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Sample records for abstract service cas

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

    NASA Astrophysics Data System (ADS)

    Kostakos, Charles N.

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

  2. Chemical Abstracts' Document Delivery Service.

    ERIC Educational Resources Information Center

    Rollins, Stephen

    1984-01-01

    The Document Delivery Service offered by Chemical Abstracts is described in terms of the DIALORDER option on the Dialog information retrieval system, mail requests, and requests transmitted through OCLC's Interlibrary Loan system. Transmission costs, success rates, delivery rates, and other considerations in utilizing the service are included.…

  3. Reference in the Age of Automation: Changes in Reference Service at Chemical Abstracts Service Library.

    ERIC Educational Resources Information Center

    Hodges, Pauline R.

    1989-01-01

    Describes the use of office automation equipment at Chemical Abstracts Services (CAS) Library Services and its impact on the reference services offered. Functions discussed include the use of electronic mail for 24-hour library access, online searching, online access to other library collections, and coordination of online searching done by CAS…

  4. 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…

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

    ERIC Educational Resources Information Center

    NACADA Journal, 1986

    1986-01-01

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

  6. New Features in the ADS Abstract Service

    NASA Technical Reports Server (NTRS)

    Eichhorn, Guenther; Accomazzi, Alberto; Grant, Carolyn S.; Kurtz, Michael J.; Henneken, Edwin A.; Thompson, Donna M.; Murray, Stephen S.

    2005-01-01

    The NASA-ADS Abstract Service provides a sophisticated search capability for the literature in Astronomy, Planetary Sciences, Physics/Geophysics, and Space Instrumentation. The ADS is funded by NASA and access to the ADS services is free to anybody world-wide without restrictions. It allows the user to search the literature by author, title, and abstract text.

  7. 40 CFR 799.5000 - Testing consent orders for substances and mixtures with Chemical Abstract Service Registry Numbers.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... adopted under 40 CFR part 790. Listed below in Chemical Abstract Service (CAS) Registry Number order are...) glycidyl ether 1 Health effects June 11, 1996. 68515-47-9 Ditridecyl phthalate (mixed isomers) Chemical fate January 9, 1989. 68515-49-1 Diisodecyl phthalate (mixed isomers) Chemical fate January 9,...

  8. 40 CFR 799.5000 - Testing consent orders for substances and mixtures with Chemical Abstract Service Registry Numbers.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... adopted under 40 CFR part 790. Listed below in Chemical Abstract Service (CAS) Registry Number order are...) glycidyl ether 1 Health effects June 11, 1996. 68515-47-9 Ditridecyl phthalate (mixed isomers) Chemical fate January 9, 1989. 68515-49-1 Diisodecyl phthalate (mixed isomers) Chemical fate January 9,...

  9. 40 CFR 799.5000 - Testing consent orders for substances and mixtures with Chemical Abstract Service Registry Numbers.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... adopted under 40 CFR part 790. Listed below in Chemical Abstract Service (CAS) Registry Number order are.... 68515-47-9 Ditridecyl phthalate (mixed isomers) Chemical fate January 9, 1989. 68515-49-1 Diisodecyl phthalate (mixed isomers) Chemical fate January 9, 1989. 68515-50-4 Dihexyl phthalate (mixed...

  10. 40 CFR 799.5000 - Testing consent orders for substances and mixtures with Chemical Abstract Service Registry Numbers.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... adopted under 40 CFR part 790. Listed below in Chemical Abstract Service (CAS) Registry Number order are...) glycidyl ether 1 Health effects June 11, 1996. 68515-47-9 Ditridecyl phthalate (mixed isomers) Chemical fate January 9, 1989. 68515-49-1 Diisodecyl phthalate (mixed isomers) Chemical fate January 9,...

  11. 40 CFR 799.5000 - Testing consent orders for substances and mixtures with Chemical Abstract Service Registry Numbers.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... adopted under 40 CFR part 790. Listed below in Chemical Abstract Service (CAS) Registry Number order are...) glycidyl ether 1 Health effects June 11, 1996. 68515-47-9 Ditridecyl phthalate (mixed isomers) Chemical fate January 9, 1989. 68515-49-1 Diisodecyl phthalate (mixed isomers) Chemical fate January 9,...

  12. 41 CFR 102-33.95 - What is the process for budgeting to acquire commercial aviation services (CAS)?

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... budgeting to acquire commercial aviation services (CAS)? 102-33.95 Section 102-33.95 Public Contracts and... budgeting to acquire commercial aviation services (CAS)? Except for leases and lease-purchases, for which... fund your commercial aviation services (CAS) hires out of your agency's operating budget....

  13. 41 CFR 102-33.95 - What is the process for budgeting to acquire commercial aviation services (CAS)?

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... budgeting to acquire commercial aviation services (CAS)? 102-33.95 Section 102-33.95 Public Contracts and... budgeting to acquire commercial aviation services (CAS)? Except for leases and lease-purchases, for which... fund your commercial aviation services (CAS) hires out of your agency's operating budget....

  14. 41 CFR 102-33.95 - What is the process for budgeting to acquire commercial aviation services (CAS)?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... budgeting to acquire commercial aviation services (CAS)? 102-33.95 Section 102-33.95 Public Contracts and... budgeting to acquire commercial aviation services (CAS)? Except for leases and lease-purchases, for which... fund your commercial aviation services (CAS) hires out of your agency's operating budget....

  15. 41 CFR 102-33.95 - What is the process for budgeting to acquire commercial aviation services (CAS)?

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... budgeting to acquire commercial aviation services (CAS)? 102-33.95 Section 102-33.95 Public Contracts and... budgeting to acquire commercial aviation services (CAS)? Except for leases and lease-purchases, for which... fund your commercial aviation services (CAS) hires out of your agency's operating budget....

  16. 41 CFR 102-33.95 - What is the process for budgeting to acquire commercial aviation services (CAS)?

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... budgeting to acquire commercial aviation services (CAS)? 102-33.95 Section 102-33.95 Public Contracts and... budgeting to acquire commercial aviation services (CAS)? Except for leases and lease-purchases, for which... fund your commercial aviation services (CAS) hires out of your agency's operating budget....

  17. 78 FR 20101 - Access to Confidential Business Information by Chemical Abstract Services

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-04-03

    ... Services (CAS) of Columbus, Ohio, to access information which has been submitted to EPA under sections 5... is expected under EPA Contract Number EP-W-13-008, contractor CAS of 2540 Olentangy River Rd., P.O... under Contract Number EP-W- 06-011. This is a renewal of a long-existing contract with CAS....

  18. Choosing a Database for Social Work: A Comparison of Social Work Abstracts and Social Service Abstracts

    ERIC Educational Resources Information Center

    Flatley, Robert K.; Lilla, Rick; Widner, Jack

    2007-01-01

    This study compared Social Work Abstracts and Social Services Abstracts databases in terms of indexing, journal coverage, and searches. The authors interviewed editors, analyzed journal coverage, and compared searches. It was determined that the databases complement one another more than compete. The authors conclude with some considerations.

  19. Journal of Human Services Abstracts. Volume 8. Number 3.

    ERIC Educational Resources Information Center

    Journal of Human Services Abstracts, 1983

    1983-01-01

    This quarterly journal contains abstracts of 150 of the documents recently included in Project SHARE automated database. (Project SHARE is a national clearinghouse for improving the management of human services.) The journal is divided into four parts: the abstracts themselves arranged in alphabetical order by title, a list of personal and…

  20. The abstract model of dynamic evolution based on services

    NASA Astrophysics Data System (ADS)

    Qian, Ye; Li, Tong; Li, Yunfei; Gu, Hongxing

    2012-01-01

    Service-oriented software system is facing a challenge to regulate itself promptly because of the evolving Internet environment and user requirements In this paper, a new way that describe the dynamic evolution of services according to 3C mode(Will 1990) is proposed, and Extended workflow net is utilized to describe the abstract model of dynamic evolution of services from specific-functional-domain which is defined in this paper to the whole system.

  1. The abstract model of dynamic evolution based on services

    NASA Astrophysics Data System (ADS)

    Qian, Ye; Li, Tong; Li, Yunfei; Gu, Hongxing

    2011-12-01

    Service-oriented software system is facing a challenge to regulate itself promptly because of the evolving Internet environment and user requirements In this paper, a new way that describe the dynamic evolution of services according to 3C mode(Will 1990) is proposed, and Extended workflow net is utilized to describe the abstract model of dynamic evolution of services from specific-functional-domain which is defined in this paper to the whole system.

  2. Chemical Abstracts Service Chemical Registry System: History, Scope, and Impacts.

    ERIC Educational Resources Information Center

    Weisgerber, David W.

    1997-01-01

    Describes the history, scope, and applications of the Chemical Abstracts Service Chemical Registry System, a computerized database that uniquely identifies chemical substances on the basis of their molecular structures. Explains searching the system is and discusses its use as an international resource. (66 references) (Author/LRW)

  3. Identification of "Known Unknowns" Utilizing Accurate Mass Data and Chemical Abstracts Service Databases

    NASA Astrophysics Data System (ADS)

    Little, James L.; Cleven, Curtis D.; Brown, Stacy D.

    2011-02-01

    In many cases, an unknown to an investigator is actually known in the chemical literature. We refer to these types of compounds as "known unknowns." Chemical Abstracts Service (CAS) Registry is a particularly good source of these substances as it contains over 54 million entries. Accurate mass measurements can be used to query the CAS Registry by either molecular formulae or average molecular weights. Searching the database by the web-based version of SciFinder is the preferred approach when molecular formulae are available. However, if a definitive molecular formula cannot be ascertained, searching the database with STN Express by average molecular weights is a viable alternative. The results from either approach are refined by employing the number of associated references or minimal sample history as orthogonal filters. These approaches were shown to be successful in identifying "known unknowns" noted in LC-MS and even GC-MS analyses in our laboratory. In addition, they were demonstrated in the identification of a variety of compounds of interest to others.

  4. Introduction to the CAS "Standards and Guidelines for Commuter Student Programs and Services."

    ERIC Educational Resources Information Center

    Jacoby, Barbara; Thomas, William L., Jr.

