas loci. The reported collection of dual-RNA:Cas9 with associated PAMs expands the possibilities for multiplex genome editing and could provide means to improve the specificity of the RNA-programmable Cas9 tool. PMID:24270795

  16. 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

  17. 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...

  18. 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

  19. CRISPR-Cas: biology, mechanisms and relevance

    PubMed Central

    Hille, Frank

    2016-01-01

    Prokaryotes have evolved several defence mechanisms to protect themselves from viral predators. Clustered regularly interspaced short palindromic repeats (CRISPR) and their associated proteins (Cas) display a prokaryotic adaptive immune system that memorizes previous infections by integrating short sequences of invading genomes—termed spacers—into the CRISPR locus. The spacers interspaced with repeats are expressed as small guide CRISPR RNAs (crRNAs) that are employed by Cas proteins to target invaders sequence-specifically upon a reoccurring infection. The ability of the minimal CRISPR-Cas9 system to target DNA sequences using programmable RNAs has opened new avenues in genome editing in a broad range of cells and organisms with high potential in therapeutical applications. While numerous scientific studies have shed light on the biochemical processes behind CRISPR-Cas systems, several aspects of the immunity steps, however, still lack sufficient understanding. This review summarizes major discoveries in the CRISPR-Cas field, discusses the role of CRISPR-Cas in prokaryotic immunity and other physiological properties, and describes applications of the system as a DNA editing technology and antimicrobial agent. This article is part of the themed issue ‘The new bacteriology’. PMID:27672148

  20. CRISPR-Cas: biology, mechanisms and relevance.

    PubMed

    Hille, Frank; Charpentier, Emmanuelle

    2016-11-01

    Prokaryotes have evolved several defence mechanisms to protect themselves from viral predators. Clustered regularly interspaced short palindromic repeats (CRISPR) and their associated proteins (Cas) display a prokaryotic adaptive immune system that memorizes previous infections by integrating short sequences of invading genomes-termed spacers-into the CRISPR locus. The spacers interspaced with repeats are expressed as small guide CRISPR RNAs (crRNAs) that are employed by Cas proteins to target invaders sequence-specifically upon a reoccurring infection. The ability of the minimal CRISPR-Cas9 system to target DNA sequences using programmable RNAs has opened new avenues in genome editing in a broad range of cells and organisms with high potential in therapeutical applications. While numerous scientific studies have shed light on the biochemical processes behind CRISPR-Cas systems, several aspects of the immunity steps, however, still lack sufficient understanding. This review summarizes major discoveries in the CRISPR-Cas field, discusses the role of CRISPR-Cas in prokaryotic immunity and other physiological properties, and describes applications of the system as a DNA editing technology and antimicrobial agent.This article is part of the themed issue 'The new bacteriology'.

  1. CRISPR-Cas: biology, mechanisms and relevance.

    PubMed

    Hille, Frank; Charpentier, Emmanuelle

    2016-11-01

    Prokaryotes have evolved several defence mechanisms to protect themselves from viral predators. Clustered regularly interspaced short palindromic repeats (CRISPR) and their associated proteins (Cas) display a prokaryotic adaptive immune system that memorizes previous infections by integrating short sequences of invading genomes-termed spacers-into the CRISPR locus. The spacers interspaced with repeats are expressed as small guide CRISPR RNAs (crRNAs) that are employed by Cas proteins to target invaders sequence-specifically upon a reoccurring infection. The ability of the minimal CRISPR-Cas9 system to target DNA sequences using programmable RNAs has opened new avenues in genome editing in a broad range of cells and organisms with high potential in therapeutical applications. While numerous scientific studies have shed light on the biochemical processes behind CRISPR-Cas systems, several aspects of the immunity steps, however, still lack sufficient understanding. This review summarizes major discoveries in the CRISPR-Cas field, discusses the role of CRISPR-Cas in prokaryotic immunity and other physiological properties, and describes applications of the system as a DNA editing technology and antimicrobial agent.This article is part of the themed issue 'The new bacteriology'. PMID:27672148

  2. 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

  3. 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.

  4. 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.

  5. 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.

  6. 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.

  7. CRISPR/Cas9-mediated genome editing of Epstein-Barr virus in human cells.

    PubMed

    Yuen, Kit-San; Chan, Chi-Ping; Wong, Nok-Hei Mickey; Ho, Chau-Ha; Ho, Ting-Hin; Lei, Ting; Deng, Wen; Tsao, Sai Wah; Chen, Honglin; Kok, Kin-Hang; Jin, Dong-Yan

    2015-03-01

    The CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR-associated 9) system is a highly efficient and powerful tool for RNA-guided editing of the cellular genome. Whether CRISPR/Cas9 can also cleave the genome of DNA viruses such as Epstein-Barr virus (EBV), which undergo episomal replication in human cells, remains to be established. Here, we reported on CRISPR/Cas9-mediated editing of the EBV genome in human cells. Two guide RNAs (gRNAs) were used to direct a targeted deletion of 558 bp in the promoter region of BART (BamHI A rightward transcript) which encodes viral microRNAs (miRNAs). Targeted editing was achieved in several human epithelial cell lines latently infected with EBV, including nasopharyngeal carcinoma C666-1 cells. CRISPR/Cas9-mediated editing of the EBV genome was efficient. A recombinant virus with the desired deletion was obtained after puromycin selection of cells expressing Cas9 and gRNAs. No off-target cleavage was found by deep sequencing. The loss of BART miRNA expression and activity was verified, supporting the BART promoter as the major promoter of BART RNA. Although CRISPR/Cas9-mediated editing of the multicopy episome of EBV in infected HEK293 cells was mostly incomplete, viruses could be recovered and introduced into other cells at low m.o.i. Recombinant viruses with an edited genome could be further isolated through single-cell sorting. Finally, a DsRed selectable marker was successfully introduced into the EBV genome during the course of CRISPR/Cas9-mediated editing. Taken together, our work provided not only the first genetic evidence that the BART promoter drives the expression of the BART transcript, but also a new and efficient method for targeted editing of EBV genome in human cells.

  8. 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.

  9. Work plan for upgrading the 241-A-701 compressed air system and motor control center. Revision 1

    SciTech Connect

    Carpenter, K.E.

    1995-01-17

    This work plan will outline the responsibilities associated with the 241-A-701 Compressed Air System (CAS) and Motor Control Center (MCC) upgrades. All activities required to design, install, test, and operate the modified systems are addressed in this document. Upgrades Technical Support (UTS) of TWRS Engineering is responsible for the completion of all tasks associated with this upgrade. UTS will coordinate the upgrade activities, and ensure all tasks are successfully completed on or before the scheduled dates. The primary objective of the 241-A-701 Compressor and MCC Upgrade is to provide a reliable source of process and instrument compressed air to the A, AX, AY, and AZ tank farms.

  10. 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…

  11. 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

  12. 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....

  13. 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....

  14. 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....

  15. 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

  16. 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.

  17. CRISPR-Cas immunity in prokaryotes.

    PubMed

    Marraffini, Luciano A

    2015-10-01

    Prokaryotic organisms are threatened by a large array of viruses and have developed numerous defence strategies. Among these, only clustered, regularly interspaced short palindromic repeat (CRISPR)-Cas systems provide adaptive immunity against foreign elements. Upon viral injection, a small sequence of the viral genome, known as a spacer, is integrated into the CRISPR locus to immunize the host cell. Spacers are transcribed into small RNA guides that direct the cleavage of the viral DNA by Cas nucleases. Immunization through spacer acquisition enables a unique form of evolution whereby a population not only rapidly acquires resistance to its predators but also passes this resistance mechanism vertically to its progeny.

  18. 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

  19. CRISPR-Cas: Revolutionising genome engineering.

    PubMed

    Nicholson, Samantha Anne; Pepper, Michael Sean

    2016-08-01

    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.

  20. 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...

  1. 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...

  2. 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

  3. 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....

  4. 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....

  5. 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....

  6. 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....

  7. 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...

  8. 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....

  9. 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...

  10. 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....

  11. 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....

  12. 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...

  13. 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....

  14. 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...

  15. 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....

  16. 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....

  17. 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.

  18. 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

  19. Effects of Using a Computer Algebra System (CAS) on Junior College Students' Attitudes towards CAS and Achievement in Mathematics

    ERIC Educational Resources Information Center

    Leng, Ng Wee; Choo, Kwee Tiow; Soon, Lau Hock; Yi-Huak, Koh; Sun, Yap Yew

    2005-01-01

    This study examines the effects of using Texas Instruments' Voyage 200 calculator (V200), a graphing calculator with a built-in computer algebra system (CAS), on attitudes towards CAS and achievement in mathematics of junior college students (17 year olds). Students' attitudes towards CAS were examined using a 40-item Likert-type instrument…

  20. 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.

  1. [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

  2. 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.

  3. 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.

  4. 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

  5. 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

  6. 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.

  7. Nucleosome breathing and remodeling constrain CRISPR-Cas9 function.

    PubMed

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

    2016-04-28

    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.

  8. 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...

  9. “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...

  10. 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...

  11. 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

  12. Characterization of composite phthalocyanine-fatty acid films from the air/water interface to solid supports.

    PubMed

    Giancane, G; Manno, D; Serra, A; Sgobba, V; Valli, L

    2011-12-22

    A commercial vanadyl 2,9,16,23-tetraphenoxy-29H,31H-phthalocyanine (VOPc) was dissolved in chloroform and spread on ultrapure water subphase in a Langmuir trough. The floating film was thoroughly characterized at the air-water interface by means of the Langmuir isotherm, Brewster angle microscopy, UV-vis reflection spectroscopy, and infrared measurements carried out directly at the air-water interface. All the results showed the formation of a non-uniform and aggregated floating layer, too rigid to be transferred by the Langmuir-Blodgett (LB) method. For this reason, a mixture of arachidic acid and VOPc was realized, characterized, and transferred by the LB technique on solid substrates. Interface measurements and atomic force microscopy analysis suggested the formation of a uniform arachidic acid film and a superimposed VOPc placed in prone configuration.

  13. 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

  14. 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.

  15. 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

  16. Annotation and Classification of CRISPR-Cas Systems.

    PubMed

    Makarova, Kira S; Koonin, Eugene V

    2015-01-01

    The clustered regularly interspaced short palindromic repeats (CRISPR)-Cas (CRISPR-associated proteins) is a prokaryotic adaptive immune system that is represented in most archaea and many bacteria. Among the currently known prokaryotic defense systems, the CRISPR-Cas genomic loci show unprecedented complexity and diversity. Classification of CRISPR-Cas variants that would capture their evolutionary relationships to the maximum possible extent is essential for comparative genomic and functional characterization of this theoretically and practically important system of adaptive immunity. To this end, a multipronged approach has been developed that combines phylogenetic analysis of the conserved Cas proteins with comparison of gene repertoires and arrangements in CRISPR-Cas loci. This approach led to the current classification of CRISPR-Cas systems into three distinct types and ten subtypes for each of which signature genes have been identified. Comparative genomic analysis of the CRISPR-Cas systems in new archaeal and bacterial genomes performed over the 3 years elapsed since the development of this classification makes it clear that new types and subtypes of CRISPR-Cas need to be introduced. Moreover, this classification system captures only part of the complexity of CRISPR-Cas organization and evolution, due to the intrinsic modularity and evolutionary mobility of these immunity systems, resulting in numerous recombinant variants. Moreover, most of the cas genes evolve rapidly, complicating the family assignment for many Cas proteins and the use of family profiles for the recognition of CRISPR-Cas subtype signatures. Further progress in the comparative analysis of CRISPR-Cas systems requires integration of the most sensitive sequence comparison tools, protein structure comparison, and refined approaches for comparison of gene neighborhoods.

  17. 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

  18. Scalable air cathode microbial fuel cells using glass fiber separators, plastic mesh supporters, and graphite fiber brush anodes.

    PubMed

    Zhang, Xiaoyuan; Cheng, Shaoan; Liang, Peng; Huang, Xia; Logan, Bruce E

    2011-01-01

    The combined use of brush anodes and glass fiber (GF1) separators, and plastic mesh supporters were used here for the first time to create a scalable microbial fuel cell architecture. Separators prevented short circuiting of closely-spaced electrodes, and cathode supporters were used to avoid water gaps between the separator and cathode that can reduce power production. The maximum power density with a separator and supporter and a single cathode was 75 ± 1 W/m(3). Removing the separator decreased power by 8%. Adding a second cathode increased power to 154 ± 1 W/m(3). Current was increased by connecting two MFCs connected in parallel. These results show that brush anodes, combined with a glass fiber separator and a plastic mesh supporter, produce a useful MFC architecture that is inherently scalable due to good insulation between the electrodes and a compact architecture. PMID:20566288

  19. Scalable air cathode microbial fuel cells using glass fiber separators, plastic mesh supporters, and graphite fiber brush anodes.

    PubMed

    Zhang, Xiaoyuan; Cheng, Shaoan; Liang, Peng; Huang, Xia; Logan, Bruce E

    2011-01-01

    The combined use of brush anodes and glass fiber (GF1) separators, and plastic mesh supporters were used here for the first time to create a scalable microbial fuel cell architecture. Separators prevented short circuiting of closely-spaced electrodes, and cathode supporters were used to avoid water gaps between the separator and cathode that can reduce power production. The maximum power density with a separator and supporter and a single cathode was 75 ± 1 W/m(3). Removing the separator decreased power by 8%. Adding a second cathode increased power to 154 ± 1 W/m(3). Current was increased by connecting two MFCs connected in parallel. These results show that brush anodes, combined with a glass fiber separator and a plastic mesh supporter, produce a useful MFC architecture that is inherently scalable due to good insulation between the electrodes and a compact architecture.

  20. 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.

  1. 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.

  2. 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.

  3. 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.

  4. 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.

  5. Successful trans-Atlantic air ambulance transfer of a patient supported by a bi-ventricular assist device.

    PubMed

    McLean, Neilson; Copeland, Ryan; Casey, Neil; Samoukovic, Gordon; Quigley, Robert

    2011-08-01

    The ventricular assist device (VAD) is a hemodynamic support device that augments cardiac output for patients with severe ventricular dysfunction. With improved reliability and technological advances, the use of VADs to support patients is increasing. Many VAD-dependent patients ultimately require heart transplants that are only available in specialized centers, necessitating an interhospital transfer. To date there are few reports of long-distance fixed wing aeromedical transport of patients dependent on a VAD. Here we describe the successful transfer of a patient supported by a biventricular assist device (BiVAD) from Cambridge, UK, to Durham, NC, via fixed-wing jet aircraft. During this transfer, we observed hemodynamic alterations secondary to gravitational forces, which should be anticipated and may be mitigated with simple maneuvers. With high-level logistical planning and appropriate medical oversight, patients dependant on BiVADs can be safely transported by fixed wing aircraft over long distances.

  6. 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

  7. 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…

  8. 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

  9. Microoxic Niches within the Thylakoid Stroma of Air-Grown Chlamydomonas reinhardtii Protect [FeFe]-Hydrogenase and Support Hydrogen Production under Fully Aerobic Environment1[OPEN

    PubMed Central

    Liran, Oded; Milrad, Yuval; Eilenberg, Haviva; Weiner, Iddo

    2016-01-01

    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, 18O2 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

  10. 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

  11. 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

  12. 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.