    1986-01-01

    The Council for the Advancement of Standards (CAS) has put forth uniform national criteria for assessing educational effectiveness through the publication of "Standards and Guidelines for Student Services/Development Programs." This document is described briefly, and guidelines for interpretation are offered. (KS)

  5. New Data in the ADS Abstract and Article Service

    NASA Astrophysics Data System (ADS)

    Eichhorn, G.; Accomazzi, A.; Grant, C. S.; Kurtz, M. J.; Murray, S. S.

    1996-05-01

    In the last few months the data holdings in the ADS have been considerably expanded. In the abstracts databases we have included over 50,000 abstracts from SPIE conference proceedings (provided by SPIE), a complete set of references for lunar and planetary sciences, and abstracts from recent Lunar and Planetary Institute sponsored conferences (both provided by the Lunar and Planetary Institute). We also extended our cooperation with the CDS in Strasbourg, France by providing a link to the list of objects that are in the SIMBAD database for each reference. The ADS article service now holds full-text articles for 20 years of the Astrophysical Journal Letters, the Astronomical Journal, and the Publications of the Astronomical Society of the Pacific, and 5 years of the Astrophysical Journal on-line. The following journals are being processed and some may be on-line for this AAS meeting: Astrophysical Journal (1975-1989), Astronomy and Astrophysics, Proceedings of the Astronomical Society of Australia, Publications of the Astronomical Society of Japan, Revista Mexicana, Bulletin of the Astronomical Society of India, Obs. Reports of Skalnate Pleso, and Baltic Astronomy. We are now working with two scanning companies to speed up the scanning process and hope that by the end of the year we have all of these journals completely on-line for at least the period 1975 - 1995. Usage statistics for Jan - March 1996: Users Queries References Full Abstracts retrieved retrieved ------------------------------------------- 13,823 365,812 7,953,930 309,866

  6. Comparison of Printed Bibliographic Descriptions Distributed by BIOSIS, CAS, and Ei

    ERIC Educational Resources Information Center

    Larson, Julian R.; And Others

    1976-01-01

    Printed bibliographic descriptions of journal articles cited by BioSciences Information Service of Biological Abstracts (BIOSIS), Chemical Abstracts Service (CAS), and Engineering Index (Ei) were compared to identify similarities and differences in form, format, and data content. (Author)

  7. New Capabilities of the ADS Abstract and Article Service

    NASA Astrophysics Data System (ADS)

    Eichhorn, G.; Accomazzi, A.; Grant, C. S.; Kurtz, M. J.; Murray, S. S.

    The ADS abstract service at: http://adswww.harvard.edu has been updated considerably in the last year. New capabilities in the search engine include searching for multi-word phrases and searching for various logical combinations of search terms. Through optimization of the custom built search software, the search times were decreased by a factor of 4 in the last year. The WWW interface now uses WWW cookies to store and retrieve individual user preferences. This allows our users to set preferences for printing, accessing mirror sites, fonts, colors, etc. Information about most recently accessed references allows customized retrieval of the most recent unread volume of selected journals. The information stored in these preferences is kept completely confidential and is not used for any other purposes. Two mirror sites (at the CDS in Strasbourg, France and at NAO in Tokyo, Japan) provide faster access for our European and Asian users. To include new information in the ADS as fast as possible, new indexing and search software was developed to allow updating the index data files within minutes of receipt of time critical information (e.g., IAU Circulars which report on supernova and comet discoveries). The ADS is currently used by over 10,000 users per month, which retrieve over 4.5 million references and over 250,000 full article pages each month.

  8. NASA's Astrophysics Data System: new abstract service and article service features.

    NASA Astrophysics Data System (ADS)

    Eichhorn, G.; Accomazzi, A.; Grant, C. S.; Kurtz, M. J.

    1997-09-01

    The NASA Astrophysics Data System provides access to about 1 million abstracts and 50,000 journal articles. This service is funded by NASA and is accessible world-wide through the World Wide Web free without restrictions at: http://adswww.harvard.edu The ADS abstract service allows the searching of a database of abstracts. The abstracts can be searched through a sophisticated search engine by author, astronomical object, title, and text. Various logical combinations of the search terms can be selected. Search results can be filtered by journal, publication date, and other criteria. The search returns a list of references with titles, author lists, and scores that indicate how closely the reference matches the query. The returned references also include hyperlinks to other information where available (e.g. abstracts, full journal articles, data tables, electronic on-line publications, references, citations, lists of astronomical objects, etc.) The ADS article service provides access to full journal articles for all major and many smaller journals. We scan the journals that give us permission at a resolution of 600 dpi. The scans can then be retrieved either for viewing on-screen in gif format, or for printing in Postscript or PCL format. We currently have articles back to 1975 on-line. We plan to eventually scan most of the historical astronomical literature as well. We have recently included the preprints from the Los Alamos preprint server. they are available in a separate database through the same search engine as the other abstracts. Preprints are kept on-line for 6 months. The American Astronomical Society has purchased citation information for 100,000 articles from the Institute for Scientific Information (ISI). This allows us to include links to lists of references and citations in our results page for articles in this set. In order to improve access times for non-US users, we are mirroring the abstract service. We currently have mirrors at the CDS in

  9. CAS Online: A New Source of Substance Information from Chemical Abstracts Service.

    ERIC Educational Resources Information Center

    Farmer, Nick A.; O'Hara, Michael P.

    1980-01-01

    Describes features of a new type of chemical database which provides the ability to search for substances sharing particular structural characteristics. Search concepts are examined in detail, and menu selection of "screens," system commands, and interpretation of results (which appear in the form of chemical bond structures) are explained. (SW)

  10. New Data and Search Features in the NASA ADS Abstract Service

    NASA Technical Reports Server (NTRS)

    Eichhorn, G.; Accomazzi, A.; Grant, C. S.; Kurtz, M. J.; Bacaicoa, V. Rey; Murray, S. S.

    2002-01-01

    The NASA-ADS Abstract Service provides search capability for literature in Astronomy/Planetary Sciences, Physics/Geophysics, and Instrumentation. Scanned articles are available for many journals and proceedings. The ADS service is free for anybody. Additional information is contained in the original extended abstract.

  11. Sophisticated Search Capabilities in the ADS Abstract Service

    NASA Astrophysics Data System (ADS)

    Eichhorn, G.; Accomazzi, A.; Grant, C. S.; Henneken, E.; Kurtz, M. J.; Murray, S. S.

    2003-12-01

    The ADS provides access to over 940,000 references from astronomy and planetary sciences publications and 1.5 million records from physics publications. It is funded by NASA and provides free access to these references, as well as to 2.4 million scanned pages from the astronomical literature. These include most of the major astronomy and several planetary sciences journals, as well as many historical observatory publications. The references now include the abstracts from all volumes of the Journal of Geophysical Research (JGR) since the beginning of 2002. We get these abstracts on a regular basis. The Kluwer journal Solar Physics has been scanned back to volume 1 and is available through the ADS. We have extracted the reference lists from this and many other journals and included them in the reference and citation database of the ADS. We have recently scanning Earth, Moon and Planets, another Kluwer journal, and will scan other Kluwer journals in the future as well. We plan on extracting references from these journals as well in the near future. The ADS has many sophisticated query features. These allow the user to formulate complex queries. Using results lists to get further information about the selected articles provide the means to quickly find important and relevant articles from the database. Three advanced feedback queries are available from the bottom of the ADS results list (in addition to regular feedback queries already available from the abstract page and from the bottom of the results list): 1. Get reference list for selected articles: This query returns all known references for the selected articles (or for all articles in the first list). The resulting list will be ranked according to how often each article is referred to and will show the most referenced articles in the field of study that created the first list. It presumably shows the most important articles in that field. 2. Get citation list for selected articles: This returns all known articles

  12. Emergency Medical Services for Children: Abstracts of Active Projects FY 1996.

    ERIC Educational Resources Information Center

    National Center for Education in Maternal and Child Health, Arlington, VA.

    This publication provides abstracts of 58 active and 42 completed projects designed to improve emergency medical services for children (EMSC). The projects were funded by the United States Department of Health and Human Services' Maternal and Child Health Bureau, in collaboration with the United States Department of Transportation's National…

  13. [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…

  14. Development of a Computerized Current Awareness Service Using "Chemical Abstracts" Condensates

    ERIC Educational Resources Information Center

    Roberts, Anita B.; And Others

    1972-01-01

    The experiences in developing current awareness services for selective dissemination of information from Chemical Abstracts'' Condensates data base are described. File standardization, the weighted-term method of searching, and the algorithm used to perform the search on the CDC 6600 computer and a user survey are discussed. (2 references)…

  15. Full Text Searching and Customization in the NASA ADS Abstract Service

    NASA Technical Reports Server (NTRS)

    Eichhorn, G.; Accomazzi, A.; Grant, C. S.; Kurtz, M. J.; Henneken, E. A.; Thompson, D. M.; Murray, S. S.

    2004-01-01

    The NASA-ADS Abstract Service provides a sophisticated search capability for the literature in Astronomy, Planetary Sciences, Physics/Geophysics, and Space Instrumentation. The ADS is funded by NASA and access to the ADS services is free to anybody worldwide without restrictions. It allows the user to search the literature by author, title, and abstract text. The ADS database contains over 3.6 million references, with 965,000 in the Astronomy/Planetary Sciences database, and 1.6 million in the Physics/Geophysics database. 2/3 of the records have full abstracts, the rest are table of contents entries (titles and author lists only). The coverage for the Astronomy literature is better than 95% from 1975. Before that we cover all major journals and many smaller ones. Most of the journal literature is covered back to volume 1. We now get abstracts on a regular basis from most journals. Over the last year we have entered basically all conference proceedings tables of contents that are available at the Harvard Smithsonian Center for Astrophysics library. This has greatly increased the coverage of conference proceedings in the ADS. The ADS also covers the ArXiv Preprints. We download these preprints every night and index all the preprints. They can be searched either together with the other abstracts or separately. There are currently about 260,000 preprints in that database. In January 2004 we have introduced two new services, full text searching and a personal notification service called "myADS". As all other ADS services, these are free to use for anybody.

  16. 40 CFR 799.5025 - Testing consent orders for mixtures without Chemical Abstracts Service Registry Numbers.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... consent orders adopted under 40 CFR part 790. Listed below are the mixtures which are the subject of these... substances: (1) diheptyl phthalate (branched and linear isomers), CAS No. 68515-44-6 Environmental effects. January 9, 1989. (2) dinonyl phthalate (branched and linear isomers), CAS No. 68515-45-7 ......do Do....