  13. Identifying inequitable exposure to toxic air pollution in racialized and low-income neighbourhoods to support pollution prevention.

    PubMed

    Kershaw, Suzanne; Gower, Stephanie; Rinner, Claus; Campbell, Monica

    2013-05-01

    Numerous environmental justice studies have confirmed a relationship between population characteristics such as low-income or minority status and the location of environmental health hazards. However, studies of the health risks from exposure to harmful substances often do not consider their toxicological characteristics. We used two different methods, the unit-hazard and the distance-based approach, to evaluate demographic and socio-economic characteristics of the population residing near industrial facilities in the City of Toronto, Canada. In addition to the mass of air emissions obtained from the national pollutant release inventory (NPRI), we also considered their toxicity using toxic equivalency potential (TEP) scores. Results from the unit-hazard approach indicate no significant difference in the proportion of low-income individuals living in host versus non-host census tracts (t(107) = 0.3, P = 0.735). However, using the distance-based approach, the proportion of low-income individuals was significantly higher (+5.1%, t(522) = 6.0, P <0.001) in host tracts, while the indicator for "racialized" communities ("visible minority") was 16.1% greater (t(521) = 7.2, P <0.001) within 2 km of a NPRI facility. When the most toxic facilities by non-carcinogenic TEP score were selected, the rate of visible minorities living near the most toxic NPRI facilities was significantly higher (+12.9%, t(352) = 3.5, P = 0.001) than near all other NPRI facilities. TEP scores were also used to identify areas in Toronto that face a double burden of poverty and air toxics exposure in order to prioritise pollution prevention.

  14. 41 CFR 102-33.440 - Who must report CAS cost and utilization data?

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... (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...

  15. 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

  16. 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.

  17. 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.

  18. 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

  19. 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

  20. Leiomyoblastome gastrique: à propos de trois cas

    PubMed Central

    Moujahid, Mountassir; Ennafaa, Issam; EL Rhari, Ahmed; Serghini, Issam; Chekoura, Khalid; Tahiri, Moulay Hassan

    2015-01-01

    Le leiomyoblastome gastrique est une tumeur musculaire rare qui touche essentiellement l'adulte. Son développement est souvent exogastrique. Le diagnostic positif repose sur l'histologie et le traitement est basé sur la chirurgie. Nous rapportons trois cas de leiomyoblastome gastrique colligés dans le service de chirurgie générale au 5ème Hôpital Militaire. L’âge moyen des patients est de 47 ans; le motif de consultation était représenté par une hémorragie digestive et l'imagerie médicale a posé le diagnostic de masse tumorale dans tous les cas. Le traitement chirurgical consistait en une gastrectomie partielle et le compte rendu anatomopathologique a confirmé le leiomyoblastome gastrique dans les trois cas. Le siège de la tumeur a été posé par la fibroscopie oeso gastroduodénale, le traitement était chirurgical et les suites post opératoires étaient simples avec un contrôle par des fibroscopies répétitives sans aucun signe de récidive. Le leiomyoblastome gastrique est une tumeur rare. L’écho endoscopie joue un rôle primordial dans le diagnostic positif ainsi que dans l’évaluation de l'extension pariétale de ces tumeurs. Le traitement est essentiellement chirurgical. PMID:26090000

  1. 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

  2. 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

  3. 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

    2015-12-08

    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.

  4. 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.

  5. Using CAS to Solve a Mathematics Task: A Deconstruction

    ERIC Educational Resources Information Center

    Berger, Margot

    2010-01-01

    I investigate how and whether a heterogeneous group of first-year university mathematics students in South Africa harness the potential power of a computer algebra system (CAS) when doing a specific mathematics task. In order to do this, I develop a framework for deconstructing a mathematics task requiring the use of CAS, into its primary…

  6. 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…

  7. 48 CFR 9903.201-2 - Types of CAS coverage.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ..., OFFICE OF FEDERAL PROCUREMENT POLICY, OFFICE OF MANAGEMENT AND BUDGET PROCUREMENT PRACTICES AND COST... coverage. Full coverage requires that the business unit comply with all of the CAS specified in part 9904... later award of a CAS-covered contract. Full coverage applies to contractor business units that—...

  8. 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...

  9. 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...

  10. 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...

  11. Diversity of CRISPR-Cas immune systems and molecular machines.

    PubMed

    Barrangou, Rodolphe

    2015-01-01

    Bacterial adaptive immunity hinges on CRISPR-Cas systems that provide DNA-encoded, RNA-mediated targeting of exogenous nucleic acids. A plethora of CRISPR molecular machines occur broadly in prokaryotic genomes, with a diversity of Cas nucleases that can be repurposed for various applications.

  12. 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…

  13. Cas Proteins in Normal and Pathological Cell Growth Control

    PubMed Central

    Tikhmyanova, Nadezhda; Little, Joy L.; Golemis, Erica A.

    2009-01-01

    Proteins of the CAS (Crk-Associated Substrate) family (BCAR1/p130Cas, NEDD9/HEF1/Cas-L, EFS/SIN and CASS4/HEPL) are integral players in normal and pathological cell biology. CAS proteins act as scaffolds to regulate protein complexes controlling migration and chemotaxis, apoptosis, cell cycle, and differentiation, and have more recently been linked to a role in progenitor cell function. Reflecting these complex functions, over-expression of CAS proteins has now been strongly linked to poor prognosis and increased metastasis in cancer, as well as resistance to first-line chemotherapeutics in multiple tumor types including breast and lung cancers, glioblastoma, and melanoma. Further, CAS proteins have also been linked to additional pathological conditions including inflammatory disorders, Alzheimer’s and Parkinson’s disease, as well as developmental defects. This review will explore the roles of the CAS proteins in normal and pathological states in the context of the many mechanistic insights into CAS protein function that have emerged in the past decade. PMID:19937461

  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.

  15. 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.

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

    PubMed

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

    2016-06-01

    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.

  17. 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…

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

    PubMed

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

    2015-05-12

    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.

  19. Adaptive optics at the IOE, CAS

    NASA Astrophysics Data System (ADS)

    Jiang, Wenhan; Rao, Changhui; Zhang, Yudong; Ling, Ning; Guan, Chunlin

    2009-02-01

    R&D on Adaptive Optics in the Institute of Optics & Electronics (IOE), Chinese Academy of Sciences (CAS) began in 1979. In this paper, several recent achievements will be reported: 1. AO for astronomical telescopes. AO system for 1.2 m telescope at Yunnan Astronomical Observatory was built in 1998. It was updated in 2004, and high resolution images approaching diffraction limit were obtained. A new AO telescope with 1.8m aperture is being built, and a 4m AO telescope is being planned. 2. AO for ICF facility. A 19-element AO system with hill-climbing control algorithm for "Shenguang I" ICF facility was built in 1985, which was the first AO system used in ICF facility in the world. A set of 8 AO systems was installed in a bundle of ICF prototype for "Shenguang III" ICF facility. A new system is being built for further development of ICF facility. 3. AO for retinal imaging. In 1999, the first AO system with 19-element DM was built for retinal imaging. Several AO systems with 37-element deformable mirror (DM) were built and used for vision research and clinical inspection. It is being integrated with an OCT system for high resolution retinal imaging. All of the main subsystems, such as DMs, wavefront sensors, and high speed processors, were built in the Laboratory on Adaptive Optics of IOE, CAS.

  20. 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.

  1. 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.

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

    PubMed

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

    2015-10-15

    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.

  4. 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.

  5. 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.…

  6. Crystal Structure of Streptococcus pyogenes Cas1 and Its Interaction with Csn2 in the Type II CRISPR-Cas System.

    PubMed

    Ka, Donghyun; Lee, Hasup; Jung, Yi-Deun; Kim, Kyunggon; Seok, Chaok; Suh, Nayoung; Bae, Euiyoung

    2016-01-01

    CRISPRs and Cas proteins constitute an RNA-guided microbial immune system against invading nucleic acids. Cas1 is a universal Cas protein found in all three types of CRISPR-Cas systems, and its role is implicated in new spacer acquisition during CRISPR-mediated adaptive immunity. Here, we report the crystal structure of Streptococcus pyogenes Cas1 (SpCas1) in a type II CRISPR-Cas system and characterize its interaction with S. pyogenes Csn2 (SpCsn2). The SpCas1 structure reveals a unique conformational state distinct from type I Cas1 structures, resulting in a more extensive dimerization interface, a more globular overall structure, and a disruption of potential metal-binding sites for catalysis. We demonstrate that SpCas1 directly interacts with SpCsn2, and identify the binding interface and key residues for Cas complex formation. These results provide structural information for a type II Cas1 protein, and lay a foundation for studying multiprotein Cas complexes functioning in type II CRISPR-Cas systems.

  7. 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

  8. Genomic Disruption of VEGF-A Expression in Human Retinal Pigment Epithelial Cells Using CRISPR-Cas9 Endonuclease

    PubMed Central

    Yiu, Glenn; Tieu, Eric; Nguyen, Anthony T.; Wong, Brittany; Smit-McBride, Zeljka

    2016-01-01

    Purpose To employ type II clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 endonuclease to suppress ocular angiogenesis by genomic disruption of VEGF-A in human RPE cells. Methods CRISPR sequences targeting exon 1 of human VEGF-A were computationally identified based on predicted Cas9 on- and off-target probabilities. Single guide RNA (gRNA) cassettes with these target sequences were cloned into lentiviral vectors encoding the Streptococcus pyogenes Cas9 endonuclease (SpCas9) gene. The lentiviral vectors were used to infect ARPE-19 cells, a human RPE cell line. Frequency of insertion or deletion (indel) mutations was assessed by T7 endonuclease 1 mismatch detection assay; mRNA levels were assessed with quantitative real-time PCR; and VEGF-A protein levels were determined by ELISA. In vitro angiogenesis was measured using an endothelial cell tube formation assay. Results Five gRNAs targeting VEGF-A were selected based on the highest predicted on-target probabilities, lowest off-target probabilities, or combined average of both scores. Lentiviral delivery of the top-scoring gRNAs with SpCas9 resulted in indel formation in the VEGF-A gene at frequencies up to 37.0% ± 4.0% with corresponding decreases in secreted VEGF-A protein up to 41.2% ± 7.4% (P < 0.001), and reduction of endothelial tube formation up to 39.4% ± 9.8% (P = 0.02). No significant indel formation in the top three putative off-target sites tested was detected. Conclusions The CRISPR-Cas9 endonuclease system may reduce VEGF-A secretion from human RPE cells and suppress angiogenesis, supporting the possibility of employing gene editing for antiangiogenesis therapy in ocular diseases. PMID:27768202

  9. Light Curves and Analyses of the Eclipsing Binaries EG Cas and EP Cas

    NASA Astrophysics Data System (ADS)

    Bradstreet, David H.; Sanders, S. J.; McClain, T. R.

    2010-01-01

    New precision V & Rc light curves of the eclipsing binaries EG Cas and EP Cas have been obtained using the 41-cm telescope at the Eastern University Observatory equipped with an SBIG ST-10XME CCD. EG Cas (P = 0.6115 days, Vmax = 12.9) has no published light curves and only a few dozen (mostly visual) timings of minimum light. The system is being observed throughout the fall of 2009 and the current light curves distinctly show that the system is a totally eclipsing overcontact binary. The light curves are also significantly asymmetric (strong O'Connell effect) indicating the presence of large, cool starspots, most likely on both stars. Preliminary analysis indicates that the binary is an A-type (the larger, more massive star is the hotter component), has a mass ratio of 0.32, very large temperature difference between the stars greater than 1600 K, and a fillout of 0.26. The modern timings of minimum light combined with those in the literature indicate that the binary's period is decreasing. The large temperature difference coupled to a significant fillout factor seems contradictory, and further data acquisition and analysis will hopefully resolve this seeming enigma. EP Cas (P = 0.8134 days, Vmax = 11.2) is a partially eclipsing detached system with a relatively deep primary eclipse of 1.0 mag in Rc. No published light curves exist for this system although many timings of minimum light have been published. The O-C curve indicates that the period for the binary has remained relatively constant since observations were first published in 1936. Preliminary light curve models indicate a partially eclipsing system consisting of detached but tidally distorted stars. The complete light curve analyses as well as a period study of all published times of minimum light will be presented for both systems.

  10. Performance of the Cas9 nickase system in Drosophila melanogaster.

    PubMed

    Ren, Xingjie; Yang, Zhihao; Mao, Decai; Chang, Zai; Qiao, Huan-Huan; Wang, Xia; Sun, Jin; Hu, Qun; Cui, Yan; Liu, Lu-Ping; Ji, Jun-Yuan; Xu, Jiang; Ni, Jian-Quan

    2014-10-01

    Recent studies of the Cas9/sgRNA system in Drosophila melanogaster genome editing have opened new opportunities to generate site-specific mutant collections in a high-throughput manner. However, off-target effects of the system are still a major concern when analyzing mutant phenotypes. Mutations converting Cas9 to a DNA nickase have great potential for reducing off-target effects in vitro. Here, we demonstrated that injection of two plasmids encoding neighboring offset sgRNAs into transgenic Cas9(D10A) nickase flies efficiently produces heritable indel mutants. We then determined the effective distance between the two sgRNA targets and their orientations that affected the ability of the sgRNA pairs to generate mutations when expressed in the transgenic nickase flies. Interestingly, Cas9 nickase greatly reduces the ability to generate mutants with one sgRNA, suggesting that the application of Cas9 nickase and sgRNA pairs can almost avoid off-target effects when generating indel mutants. Finally, a defined piwi mutant allele is generated with this system through homology-directed repair. However, Cas9(D10A) is not as effective as Cas9 in replacing the entire coding sequence of piwi with two sgRNAs. PMID:25128437

  11. 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

  12. 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.

  13. 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...

  14. 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...

  15. 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...

  16. 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...

  17. 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...

  18. 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...

  19. 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...

  20. 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...

  1. 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.

  2. 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.

  3. 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

  4. Orbital period determination in an eclipsing dwarf nova HT Cas

    NASA Astrophysics Data System (ADS)

    Bąkowska, Karolina; Olech, Arkadiusz

    2014-09-01

    HT Cassiopeiae was discovered over seventy years ago (Hoffmeister 1943). Unfortunately, for 35 years this object did not receive any attention, until the eclipses of HT Cas were observed by Bond. After a first analysis, Patterson (1981) called HT Cas "a Rosetta stone among dwarf novae". Since then, the literature on this star is still growing, reaching several dozens of publications. We present an orbital period determination of HT Cas during the November 2010 super-outburst, but also during a longer time span, to check its stability.

  5. CRISPR/Cas9-Mediated Correction of the Sickle Mutation in Human CD34+ cells.

    PubMed

    Hoban, Megan D; Lumaquin, Dianne; Kuo, Caroline Y; Romero, Zulema; Long, Joseph; Ho, Michelle; Young, Courtney S; Mojadidi, Michelle; Fitz-Gibbon, Sorel; Cooper, Aaron R; Lill, Georgia R; Urbinati, Fabrizia; Campo-Fernandez, Beatriz; Bjurstrom, Carmen F; Pellegrini, Matteo; Hollis, Roger P; Kohn, Donald B

    2016-09-01

    Targeted genome editing technology can correct the sickle cell disease mutation of the β-globin gene in hematopoietic stem cells. This correction supports production of red blood cells that synthesize normal hemoglobin proteins. Here, we demonstrate that Transcription Activator-Like Effector Nucleases (TALENs) and the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 nuclease system can target DNA sequences around the sickle-cell mutation in the β-globin gene for site-specific cleavage and facilitate precise correction when a homologous donor template is codelivered. Several pairs of TALENs and multiple CRISPR guide RNAs were evaluated for both on-target and off-target cleavage rates. Delivery of the CRISPR/Cas9 components to CD34+ cells led to over 18% gene modification in vitro. Additionally, we demonstrate the correction of the sickle cell disease mutation in bone marrow derived CD34+ hematopoietic stem and progenitor cells from sickle cell disease patients, leading to the production of wild-type hemoglobin. These results demonstrate correction of the sickle mutation in patient-derived CD34+ cells using CRISPR/Cas9 technology.

  6. Fibronectin Rigidity Response through Fyn and p130Cas Recruitment to the Leading Edge

    PubMed Central

    Kostic, Ana

    2006-01-01

    Cell motility on extracellular matrices critically depends on matrix rigidity, which affects cell adhesion and formation of focal contacts. Receptor-like protein tyrosine phosphatase alpha (RPTPα) and the αvβ3 integrin form a rigidity-responsive complex at the leading edge. Here we show that the rigidity response through increased spreading and growth correlates with leading edge recruitment of Fyn, but not endogenous c-Src. Recruitment of Fyn requires the palmitoylation site near the N-terminus and addition of that site to c-Src enables it to support a rigidity response. In all cases, the rigidity response correlates with the recruitment of the Src family kinase to early adhesions. The stretch-activated substrate of Fyn and c-Src, p130Cas, is also required for a rigidity response and it is phosphorylated at the leading edge in a Fyn-dependent process. A possible mechanism for the fibronectin rigidity response involves force-dependent Fyn phosphorylation of p130Cas with rigidity-dependent displacement. With the greater displacement of Fyn from p130Cas on softer surfaces, there will be less phosphorylation. These studies emphasize the importance of force and nanometer-level movements in cell growth and function. PMID:16597701

  7. Study of Eclipsing Binary and Multiple Systems in OB Associations IV: Cas OB6 Member DN Cas

    NASA Astrophysics Data System (ADS)

    Bakış, V.; Bakış, H.; Bilir, S.; Eker, Z.

    2016-09-01

    An early-type, massive, short-period (Porb=2d.310951) eclipsing spectroscopic binary DN Cas has been re-visited with new spectral and photometric data. The masses and radii of the components have been obtained as M1=19.04± 0.07 M⊙, M2=13.73± 0.05 M⊙ and R1=7.22± 0.06 R⊙, R2=5.79± 0.06 R⊙, respectively. Both components present synchronous rotation (Vrot1=160 km s-1, Vrot2=130 km s-1) with their orbit. Orbital period analysis yielded a physically bound additional component in the system with a minimum mass of M3=0.88 M⊙ orbiting in an eccentric orbit (e = 0.37 ± 0.2) with an orbital period of P 12 = 42 ± 9 yr. High precision absolute parameters of the system allowed us to derive a distance to DN Cas as 1.7 ± 0.2 kpc which locates the system within the borders of the Cas OB6 association (d = 1.8 kpc). The space velocities and the age of DN Cas are in agreement with those of Cas OB6. The age of DN Cas (τ = 3-5 Myr) is found to be 1-2 Myr older than the embedded clusters (IC 1795, IC 1805, and IC 1848) in the Cas OB6 association, which implies a sequential star formation in the association.

  8. 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.

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

    PubMed

    Kim, Hyun-Min; Colaiácovo, Monica P

    2016-07-01

    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.

  10. 48 CFR 970.3002-1 - CAS applicability.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... SUPPLEMENTARY REGULATIONS DOE MANAGEMENT AND OPERATING CONTRACTS Cost Accounting Standards Administration 970.3002-1 CAS applicability. The provisions of 48 CFR part 30 and 48 CFR chapter 99 (FAR Appendix)...

  11. 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

  12. 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

  13. Editing plant genomes with CRISPR/Cas9.

    PubMed

    Belhaj, Khaoula; Chaparro-Garcia, Angela; Kamoun, Sophien; Patron, Nicola J; Nekrasov, Vladimir

    2015-04-01

    CRISPR/Cas9 is a rapidly developing genome editing technology that has been successfully applied in many organisms, including model and crop plants. Cas9, an RNA-guided DNA endonuclease, can be targeted to specific genomic sequences by engineering a separately encoded guide RNA with which it forms a complex. As only a short RNA sequence must be synthesized to confer recognition of a new target, CRISPR/Cas9 is a relatively cheap and easy to implement technology that has proven to be extremely versatile. Remarkably, in some plant species, homozygous knockout mutants can be produced in a single generation. Together with other sequence-specific nucleases, CRISPR/Cas9 is a game-changing technology that is poised to revolutionise basic research and plant breeding.

  14. 48 CFR 9903.201-2 - Types of CAS coverage.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... to the same cost accounting practices. The term includes Government-owned contractor-operated (GOCO...), as applicable, shall be incorporated. (6) Continuity in fully CAS-covered contracts. Where...

  15. 48 CFR 9903.201-2 - Types of CAS coverage.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... to the same cost accounting practices. The term includes Government-owned contractor-operated (GOCO...), as applicable, shall be incorporated. (6) Continuity in fully CAS-covered contracts. Where...

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

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... to the same cost accounting practices. The term includes Government-owned contractor-operated (GOCO...), as applicable, shall be incorporated. (6) Continuity in fully CAS-covered contracts. Where...

  17. 48 CFR 970.3002-1 - CAS applicability.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... SUPPLEMENTARY REGULATIONS DOE MANAGEMENT AND OPERATING CONTRACTS Cost Accounting Standards Administration 970.3002-1 CAS applicability. The provisions of 48 CFR part 30 and 48 CFR chapter 99 (FAR Appendix)...

  18. 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

  19. 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

  20. 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.

  1. Calibrated Ancillary System (CAS) user's guide, volume 6

    NASA Technical Reports Server (NTRS)

    1986-01-01

    The Calibrated Ancillary System (CAS) provides real-time calibrated parameters from the orbiter downlink (ancillary data) to the Goddard Space Flight Center (GSFC). This user's guide contains the introduction to the equipment, operation, general procedures, and specific procedures of CAS. Volume 6 describes ancillary products procedures, enhancement menu and processing task procedures for SDT/TDT (shuttle data tape/telemetry descriptor tape), database errors and network data driver (NDD) product menu procedures, and utility menu procedures.

  2. 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.

  3. 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

  4. 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

    2014-12-09

    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.

  5. 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.

  6. 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.

  7. Rational design of a split-Cas9 enzyme complex

    DOE PAGES

    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

  8. 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.

  9. 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

  10. 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

  11. 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

  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.

  13. 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

  14. 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.

  15. 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.

  16. 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

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

    PubMed

    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.

  18. 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

  19. Soft modes and superconductivity in the layered hexagonal carbides V2CAs, Nb2CAs, and Nb2CS

    NASA Astrophysics Data System (ADS)

    Halilov, S. V.; Singh, D. J.; Papaconstantopoulos, D. A.

    2002-05-01

    The electronic structure, selected phonon frequencies, and electron phonon coupling of the hexagonal layered compounds Nb2CS, Nb2CAs, and V2CAs are elucidated using density functional calculations. All materials are good three-dimensional metals with moderate values of the density of states, N(EF). The electronic structure of Nb2CS, the only superconductor within this family of materials, consists of well-hybridized Nb d-S p-C p derived bands characteristic around the Fermi energy. The S is more ionic in this compound than the As in the related arsenides Nb2CAs or V2CAs. This results in a relative softness for phonon modes involving S. Rigid muffin tin approximation calculations show moderate electron phonon couplings consistent with low-temperature superconductivity. The key difference from the carbo-arsenides is the presence of soft moderately coupled S modes.

  20. 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

  1. 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

  2. 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.

  3. 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.

  4. 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

  5. A laboratory study examining the impact of linen use on low-air-loss support surface heat and water vapor transmission rates.

    PubMed

    Williamson, Rachel; Lachenbruch, Charlie; VanGilder, Catherine

    2013-08-01

    Layers of linens are frequently placed under patients to manage moisture and/or assist with positioning immobile patients, including persons placed on a therapeutic surface because they are at risk for developing pressure ulcers. Because skin microclimate is believed to affect pressure ulcer risk, some therapeutic surfaces are designed to manage skin temperature and humidity (microclimate management). The purpose of this study was to measure the effects of linens and underpads on a low-air-loss (LAL) surface's ability to disperse heat and evaporate moisture. Underpads and transfer sheet combinations (grouped by three common linen functions: immobility, moisture management, and immobility and moisture management) were tested using the sweating guarded hot plate method, which allows for the measurement of the evaporative capacity (g H2O/m2*hour) and the total rate of heat withdrawal (Watts/m2) associated with nine different linen configurations placed on the support surface. Total heat withdrawal and evaporative capacity of the LAL surface with a fitted sheet only was used for comparison (P <0.05) Compared with fitted sheet only, heat withdrawal was significantly reduced by five of eight combinations, and evaporative moisture reduction was significantly reduced by six of eight linen combinations (P <0.05). All combinations that included plastic-containing underpads significantly reduced the surface's ability to dissipate heat and evaporate moisture, and use of the maximum number of layers (nine) reduced heat withdrawal to the level of a static, nonLAL surface. The results of this study suggest that putting additional linens or underpads on LAL surfaces may adversely affect skin temperature and moisture, thereby reducing the pressure ulcer prevention potential of these surfaces. Additional studies to examine the effect of linens and underpads as well as microclimate management strategies on pressure ulcer risk are needed.

  6. 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).

  7. 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

  8. 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

  9. 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

  10. 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 Central

    Yong, HH; Foong, K; Borland, R; Omar, M; Hamann, S; Sirirassamee, B; Fong, GT; Fotuhi, O; Hyland, A

    2015-01-01

    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% versus 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

  11. 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.

  12. Repurposing CRISPR/Cas9 for in situ functional assays.

    PubMed

    Malina, Abba; Mills, John R; Cencic, Regina; Yan, Yifei; Fraser, James; Schippers, Laura M; Paquet, Marilène; Dostie, Josée; Pelletier, Jerry

    2013-12-01

    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.

  13. 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.

  14. SD-CAS: Spin Dynamics by Computer Algebra System

    NASA Astrophysics Data System (ADS)

    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.

  15. 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.

  16. 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...

  17. CRISPR/Cas9 mediates efficient conditional mutagenesis in Drosophila.

    PubMed

    Xue, Zhaoyu; Wu, Menghua; Wen, Kejia; Ren, Menda; Long, Li; Zhang, Xuedi; Gao, Guanjun

    2014-11-01

    Existing transgenic RNA interference (RNAi) methods greatly facilitate functional genome studies via controlled silencing of targeted mRNA in Drosophila. Although the RNAi approach is extremely powerful, concerns still linger about its low efficiency. Here, we developed a CRISPR/Cas9-mediated conditional mutagenesis system by combining tissue-specific expression of Cas9 driven by the Gal4/upstream activating site system with various ubiquitously expressed guide RNA transgenes to effectively inactivate gene expression in a temporally and spatially controlled manner. Furthermore, by including multiple guide RNAs in a transgenic vector to target a single gene, we achieved a high degree of gene mutagenesis in specific tissues. The CRISPR/Cas9-mediated conditional mutagenesis system provides a simple and effective tool for gene function analysis, and complements the existing RNAi approach. PMID:25193494

  18. CRISPR/Cas9 Genome Editing in Embryonic Stem Cells.

    PubMed

    Andrey, Guillaume; Spielmann, Malte

    2017-01-01

    Targeted mutagenesis is required to evaluate the function of DNA segments across the genome. In recent years the CRISPR/Cas9 technology has been widely used for functional genome studies and is partially replacing classical homologous recombination methods in different aspects. CRISPR/Cas9-derived tools indeed allow the production of a wide-range of engineered mutations: from point mutations to large chromosomal rearrangements such as deletions, duplications and inversions. Here we present a protocol to engineer Embryonic Stem Cells (ESC) with desired mutations using transfection of custom-made CRISPR/Cas9 vectors. These methods allow the in vivo modeling of congenital mutations and the functional interrogation of DNA sequences. PMID:27662879

  19. Genome engineering using CRISPR-Cas9 system.

    PubMed

    Cong, Le; Zhang, Feng

    2015-01-01

    The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-Cas9 system is an adaptive immune system that exists in a variety of microbes. It could be engineered to function in eukaryotic cells as a fast, low-cost, efficient, and scalable tool for manipulating genomic sequences. In this chapter, detailed protocols are described for harnessing the CRISPR-Cas9 system from Streptococcus pyogenes to enable RNA-guided genome engineering applications in mammalian cells. We present all relevant methods including the initial site selection, molecular cloning, delivery of guide RNAs (gRNAs) and Cas9 into mammalian cells, verification of target cleavage, and assays for detecting genomic modification including indels and homologous recombination. These tools provide researchers with new instruments that accelerate both forward and reverse genetics efforts.

  20. CRISPR-Cas adaptation: insights into the mechanism of action.

    PubMed

    Amitai, Gil; Sorek, Rotem

    2016-02-01

    Since the first demonstration that CRISPR-Cas systems provide bacteria and archaea with adaptive immunity against phages and plasmids, numerous studies have yielded key insights into the molecular mechanisms governing how these systems attack and degrade foreign DNA. However, the molecular mechanisms underlying the adaptation stage, in which new immunological memory is formed, have until recently represented a major unresolved question. In this Progress article, we discuss recent discoveries that have shown both how foreign DNA is identified by the CRISPR-Cas adaptation machinery and the molecular basis for its integration into the chromosome to form an immunological memory. Furthermore, we describe the roles of each of the specific CRISPR-Cas components that are involved in memory formation, and consider current models for their evolutionary origin.

  1. Harnessing CRISPR-Cas systems for bacterial genome editing.

    PubMed

    Selle, Kurt; Barrangou, Rodolphe

    2015-04-01

    Manipulation of genomic sequences facilitates the identification and characterization of key genetic determinants in the investigation of biological processes. Genome editing via clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated (Cas) constitutes a next-generation method for programmable and high-throughput functional genomics. CRISPR-Cas systems are readily reprogrammed to induce sequence-specific DNA breaks at target loci, resulting in fixed mutations via host-dependent DNA repair mechanisms. Although bacterial genome editing is a relatively unexplored and underrepresented application of CRISPR-Cas systems, recent studies provide valuable insights for the widespread future implementation of this technology. This review summarizes recent progress in bacterial genome editing and identifies fundamental genetic and phenotypic outcomes of CRISPR targeting in bacteria, in the context of tool development, genome homeostasis, and DNA repair.