  17. 40 CFR 799.5025 - Testing consent orders for mixtures without Chemical Abstracts Service Registry Numbers.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... consent orders adopted under 40 CFR part 790. Listed below are the mixtures which are the subject of these... substances: (1) diheptyl phthalate (branched and linear isomers), CAS No. 68515-44-6 Environmental effects. January 9, 1989. (2) dinonyl phthalate (branched and linear isomers), CAS No. 68515-45-7 ......do Do....

  18. 40 CFR 799.5025 - Testing consent orders for mixtures without Chemical Abstracts Service Registry Numbers.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... consent orders adopted under 40 CFR part 790. Listed below are the mixtures which are the subject of these... substances: (1) diheptyl phthalate (branched and linear isomers), CAS No. 68515-44-6 Environmental effects. January 9, 1989. (2) dinonyl phthalate (branched and linear isomers), CAS No. 68515-45-7 ......do Do....

  19. 40 CFR 799.5025 - Testing consent orders for mixtures without Chemical Abstracts Service Registry Numbers.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... consent orders adopted under 40 CFR part 790. Listed below are the mixtures which are the subject of these... substances: (1) diheptyl phthalate (branched and linear isomers), CAS No. 68515-44-6 Environmental effects. January 9, 1989. (2) dinonyl phthalate (branched and linear isomers), CAS No. 68515-45-7 ......do Do....

  20. 40 CFR 799.5025 - Testing consent orders for mixtures without Chemical Abstracts Service Registry Numbers.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... consent orders adopted under 40 CFR part 790. Listed below are the mixtures which are the subject of these... substances: (1) diheptyl phthalate (branched and linear isomers), CAS No. 68515-44-6 Environmental effects. January 9, 1989. (2) dinonyl phthalate (branched and linear isomers), CAS No. 68515-45-7 ......do Do....

  1. The UKCIS Advisory Service.

    ERIC Educational Resources Information Center

    Bingham, Archie

    1979-01-01

    Describes the free advisory service available to both users and potential users of chemical and biological databases in the United Kingdom and Ireland. Three specific areas are discussed in which queries about Chemical Abstracts Service (CAS) Registry Numbers have been received: isomers, replacing registry numbers, and mixed compounds. (JD)

  2. Les infections à Pseudomonas aeruginosa au service des maladies infectieuses du CHU YO, Burkina Faso: à propos deux cas

    PubMed Central

    Mamoudou, Savadogo; Lassina, Dao; Fla, Koueta

    2015-01-01

    Nous rapportons deux cas d'infection à Pseudomonas aeruginosa: un cas de méningite et un cas d'infection urinaire. Les auteurs rappellent qu’à côté des étiologies classiques des méningites et des infections urinaires, des germes résistants comme Pseudomonas aeruginosa peuvent être responsables d'infections à localisation méningées et urinaires et dont il faut connaître pour une bonne prise en charge. Le traitement de ces infections requiert un antibiogramme au regard de la grande capacité de résistance de Pseudomonas aeruginosa en milieu hospitalier. La limitation des gestes invasifs et l'application rigoureuse des mesures de prévention des infections en milieu hospitalier contribueront à lutter efficacement contre ces infections en milieu de soins. PMID:26491521

  3. A Comparison of the Guides to Abstracting and Indexing Services Provided by Katz, Chicorel, and Ulrich

    ERIC Educational Resources Information Center

    Swenk, Cynthia; Robinson, Wendy

    1978-01-01

    This study investigated the accuracy of these services: whether they excluded sources in which a journal was actually indexed; and whether the journals accurately reported which services indexed them. (Author/JAB)

  4. Health Services Utilization between Older and Younger Homeless Adults.(author Abstract)

    ERIC Educational Resources Information Center

    Nakonezny, Paul A.; Ojeda, Michael

    2005-01-01

    Purpose: Our purpose in the current study was to examine the relationship between health services utilization delivered by means of the Homeless Outreach Medical Services (HOMES) program and health services utilization delivered by means of the Parkland emergency room and inpatient units among a sample of older and younger homeless adults being…

  5. Final Report for 'An Abstract Job Handling Grid Service for Dataset Analysis'

    SciTech Connect

    David A Alexander

    2005-07-11

    For Phase I of the Job Handling project, Tech-X has built a Grid service for processing analysis requests, as well as a Graphical User Interface (GUI) client that uses the service. The service is designed to generically support High-Energy Physics (HEP) experimental analysis tasks. It has an extensible, flexible, open architecture and language. The service uses the Solenoidal Tracker At RHIC (STAR) experiment as a working example. STAR is an experiment at the Relativistic Heavy Ion Collider (RHIC) at the Brookhaven National Laboratory (BNL). STAR and other experiments at BNL generate multiple Petabytes of HEP data. The raw data is captured as millions of input files stored in a distributed data catalog. Potentially using thousands of files as input, analysis requests are submitted to a processing environment containing thousands of nodes. The Grid service provides a standard interface to the processing farm. It enables researchers to run large-scale, massively parallel analysis tasks, regardless of the computational resources available in their location.

  6. Peak Lead Levels and Diagnostics in Lead Service Lines Dominated by PbO2 - abstract

    EPA Science Inventory

    Multiple studies have presented “profiles” of water lead levels from tap to main through lead service lines (LSLs), in systems where the LSLs were coated with common Pb(II) corrosion solids. These Pb(II) solids were either actual Pb(II) minerals or Pb(II) sorbed onto other pipe ...

  7. The Pricing of Information--A Search-Based Approach to Pricing an Online Search Service.

    ERIC Educational Resources Information Center

    Boyle, Harry F.

    1982-01-01

    Describes innovative pricing structure consisting of low connect time fee, print fees, and search fees, offered by Chemical Abstracts Service (CAS) ONLINE--an online searching system used to locate chemical substances. Pricing options considered by CAS, the search-based pricing approach, and users' reactions to pricing structures are noted. (EJS)

  8. 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...

  9. 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...

  10. 41 CFR 102-33.130 - If we hire CAS, what are our management responsibilities?

    Code of Federal Regulations, 2011 CFR

    2011-01-01

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

  11. 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...

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

    Code of Federal Regulations, 2010 CFR

    2010-07-01

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

  13. 41 CFR 102-33.130 - If we hire CAS, what are our management responsibilities?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

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

  14. 41 CFR 102-33.435 - What CAS cost and utilization data must we report?

    Code of Federal Regulations, 2013 CFR

    2013-07-01

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

  15. 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...

  16. 41 CFR 102-33.435 - What CAS cost and utilization data must we report?

    Code of Federal Regulations, 2011 CFR

    2011-01-01

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

  17. 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...

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

    ERIC Educational Resources Information Center

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

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

  19. Efficacy of Indexing and Abstracting Services for the Dissemination of Agricultural Information Resources in the Institure for Agricultural Research Library, Ahmadu Bello University, Zaria

    NASA Astrophysics Data System (ADS)

    Gabriel, KASA, M.

    2012-10-01

    The efficacy of Indexing and Abstracting service for effective organization, storage and retrieval of information resources for agricultural research in Ahmadu Bello University, Zaria necessitated examining the situation in Agricultural Library, Institute for Agricultural Research, Samaru. The study examines the processes, awareness and problems militating against the effective exploitation of the indexing and abstracting services in the Agricultural library established in 1975. The study was conducted ex post facto, data collected span from 2006 ñ 2010. Total sample sizes of 752 patrons and 20,236 intellectually indexed and abstracted resources were involved in the study. Data collected were subjected to descriptive and inferential statistics. The result revealed that a total of 644 articles were indexed and abstracted, 35% of these was done in 2010. Results for awareness show 452 (60.11%) to be aware in 2008. A total 584 articles were indexed and abstracted from which 167 (28.59%) was retrieved in 2006. Patrons, 270 (35.90%) attributed the poor use of the service to assumption it is a referral unit. The hypothesis testing revealed that there is significant association between articles indexed and abstracted with information consulted by patrons (?2cal,100.31>?2tab,9.488) at 5% level of probability and df, 4. In conclusion, enormous documents on Nigerian agriculture are indexed and abstracted in the unit, implying that the service is desirous and consistent. The study recommends that the unit should explore the use of modern technology, employ a permanent subject specialist, train and retrain the unit staff as well as intensify it general orientation campaigns to focus on awareness and use of the indexing and abstracting services.

  20. Ponction biopsies rénales dans le Service de Néphrologie de Fès: indications et résultats: à propos de 522 cas

    PubMed Central

    Mbarki, Houda; Belghiti, Khadija Alaoui; Harmouch, Taoufiq; Najdi, Adil; Arrayhani, Mohamed; Sqalli, Tarik

    2016-01-01

    L'apport de la ponction biopsie rénale (PBR) dans le diagnostic, le choix thérapeutique et l’évaluation pronostique des néphropathies est considérable. Aucune étude marocaine n'a évalué la pratique et l'apport de la PBR. Notre objectif est d’étudier les indications de la PBR, déterminer la fréquence des maladies rénales identifiées par PBR dans notre région et de faire une confrontation entre les données clinico-biologiques et le diagnostic historique. Notre étude menée entre Janvier 2009 et Décembre 2012, est rétrospective. Nous avons inclus tous les patients du service de Néphrologie du CHU Hassan II de Fès ayant bénéficié d'une biopsie de reins natifs. 522 PBR ont été réalisées. Nous avons exclu 8 biopsies devant le manque de renseignements et avons donc retenu 514. L’âge moyen des patients au moment de la PBR est de 39 ±17 ans (3-82 ans). Le sex ratio est de 0,9. Le syndrome néphrotique est le diagnostic clinique le plus fréquent à tous les âges (58,2%). Les néphropathies glomérulaires représentent 94,2% des maladies rénales diagnostiquées, leur distribution varie selon l’âge des patients. La PBR a confirmé le premier diagnostic suspecté cliniquement dans 40,65% des cas, alors qu'elle a révélé un diagnostic inattendu chez 22,5% d'entre eux. Le diagnostic syndromique permet d'orienter vers la maladie rénale la plus probable et de guider les thérapeutiques urgentes en attendant les résultats de la PBR. Mais il ne peut en aucun remplacer la PBR qui reste le gold standard.