  2. Structure Principles of CRISPR-Cas Surveillance and Effector Complexes.

    PubMed

    Tsui, Tsz Kin Martin; Li, Hong

    2015-01-01

    The pathway of CRISPR-Cas immunity redefines the roles of RNA in the flow of genetic information and ignites excitement for next-generation gene therapy tools. CRISPR-Cas machineries offer a fascinating set of new enzyme assemblies from which one can learn principles of molecular interactions and chemical activities. The interference step of the CRISPR-Cas immunity pathway congregates proteins, RNA, and DNA into a single molecular entity that selectively destroys invading nucleic acids. Although much remains to be discovered, a picture of how the interference process takes place is emerging. This review focuses on the current structural data for the three known types of RNA-guided nucleic acid interference mechanisms. In it, we describe key features of individual complexes and we emphasize comparisons across types and along functional stages. We aim to provide readers with a set of core principles learned from the three types of interference complexes and a deep appreciation of the diversity among them.

  3. CRISPR-Cas9: A Revolutionary Tool for Cancer Modelling.

    PubMed

    Torres-Ruiz, Raul; Rodriguez-Perales, Sandra

    2015-09-14

    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.

  4. 41 CFR 102-33.130 - If we hire CAS, what are our management responsibilities?

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... responsible for— (a) Establishing agency-specific Flight Program Standards, as defined in §§ 102-33.140... your aircraft hired as CAS; and (d) Reporting the cost and usage data for your CAS hires (for...

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

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... responsible for— (a) Establishing agency-specific Flight Program Standards, as defined in §§ 102-33.140... your aircraft hired as CAS; and (d) Reporting the cost and usage data for your CAS hires (for...

  6. The use of heteroduplex analysis of polymerase chain reaction products to support the possible transmission of Legionella pneumophila from a malfunctioning automobile air conditioner.

    PubMed

    Pinar, Ahmet; Ramirez, Julio A; Schindler, Laura L; Miller, Richard D; Summersgill, James T

    2002-03-01

    Air conditioner condensates have not been previously associated with cases of Legionnaires' disease. We report the possible transmission of Legionella pneumophila serogroup 1 from a malfunctioning automobile air conditioning system's leaking water onto the floorboard of a car driven for a long distance by the patient. Heteroduplex analysis of polymerase chain reaction products was used to help establish an epidemiologic link between the water specimen and the patient. PMID:11918120

  7. 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

  8. 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

  9. 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

  10. Expanding the Biologist's Toolkit with CRISPR-Cas9.

    PubMed

    Sternberg, Samuel H; Doudna, Jennifer A

    2015-05-21

    Few discoveries transform a discipline overnight, but biologists today can manipulate cells in ways never possible before, thanks to a peculiar form of prokaryotic adaptive immunity mediated by clustered regularly interspaced short palindromic repeats (CRISPR). From elegant studies that deciphered how these immune systems function in bacteria, researchers quickly uncovered the technological potential of Cas9, an RNA-guided DNA cleaving enzyme, for genome engineering. Here we highlight the recent explosion in visionary applications of CRISPR-Cas9 that promises to usher in a new era of biological understanding and control.

  11. Applications of CRISPR-Cas systems in neuroscience.

    PubMed

    Heidenreich, Matthias; Zhang, Feng

    2016-01-01

    Genome-editing tools, and in particular those based on CRISPR-Cas (clustered regularly interspaced short palindromic repeat (CRISPR)-CRISPR-associated protein) systems, are accelerating the pace of biological research and enabling targeted genetic interrogation in almost 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 models 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.

  12. 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

  13. Applications of CRISPR-Cas9 mediated genome engineering.

    PubMed

    Yang, Xiao

    2015-01-01

    Targeted mutagenesis based on homologous recombination has been a powerful tool for understanding the mechanisms underlying development, normal physiology, and disease. A recent breakthrough in genome engineering technology based on the class of RNA-guided endonucleases, such as clustered regularly interspaced short palindromic repeats (CRISPR)-associated Cas9, is further revolutionizing biology and medical studies. The simplicity of the CRISPR-Cas9 system has enabled its widespread applications in generating germline animal models, somatic genome engineering, and functional genomic screening and in treating genetic and infectious diseases. This technology will likely be used in all fields of biomedicine, ranging from basic research to human gene therapy.

  14. 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

  15. 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

  16. 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.

  17. 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.

  18. 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

  19. VizieR Online Data Catalog: γ Cas radial velocity curve (Smith+, 2012)

    NASA Astrophysics Data System (ADS)

    Smith, M. A.; Lopes de Oliveira, R.; Motch, C.; Henry, G. W.; Richardson, N. D.; Bjorkman, K. S.; Stee, P.; Mourard, D.; Monnier, J. D.; Che, X.; Buecke, R.; Pollmann, E.; Gies, D. R.; Schaefer, G. H.; ten Brummelaar, T.; McAlister, H. A.; Turner, N. H.; Sturmann, J.; Sturmann, L.; Ridgway, S. T.

    2012-05-01

    Our LBOI observations were conducted in the infrared using Georgia State University's Center for High Angular Resolution Astronomy (CHARA) array facility on Mt. Wilson. CHARA is an interferometric array of six 1-m telescopes configured in a Y-shaped pattern with baselines of up to 330m for maximum spatial resolution. Our observations utilized two new-generation instruments for imaging of γ Cas's decretion disk: the Michigan Infrared Combiner (MIRC) in the H-band (1.64um) and the Visible spEctroGraph and polArimeter (VEGA), which became operational in 2009. Our optical continuum observations of γ Cas were made in the standard Johnson V and B band filters on every available photometric night (sometimes several times per night) using the 0.4m T3 Automated Photometric Telescope (APT) located at Fairborn Observatory in the Patagonia mountains in southern Arizona. To support this program we made use of 306 echelle spectra of γ Cas taken in a long-term monitoring program with the spectrograph fiber-fed from the Cassegrain focus of the 1.06-m telescope at Ritter Observatory (administered by the University of Toledo) in 2009 December and the second half of 2010. We conducted four XMM-Newton observations under the auspices of Guest Investigator cycle A09. These were carried out on 2010 July 7 (ObsID 0651670201; referred to here as OBS1), July 24 (0651670301; OBS2), August 2 (0651670401; OBS3), and August 24 (0651670501; OBS4), beginning on MJD dates 55384.30, 55401.15, 55410.77, and 55428.09, respectively. (1 data file).

  20. 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

  1. 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

  2. How Do Traditional Examination Questions Fare in the Presence of a Computer Algebra System (CAS)?

    ERIC Educational Resources Information Center

    Malabar, Ian; Pountney, Dave

    2001-01-01

    Describes the outcomes and discusses possible implications for the development of assessment with a Computer Algebra System (CAS) when a group of undergraduate mathematics students, familiar with using a CAS in examinations, tackled an assortment of traditional (i.e., non-CAS type) questions. (Author/MM)

  3. 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...

  4. 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...

  5. 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...

  6. 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...

  7. 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...

  8. 76 FR 40817 - Cost Accounting Standards: Change to the CAS Applicability Threshold for the Inflation Adjustment...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-07-12

    ... to the CAS Applicability Threshold for the Inflation Adjustment to the Truth in Negotiations Act... revising the threshold for the application of CAS from ``$650,000'' to ``the Truth in Negotiations Act... are open for comment. Simply type ``CAS-TINA Threshold'' (without quotation marks) in the Comment...

  9. 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…

  10. 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…

  11. 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…

  12. 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

  13. An Overview of the Summer 2014 Airborne Study of Oil Sands Air Pollutants in Support of the Joint Oil Sands Monitoring Plan

    NASA Astrophysics Data System (ADS)

    Li, S. M.; Hayden, K. L.; Cober, S.; Wolde, M.; Liggio, J.; Liu, P.; Leithead, A.; O'brien, J.; Wang, D. K.; Moussa, S. G.; Gordon, M.; Darlington, A. L.; McLaren, R.; Makar, P.; Stroud, C.; Wentzell, J. J. B.; Brook, J.; Narayan, J.; Elford, A.; Sung, K.; Sheppard, A.

    2014-12-01

    A short term airborne study of oil sands air pollutants was carried out in August and September 2013. The study had three objectives: 1. to validate emissions of criteria air contaminants (CACs) and other air pollutants from surface mining facilities in the Athabasca oil sands region, using airborne ambient air measurements; 2. to understand the transport and transformation of primary pollutants; and 3. to provide data for model and satellite retrieval validation. The data will be used for the evaluation and improvement of high-resolution air quality models for eventual application in determining the fates of these pollutants and their deposition to the downwind ecosystems. Various chemical species were measured from the National Research Council of Canada Convair-580 aircraft at high time resolution of 1-10 seconds. A total of 22 flights were flown, with 14 flights dedicated to emission validation, 5 flights to transport and transformation of oil sands pollutants, and 5 flights to satellite data validation. An algorithm will be shown to demonstrate how the data from the emission flights can be used to derive a top-down estimate of SO2 emission rates.

  14. 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.

  15. CRISPR-Cas9 Genome Editing in Drosophila.

    PubMed

    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 (NHEJ) 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. 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…

  17. 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…

  18. 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 SUPPLEMENTARY REGULATIONS DOE MANAGEMENT AND OPERATING CONTRACTS Cost Accounting Standards Administration...

  19. Cas9-based genome editing in Arabidopsis and tobacco.

    PubMed

    Li, Jian-Feng; Zhang, Dandan; Sheen, Jen

    2014-01-01

    Targeted modification of plant genome is key to elucidating and manipulating gene functions in plant research and biotechnology. The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) technology is emerging as a powerful genome-editing method in diverse plants that traditionally lacked facile and versatile tools for targeted genetic engineering. This technology utilizes easily reprogrammable guide RNAs (sgRNAs) to direct Streptococcus pyogenes Cas9 endonuclease to generate DNA double-stranded breaks in targeted genome sequences, which facilitates efficient mutagenesis by error-prone nonhomologous end-joining (NHEJ) or sequence replacement by homology-directed repair (HDR). In this chapter, we describe the procedure to design and evaluate dual sgRNAs for plant codon-optimized Cas9-mediated genome editing using mesophyll protoplasts as model cell systems in Arabidopsis thaliana and Nicotiana benthamiana. We also discuss future directions in sgRNA/Cas9 applications for generating targeted genome modifications and gene regulations in plants.

  20. 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

  1. 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…

  2. 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…

  3. Toward Whole-Transcriptome Editing with CRISPR-Cas9.

    PubMed

    Heckl, Dirk; Charpentier, Emmanuelle

    2015-05-21

    Targeted regulation of gene expression holds huge promise for biomedical research. In a series of recent publications (Gilbert et al., 2014; Konermann et al., 2015; Zalatan et al., 2015), sophisticated, multiplex-compatible transcriptional activator systems based on the CRISPR-Cas9 technology and genome-scale libraries advance the field toward whole-transcriptome control. PMID:26000839

  4. 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.

  5. 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 SUPPLEMENTARY REGULATIONS DOE MANAGEMENT AND OPERATING CONTRACTS Cost Accounting Standards Administration...

  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. 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.

  8. Medical marijuana: CAS releases report, government cuts research funding.

    PubMed

    Betteridge, Glenn

    2006-12-01

    In June 2006, the Canadian AIDS Society (CAS) released a comprehensive report with recommendations to overcome barriers to the use of cannabis for medical purposes faced by people living with HIV/AIDS in Canada. On 25 September 2006, as part of package of spending cuts, the federal government announced plans to eliminate its marijuana medical research program. PMID:17373064

  9. 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…

  10. Les tuberculomes intracraniens: à propos de 125 cas

    PubMed Central

    Moufid, Faycal; Oulali, Noureddine; El Fatemi, Nizare; Gana, Rachid; Maaqili, Rachid; Bellakhdar, Fouad

    2012-01-01

    Les tuberculomes intracrâniens représentent l'une des localisations les plus graves de la tuberculose, leur incidence varie en fonction du contexte représentant 0,2% des processus intracrâniens dans les pays occidentaux et 5 à 10% des masses intracrâniennes dans les pays en voie de développement. Nous rapportons une étude rétrospective de 125 cas. L'hypertension intracrânienne (45%) et le déficit neurologique (36%) sont les signes cliniques les plus fréquents. La lésion était localisée dans 60% des cas en sus-tentoriel et dans 40% des cas en sous-tentoriel. L'approche thérapeutique a consisté en un abord direct du tuberculome dans 67 cas (53%), une biopsie stéréotaxique dans 32 cas (25%), le traitement médical en première intention sans confirmation histologique dans 26 cas (20%). Avant 1993 notre service ne disposait pas de cadre de stéréotaxie, notre attitude thérapeutique consistait soit en un abord direct de la lésion dans 70% des cas, soit un traitement antituberculeux en première intention sans confirmation histologique (30%). Cette attitude était corrélée à une mortalité et morbidité non négligeables respectivement 3% et 10%. Après 1993; le taux d'abords direct a chuté a 38%, avec 47% de biopsies stéréotaxiques et seulement 13% des patients traités par antibacillaires sans preuve histologique. Ceci s'est accompagné d'une réduction significative de mortalité a 1,4% (p = 0,0003) et de morbidité a 2% (p = 0,0027). PMID:22937196

  11. 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

  12. 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 ...

  13. CRISPR/Cas9-mediated targeted mutagenesis in Nicotiana tabacum.

    PubMed

    Gao, Junping; Wang, Genhong; Ma, Sanyuan; Xie, Xiaodong; Wu, Xiangwei; Zhang, Xingtan; Wu, Yuqian; Zhao, Ping; Xia, Qingyou

    2015-01-01

    Genome editing is one of the most powerful tools for revealing gene function and improving crop plants. Recently, RNA-guided genome editing using the type II clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein (Cas) system has been used as a powerful and efficient tool for genome editing in various organisms. Here, we report genome editing in tobacco (Nicotiana tabacum) mediated by the CRISPR/Cas9 system. Two genes, NtPDS and NtPDR6, were used for targeted mutagenesis. First, we examined the transient genome editing activity of this system in tobacco protoplasts, insertion and deletion (indel) mutations were observed with frequencies of 16.2-20.3% after transfecting guide RNA (gRNA) and the nuclease Cas9 in tobacco protoplasts. The two genes were also mutated using multiplexing gRNA at a time. Additionally, targeted deletions and inversions of a 1.8-kb fragment between two target sites in the NtPDS locus were demonstrated, while indel mutations were also detected at both the sites. Second, we obtained transgenic tobacco plants with NtPDS and NtPDR6 mutations induced by Cas9/gRNA. The mutation percentage was 81.8% for NtPDS gRNA4 and 87.5% for NtPDR6 gRNA2. Obvious phenotypes were observed, etiolated leaves for the psd mutant and more branches for the pdr6 mutant, indicating that highly efficient biallelic mutations occurred in both transgenic lines. No significant off-target mutations were obtained. Our results show that the CRISPR/Cas9 system is a useful tool for targeted mutagenesis of the tobacco genome.

  14. 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

  15. 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.

  16. Primary processing of CRISPR RNA by the endonuclease Cas6 in Staphylococcus epidermidis.

    PubMed

    Wakefield, Noelle; Rajan, Rakhi; Sontheimer, Erik J

    2015-10-01

    In many bacteria and archaea, an adaptive immune system (CRISPR-Cas) provides immunity against foreign genetic elements. This system uses CRISPR RNAs (crRNAs) derived from the CRISPR array, along with CRISPR-associated (Cas) proteins, to target foreign nucleic acids. In most CRISPR systems, endonucleolytic processing of crRNA precursors (pre-crRNAs) is essential for the pathway. Here we study the Cas6 endonuclease responsible for crRNA processing in the Type III-A CRISPR-Cas system from Staphylococcus epidermidis RP62a, a model for Type III-A CRISPR-Cas systems, and define substrate requirements for SeCas6 activity. We find that SeCas6 is necessary and sufficient for full-length crRNA biogenesis in vitro, and that it relies on both sequence and stem-loop structure in the 3' half of the CRISPR repeat for recognition and processing.