  1. Abstract Painting

    ERIC Educational Resources Information Center

    Henkes, Robert

    1978-01-01

    Abstract art provokes numerous interpretations, and as many misunderstandings. The adolescent reaction is no exception. The procedure described here can help the student to understand the abstract from at least one direction. (Author/RK)

  2. Library Programs; Library Services for Indian Tribes and Hawaiian Natives Program. Abstracts of Funded Projects, 1987. LSCA Title IV.

    ERIC Educational Resources Information Center

    Office of Educational Research and Improvement (ED), Washington, DC.

    This booklet provides information on the third year of funding (fiscal year 1987) of the Library Services and Construction Act (LSCA), Title IV grant programs, which provides appropriations for Indian tribes and Hawaiian natives under two program cycles: Basic Grants and Special Projects. An explanation of the Basic Grants program is presented,…

  3. Abstract Constructions.

    ERIC Educational Resources Information Center

    Pietropola, Anne

    1998-01-01

    Describes a lesson designed to culminate a year of eighth-grade art classes in which students explore elements of design and space by creating 3-D abstract constructions. Outlines the process of using foam board and markers to create various shapes and optical effects. (DSK)

  4. 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.

  5. 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

  6. Cas9 Functionally Opens Chromatin

    PubMed Central

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

    2016-01-01

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

  7. INVENTORY ABSTRACTION

    SciTech Connect

    G. Ragan

    2001-12-19

    The purpose of the inventory abstraction, which has been prepared in accordance with a technical work plan (CRWMS M&O 2000e for ICN 02 of the present analysis, and BSC 2001e for ICN 03 of the present analysis), is to: (1) Interpret the results of a series of relative dose calculations (CRWMS M&O 2000c, 2000f). (2) Recommend, including a basis thereof, a set of radionuclides that should be modeled in the Total System Performance Assessment in Support of the Site Recommendation (TSPA-SR) and the Total System Performance Assessment in Support of the Final Environmental Impact Statement (TSPA-FEIS). (3) Provide initial radionuclide inventories for the TSPA-SR and TSPA-FEIS models. (4) Answer the U.S. Nuclear Regulatory Commission (NRC)'s Issue Resolution Status Report ''Key Technical Issue: Container Life and Source Term'' (CLST IRSR) key technical issue (KTI): ''The rate at which radionuclides in SNF [spent nuclear fuel] are released from the EBS [engineered barrier system] through the oxidation and dissolution of spent fuel'' (NRC 1999, Subissue 3). The scope of the radionuclide screening analysis encompasses the period from 100 years to 10,000 years after the potential repository at Yucca Mountain is sealed for scenarios involving the breach of a waste package and subsequent degradation of the waste form as required for the TSPA-SR calculations. By extending the time period considered to one million years after repository closure, recommendations are made for the TSPA-FEIS. The waste forms included in the inventory abstraction are Commercial Spent Nuclear Fuel (CSNF), DOE Spent Nuclear Fuel (DSNF), High-Level Waste (HLW), naval Spent Nuclear Fuel (SNF), and U.S. Department of Energy (DOE) plutonium waste. The intended use of this analysis is in TSPA-SR and TSPA-FEIS. Based on the recommendations made here, models for release, transport, and possibly exposure will be developed for the isotopes that would be the highest contributors to the dose given a release to the

  8. Abstracts of SIG Sessions.

    ERIC Educational Resources Information Center

    Proceedings of the ASIS Annual Meeting, 1995

    1995-01-01

    Presents abstracts of 15 special interest group (SIG) sessions. Topics include navigation and information utilization in the Internet, natural language processing, automatic indexing, image indexing, classification, users' models of database searching, online public access catalogs, education for information professions, information services,…

  9. Abstracting and indexing guide

    USGS Publications Warehouse

    U.S. Department of the Interior; Office of Water Resources Research

    1974-01-01

    These instructions have been prepared for those who abstract and index scientific and technical documents for the Water Resources Scientific Information Center (WRSIC). With the recent publication growth in all fields, information centers have undertaken the task of keeping the various scientific communities aware of current and past developments. An abstract with carefully selected index terms offers the user of WRSIC services a more rapid means for deciding whether a document is pertinent to his needs and professional interests, thus saving him the time necessary to scan the complete work. These means also provide WRSIC with a document representation or surrogate which is more easily stored and manipulated to produce various services. Authors are asked to accept the responsibility for preparing abstracts of their own papers to facilitate quick evaluation, announcement, and dissemination to the scientific community.

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

    PubMed Central

    Zhang, Wen-Wei

    2015-01-01

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

  11. La Greffe de Peau dans le Traitement des Sequelles de la Main Brulee. A Propos de 152 Cas - Experience du Service de Chirurgie Plastique du Centre Hospitalier Universitaire Ibn-Sina, Rabat, Maroc

    PubMed Central

    El Mazouz, S.; Fejjal, N.; Hafidi, J.; Cherkab, L.; Mejjati, H.; Belfqih, R.; Gharib, N.; Abbassi, A.

    2010-01-01

    Summary La main est fréquemment exposée aux brûlures, entraînant des séquelles esthétiques et fonctionnelles. Le traitement de ces séquelles est surtout chirurgical et consiste en la greffe de peau, dont le type dépend de la localisation de la brûlure et du type des séquelles. Dans ce travail rétrospectif, nous rapportons une série de 152 cas de brûlures des mains colligés au service de chirurgie plastique du Centre Hospitalier Universitaire Ibn-Sina de Rabat sur une période de dix ans, allant de 1998 à 2007. Les indications thérapeutiques dépendent du type de séquelles et de la localisation de la brûlure. En tout, 97 patients ont bénéficié d'une greffe cutanée, dont 76% par greffe de peau totale, 21% par greffe de peau demi-épaisse et 3% par peau fine. Les séquelles des brûlures des mains posent un problème thérapeutique majeur, malgré la diversité des procédés chirurgicaux, d'où l'intérêt de la prévention. PMID:21991196

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

    PubMed

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

    2016-04-01

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

  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. PMID:20843716

  14. Differences between Beilstein and the CAS Databases.

    ERIC Educational Resources Information Center

    Heller, Stephen R.

    1987-01-01

    Describes the different approaches taken by the Chemical Abstracts Services database, which abstracts and indexes chemical publications, and the Belstein Handbook of Organic Chemistry database, which produces a collection of critical reviews. The resulting content of the databases and their ability to meet the needs of different users are…

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

    PubMed Central

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

    2016-01-01

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

  17. 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.

  18. 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

  19. Workshop on seedling physiology and growth problems in oak plantings (5th) (abstracts). Held in Ames, Iowa on March 4-5, 1992. Forest Service general technical report

    SciTech Connect

    Thompson, J.R.; Schultz, R.C.; Van Sambeek, J.W.

    1993-01-01

    The fifth workshop on seedling physiology and growth problems in oak plantings was held at the Holiday Inn Gateway Center in Ames, Iowa on March 4 and 5, 1992 with more than 45 participants. The workshop continues to serve as an informal forum for researchers to exchange ideas and research results. Papers were divided into four general subject areas: (1) field performance of planted oaks, (2) seedling propagation and production, (3) oak physiology and genetics, and (4) natural and acorn germination regeneration. All abstracts prepared for the workshop are included in the technical report.

  20. Exploiting CRISPR/Cas systems for biotechnology

    PubMed Central

    Sampson, Timothy R.; Weiss, David S.

    2015-01-01

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

  1. ISC, a Novel Group of Bacterial and Archaeal DNA Transposons That Encode Cas9 Homologs

    PubMed Central

    Kapitonov, Vladimir V.; Makarova, Kira S.

    2015-01-01

    ABSTRACT Bacterial genomes encode numerous homologs of Cas9, the effector protein of the type II CRISPR-Cas systems. The homology region includes the arginine-rich helix and the HNH nuclease domain that is inserted into the RuvC-like nuclease domain. These genes, however, are not linked to cas genes or CRISPR. Here, we show that Cas9 homologs represent a distinct group of nonautonomous transposons, which we denote ISC (insertion sequences Cas9-like). We identify many diverse families of full-length ISC transposons and demonstrate that their terminal sequences (particularly 3′ termini) are similar to those of IS605 superfamily transposons that are mobilized by the Y1 tyrosine transposase encoded by the TnpA gene and often also encode the TnpB protein containing the RuvC-like endonuclease domain. The terminal regions of the ISC and IS605 transposons contain palindromic structures that are likely recognized by the Y1 transposase. The transposons from these two groups are inserted either exactly in the middle or upstream of specific 4-bp target sites, without target site duplication. We also identify autonomous ISC transposons that encode TnpA-like Y1 transposases. Thus, the nonautonomous ISC transposons could be mobilized in trans either by Y1 transposases of other, autonomous ISC transposons or by Y1 transposases of the more abundant IS605 transposons. These findings imply an evolutionary scenario in which the ISC transposons evolved from IS605 family transposons, possibly via insertion of a mobile group II intron encoding the HNH domain, and Cas9 subsequently evolved via immobilization of an ISC transposon. IMPORTANCE Cas9 endonucleases, the effectors of type II CRISPR-Cas systems, represent the new generation of genome-engineering tools. Here, we describe in detail a novel family of transposable elements that encode the likely ancestors of Cas9 and outline the evolutionary scenario connecting different varieties of these transposons and Cas9. PMID:26712934

  2. Piaget on Abstraction.

    ERIC Educational Resources Information Center

    Moessinger, Pierre; Poulin-Dubois, Diane

    1981-01-01

    Reviews and discusses Piaget's recent work on abstract reasoning. Piaget's distinction between empirical and reflective abstraction is presented; his hypotheses are considered to be metaphorical. (Author/DB)

  3. Interactive Intergenerational Learning in the Context of CAS in the IB Diploma Programme: A UK Case Study

    ERIC Educational Resources Information Center

    Cambridge, James; Simandiraki, Anna

    2006-01-01

    Creativity, Action, Service (CAS) forms a core component of the International Baccalaureate Diploma Programme. CAS frequently includes intergenerational practice involving school students and older adults. This article reports the development of a typology to describe and analyse learning that arises from intergenerational practice in three…

  4. Condition Assessment survey (CAS) Program. Deficiency standards and inspections methods manual: Volume 9, 0.09 Electrical, Book 1

    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; and system work breakdown structure. Deficiency standards are presented for service & distribution; lighting; and special systems.