  17. Structures of Cas9 Endonucleases Reveal RNA-Mediated Conformational Activation

    PubMed Central

    Jinek, Martin; Jiang, Fuguo; Taylor, David W.; Sternberg, Samuel H.; Kaya, Emine; Ma, Enbo; Anders, Carolin; Hauer, Michael; Zhou, Kaihong; Lin, Steven; Kaplan, Matias; Iavarone, Anthony T.; Charpentier, Emmanuelle; Nogales, Eva; Doudna, Jennifer A.

    2014-01-01

    Type II CRISPR (clustered regularly interspaced short palindromic repeats)–Cas (CRISPR-associated) systems use an RNA-guided DNA endonuclease, Cas9, to generate double-strand breaks in invasive DNA during an adaptive bacterial immune response. Cas9 has been harnessed as a powerful tool for genome editing and gene regulation in many eukaryotic organisms. We report 2.6 and 2.2 angstrom resolution crystal structures of two major Cas9 enzyme subtypes, revealing the structural core shared by all Cas9 family members. The architectures of Cas9 enzymes define nucleic acid binding clefts, and single-particle electron microscopy reconstructions show that the two structural lobes harboring these clefts undergo guide RNA–induced reorientation to form a central channel where DNA substrates are bound. The observation that extensive structural rearrangements occur before target DNA duplex binding implicates guide RNA loading as a key step in Cas9 activation. PMID:24505130

  18. CRISPR/Cas9 for genome editing: progress, implications and challenges.

    PubMed

    Zhang, Feng; Wen, Yan; Guo, Xiong

    2014-09-15

    Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) protein 9 system provides a robust and multiplexable genome editing tool, enabling researchers to precisely manipulate specific genomic elements, and facilitating the elucidation of target gene function in biology and diseases. CRISPR/Cas9 comprises of a nonspecific Cas9 nuclease and a set of programmable sequence-specific CRISPR RNA (crRNA), which can guide Cas9 to cleave DNA and generate double-strand breaks at target sites. Subsequent cellular DNA repair process leads to desired insertions, deletions or substitutions at target sites. The specificity of CRISPR/Cas9-mediated DNA cleavage requires target sequences matching crRNA and a protospacer adjacent motif locating at downstream of target sequences. Here, we review the molecular mechanism, applications and challenges of CRISPR/Cas9-mediated genome editing and clinical therapeutic potential of CRISPR/Cas9 in future.

  19. A simplified and efficient germline-specific CRISPR/Cas9 system for Drosophila genomic engineering.

    PubMed

    Sebo, Zachary L; Lee, Han B; Peng, Ying; Guo, Yi

    2014-01-01

    The type II CRISPR/Cas9 system (clustered regularly interspaced short palindromic repeats/CRISPR-associated) has recently emerged as an efficient and simple tool for site-specific engineering of eukaryotic genomes. To improve its applications in Drosophila genome engineering, we simplified the standard two-component CRISPR/Cas9 system by generating a stable transgenic fly line expressing the Cas9 endonuclease in the germline (Vasa-Cas9 line). By injecting vectors expressing engineered target-specific guide RNAs into Vasa-Cas9 fly embryos, mutations were generated from site-specific DNA cleavages and efficiently transmitted into progenies. Because Cas9 endonuclease is the universal component of the type II CRISPR/Cas9 system, site-specific genomic engineering based on this improved platform can be achieved with lower complexity and toxicity, greater consistency, and excellent versatility.

  20. CRISPR/Cas9 Platforms for Genome Editing in Plants: Developments and Applications.

    PubMed

    Ma, Xingliang; Zhu, Qinlong; Chen, Yuanling; Liu, Yao-Guang

    2016-07-01

    The clustered regularly interspaced short palindromic repeat (CRISPR)-associated protein9 (Cas9) genome editing system (CRISPR/Cas9) is adapted from the prokaryotic type II adaptive immunity system. The CRISPR/Cas9 tool surpasses other programmable nucleases, such as ZFNs and TALENs, for its simplicity and high efficiency. Various plant-specific CRISPR/Cas9 vector systems have been established for adaption of this technology to many plant species. In this review, we present an overview of current advances on applications of this technology in plants, emphasizing general considerations for establishment of CRISPR/Cas9 vector platforms, strategies for multiplex editing, methods for analyzing the induced mutations, factors affecting editing efficiency and specificity, and features of the induced mutations and applications of the CRISPR/Cas9 system in plants. In addition, we provide a perspective on the challenges of CRISPR/Cas9 technology and its significance for basic plant research and crop genetic improvement.

  1. 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

  2. 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.

  3. 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

  4. 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.

  5. 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

  6. 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

  7. 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

  8. Insights into the CRISPR/Cas system of Gardnerella vaginalis

    PubMed Central

    2012-01-01

    Background Gardnerella vaginalis is identified as the predominant colonist of the vaginal tracts of women diagnosed with bacterial vaginosis (BV). G. vaginalis can be isolated from healthy women, and an asymptomatic BV state is also recognised. The association of G. vaginalis with different clinical phenotypes could be explained by different cytotoxicity of the strains, presumably based on disparate gene content. The contribution of horizontal gene transfer to shaping the genomes of G. vaginalis is acknowledged. The CRISPR loci of the recently discovered CRISPR/Cas microbial defence system provide a historical view of the exposure of prokaryotes to a variety of foreign genetic elements. Results The CRISPR/Cas loci were analysed using available sequence data from three G. vaginalis complete genomes and 18 G. vaginalis draft genomes in the NCBI database, as well as PCR amplicons of the genomic DNA of 17 clinical isolates. The cas genes in the CRISPR/Cas loci of G. vaginalis belong to the E. coli subtype. Approximately 20% of the spacers had matches in the GenBank database. Sequence analysis of the CRISPR arrays revealed that nearly half of the spacers matched G. vaginalis chromosomal sequences. The spacers that matched G. vaginalis chromosomal sequences were determined to not be self-targeting and were presumably neither constituents of mobile-element-associated genes nor derived from plasmids/viruses. The protospacers targeted by these spacers displayed conserved protospacer-adjacent motifs. Conclusions The CRISPR/Cas system has been identified in about one half of the analysed G. vaginalis strains. Our analysis of CRISPR sequences did not reveal a potential link between their presence and the virulence of the G. vaginalis strains. Based on the origins of the spacers found in the G. vaginalis CRISPR arrays, we hypothesise that the transfer of genetic material among G. vaginalis strains could be regulated by the CRISPR/Cas mechanism. The present study is the first

  9. 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

  10. 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

  11. 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

  12. 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

  13. 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

  14. High-fidelity CRISPR-Cas9 variants with undetectable genome-wide off-targets

    PubMed Central

    Kleinstiver, Benjamin P.; Pattanayak, Vikram; Prew, Michelle S.; Tsai, Shengdar Q.; Nguyen, Nhu; Zheng, Zongli; Joung, J. Keith

    2015-01-01

    CRISPR-Cas9 nucleases are widely used for genome editing but can induce unwanted off-target mutations. Existing strategies for reducing genome-wide off-targets of the broadly used Streptococcus pyogenes Cas9 (SpCas9) are imperfect, possessing only partial or unproven efficacies and other limitations that constrain their use. Here we describe SpCas9-HF1, a high-fidelity variant harboring alterations designed to reduce non-specific DNA contacts. SpCas9-HF1 retains on-target activities comparable to wild-type SpCas9 with >85% of single-guide RNAs (sgRNAs) tested in human cells. Strikingly, with sgRNAs targeted to standard non-repetitive sequences, SpCas9-HF1 rendered all or nearly all off-target events undetectable by genome-wide break capture and targeted sequencing methods. Even for atypical, repetitive target sites, the vast majority of off-targets induced by SpCas9-HF1 were not detected. With its exceptional precision, SpCas9-HF1 provides an alternative to wild-type SpCas9 for research and therapeutic applications. More broadly, our results suggest a general strategy for optimizing genome-wide specificities of other RNA-guided nucleases. PMID:26735016

  15. 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

  16. 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

  17. 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.

  18. 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.

  19. Adapting CRISPR/Cas9 for functional genomics screens.

    PubMed

    Malina, Abba; Katigbak, Alexandra; Cencic, Regina; Maïga, Rayelle Itoua; Robert, Francis; Miura, Hisashi; Pelletier, Jerry

    2014-01-01

    The use of CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR-associated protein) for targeted genome editing has been widely adopted and is considered a "game changing" technology. The ease and rapidity by which this approach can be used to modify endogenous loci in a wide spectrum of cell types and organisms makes it a powerful tool for customizable genetic modifications as well as for large-scale functional genomics. The development of retrovirus-based expression platforms to simultaneously deliver the Cas9 nuclease and single guide (sg) RNAs provides unique opportunities by which to ensure stable and reproducible expression of the editing tools and a broad cell targeting spectrum, while remaining compatible with in vivo genetic screens. Here, we describe methods and highlight considerations for designing and generating sgRNA libraries in all-in-one retroviral vectors for such applications.

  20. 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.

  1. 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.

  2. 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.

  3. 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

  4. 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

  5. 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

  6. CRISPR/Cas-mediated targeted mutagenesis in Daphnia magna.

    PubMed

    Nakanishi, Takashi; Kato, Yasuhiko; Matsuura, Tomoaki; Watanabe, Hajime

    2014-01-01

    The water flea Daphnia magna has been used as an animal model in ecology, evolution, and environmental sciences. Thanks to the recent progress in Daphnia genomics, genetic information such as the draft genome sequence and expressed sequence tags (ESTs) is now available. To investigate the relationship between phenotypes and the available genetic information about Daphnia, some gene manipulation methods have been developed. However, a technique to induce targeted mutagenesis into Daphnia genome remains elusive. To overcome this problem, we focused on an emerging genome editing technique mediated by the clustered regularly interspaced short palindromic repeats/CRISPR-associated (CRISPR/Cas) system to introduce genomic mutations. In this study, we targeted a functionally conserved regulator of eye development, the eyeless gene in D. magna. When we injected Cas9 mRNAs and eyeless-targeting guide RNAs into eggs, 18-47% of the survived juveniles exhibited abnormal eye morphology. After maturation, up to 8.2% of the adults produced progenies with deformed eyes, which carried mutations in the eyeless loci. These results showed that CRISPR/Cas system could introduce heritable mutations into the endogenous eyeless gene in D. magna. This is the first report of a targeted gene knockout technique in Daphnia and will be useful in uncovering Daphnia gene functions.

  7. 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

  8. CRISPR-Cas systems for editing, regulating and targeting genomes.

    PubMed

    Sander, Jeffry D; Joung, J Keith

    2014-04-01

    Targeted genome editing using engineered nucleases has rapidly gone from being 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 approach for generating RNA-guided nucleases, such as Cas9, with customizable specificities. Genome editing mediated by these nucleases has been used to rapidly, easily and efficiently modify endogenous genes in a wide variety of biomedically important cell types and in 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 power 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.

  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 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

  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

    Liu, Hai-Ying; Skjetne, Erik; Kobernus, Mike

    2013-11-04

    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.

  11. 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 ...

  12. Integration and exchange of split dCas9 domains for transcriptional controls in mammalian cells

    PubMed Central

    Ma, Dacheng; Peng, Shuguang; Xie, Zhen

    2016-01-01

    Programmable and precise regulation of dCas9 functions in response to multiple molecular signals by using synthetic gene circuits will expand the application of the CRISPR-Cas technology. However, the application of CRISPR-Cas therapeutic circuits is still challenging due to the restrictive cargo size of existing viral delivery vehicles. Here, we construct logic AND circuits by integrating multiple split dCas9 domains, which is useful to reduce the size of synthetic circuits. In addition, we engineer sensory switches by exchanging split dCas9 domains, allowing differential regulations on one gene, or activating two different genes in response to cell-type specific microRNAs. Therefore, we provide a valuable split-dCas9 toolkit to engineer complex transcription controls, which may inspire new biomedical applications. PMID:27694915

  13. 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

  14. 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

  15. Direct CRISPR spacer acquisition from RNA by a natural reverse-transcriptase-Cas1 fusion protein

    PubMed Central

    Sidote, David J.; Markham, Laura M.; Sanchez-Amat, Antonio; Bhaya, Devaki; Lambowitz, Alan M.; Fire, Andrew Z.

    2016-01-01

    CRISPR (Clustered Regularly Interspaced Short Palindromic Repeat) 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 show that an RT-Cas1 fusion enables the acquisition of RNA spacers in vivo in an RT-dependent manner. In vitro, the MMB-1 RT-Cas1 and Cas2 proteins catalyze ligation of RNA segments into the CRISPR array, followed by reverse transcription. These observations outline a host-mediated mechanism for reverse information flow from RNA to DNA. PMID:26917774

  16. 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

  17. [Application of CRISPR/Cas9 mediated genome editing in farm animals].

    PubMed

    Yuyun, Xing; Qiang, Yang; Jun, Ren

    2016-03-01

    CRISPR (Clustered regularly interspaced short palindromic repeats)/Cas (CRISPR associated proteins) is an acquired immune system found in bacteria and archaea that fight against invasion of viruses or plasmids. CRISPR/Cas systems are currently classified into three main types: I, II and III, of which type II has relatively simple components. The CRISPR/Cas9 technology modified from type II CRISPR/Cas system has been developed as an efficient genome editing tool. Since the initial application of the CRISPR/Cas9 technology in mammals in 2013, the reports of this system for genomic editing has skyrocketed. Farm animals are not only economically important animals, but also ideal animal models for human diseases and biomedical studies. In this review, we summarize the applications of CRISPR/Cas9 in farm animals, briefly describe the off-target effects and the main solutions, and finally highlight the future perspectives of this technology.

  18. 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.

  19. 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.

  20. 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

  1. 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.

  2. 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

  3. Successful transient expression of Cas9 and single guide RNA genes in Chlamydomonas reinhardtii.

    PubMed

    Jiang, Wenzhi; Brueggeman, Andrew J; Horken, Kempton M; Plucinak, Thomas M; Weeks, Donald P

    2014-11-01

    The clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 system has become a powerful and precise tool for targeted gene modification (e.g., gene knockout and gene replacement) in numerous eukaryotic organisms. Initial attempts to apply this technology to a model, the single-cell alga, Chlamydomonas reinhardtii, failed to yield cells containing edited genes. To determine if the Cas9 and single guide RNA (sgRNA) genes were functional in C. reinhardtii, we tested the ability of a codon-optimized Cas9 gene along with one of four different sgRNAs to cause targeted gene disruption during a 24-h period immediately following transformation. All three exogenously supplied gene targets as well as the endogenous FKB12 (rapamycin sensitivity) gene of C. reinhardtii displayed distinct Cas9/sgRNA-mediated target site modifications as determined by DNA sequencing of cloned PCR amplicons of the target site region. Success in transient expression of Cas9 and sgRNA genes contrasted with the recovery of only a single rapamycin-resistant colony bearing an appropriately modified FKB12 target site in 16 independent transformation experiments involving >10(9) cells. Failure to recover transformants with intact or expressed Cas9 genes following transformation with the Cas9 gene alone (or even with a gene encoding a Cas9 lacking nuclease activity) provided strong suggestive evidence for Cas9 toxicity when Cas9 is produced constitutively in C. reinhardtii. The present results provide compelling evidence that Cas9 and sgRNA genes function properly in C. reinhardtii to cause targeted gene modifications and point to the need for a focus on development of methods to properly stem Cas9 production and/or activity following gene editing.

  4. 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

  5. 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

  6. Air Pollution Primer. Revised Education.

    ERIC Educational Resources Information Center

    Corman, Rena

    This revised and updated book is written to inform the citizens on the nature, causes, and effects of air pollution. It is written in terms familiar to the layman with the purpose of providing knowledge and motivation to spur community action on clean air policies. Numerous charts and drawings are provided to support discussion of air pollution…

  7. 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.

  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. 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

  10. 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

  11. 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

  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. 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

    2016-03-01

    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.

  14. 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

    2015-01-01

    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

  15. 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

  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

    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.

  18. Tribal Air Quality Monitoring.

    ERIC Educational Resources Information Center

    Wall, Dennis

    2001-01-01

    The Institute for Tribal Environmental Professionals (ITEP) (Flagstaff, Arizona) provides training and support for tribal professionals in the technical job skills needed for air quality monitoring and other environmental management tasks. ITEP also arranges internships, job placements, and hands-on training opportunities and supports an…

  19. MacSyFinder: A Program to Mine Genomes for Molecular Systems with an Application to CRISPR-Cas Systems

    PubMed Central

    Abby, Sophie S.; Néron, Bertrand; Ménager, Hervé; Touchon, Marie; Rocha, Eduardo P. C.

    2014-01-01

    Motivation Biologists often wish to use their knowledge on a few experimental models of a given molecular system to identify homologs in genomic data. We developed a generic tool for this purpose. Results Macromolecular System Finder (MacSyFinder) provides a flexible framework to model the properties of molecular systems (cellular machinery or pathway) including their components, evolutionary associations with other systems and genetic architecture. Modelled features also include functional analogs, and the multiple uses of a same component by different systems. Models are used to search for molecular systems in complete genomes or in unstructured data like metagenomes. The components of the systems are searched by sequence similarity using Hidden Markov model (HMM) protein profiles. The assignment of hits to a given system is decided based on compliance with the content and organization of the system model. A graphical interface, MacSyView, facilitates the analysis of the results by showing overviews of component content and genomic context. To exemplify the use of MacSyFinder we built models to detect and class CRISPR-Cas systems following a previously established classification. We show that MacSyFinder allows to easily define an accurate “Cas-finder” using publicly available protein profiles. Availability and Implementation MacSyFinder is a standalone application implemented in Python. It requires Python 2.7, Hmmer and makeblastdb (version 2.2.28 or higher). It is freely available with its source code under a GPLv3 license at https://github.com/gem-pasteur/macsyfinder. It is compatible with all platforms supporting Python and Hmmer/makeblastdb. The “Cas-finder” (models and HMM profiles) is distributed as a compressed tarball archive as Supporting Information. PMID:25330359

  20. Air quality management in Mexico.

    PubMed

    Fernández-Bremauntz, Adrián

    2008-01-01

    Several significant program and policy measures have been implemented in Mexico over the past 15 yr to improve air quality. This article provides an overview of air quality management strategies in Mexico, including (1) policy initiatives such as vehicle use restrictions, air quality standards, vehicle emissions, and fuel quality standards, and (2) supporting programs including establishment of a national emission inventory, an air pollution episodes program, and the implementation of exposure and health effects studies. Trends in air pollution episodes and ambient air pollutant concentrations are described.

  1. Fibromatose desmoïde du sein: à propos d'un cas et une revue de la literature

    PubMed Central

    Amourak, Sarah; Alaoui, Fatimazahra Fdili; Jayi, Sofia; Chaara, Hikmat; Melhouf, Moulay Abdelilah

    2015-01-01

    La fibromatose desmoïde mammaire est une entité rare, mimant sur le plan clinique et échographique un cancer du sein. Seule l'histologie apportera le diagnostic en objectivant une prolifération de cellule fusiforme (fibro et myofibroblastique sans atypies nucléaires), agencée en faisceaux, mêlés à des bandes de collagène, sans composante épithéliale. Le diagnostic différentiel se pose essentiellement avec le carcinome métaplasique à cellules fusiformes. L’évolution est strictement locale, avec un grand pouvoir récidivant. L'exérèse chirurgicale complète avec des marges saines (jusqu’à 3cm) est le traitement de choix, la radiothérapie reste une option thérapeutique en complément de la chirurgie dans les exérèses incomplètes et en cas de récidives multiples. A travers notre cas et une revue de la littérature, nous essayerons de mettre le point sur le diagnostic de cette entité rare et de sa prise en charge puisqu'elle va conditionner le pronostic. PMID:26491531

  2. Aspects epidemiologiques, cliniques et therapeutiques des otites externes: à propos de 801 cas

    PubMed Central

    Bathokedeou, Amana; Essobozou, Pegbessou; Akouda, Patassi; Essohanam, Boko; Eyawelohn, Kpémissi

    2014-01-01

    L'objectif de ce travail etait de déterminer l’épidémiologie, la clinique et la thérapeutique des otites externes (OE). Il s'agissait d'une étude rétrospective d'une année menée du 1er janvier au 31 décembre 2006 dans le service ORL du CHU-Tokoin. Huit cent un cas d'otite externe (OE) soit 11,9% des consultations étaient recensés. Le sexe féminin représentait 476 cas (59,42%). Le sex ratio était de 0,68. L’âge moyen des patients était de 25,4 ans avec des extrêmes de 05 mois et 81 ans. La tranche d’âge de 0-15 ans était la plus fréquente avec 360 cas (45%). L'allergie dans 74 cas (60,66%), la lésion de grattage dans 24 cas (19,67%), les corps étrangers du conduit auditif externe dans 18 cas (14,75%) et la natation dans 6 cas (4,92%) étaient les facteurs favorisants. L'otalgie dans 638 cas (79,65%) était le symptôme le plus fréquent. Les différentes formes cliniques des otites externes se répartissaient comme suit: otite externe diffuse dans 612 cas (76,40%), furoncle du CAE 126 dans cas (15,73%), otomycose dans 58 cas (7,24%), zona du conduit auditif externe dans 3 cas (0,37%) et otite externe nécrosante dans 2 cas (0,25%). Les gouttes auriculaires étaient administrées à tous les patients. L’évolution avait noté 799 patients (99,75%) guéris sans complication, un cas de décès et un cas de guérison avec séquelle. Traitée correctement, l'otite externe guérit sans complication. Son éviction passe par une sensibilisation des populations sur l'entretien du conduit auditif externe. PMID:25379111

  3. Utilization of the CRISPR/Cas9 system for the efficient production of mutant mice using crRNA/tracrRNA with Cas9 nickase and FokI-dCas9

    PubMed Central

    Terao, Miho; Tamano, Moe; Hara, Satoshi; Kato, Tomoko; Kinoshita, Masato; Takada, Shuji

    2016-01-01

    The CRISPR/Cas9 system is a powerful genome editing tool for the production of genetically modified animals. To produce mutant mice, chimeric single-guide RNA (sgRNA) is cloned in a plasmid vector and a mixture of sgRNA and Cas9 are microinjected into the fertilized eggs. An issue associated with gene manipulation using the CRISPR/Cas9 system is that there can be off-target effects. To simplify the production of mutant mice with low risks of off-target effects caused by the CRISPR/Cas9 system, we demonstrated that genetically modified mice can be efficiently obtained using chemically synthesized CRISPR RNA (crRNA), trans-activating crRNA (tracrRNA), and modified Cas9s, such as the nickase version and FokI-fused catalytically inactive Cas9, by microinjection into fertilized eggs. Using this method, it is no longer necessary to clone sgRNA into a plasmid vector, and this enables high-throughput production of mutant mice. PMID:26972821

  4. The RNA- and DNA-targeting CRISPR–Cas immune systems of Pyrococcus furiosus

    PubMed Central

    Terns, Rebecca M.; Terns, Michael P.

    2014-01-01

    Using the hyperthermophile Pyrococcus furiosus, we have delineated several key steps in CRISPR (clustered regularly interspaced short palindromic repeats)–Cas (CRISPR-associated) invader defence pathways. P. furiosus has seven transcriptionally active CRISPR loci that together encode a total of 200 crRNAs (CRISPR RNAs). The 27 Cas proteins in this organism represent three distinct pathways and are primarily encoded in two large gene clusters. The Cas6 protein dices CRISPR locus transcripts to generate individual invader-targeting crRNAs. The mature crRNAs include a signature sequence element (the 5′ tag) derived from the CRISPR locus repeat sequence that is important for function. crRNAs are tailored into distinct species and integrated into three distinct crRNA–Cas protein complexes that are all candidate effector complexes. The complex formed by the Cmr [Cas module RAMP (repeat-associated mysterious proteins)] (subtype III-B) proteins cleaves complementary target RNAs and can be programmed to cleave novel target RNAs in a prokaryotic RNAi-like manner. Evidence suggests that the other two CRISPR–Cas systems in P. furiosus, Csa (Cas subtype Apern) (subtype I-A) and Cst (Cas subtype Tneap) (subtype I-B), target invaders at the DNA level. Studies of the CRISPR–Cas systems from P. furiosus are yielding fundamental knowledge of mechanisms of crRNA biogenesis and silencing for three of the diverse CRISPR–Cas pathways, and reveal that organisms such as P. furiosus possess an arsenal of multiple RNA-guided mechanisms to resist diverse invaders. Our knowledge of the fascinating CRISPR–Cas pathways is leading in turn to our ability to co-opt these systems for exciting new biomedical and biotechnological applications. PMID:24256230

  5. CRISPR-Cas: evolution of an RNA-based adaptive immunity system in prokaryotes.

    PubMed

    Koonin, Eugene V; Makarova, Kira S

    2013-05-01

    The CRISPR-Cas (clustered regularly interspaced short palindromic repeats, CRISPR-associated genes) is an adaptive immunity system in bacteria and archaea that functions via a distinct self-non-self recognition mechanism that is partially analogous to the mechanism of eukaryotic RNA interference (RNAi). The CRISPR-Cas system incorporates fragments of virus or plasmid DNA into the CRISPR repeat cassettes and employs the processed transcripts of these spacers as guide RNAs to cleave the cognate foreign DNA or RNA. The Cas proteins, however, are not homologous to the proteins involved in RNAi and comprise numerous, highly diverged families. The majority of the Cas proteins contain diverse variants of the RNA recognition motif (RRM), a widespread RNA-binding domain. Despite the fast evolution that is typical of the cas genes, the presence of diverse versions of the RRM in most Cas proteins provides for a simple scenario for the evolution of the three distinct types of CRISPR-cas systems. In addition to several proteins that are directly implicated in the immune response, the cas genes encode a variety of proteins that are homologous to prokaryotic toxins that typically possess nuclease activity. The predicted toxins associated with CRISPR-Cas systems include the essential Cas2 protein, proteins of COG1517 that, in addition to a ligand-binding domain and a helix-turn-helix domain, typically contain different nuclease domains and several other predicted nucleases. The tight association of the CRISPR-Cas immunity systems with predicted toxins that, upon activation, would induce dormancy or cell death suggests that adaptive immunity and dormancy/suicide response are functionally coupled. Such coupling could manifest in the persistence state being induced and potentially providing conditions for more effective action of the immune system or in cell death being triggered when immunity fails.

  6. Spectroscopic studies of the small-amplitude Cepheid SU Cas

    NASA Astrophysics Data System (ADS)

    Usenko, I. A.; Klochkova, V. G.; Tavolzhanskaya, N. S.

    2013-09-01

    A new set of 16 high-resolution spectra for the small-amplitude Cepheid SU Cas obtained in 2007-2009 has allowed us to determine its atmospheric parameters ( T eff = 6345 ± 30 K, log g = 2.40, V t = 3.25 km s-1) and to measure its radial velocities. The latter were added to the general list of radial velocities (375 estimates) obtained in the last 90 years. Using a frequency analysis, we have refined the pulsation and orbital periods of the Cepheid. Apart from the well-known fundamental pulsation period [Figure not available: see fulltext.], we have detected a possible secondary period of [Figure not available: see fulltext.]. Their ratio of 0.96 suggests the existence of nonradial pulsations in the Cepheid's atmosphere. Based on photoelectric photometry in the last 60 years, we have shown that the effective temperature undergoes cyclic secular changes of ±200 K with an unknown period. The mean effective temperature T eff = 6395 ± 52 K estimated from photometric data agrees well with our estimate from spectroscopic data. The variations of the mean color index, effective temperature, and γ-velocity (in 90 years of observations) point to a possible orbital motion of the well-known hot companion with the most probable periods of [Figure not available: see fulltext.], [Figure not available: see fulltext.], and [Figure not available: see fulltext.]. The elemental abundances in the atmosphere of SU Cas confirm the conclusion that this Cepheid is a typical yellow supergiant after the first dredge-up. Our T eff estimate gives a radius of 32 R ⊙ and a distance of 455 pc for it, which is inconsistent with its membership in the open cluster Alessi 95. The question about the pulsation mode of SU Cas still remains open.

  7. Textilome abdominal, à propos d'un cas

    PubMed Central

    Erguibi, Driss; Hassan, Robleh; Ajbal, Mohamed; Kadiri, Bouchaib

    2015-01-01

    Le textilome, également appelé gossybipomas, est une complication postopératoire très rare. Il peut s'agir d'un corps étranger composé de compresse(s) ou champ(s) chirurgicaux oubliés au niveau d'un foyer opératoire. Ils sont plus souvent asymptomatiques, et difficile à diagnostiquer. En particulier, les cas chroniques ne présentent pas de signes cliniques et radiologiques spécifiques pour le diagnostic différentiel. L'anamnèse est donc indispensable pour le diagnostic vu que les signes cliniques ne sont pas concluants. Le cliché d'abdomen sans préparation est peu contributif, l’échographie est fiable. La tomodensitométrie permet un diagnostic topographique précis, mais ce n'est pas toujours le cas. Certaines équipes proposent des explorations par IRM. Nous rapportons un cas de textilome intra abdominal, chez une patiente de 31 ans opérée il y a 8 ans pour grossesse extra-utérine, chez qui la TDM abdomino-pelvienne a évoqué un kyste hydatique péritonéale sans localisation du foie. Traitée par extrait d'un petit champ de 25x15cm et adhérant au sigmoïde. Le but de ce travail est de mettre en évidence le problème de diagnostic de cette pathologie et l'importance de la laparotomie exploratrice. PMID:26523184

  8. Cas9-based genome editing in Xenopus tropicalis.

    PubMed

    Nakayama, Takuya; Blitz, Ira L; Fish, Margaret B; Odeleye, Akinleye O; Manohar, Sumanth; Cho, Ken W Y; Grainger, Robert M

    2014-01-01

    Xenopus tropicalis has been developed as a model organism for developmental biology, providing a system offering both modern genetics and classical embryology. Recently, the Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated (CRISPR/Cas) system for genome modification has provided an additional tool for Xenopus researchers to achieve simple and efficient targeted mutagenesis. Here, we provide insights into experimental design and procedures permitting successful application of this technique to Xenopus researchers, and offer a general strategy for performing loss-of-function assays in F0 and subsequently F1 embryos.