  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. "CAS" Statement of Shared Ethical Principles

    ERIC Educational Resources Information Center

    Council for the Advancement of Standards in Higher Education, 2006

    2006-01-01

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

  7. Manpower Management Studies: Selected Abstracts.

    ERIC Educational Resources Information Center

    Ryerson, William R., Comp.

    This bibliography contains 58 selected abstracts of research reports dating back to 1964 on the general subject of manpower management. It was prepared from a search of the National Technical Information Service data base of more than 300,000 documents submitted by agencies of the Federal Government and also by private organizations or individuals…

  8. Annual Conference Abstracts

    ERIC Educational Resources Information Center

    Engineering Education, 1975

    1975-01-01

    Papers abstracted represent those submitted to the distribution center at the 83rd American Society for Engineering Education Convention. Abstracts are grouped under headings corresponding to the main topic of the paper. (Editor/CP)

  9. Abstraction and Consolidation

    ERIC Educational Resources Information Center

    Monaghan, John; Ozmantar, Mehmet Fatih

    2006-01-01

    The framework for this paper is a recently developed theory of abstraction in context. The paper reports on data collected from one student working on tasks concerned with absolute value functions. It examines the relationship between mathematical constructions and abstractions. It argues that an abstraction is a consolidated construction that can…

  10. Abstraction and Problem Reformulation

    NASA Technical Reports Server (NTRS)

    Giunchiglia, Fausto

    1992-01-01

    In work done jointly with Toby Walsh, the author has provided a sound theoretical foundation to the process of reasoning with abstraction (GW90c, GWS9, GW9Ob, GW90a). The notion of abstraction formalized in this work can be informally described as: (property 1), the process of mapping a representation of a problem, called (following historical convention (Sac74)) the 'ground' representation, onto a new representation, called the 'abstract' representation, which, (property 2) helps deal with the problem in the original search space by preserving certain desirable properties and (property 3) is simpler to handle as it is constructed from the ground representation by "throwing away details". One desirable property preserved by an abstraction is provability; often there is a relationship between provability in the ground representation and provability in the abstract representation. Another can be deduction or, possibly inconsistency. By 'throwing away details' we usually mean that the problem is described in a language with a smaller search space (for instance a propositional language or a language without variables) in which formulae of the abstract representation are obtained from the formulae of the ground representation by the use of some terminating rewriting technique. Often we require that the use of abstraction results in more efficient .reasoning. However, it might simply increase the number of facts asserted (eg. by allowing, in practice, the exploration of deeper search spaces or by implementing some form of learning). Among all abstractions, three very important classes have been identified. They relate the set of facts provable in the ground space to those provable in the abstract space. We call: TI abstractions all those abstractions where the abstractions of all the provable facts of the ground space are provable in the abstract space; TD abstractions all those abstractions wllere the 'unabstractions' of all the provable facts of the abstract space are

  11. CAS-1 lunar soil simulant

    NASA Astrophysics Data System (ADS)

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

    2009-02-01

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

  12. Abstraction in mathematics.

    PubMed

    Ferrari, Pier Luigi

    2003-07-29

    Some current interpretations of abstraction in mathematical settings are examined from different perspectives, including history and learning. It is argued that abstraction is a complex concept and that it cannot be reduced to generalization or decontextualization only. In particular, the links between abstraction processes and the emergence of new objects are shown. The role that representations have in abstraction is discussed, taking into account both the historical and the educational perspectives. As languages play a major role in mathematics, some ideas from functional linguistics are applied to explain to what extent mathematical notations are to be considered abstract. Finally, abstraction is examined from the perspective of mathematics education, to show that the teaching ideas resulting from one-dimensional interpretations of abstraction have proved utterly unsuccessful. PMID:12903658

  13. 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

  14. 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

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

    PubMed

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

    2016-01-01

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

  16. The Impact of CAS Standards on the Practice of Student Affairs.

    ERIC Educational Resources Information Center

    Mann, Barbara A.; And Others

    1991-01-01

    Studied impact of Council for the Advancement of Standards (CAS) for Student Services/Development Programs'"Standards and Guidelines for Student Services/Development Programs." Findings from 130 chief student affairs officers revealed that, although emphasis on assessment, student involvement, and holistic development suggested by standards was…

  17. In Vitro CRISPR/Cas9 System for Efficient Targeted DNA Editing

    PubMed Central

    Liu, Yunkun; Tao, Weixin; Wen, Shishi; Li, Zhengyuan; Yang, Anna; Deng, Zixin

    2015-01-01

    ABSTRACT The clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9) system, an RNA-guided nuclease for specific genome editing in vivo, has been adopted in a wide variety of organisms. In contrast, the in vitro application of the CRISPR/Cas9 system has rarely been reported. We present here a highly efficient in vitro CRISPR/Cas9-mediated editing (ICE) system that allows specific refactoring of biosynthetic gene clusters in Streptomyces bacteria and other large DNA fragments. Cleavage by Cas9 of circular pUC18 DNA was investigated here as a simple model, revealing that the 3′→5′ exonuclease activity of Cas9 generates errors with 5 to 14 nucleotides (nt) randomly missing at the editing joint. T4 DNA polymerase was then used to repair the Cas9-generated sticky ends, giving substantial improvement in editing accuracy. Plasmid pYH285 and cosmid 10A3, harboring a complete biosynthetic gene cluster for the antibiotics RK-682 and holomycin, respectively, were subjected to the ICE system to delete the rkD and homE genes in frame. Specific insertion of the ampicillin resistance gene (bla) into pYH285 was also successfully performed. These results reveal the ICE system to be a rapid, seamless, and highly efficient way to edit DNA fragments, and a powerful new tool for investigating and engineering biosynthetic gene clusters. PMID:26556277

  18. Loving Those Abstracts

    ERIC Educational Resources Information Center

    Stevens, Lori

    2004-01-01

    The author describes a lesson she did on abstract art with her high school art classes. She passed out a required step-by-step outline of the project process. She asked each of them to look at abstract art. They were to list five or six abstract artists they thought were interesting, narrow their list down to the one most personally intriguing,…

  19. Targeted mutagenesis using CRISPR/Cas system in medaka

    PubMed Central

    Ansai, Satoshi; Kinoshita, Masato

    2014-01-01

    ABSTRACT Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) system-based RNA-guided endonuclease (RGEN) has recently emerged as a simple and efficient tool for targeted genome editing. In this study, we showed successful targeted mutagenesis using RGENs in medaka, Oryzias latipes. Somatic and heritable mutations were induced with high efficiency at the targeted genomic sequence on the DJ-1 gene in embryos that had been injected with the single guide RNA (sgRNA) transcribed by a T7 promoter and capped RNA encoding a Cas9 nuclease. The sgRNAs that were designed for the target genomic sequences without the 5′ end of GG required by the T7 promoter induced the targeted mutations. This suggests that the RGEN can target any sequence adjacent to an NGG protospacer adjacent motif (PAM) sequence, which occurs once every 8 bp. The off-target alterations at 2 genomic loci harboring double mismatches in the 18-bp targeting sequences were induced in the RGEN-injected embryos. However, we also found that the off-target effects could be reduced by lower dosages of sgRNA. Taken together, our results suggest that CRISPR/Cas-mediated RGENs may be an efficient and flexible tool for genome editing in medaka. PMID:24728957

  20. Community Development Abstracts.

    ERIC Educational Resources Information Center

    Agency for International Development (Dept. of State), Washington, DC.

    This volume of 1,108 abstracts summarizes the majority of important works on community development during the last ten years. Part I contains abstracts of periodical literature and is classified into 19 sections, including general history, communications, community and area studies, decision-making, leadership, migration and settlement, social…

  1. Leadership Abstracts, Volume 10.

    ERIC Educational Resources Information Center

    Milliron, Mark D., Ed.

    1997-01-01

    The abstracts in this series provide brief discussions of issues related to leadership, administration, professional development, technology, and education in community colleges. Volume 10 for 1997 contains the following 12 abstracts: (1) "On Community College Renewal" (Nathan L. Hodges and Mark D. Milliron); (2) "The Community College Niche in a…

  2. Has Abstractness Been Resolved?

    ERIC Educational Resources Information Center

    Al-Omoush, Ahmad

    1989-01-01

    A discussion focusing on the abstractness of analysis in phonology, debated since the 1960s, describes the issue, reviews the literature on the subject, cites specific natural language examples, and examines the extent to which the issue has been resolved. An underlying representation is said to be abstract if it is different from the derived one,…

  3. Designing for Mathematical Abstraction

    ERIC Educational Resources Information Center

    Pratt, Dave; Noss, Richard

    2010-01-01

    Our focus is on the design of systems (pedagogical, technical, social) that encourage mathematical abstraction, a process we refer to as "designing for abstraction." In this paper, we draw on detailed design experiments from our research on children's understanding about chance and distribution to re-present this work as a case study in designing…

  4. Knowledge-Based Abstracting.

    ERIC Educational Resources Information Center

    Black, William J.

    1990-01-01

    Discussion of automatic abstracting of technical papers focuses on a knowledge-based method that uses two sets of rules. Topics discussed include anaphora; text structure and discourse; abstracting techniques, including the keyword method and the indicator phrase method; and tools for text skimming. (27 references) (LRW)

  5. Leadership Abstracts, 1995.

    ERIC Educational Resources Information Center

    Johnson, Larry, Ed.

    1995-01-01

    The abstracts in this series provide two-page discussions of issues related to leadership, administration, and teaching in community colleges. The 12 abstracts for Volume 8, 1995, are: (1) "Redesigning the System To Meet the Workforce Training Needs of the Nation," by Larry Warford; (2) "The College President, the Board, and the Board Chair: A…

  6. Paper Abstract Animals

    ERIC Educational Resources Information Center

    Sutley, Jane

    2010-01-01

    Abstraction is, in effect, a simplification and reduction of shapes with an absence of detail designed to comprise the essence of the more naturalistic images being depicted. Without even intending to, young children consistently create interesting, and sometimes beautiful, abstract compositions. A child's creations, moreover, will always seem to…

  7. Is It Really Abstract?

    ERIC Educational Resources Information Center

    Kernan, Christine

    2011-01-01

    For this author, one of the most enjoyable aspects of teaching elementary art is the willingness of students to embrace the different styles of art introduced to them. In this article, she describes a project that allows upper-elementary students to learn about abstract art and the lives of some of the master abstract artists, implement the idea…

  8. Journalism Abstracts. Vol. 15.

    ERIC Educational Resources Information Center

    Popovich, Mark N., Ed.