  9. SSFinder: high throughput CRISPR-Cas target sites prediction tool.

    PubMed

    Upadhyay, Santosh Kumar; Sharma, Shailesh

    2014-01-01

    Clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein (Cas) system facilitates targeted genome editing in organisms. Despite high demand of this system, finding a reliable tool for the determination of specific target sites in large genomic data remained challenging. Here, we report SSFinder, a python script to perform high throughput detection of specific target sites in large nucleotide datasets. The SSFinder is a user-friendly tool, compatible with Windows, Mac OS, and Linux operating systems, and freely available online. PMID:25089276

  10. Tuberculose orbitaire: à propos d'un cas

    PubMed Central

    Khrifi, Zineb; Abdellaoui, Meriem; Alaoui, Abdellah; Benatiya, Idriss Andaloussi; Tahri, Hicham

    2014-01-01

    La tuberculose sévit à l’état endémique au Maroc, l'atteinte orbitaire est rare et peut se faire par voie hématogène ou par contigüité à partir d'un foyer de voisinage. Nous rapportons le cas d'une patiente de 42 ans qui présente une exophtalmie droite dont l'enquête étiologique révèle une tuberculose orbitaire. PMID:25018814

  11. New spectral observations of the EBS star UU Cas

    NASA Astrophysics Data System (ADS)

    Markov, H.; Markova, N.; Vince, I.; Jurasevich, G.

    High-resolution (R = 30,000) spectroscopy of the Eclipsing Binary System UU Cas obtained with the Coude spectrograph of the 2m telescope of the Bulgarian NAO is presented. The observations cover three spectral regions: the one centred on λ=4520 Å, the other one centred on λ=4720 Å, and the region around Hβ. In each region detailed line identification and radial velocity measurements were performed. The new data together with those published by Markov et al. te{Markov} are used to obtain a better solution of the RV curve and to get some insight into the nature of the system.

  12. Modeling disease in vivo with CRISPR/Cas9

    PubMed Central

    Dow, Lukas E.

    2015-01-01

    The recent advent of CRISPR/Cas9-mediated genome editing has created a wave of excitement across the scientific research community, carrying the promise of simple and effective genomic manipulation of nearly any cell type. CRISPR has quickly become the preferred tool for genetic manipulation, and shows incredible promise as a platform for studying gene function in vivo. Here, I discuss the current application of CRISPR technology to create new in vivo disease models, with a particular focus on how these tools, derived from an adaptive bacterial immune system, are helping us better model the complexity of human cancer. PMID:26432018

  13. [Job placement for people with severe mental illness. The group experience and employment support in a community mental health institution in Buenos Aires].

    PubMed

    Hönig, Adriana

    2015-01-01

    Starting from the depiction of the experience of a reflection group dedicated to job search and job placement for people with severe mental illness, social and working inclusion problem is analyzed. Users' and work receptors' perspectives are analyzed after considering work as a transforming tool of their subjectivity. This experience is based on the concept of employability. Supported employment is highlighted. Emotional moving and subjective affectation within participants are specifically taken into account, and individual pathways towards working insertion valued and respected. This reflection group is analyzed in terms of mental health services' imaginary transformation, as well as professionals, firms and community's attitudes towards people with mental illness, and the concept of recovery and stigma associated to them.

  14. [Job placement for people with severe mental illness. The group experience and employment support in a community mental health institution in Buenos Aires].

    PubMed

    Hönig, Adriana

    2015-01-01

    Starting from the depiction of the experience of a reflection group dedicated to job search and job placement for people with severe mental illness, social and working inclusion problem is analyzed. Users' and work receptors' perspectives are analyzed after considering work as a transforming tool of their subjectivity. This experience is based on the concept of employability. Supported employment is highlighted. Emotional moving and subjective affectation within participants are specifically taken into account, and individual pathways towards working insertion valued and respected. This reflection group is analyzed in terms of mental health services' imaginary transformation, as well as professionals, firms and community's attitudes towards people with mental illness, and the concept of recovery and stigma associated to them. PMID:27089164

  15. Prenatal Air Pollution Exposure and Early Cardiovascular Phenotypes in Young Adults.

    PubMed

    Breton, Carrie V; Mack, Wendy J; Yao, Jin; Berhane, Kiros; Amadeus, Milena; Lurmann, Fred; Gilliland, Frank; McConnell, Rob; Hodis, Howard N; Künzli, Nino; Avol, Ed

    2016-01-01

    Exposure to ambient air pollutants increases risk for adverse cardiovascular health outcomes in adults. We aimed to evaluate the contribution of prenatal air pollutant exposure to cardiovascular health, which has not been thoroughly evaluated. The Testing Responses on Youth (TROY) study consists of 768 college students recruited from the University of Southern California in 2007-2009. Participants attended one study visit during which blood pressure, heart rate and carotid artery arterial stiffness (CAS) and carotid artery intima-media thickness (CIMT) were assessed. Prenatal residential addresses were geocoded and used to assign prenatal and postnatal air pollutant exposure estimates using the U.S. Environmental Protection Agency's Air Quality System (AQS) database. The associations between CAS, CIMT and air pollutants were assessed using linear regression analysis. Prenatal PM10 and PM2.5 exposures were associated with increased CAS. For example, a 2 SD increase in prenatal PM2.5 was associated with CAS indices, including a 5% increase (β = 1.05, 95% CI 1.00-1.10) in carotid stiffness index beta, a 5% increase (β = 1.05, 95% CI 1.01-1.10) in Young's elastic modulus and a 5% decrease (β = 0.95, 95% CI 0.91-0.99) in distensibility. Mutually adjusted models of pre- and postnatal PM2.5 further suggested the prenatal exposure was most relevant exposure period for CAS. No associations were observed for CIMT. In conclusion, prenatal exposure to elevated air pollutants may increase carotid arterial stiffness in a young adult population of college students. Efforts aimed at limiting prenatal exposures are important public health goals.

  16. RNA-guided genome editing in Drosophila with the purified Cas9 protein.

    PubMed

    Lee, Jeong-Soo; Kwak, Su-Jin; Kim, Jungeun; Kim, Ae-Kyeong; Noh, Hae Min; Kim, Jin-Soo; Yu, Kweon

    2014-07-01

    We report a method for generating Drosophila germline mutants effectively via injection of the complex of the purified Cas9 protein, tracrRNA, and gene-specific crRNAs, which may reduce delayed mutations because of the transient activity of the Cas9 protein, combined with the simple mutation detection in GO founders by the T7E1 assay. PMID:24875628

  17. Host Double Strand Break Repair Generates HIV-1 Strains Resistant to CRISPR/Cas9.

    PubMed

    Yoder, Kristine E; Bundschuh, Ralf

    2016-01-01

    CRISPR/Cas9 genome editing has been proposed as a therapeutic treatment for HIV-1 infection. CRISPR/Cas9 induced double strand breaks (DSBs) targeted to the integrated viral genome have been shown to decrease production of progeny virus. Unfortunately HIV-1 evolves rapidly and may readily produce CRISPR/Cas9 resistant strains. Here we used next-generation sequencing to characterize HIV-1 strains that developed resistance to six different CRISPR/Cas9 guide RNAs (gRNAs). Reverse transcriptase (RT) derived base substitution mutations were commonly found at sites encoding unpaired bases of RNA stem-loop structures. In addition to RT mutations, insertion and/or deletion (indel) mutations were common. Indels localized to the CRISPR/Cas9 cleavage site were major contributors to CRISPR gRNA resistance. While most indels at non-coding regions were a single base pair, 3 base pair indels were observed when a coding region of HIV-1 was targeted. The DSB repair event may preserve the HIV-1 reading frame, while destroying CRISPR gRNA homology. HIV-1 may be successfully edited by CRISPR/Cas9, but the virus remains competent for replication and resistant to further CRISPR/Cas9 targeting at that site. These observations strongly suggest that host DSB repair at CRISPR/Cas9 cleavage sites is a novel and important pathway that may contribute to HIV-1 therapeutic resistance. PMID:27404981

  18. A New Practice Evolving in Learning Mathematics: Differences in Students' Written Records with CAS

    ERIC Educational Resources Information Center

    Ball, Lynda; Stacey, Kaye

    2004-01-01

    Students who learn mathematics with CAS calculators are likely to develop new practices for doing and recording mathematics. Students discussed in this paper were able to use CAS calculators in examinations, making their own decisions about what to document as written records for solutions to problems. A comparison of some features of written…

  19. An Undergraduate Laboratory Class Using CRISPR/Cas9 Technology to Mutate Drosophila Genes

    ERIC Educational Resources Information Center

    Adame, Vanesa; Chapapas, Holly; Cisneros, Marilyn; Deaton, Carol; Deichmann, Sophia; Gadek, Chauncey; Lovato, TyAnna L.; Chechenova, Maria B.; Guerin, Paul; Cripps, Richard M.

    2016-01-01

    CRISPR/Cas9 genome editing technology is used in the manipulation of genome sequences and gene expression. Because of the ease and rapidity with which genes can be mutated using CRISPR/Cas9, we sought to determine if a single-semester undergraduate class could be successfully taught, wherein students isolate mutants for specific genes using…

  20. Host Double Strand Break Repair Generates HIV-1 Strains Resistant to CRISPR/Cas9

    PubMed Central

    Yoder, Kristine E.; Bundschuh, Ralf

    2016-01-01

    CRISPR/Cas9 genome editing has been proposed as a therapeutic treatment for HIV-1 infection. CRISPR/Cas9 induced double strand breaks (DSBs) targeted to the integrated viral genome have been shown to decrease production of progeny virus. Unfortunately HIV-1 evolves rapidly and may readily produce CRISPR/Cas9 resistant strains. Here we used next-generation sequencing to characterize HIV-1 strains that developed resistance to six different CRISPR/Cas9 guide RNAs (gRNAs). Reverse transcriptase (RT) derived base substitution mutations were commonly found at sites encoding unpaired bases of RNA stem-loop structures. In addition to RT mutations, insertion and/or deletion (indel) mutations were common. Indels localized to the CRISPR/Cas9 cleavage site were major contributors to CRISPR gRNA resistance. While most indels at non-coding regions were a single base pair, 3 base pair indels were observed when a coding region of HIV-1 was targeted. The DSB repair event may preserve the HIV-1 reading frame, while destroying CRISPR gRNA homology. HIV-1 may be successfully edited by CRISPR/Cas9, but the virus remains competent for replication and resistant to further CRISPR/Cas9 targeting at that site. These observations strongly suggest that host DSB repair at CRISPR/Cas9 cleavage sites is a novel and important pathway that may contribute to HIV-1 therapeutic resistance. PMID:27404981

  1. Lentivirus pre-packed with Cas9 protein for safer gene editing.

    PubMed

    Choi, J G; Dang, Y; Abraham, S; Ma, H; Zhang, J; Guo, H; Cai, Y; Mikkelsen, J G; Wu, H; Shankar, P; Manjunath, N

    2016-07-01

    The CRISPR/Cas9 system provides an easy way to edit specific site/s in the genome and thus offers tremendous opportunity for human gene therapy for a wide range of diseases. However, one major concern is off-target effects, particularly with long-term expression of Cas9 nuclease when traditional expression methods such as via plasmid/viral vectors are used. To overcome this limitation, we pre-packaged Cas9 protein (Cas9P LV) in lentiviral particles for transient exposure and showed its effectiveness for gene disruption in cells, including primary T cells expressing specific single guide RNAs (sgRNAs). We then constructed an 'all in one virus' to express sgRNAs in association with pre-packaged Cas9 protein (sgRNA/Cas9P LV). We successfully edited CCR5 in TZM-bl cells by this approach. Using an sgRNA-targeting HIV long terminal repeat, we also were able to disrupt HIV provirus in the J-LAT model of viral latency. Moreover, we also found that pre-packaging Cas9 protein in LV particle reduced off-target editing of chromosome 4:-29134166 locus by CCR5 sgRNA, compared with continued expression from the vector. These results show that sgRNA/Cas9P LV can be used as a safer approach for human gene therapy applications. PMID:27052803

  2. Appropriate Use of a CAS in the Teaching and Learning of Mathematics.

    ERIC Educational Resources Information Center

    Leinbach, Carl; Pountney, David C.; Etchells, Terence

    2002-01-01

    Links the use of Computer Algebra Systems (CAS) to an existing classification scheme for mathematical tasks called the MATH Taxonomy. Illustrates through concrete examples how the goals of teaching and learning mathematics can be set using this classification together with the CAS. (Author/MM)

  3. Students' use of CAS calculators - effects on the trustworthiness and fairness of mathematics assessments

    NASA Astrophysics Data System (ADS)

    Lind Pantzare, Anna

    2012-10-01

    Calculators with computer algebra systems (CAS) are powerful tools when working with equations and algebraic expressions in mathematics. When calculators are allowed to be used during assessments but are not available or provided to every student, they may cause bias. The CAS calculators may also have an impact on the trustworthiness of results. In this study students' use of the CAS calculator in their work with released assessment items from TIMSS Advanced 2008 is studied using two approaches. Eight students familiar with CAS, from two mathematics classes in the 12th form, were video filmed when encouraged to think aloud during their work with the items. In addition, a questionnaire was distributed to all 33 students in the two classes who had been working with a CAS. The main finding is that even if the students are used to working with the CAS calculator, they are not using the calculator to a large extent. The analysis indicates that the difference in performance between the high- and low-achieving students has slightly increased due to the use of the calculator. From a validity perspective one could therefore argue that the CAS calculator is no major threat to the trustworthiness of the assessment. Nevertheless, the result indicates that those students in the study, mainly high achieving, who know how to use the CAS calculator, get an additional advantage. The advantage brings an amount of unfairness into the assessment and could be a threat to the trustworthiness and fairness.