    This book, the fifteenth volume of an annual publication, contains 373 abstracts of 52 doctoral and 321 master's theses from 50 colleges and universities. The abstracts are arranged alphabetically by author, with the doctoral dissertations appearing first. These cover such topics as advertising, audience analysis, content analysis of news issues…

  9. Leadership Abstracts, 1996.

    ERIC Educational Resources Information Center

    Johnson, Larry, Ed.

    1996-01-01

    The abstracts in this series provide two-page discussions of issues related to leadership, administration, professional development, technology, and education in community colleges. Volume 9 for 1996 includes the following 12 abstracts: (1) "Tech-Prep + School-To-Work: Working Together To Foster Educational Reform," (Roderick F. Beaumont); (2)…

  10. Mathematical Abstraction through Scaffolding

    ERIC Educational Resources Information Center

    Ozmantar, Mehmet Fatih; Roper, Tom

    2004-01-01

    This paper examines the role of scaffolding in the process of abstraction. An activity-theoretic approach to abstraction in context is taken. This examination is carried out with reference to verbal protocols of two 17 year-old students working together on a task connected to sketching the graph of |f|x|)|. Examination of the data suggests that…

  11. Abstract coherent categories.

    PubMed

    Rehder, B; Ross, B H

    2001-09-01

    Many studies have demonstrated the importance of the knowledge that interrelates features in people's mental representation of categories and that makes our conception of categories coherent. This article focuses on abstract coherent categories, coherent categories that are also abstract because they are defined by relations independently of any features. Four experiments demonstrate that abstract coherent categories are learned more easily than control categories with identical features and statistical structure, and also that participants induced an abstract representation of the category by granting category membership to exemplars with completely novel features. The authors argue that the human conceptual system is heavily populated with abstract coherent concepts, including conceptions of social groups, societal institutions, legal, political, and military scenarios, and many superordinate categories, such as classes of natural kinds. PMID:11550753

  12. Abstract Datatypes in PVS

    NASA Technical Reports Server (NTRS)

    Owre, Sam; Shankar, Natarajan

    1997-01-01

    PVS (Prototype Verification System) is a general-purpose environment for developing specifications and proofs. This document deals primarily with the abstract datatype mechanism in PVS which generates theories containing axioms and definitions for a class of recursive datatypes. The concepts underlying the abstract datatype mechanism are illustrated using ordered binary trees as an example. Binary trees are described by a PVS abstract datatype that is parametric in its value type. The type of ordered binary trees is then presented as a subtype of binary trees where the ordering relation is also taken as a parameter. We define the operations of inserting an element into, and searching for an element in an ordered binary tree; the bulk of the report is devoted to PVS proofs of some useful properties of these operations. These proofs illustrate various approaches to proving properties of abstract datatype operations. They also describe the built-in capabilities of the PVS proof checker for simplifying abstract datatype expressions.

  13. Meeting Abstracts - Annual Meeting 2016.

    PubMed

    2016-04-01

    The AMCP Abstracts program provides a forum through which authors can share their insights and outcomes of advanced managed care practice through publication in AMCP's Journal of Managed Care & Specialty Pharmacy (JMCP). Most of the reviewed and unreviewed abstracts are presented as posters so that interested AMCP meeting attendees can review findings and query authors. The Student/Resident/ Fellow poster presentation (unreviewed) is Wednesday, April 20, 2016, and the Professional poster presentation (reviewed) is Thursday, April 21. The Professional posters will also be displayed on Friday, April 22. The reviewed abstracts are published in the JMCP Meeting Abstracts supplement. The AMCP Managed Care & Specialty Pharmacy Annual Meeting 2016 in San Francisco, California, is expected to attract more than 3,500 managed care pharmacists and other health care professionals who manage and evaluate drug therapies, develop and manage networks, and work with medical managers and information specialists to improve the care of all individuals enrolled in managed care programs. Abstracts were submitted in the following categories: Research Report: describe completed original research on managed care pharmacy services or health care interventions. Examples include (but are not limited to) observational studies using administrative claims, reports of the impact of unique benefit design strategies, and analyses of the effects of innovative administrative or clinical programs. Economic Model: describe models that predict the effect of various benefit design or clinical decisions on a population. For example, an economic model could be used to predict the budget impact of a new pharmaceutical product on a health care system. Solving Problems in Managed Care: describe the specific steps taken to introduce a needed change, develop and implement a new system or program, plan and organize an administrative function, or solve other types of problems in managed care settings. These

  14. 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)

  15. Abstract Interpreters for Free

    NASA Astrophysics Data System (ADS)

    Might, Matthew

    In small-step abstract interpretations, the concrete and abstract semantics bear an uncanny resemblance. In this work, we present an analysis-design methodology that both explains and exploits that resemblance. Specifically, we present a two-step method to convert a small-step concrete semantics into a family of sound, computable abstract interpretations. The first step re-factors the concrete state-space to eliminate recursive structure; this refactoring of the state-space simultaneously determines a store-passing-style transformation on the underlying concrete semantics. The second step uses inference rules to generate an abstract state-space and a Galois connection simultaneously. The Galois connection allows the calculation of the "optimal" abstract interpretation. The two-step process is unambiguous, but nondeterministic: at each step, analysis designers face choices. Some of these choices ultimately influence properties such as flow-, field- and context-sensitivity. Thus, under the method, we can give the emergence of these properties a graph-theoretic characterization. To illustrate the method, we systematically abstract the continuation-passing style lambda calculus to arrive at two distinct families of analyses. The first is the well-known k-CFA family of analyses. The second consists of novel "environment-centric" abstract interpretations, none of which appear in the literature on static analysis of higher-order programs.

  16. 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.

  17. Abstracts of SIG Sessions.

    ERIC Educational Resources Information Center

    Proceedings of the ASIS Annual Meeting, 1997

    1997-01-01

    Presents abstracts of SIG Sessions. Highlights include digital collections; information retrieval methods; public interest/fair use; classification and indexing; electronic publication; funding; globalization; information technology projects; interface design; networking in developing countries; metadata; multilingual databases; networked…

  18. Automatic Abstraction in Planning

    NASA Technical Reports Server (NTRS)

    Christensen, J.

    1991-01-01

    Traditionally, abstraction in planning has been accomplished by either state abstraction or operator abstraction, neither of which has been fully automatic. We present a new method, predicate relaxation, for automatically performing state abstraction. PABLO, a nonlinear hierarchical planner, implements predicate relaxation. Theoretical, as well as empirical results are presented which demonstrate the potential advantages of using predicate relaxation in planning. We also present a new definition of hierarchical operators that allows us to guarantee a limited form of completeness. This new definition is shown to be, in some ways, more flexible than previous definitions of hierarchical operators. Finally, a Classical Truth Criterion is presented that is proven to be sound and complete for a planning formalism that is general enough to include most classical planning formalisms that are based on the STRIPS assumption.

  19. 1971 Annual Conference Abstracts

    ERIC Educational Resources Information Center

    Journal of Engineering Education, 1971

    1971-01-01

    Included are 112 abstracts listed under headings such as: acoustics, continuing engineering studies, educational research and methods, engineering design, libraries, liberal studies, and materials. Other areas include agricultural, electrical, mechanical, mineral, and ocean engineering. (TS)

  20. 2016 ACPA MEETING ABSTRACTS.

    PubMed

    2016-07-01

    The peer-reviewed abstracts presented at the 73rd Annual Meeting of the ACPA are published as submitted by the authors. For financial conflict of interest disclosure, please visit http://meeting.acpa-cpf.org/disclosures.html. PMID:27447885

  1. Abstracts of contributed papers

    SciTech Connect

    Not Available

    1994-08-01

    This volume contains 571 abstracts of contributed papers to be presented during the Twelfth US National Congress of Applied Mechanics. Abstracts are arranged in the order in which they fall in the program -- the main sessions are listed chronologically in the Table of Contents. The Author Index is in alphabetical order and lists each paper number (matching the schedule in the Final Program) with its corresponding page number in the book.

  2. RXTE Observations of Cas A

    NASA Technical Reports Server (NTRS)

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

    1997-01-01

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

  3. Metacognition and abstract reasoning.

    PubMed

    Markovits, Henry; Thompson, Valerie A; Brisson, Janie

    2015-05-01

    The nature of people's meta-representations of deductive reasoning is critical to understanding how people control their own reasoning processes. We conducted two studies to examine whether people have a metacognitive representation of abstract validity and whether familiarity alone acts as a separate metacognitive cue. In Study 1, participants were asked to make a series of (1) abstract conditional inferences, (2) concrete conditional inferences with premises having many potential alternative antecedents and thus specifically conducive to the production of responses consistent with conditional logic, or (3) concrete problems with premises having relatively few potential alternative antecedents. Participants gave confidence ratings after each inference. Results show that confidence ratings were positively correlated with logical performance on abstract problems and concrete problems with many potential alternatives, but not with concrete problems with content less conducive to normative responses. Confidence ratings were higher with few alternatives than for abstract content. Study 2 used a generation of contrary-to-fact alternatives task to improve levels of abstract logical performance. The resulting increase in logical performance was mirrored by increases in mean confidence ratings. Results provide evidence for a metacognitive representation based on logical validity, and show that familiarity acts as a separate metacognitive cue. PMID:25416026

  4. 48 CFR 9903.201-2 - Types of CAS coverage.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

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

  5. 48 CFR 9903.201-2 - Types of CAS coverage.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

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

  6. 48 CFR 9903.201-2 - Types of CAS coverage.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

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

  7. 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…

  8. 48 CFR 9903.201-2 - Types of CAS coverage.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

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

  9. Thyra Abstract Interface Package

    2005-09-01

    Thrya primarily defines a set of abstract C++ class interfaces needed for the development of abstract numerical atgorithms (ANAs) such as iterative linear solvers, transient solvers all the way up to optimization. At the foundation of these interfaces are abstract C++ classes for vectors, vector spaces, linear operators and multi-vectors. Also included in the Thyra package is C++ code for creating concrete vector, vector space, linear operator, and multi-vector subclasses as well as other utilitiesmore » to aid in the development of ANAs. Currently, very general and efficient concrete subclass implementations exist for serial and SPMD in-core vectors and multi-vectors. Code also currently exists for testing objects and providing composite objects such as product vectors.« less