  4. The Chloroplast Calcium Sensor CAS Is Required for Photoacclimation in Chlamydomonas reinhardtii[W

    PubMed Central

    Petroutsos, Dimitris; Busch, Andreas; Janßen, Ingrid; Trompelt, Kerstin; Bergner, Sonja Verena; Weinl, Stefan; Holtkamp, Michael; Karst, Uwe; Kudla, Jörg; Hippler, Michael

    2011-01-01

    The plant-specific calcium binding protein CAS (calcium sensor) has been localized in chloroplast thylakoid membranes of vascular plants and green algae. To elucidate the function of CAS in Chlamydomonas reinhardtii, we generated and analyzed eight independent CAS knockdown C. reinhardtii lines (cas-kd). Upon transfer to high-light (HL) growth conditions, cas-kd lines were unable to properly induce the expression of LHCSR3 protein that is crucial for nonphotochemical quenching. Prolonged exposure to HL revealed a severe light sensitivity of cas-kd lines and caused diminished activity and recovery of photosystem II (PSII). Remarkably, the induction of LHCSR3, the growth of cas-kd lines under HL, and the performance of PSII were fully rescued by increasing the calcium concentration in the growth media. Moreover, perturbing cellular Ca2+ homeostasis by application of the calmodulin antagonist W7 or the G-protein activator mastoparan impaired the induction of LHCSR3 expression in a concentration-dependent manner. Our findings demonstrate that CAS and Ca2+ are critically involved in the regulation of the HL response and particularly in the control of LHCSR3 expression. PMID:21856795

  5. Efficient fdCas9 Synthetic Endonuclease with Improved Specificity for Precise Genome Engineering

    PubMed Central

    Aouida, Mustapha; Eid, Ayman; Ali, Zahir; Cradick, Thomas; Lee, Ciaran; Deshmukh, Harshavardhan; Atef, Ahmed; AbuSamra, Dina; Gadhoum, Samah Zeineb; Merzaban, Jasmeen; Bao, Gang; Mahfouz, Magdy

    2015-01-01

    The Cas9 endonuclease is used for genome editing applications in diverse eukaryotic species. A high frequency of off-target activity has been reported in many cell types, limiting its applications to genome engineering, especially in genomic medicine. Here, we generated a synthetic chimeric protein between the catalytic domain of the FokI endonuclease and the catalytically inactive Cas9 protein (fdCas9). A pair of guide RNAs (gRNAs) that bind to sense and antisense strands with a defined spacer sequence range can be used to form a catalytically active dimeric fdCas9 protein and generate double-strand breaks (DSBs) within the spacer sequence. Our data demonstrate an improved catalytic activity of the fdCas9 endonuclease, with a spacer range of 15–39 nucleotides, on surrogate reporters and genomic targets. Furthermore, we observed no detectable fdCas9 activity at known Cas9 off-target sites. Taken together, our data suggest that the fdCas9 endonuclease variant is a superior platform for genome editing applications in eukaryotic systems including mammalian cells. PMID:26225561

  6. Attitude and CAS Use in Senior Secondary Mathematics: A Case Study of Seven Year 11 Students

    ERIC Educational Resources Information Center

    Cameron, Scott; Ball, Lynda

    2014-01-01

    This paper investigates the possible influence of attitude on seven Year 11 students' use of a Computer Algebra System (CAS) during a class activity where students could choose to use CAS or pen-and-paper in solving a range of problems. Investigation of anxiety, confidence, liking and usefulness through a survey and interview revealed that these…

  7. Students' Use of CAS Calculators--Effects on the Trustworthiness and Fairness of Mathematics Assessments

    ERIC Educational Resources Information Center

    Pantzare, Anna Lind

    2012-01-01

    Calculators with computer algebra systems (CAS) are powerful tools when working with equations and algebraic expressions in mathematics. When calculators are allowed to be used during assessments but are not available or provided to every student, they may cause bias. The CAS calculators may also have an impact on the trustworthiness of results.…

  8. 76 FR 79545 - Cost Accounting Standards: Change to the CAS Applicability Threshold for the Inflation Adjustment...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-12-22

    ... request for comment (76 FR 40817) for the purpose of revising the Cost Accounting Standards (CAS... Acquisition Regulations Council on August 30, 2010 (at 75 FR 53129). By revising the CAS applicability... Council (Councils) published a final rule in the Federal Register on August 30, 2010 (75 FR...

  9. CRISPR/Cas9: A New Tool for RNA Imaging in Live Cells.

    PubMed

    Rau, Kristina; Rentmeister, Andrea

    2016-09-15

    Cas9 can be implemented as an RNA-targeting system to track mRNA in living cells. Nuclear export is enabled by efficient targeting of GFP-fused Cas9 to an endogenous mRNA. The approach provides a new and versatile platform for RNA-targeting with applications in RNA imaging and beyond.

  10. Host Double Strand Break Repair Generates HIV-1 Strains Resistant to CRISPR/Cas9.

    PubMed

    Yoder, Kristine E; Bundschuh, Ralf

    2016-07-12

    CRISPR/Cas9 genome editing has been proposed as a therapeutic treatment for HIV-1 infection. CRISPR/Cas9 induced double strand breaks (DSBs) targeted to the integrated viral genome have been shown to decrease production of progeny virus. Unfortunately HIV-1 evolves rapidly and may readily produce CRISPR/Cas9 resistant strains. Here we used next-generation sequencing to characterize HIV-1 strains that developed resistance to six different CRISPR/Cas9 guide RNAs (gRNAs). Reverse transcriptase (RT) derived base substitution mutations were commonly found at sites encoding unpaired bases of RNA stem-loop structures. In addition to RT mutations, insertion and/or deletion (indel) mutations were common. Indels localized to the CRISPR/Cas9 cleavage site were major contributors to CRISPR gRNA resistance. While most indels at non-coding regions were a single base pair, 3 base pair indels were observed when a coding region of HIV-1 was targeted. The DSB repair event may preserve the HIV-1 reading frame, while destroying CRISPR gRNA homology. HIV-1 may be successfully edited by CRISPR/Cas9, but the virus remains competent for replication and resistant to further CRISPR/Cas9 targeting at that site. These observations strongly suggest that host DSB repair at CRISPR/Cas9 cleavage sites is a novel and important pathway that may contribute to HIV-1 therapeutic resistance.

  11. Phonological Awareness and Early Reading Development in Childhood Apraxia of Speech (CAS)

    ERIC Educational Resources Information Center

    McNeill, B. C.; Gillon, G. T.; Dodd, B.

    2009-01-01

    Background: Childhood apraxia of speech (CAS) is associated with phonological awareness, reading, and spelling deficits. Comparing literacy skills in CAS with other developmental speech disorders is critical for understanding the complexity of the disorder. Aims: This study compared the phonological awareness and reading development of children…

  12. 41 CFR 102-33.435 - What CAS cost and utilization data must we report?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... costs and contractual periods for related aviation services that you hire (i.e., by contract or through... 33-MANAGEMENT OF GOVERNMENT AIRCRAFT Reporting Information on Government Aircraft Commercial Aviation Services (cas) Cost and Utilization Data § 102-33.435 What CAS cost and utilization data must we...

  13. 41 CFR 102-33.435 - What CAS cost and utilization data must we report?

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... costs and contractual periods for related aviation services that you hire (i.e., by contract or through... 33-MANAGEMENT OF GOVERNMENT AIRCRAFT Reporting Information on Government Aircraft Commercial Aviation Services (cas) Cost and Utilization Data § 102-33.435 What CAS cost and utilization data must we...

  14. Computational Neural Modeling of Speech Motor Control in Childhood Apraxia of Speech (CAS)

    ERIC Educational Resources Information Center

    Terband, Hayo; Maassen, Ben; Guenther, Frank H.; Brumberg, Jonathan

    2009-01-01

    Purpose: Childhood apraxia of speech (CAS) has been associated with a wide variety of diagnostic descriptions and has been shown to involve different symptoms during successive stages of development. In the present study, the authors attempted to associate the symptoms of CAS in a particular developmental stage with particular…

  15. Phylogenetic Distribution of CRISPR-Cas Systems in Antibiotic-Resistant Pseudomonas aeruginosa

    PubMed Central

    van Belkum, Alex; Soriaga, Leah B.; LaFave, Matthew C.; Akella, Srividya; Veyrieras, Jean-Baptiste; Barbu, E. Magda; Shortridge, Dee; Blanc, Bernadette; Hannum, Gregory; Zambardi, Gilles; Miller, Kristofer; Enright, Mark C.; Mugnier, Nathalie; Brami, Daniel; Schicklin, Stéphane; Felderman, Martina; Schwartz, Ariel S.; Richardson, Toby H.; Peterson, Todd C.; Hubby, Bolyn

    2015-01-01

    ABSTRACT Pseudomonas aeruginosa is an antibiotic-refractory pathogen with a large genome and extensive genotypic diversity. Historically, P. aeruginosa has been a major model system for understanding the molecular mechanisms underlying type I clustered regularly interspaced short palindromic repeat (CRISPR) and CRISPR-associated protein (CRISPR-Cas)-based bacterial immune system function. However, little information on the phylogenetic distribution and potential role of these CRISPR-Cas systems in molding the P. aeruginosa accessory genome and antibiotic resistance elements is known. Computational approaches were used to identify and characterize CRISPR-Cas systems within 672 genomes, and in the process, we identified a previously unreported and putatively mobile type I-C P. aeruginosa CRISPR-Cas system. Furthermore, genomes harboring noninhibited type I-F and I-E CRISPR-Cas systems were on average ~300 kb smaller than those without a CRISPR-Cas system. In silico analysis demonstrated that the accessory genome (n = 22,036 genes) harbored the majority of identified CRISPR-Cas targets. We also assembled a global spacer library that aided the identification of difficult-to-characterize mobile genetic elements within next-generation sequencing (NGS) data and allowed CRISPR typing of a majority of P. aeruginosa strains. In summary, our analysis demonstrated that CRISPR-Cas systems play an important role in shaping the accessory genomes of globally distributed P. aeruginosa isolates. PMID:26604259

  16. Immune response of apoptosis-related cysteine peptidases from the red abalone Haliotis rufescens (HrCas8 and HrCas3): molecular characterization and transcription expression.

    PubMed

    Chávez-Mardones, Jacqueline; Gallardo-Escárate, Cristian

    2014-07-01

    Caspases play an important role in the different stages of programmed cell death, or apoptosis, which has been related to the immune response in multicellular organisms. The present study characterized an initiator caspase (HrCas8) and an effector caspase (HrCas3) from the red abalone Haliotis rufescens using the RACE method and qPCR analysis. HrCas8 showed a complete sequence of 2529 base pairs (bp) with an open-reading frame (ORF) of 1911 bp, a 5'UTR of 201 bp, and a 3'UTR of 417 bp. The estimated molecular mass for the 636 amino acids from HrCas8 was 71.5 kDa with an isoelectric point of 6.2. The HrCas8 sequence had two death-effector domains (DEDs) and the subunits p20 and p10, in addition to an active site characteristic of cysteine proteins. Meanwhile, the effector caspase HrCas3 showed a complete sequence of 1404 bp, a 5'UTR of 82 bp, and a 3'UTR of 574 bp. The ORF of this caspase had 747 bp that coded for 248 residues. Moreover, the predicted molecular mass of HrCas3 was 29.4 kDa; the theoretical isoelectric point was 5.70, and the sequence evidenced a conserved caspase recruitment domain (CARD). The distribution of the caspases in distinct tissues revealed that HrCas8 was principally expressed in the hemolymph, while HrCas3 had a higher expression in the gills. A basal level of expression was found for both caspases in muscle tissue. The immune response of caspases in H. rufescens was evaluated through an injection of Vibrio anguillarum. The results showed an increase in the transcription of HrCas8 post-challenge, as well as an activation of HrCas3, which together suggest the initiation of apoptosis as a response to bacterial infection in H. rufescens. PMID:24821426

  17. Industry Support

    NASA Technical Reports Server (NTRS)

    2003-01-01

    NASA Glenn Research Center (GRC) is responsible for the Advanced Communications for Air Traffic Management (AC/ATM) Project, a sub-element task of the Advanced Air Transportation Technologies (AATT) Project of the NASA Aviation System Capacity Program (ASC). The AC/ATM Project is developing new communications technologies and tools that will improve throughput in the U.S. Air Traffic Control System. The goal of the AC/ATM Project is to enable a communications infrastructure providing the capacity, efficiency, and flexibility necessary to realize benefits of the future mature Free-Flight environment. The capabilities and scope of communications technologies needed to accomplish this goal depend on characteristics of the future Free-Flight environment. There are many operational concepts being proposed for a future ATM system to enable user flexibility and efficiency. GRC s focus is on developing new technologies and techniques to support the digital communication of information involving airborne and ground-based users. However, the technologies and techniques must be integrated with the systems and services that industry and the Federal Aviation Administration (FAA) are developing. Thus, GRC needs to monitor and provide input to the various industry and FAA organizations and committees that are specifying new systems and services. Adoption of technologies by the FAA is partially dependent on acceptance of the technology by the aviation community. The commercial aviation community in particular would like to adopt technologies that can be used throughout the world. As a result, the adoption of common or at least compatible technologies by European countries is a key factor in getting commitments to those technologies by the US aviation community. GRC desires to keep informed of European activities that relate to aviation communication technologies, particularly those that are being supported by Eurocontrol.

  18. Technical support document: Energy efficiency standards for consumer products: Room air conditioners, water heaters, direct heating equipment, mobile home furnaces, kitchen ranges and ovens, pool heaters, fluorescent lamp ballasts and television sets. Volume 1, Methodology

    SciTech Connect

    Not Available

    1993-11-01

    The Energy Policy and Conservation Act (P.L. 94-163), as amended, establishes energy conservation standards for 12 of the 13 types of consumer products specifically covered by the Act. The legislation requires the Department of Energy (DOE) to consider new or amended standards for these and other types of products at specified times. DOE is currently considering amending standards for seven types of products: water heaters, direct heating equipment, mobile home furnaces, pool heaters, room air conditioners, kitchen ranges and ovens (including microwave ovens), and fluorescent light ballasts and is considering establishing standards for television sets. This Technical Support Document presents the methodology, data, and results from the analysis of the energy and economic impacts of the proposed standards. This volume presents a general description of the analytic approach, including the structure of the major models.

  19. Removal of ethylene from air stream by adsorption and plasma-catalytic oxidation using silver-based bimetallic catalysts supported on zeolite.

    PubMed

    Trinh, Quang Hung; Lee, Sang Baek; Mok, Young Sun

    2015-03-21

    Dynamic adsorption of ethylene on 13X zeolite-supported Ag and Ag-M(x)O(y) (M: Co, Cu, Mn, and Fe), and plasma-catalytic oxidation of the adsorbed ethylene were investigated. The experimental results showed that the incorporation of Ag into zeolite afforded a marked enhancement in the adsorptivity for ethylene. The addition of transition metal oxides was found to have a positive influence on the ethylene adsorption, except Fe(x)O(y). The presence of the additional metal oxides, however, appeared to somewhat interrupt the diffusion of ozone into the zeolite micro-pores, leading to a decrease in the plasma-catalytic oxidation efficiency of the ethylene adsorbed there. Among the second additional metal oxides, Fe(x)O(y) was able to reduce the emission of ozone during the plasma-catalytic oxidation stage while keeping a high effectiveness for the oxidative removal of the adsorbed ethylene. The periodical treatment consisting of adsorption followed by plasma-catalytic oxidation may be a promising energy-efficient ethylene abatement method.

  20. [Crispr-Cas9 Gene Editing Revolution and the Its Ethical and Legal Challenges].

    PubMed

    Bellver Capella, Vicente

    2016-01-01

    After discovering the CRISPR-Cas9 as an extraordinary method for Gene editing it is necessary to reflect on the ethical, political and legal impact of this technology. This work pretends to offer a preliminary consideration of these problems. I do not pay attention to the potential of CRISPR-Cas9 in the fields of health or environment, nor to all the ethical, legal and political challenges it involves. I principally focus the attention on the possibility of using CRISPR-Cas9 to alter the human germ line. There are some rulings on this topic delivered by intergovernmental organizations. There also are some statements from the scientific community on the matter. They are important in order to know the reasons why they propose a moratorium on the use of CRISPR-Cas9 for human germ line editing. I begin the paper with a short explanation on how CRISPR-Cas9 works. PMID:27637196