  10. Overlap in the Lists of Journals Monitored by Biosis, CAS and Ei

    ERIC Educational Resources Information Center

    Wood, James L.; And Others

    1972-01-01

    This study identifies the amount of duplication among the lists of primary journals regularly monitored by the BioSciences Information Service of Biological Abstracts, the Chemical Abstracts Service and Engineering Index, Inc. (4 references) (SJ)

  11. In vitro and in vivo growth suppression of human papillomavirus 16-positive cervical cancer cells by CRISPR/Cas9

    SciTech Connect

    Zhen, Shuai; Hua, Ling; Takahashi, Y.; Narita, S.; Liu, Yun-Hui; Li, Yan

    2014-08-08

    Highlights: • Established CRISPR/Cas9 targeting promoter of HPV 16 and targeting E6, E7 transcript. • CRISPR/Cas9 resulted in accumulation of p53 and p21, reduced the proliferation of cervical cancer cells. • Finding inhibited tumorigenesis and growth of mice incubated by cells with CRISPR/Cas9. • CRISPR/Cas9 will be a new treatment strategy, in cervical and other HPV-associated cancer therapy. - Abstract: Deregulated expression of high-risk human papillomavirus oncogenes (E6 and E7) is a pivotal event for pathogenesis and progression in cervical cancer. Both viral oncogenes are therefore regarded as ideal therapeutic targets. In the hope of developing a gene-specific therapy for HPV-related cancer, we established CRISPR/Cas9 targeting promoter of HPV 16 E6/E7 and targeting E6, E7 transcript, transduced the CRISPR/Cas9 into cervical HPV-16-positive cell line SiHa. The results showed that CRISPR/Cas9 targeting promoter, as well as targeting E6 and E7 resulted in accumulation of p53 and p21 protein, and consequently remarkably reduced the abilities of proliferation of cervical cancer cells in vitro. Then we inoculated subcutaneously cells into nude mice to establish the transplanted tumor animal models, and found dramatically inhibited tumorigenesis and growth of mice incubated by cells with CRISPR/Cas9 targeting (promoter+E6+E7)-transcript. Our results may provide evidence for application of CRISPR/Cas9 targeting HR-HPV key oncogenes, as a new treatment strategy, in cervical and other HPV-associated cancer therapy.

  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. Abstraction and art.

    PubMed Central

    Gortais, Bernard

    2003-01-01

    In a given social context, artistic creation comprises a set of processes, which relate to the activity of the artist and the activity of the spectator. Through these processes we see and understand that the world is vaster than it is said to be. Artistic processes are mediated experiences that open up the world. A successful work of art expresses a reality beyond actual reality: it suggests an unknown world using the means and the signs of the known world. Artistic practices incorporate the means of creation developed by science and technology and change forms as they change. Artists and the public follow different processes of abstraction at different levels, in the definition of the means of creation, of representation and of perception of a work of art. This paper examines how the processes of abstraction are used within the framework of the visual arts and abstract painting, which appeared during a period of growing importance for the processes of abstraction in science and technology, at the beginning of the twentieth century. The development of digital platforms and new man-machine interfaces allow multimedia creations. This is performed under the constraint of phases of multidisciplinary conceptualization using generic representation languages, which tend to abolish traditional frontiers between the arts: visual arts, drama, dance and music. PMID:12903659

  14. The SIDdatagrabber (Abstract)

    NASA Astrophysics Data System (ADS)

    Silvis, G.

    2015-12-01

    (Abstract only) The Stanford/SARA SuperSid project offers an opportunity for adding data to the AAVSO SID Monitoring project. You can now build a SID antenna and monitoring setup for about $150. And with the SIDdatagrabber application you can easily re-purpose the data collected for the AAVSO.

  15. Making the Abstract Concrete

    ERIC Educational Resources Information Center

    Potter, Lee Ann

    2005-01-01

    President Ronald Reagan nominated a woman to serve on the United States Supreme Court. He did so through a single-page form letter, completed in part by hand and in part by typewriter, announcing Sandra Day O'Connor as his nominee. While the document serves as evidence of a historic event, it is also a tangible illustration of abstract concepts…

  16. Learning Abstracts, 2001.

    ERIC Educational Resources Information Center

    Wilson, Cynthia, Ed.

    2001-01-01

    Volume 4 of the League for Innovation in the Community College's Learning Abstracts include the following: (1) "Touching Students in the Digital Age: The Move Toward Learner Relationship Management (LRM)," by Mark David Milliron, which offers an overview of an organizing concept to help community colleges navigate the intersection between digital…

  17. Leadership Abstracts, 2002.

    ERIC Educational Resources Information Center

    Wilson, Cynthia, Ed.; Milliron, Mark David, Ed.

    2002-01-01

    This 2002 volume of Leadership Abstracts contains issue numbers 1-12. Articles include: (1) "Skills Certification and Workforce Development: Partnering with Industry and Ourselves," by Jeffrey A. Cantor; (2) "Starting Again: The Brookhaven Success College," by Alice W. Villadsen; (3) "From Digital Divide to Digital Democracy," by Gerardo E. de los…

  18. Leadership Abstracts, 1993.

    ERIC Educational Resources Information Center

    Doucette, Don, Ed.

    1993-01-01

    This document includes 10 issues of Leadership Abstracts (volume 6, 1993), a newsletter published by the League for Innovation in the Community College (California). The featured articles are: (1) "Reinventing Government" by David T. Osborne; (2) "Community College Workforce Training Programs: Expanding the Mission to Meet Critical Needs" by…

  19. Abstraction through Game Play

    ERIC Educational Resources Information Center

    Avraamidou, Antri; Monaghan, John; Walker, Aisha

    2012-01-01

    This paper examines the computer game play of an 11-year-old boy. In the course of building a virtual house he developed and used, without assistance, an artefact and an accompanying strategy to ensure that his house was symmetric. We argue that the creation and use of this artefact-strategy is a mathematical abstraction. The discussion…

  20. CIRF Abstracts, Volume 12.

    ERIC Educational Resources Information Center

    International Labour Office, Geneva (Switzerland).

    The aim of the CIRF abstracts is to convey information about vocational training ideas, programs, experience, and experiments described in periodicals, books, and other publications and relating to operative personnel, supervisors, and technical and training staff in all sectors of economic activity. Information is also given on major trends in…

  1. Leadership Abstracts, 1999.

    ERIC Educational Resources Information Center

    Leadership Abstracts, 1999

    1999-01-01

    This document contains five Leadership Abstracts publications published February-December 1999. The article, "Teaching the Teachers: Meeting the National Teacher Preparation Challenge," authored by George R. Boggs and Sadie Bragg, examines the community college role and makes recommendations and a call to action for teacher education. "Chaos…

  2. Double Trouble (Abstract)

    NASA Astrophysics Data System (ADS)

    Simonsen, M.

    2015-12-01

    (Abstract only) Variable stars with close companions can be difficult to accurately measure and characterize. The companions can create misidentifications, which in turn can affect the perceived magnitudes, amplitudes, periods, and colors of the variable stars. We will show examples of these Double Trouble stars and the impact their close companions have had on our understanding of some of these variable stars.

  3. Send Me No Abstract.

    ERIC Educational Resources Information Center

    Levy, Steven

    1985-01-01

    Discusses Magazine Index's practice of assigning letter grades (sometimes inaccurate) to book, restaurant, and movie reviews, thus allowing patrons to get the point of the review from the index rather than the article itself, and argues that this situation is indicative of the larger problem of reliability of abstracts. (MBR)

  4. Annual Conference Abstracts

    ERIC Educational Resources Information Center

    Engineering Education, 1976

    1976-01-01

    Presents the abstracts of 158 papers presented at the American Society for Engineering Education's annual conference at Knoxville, Tennessee, June 14-17, 1976. Included are engineering topics covering education, aerospace, agriculture, biomedicine, chemistry, computers, electricity, acoustics, environment, mechanics, and women. (SL)

  5. Water reuse. [Lead abstract

    SciTech Connect

    Middlebrooks, E.J.

    1982-01-01

    Separate abstracts were prepared for the 31 chapters of this book which deals with all aspects of wastewater reuse. Design data, case histories, performance data, monitoring information, health information, social implications, legal and organizational structures, and background information needed to analyze the desirability of water reuse are presented. (KRM)

  6. Reasoning abstractly about resources

    NASA Technical Reports Server (NTRS)

    Clement, B.; Barrett, A.

    2001-01-01

    r describes a way to schedule high level activities before distributing them across multiple rovers in order to coordinate the resultant use of shared resources regardless of how each rover decides how to perform its activities. We present an algorithm for summarizing the metric resource requirements of an abstract activity based n the resource usages of its potential refinements.

  7. Humor, abstraction, and disbelief.

    PubMed

    Hoicka, Elena; Jutsum, Sarah; Gattis, Merideth

    2008-09-01

    We investigated humor as a context for learning about abstraction and disbelief. More specifically, we investigated how parents support humor understanding during book sharing with their toddlers. In Study 1, a corpus analysis revealed that in books aimed at 1-to 2-year-olds, humor is found more often than other forms of doing the wrong thing including mistakes, pretense, lying, false beliefs, and metaphors. In Study 2, 20 parents read a book containing humorous and non-humorous pages to their 19-to 26-month-olds. Parents used a significantly higher percentage of high abstraction extra-textual utterances (ETUs) when reading the humorous pages. In Study 3, 41 parents read either a humorous or non-humorous book to their 18-to 24-month-olds. Parents reading the humorous book made significantly more ETUs coded for a specific form of high abstraction: those encouraging disbelief of prior utterances. Sharing humorous books thus increases toddlers' exposure to high abstraction and belief-based language. PMID:21585438

  8. 2002 NASPSA Conference Abstracts.

    ERIC Educational Resources Information Center

    Journal of Sport & Exercise Psychology, 2002

    2002-01-01

    Contains abstracts from the 2002 conference of the North American Society for the Psychology of Sport and Physical Activity. The publication is divided into three sections: the preconference workshop, "Effective Teaching Methods in the Classroom;" symposia (motor development, motor learning and control, and sport psychology); and free…

  9. Annual Conference Abstracts

    ERIC Educational Resources Information Center

    Journal of Engineering Education, 1972

    1972-01-01

    Includes abstracts of papers presented at the 80th Annual Conference of the American Society for Engineering Education. The broad areas include aerospace, affiliate and associate member council, agricultural engineering, biomedical engineering, continuing engineering studies, chemical engineering, civil engineering, computers, cooperative…

  10. Learning Abstracts, 1999.

    ERIC Educational Resources Information Center

    League for Innovation in the Community Coll.

    This document contains volume two of Learning Abstracts, a bimonthly newsletter from the League for Innovation in the Community College. Articles in these seven issues include: (1) "Get on the Fast Track to Learning: An Accelerated Associate Degree Option" (Gerardo E. de los Santos and Deborah J. Cruise); (2) "The Learning College: Both Learner…

  11. Computers in Abstract Algebra

    ERIC Educational Resources Information Center

    Nwabueze, Kenneth K.

    2004-01-01

    The current emphasis on flexible modes of mathematics delivery involving new information and communication technology (ICT) at the university level is perhaps a reaction to the recent change in the objectives of education. Abstract algebra seems to be one area of mathematics virtually crying out for computer instructional support because of the…

  12. Abstract Film and Beyond.

    ERIC Educational Resources Information Center

    Le Grice, Malcolm

    A theoretical and historical account of the main preoccupations of makers of abstract films is presented in this book. The book's scope includes discussion of nonrepresentational forms as well as examination of experiments in the manipulation of time in films. The ten chapters discuss the following topics: art and cinematography, the first…

  13. 78 FR 24207 - Request for Information About Diethanolamine (CAS No. 111-42-2)

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-04-24

    ... From the Federal Register Online via the Government Publishing Office ] DEPARTMENT OF HEALTH AND HUMAN SERVICES Centers for Disease Control and Prevention Request for Information About Diethanolamine (CAS No. 111-42-2) AGENCY: National Institute for Occupational Safety and Health (NIOSH) of the...

  14. V723 Cas a borderline classical nova

    NASA Astrophysics Data System (ADS)

    Friedjung, M.; Iijima, T.

    2002-11-01

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

  15. Characterization of Cas9-Guide RNA Orthologs.

    PubMed

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

    2016-01-01

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

  16. Historical development of abstracting.

    PubMed

    Skolnik, H

    1979-11-01

    The abstract, under a multitude of names, such as hypothesis, marginalia, abridgement, extract, digest, précis, resumé, and summary, has a long history, one which is concomitant with advancing scholarship. The progression of this history from the Sumerian civilization ca. 3600 B.C., through the Egyptian and Greek civilizations, the Hellenistic period, the Dark Ages, Middle Ages, Renaissance, and into the modern period is reviewed. PMID:399482

  17. Generalized Abstract Symbolic Summaries

    NASA Technical Reports Server (NTRS)

    Person, Suzette; Dwyer, Matthew B.

    2009-01-01

    Current techniques for validating and verifying program changes often consider the entire program, even for small changes, leading to enormous V&V costs over a program s lifetime. This is due, in large part, to the use of syntactic program techniques which are necessarily imprecise. Building on recent advances in symbolic execution of heap manipulating programs, in this paper, we develop techniques for performing abstract semantic differencing of program behaviors that offer the potential for improved precision.

  18. EBS Radionuclide Transport Abstraction

    SciTech Connect

    J. Prouty

    2006-07-14

    The purpose of this report is to develop and analyze the engineered barrier system (EBS) radionuclide transport abstraction model, consistent with Level I and Level II model validation, as identified in Technical Work Plan for: Near-Field Environment and Transport: Engineered Barrier System: Radionuclide Transport Abstraction Model Report Integration (BSC 2005 [DIRS 173617]). The EBS radionuclide transport abstraction (or EBS RT Abstraction) is the conceptual model used in the total system performance assessment (TSPA) to determine the rate of radionuclide releases from the EBS to the unsaturated zone (UZ). The EBS RT Abstraction conceptual model consists of two main components: a flow model and a transport model. Both models are developed mathematically from first principles in order to show explicitly what assumptions, simplifications, and approximations are incorporated into the models used in the TSPA. The flow model defines the pathways for water flow in the EBS and specifies how the flow rate is computed in each pathway. Input to this model includes the seepage flux into a drift. The seepage flux is potentially split by the drip shield, with some (or all) of the flux being diverted by the drip shield and some passing through breaches in the drip shield that might result from corrosion or seismic damage. The flux through drip shield breaches is potentially split by the waste package, with some (or all) of the flux being diverted by the waste package and some passing through waste package breaches that might result from corrosion or seismic damage. Neither the drip shield nor the waste package survives an igneous intrusion, so the flux splitting submodel is not used in the igneous scenario class. The flow model is validated in an independent model validation technical review. The drip shield and waste package flux splitting algorithms are developed and validated using experimental data. The transport model considers advective transport and diffusive transport

  19. CRISPR-Cas9-Mediated Single-Gene and Gene Family Disruption in Trypanosoma cruzi

    PubMed Central

    Peng, Duo; Kurup, Samarchith P.; Yao, Phil Y.; Minning, Todd A.

    2014-01-01

    ABSTRACT Trypanosoma cruzi is a protozoan parasite of humans and animals, affecting 10 to 20 million people and innumerable animals, primarily in the Americas. Despite being the largest cause of infection-induced heart disease worldwide, even among the neglected tropical diseases (NTDs) T. cruzi is considered one of the least well understood and understudied. The genetic complexity of T. cruzi as well as the limited set of efficient techniques for genome engineering contribute significantly to the relative lack of progress in and understanding of this pathogen. Here, we adapted the CRISPR-Cas9 system for the genetic engineering of T. cruzi, demonstrating rapid and efficient knockout of multiple endogenous genes, including essential genes. We observed that in the absence of a template, repair of the Cas9-induced double-stranded breaks (DSBs) in T. cruzi occurs exclusively by microhomology-mediated end joining (MMEJ) with various-sized deletions. When a template for DNA repair is provided, DSB repair by homologous recombination is achieved at an efficiency several orders of magnitude higher than that in the absence of CRISPR-Cas9-induced DSBs. We also demonstrate the high multiplexing capacity of CRISPR-Cas9 in T. cruzi by knocking down expression of an enzyme gene family consisting of 65 members, resulting in a significant reduction of enzymatic product with no apparent off-target mutations. Lastly, we show that Cas9 can mediate disruption of its own coding sequence, rescuing a growth defect in stable Cas9-expressing parasites. These results establish a powerful new tool for the analysis of gene functions in T. cruzi, enabling the study of essential genes and their functions and analysis of the many large families of related genes that occupy a substantial portion of the T. cruzi genome. PMID:25550322

  20. EBS Radionuclide Transport Abstraction

    SciTech Connect

    J.D. Schreiber

    2005-08-25

    The purpose of this report is to develop and analyze the engineered barrier system (EBS) radionuclide transport abstraction model, consistent with Level I and Level II model validation, as identified in ''Technical Work Plan for: Near-Field Environment and Transport: Engineered Barrier System: Radionuclide Transport Abstraction Model Report Integration'' (BSC 2005 [DIRS 173617]). The EBS radionuclide transport abstraction (or EBS RT Abstraction) is the conceptual model used in the total system performance assessment for the license application (TSPA-LA) to determine the rate of radionuclide releases from the EBS to the unsaturated zone (UZ). The EBS RT Abstraction conceptual model consists of two main components: a flow model and a transport model. Both models are developed mathematically from first principles in order to show explicitly what assumptions, simplifications, and approximations are incorporated into the models used in the TSPA-LA. The flow model defines the pathways for water flow in the EBS and specifies how the flow rate is computed in each pathway. Input to this model includes the seepage flux into a drift. The seepage flux is potentially split by the drip shield, with some (or all) of the flux being diverted by the drip shield and some passing through breaches in the drip shield that might result from corrosion or seismic damage. The flux through drip shield breaches is potentially split by the waste package, with some (or all) of the flux being diverted by the waste package and some passing through waste package breaches that might result from corrosion or seismic damage. Neither the drip shield nor the waste package survives an igneous intrusion, so the flux splitting submodel is not used in the igneous scenario class. The flow model is validated in an independent model validation technical review. The drip shield and waste package flux splitting algorithms are developed and validated using experimental data. The transport model considers

  1. A LARI Experience (Abstract)

    NASA Astrophysics Data System (ADS)

    Cook, M.

    2015-12-01

    (Abstract only) In 2012, Lowell Observatory launched The Lowell Amateur Research Initiative (LARI) to formally involve amateur astronomers in scientific research by bringing them to the attention of and helping professional astronomers with their astronomical research. One of the LARI projects is the BVRI photometric monitoring of Young Stellar Objects (YSOs), wherein amateurs obtain observations to search for new outburst events and characterize the colour evolution of previously identified outbursters. A summary of the scientific and organizational aspects of this LARI project, including its goals and science motivation, the process for getting involved with the project, a description of the team members, their equipment and methods of collaboration, and an overview of the programme stars, preliminary findings, and lessons learned is presented.

  2. IEEE conference record -- Abstracts

    SciTech Connect

    Not Available

    1994-01-01

    This conference covers the following areas: computational plasma physics; vacuum electronic; basic phenomena in fully ionized plasmas; plasma, electron, and ion sources; environmental/energy issues in plasma science; space plasmas; plasma processing; ball lightning/spherical plasma configurations; plasma processing; fast wave devices; magnetic fusion; basic phenomena in partially ionized plasma; dense plasma focus; plasma diagnostics; basic phenomena in weakly ionized gases; fast opening switches; MHD; fast z-pinches and x-ray lasers; intense ion and electron beams; laser-produced plasmas; microwave plasma interactions; EM and ETH launchers; solid state plasmas and switches; intense beam microwaves; and plasmas for lighting. Separate abstracts were prepared for 416 papers in this conference.

  3. Les tumeurs conjonctives cutanées: à propos de 121 cas

    PubMed Central

    Hazmiri, Fatima Ezzahra; Fakhri, Anas; Rais, Hanane; Akhdari, Nadia; Amal, Said; Belaabidia, Badia

    2014-01-01

    Les tumeurs conjonctives cutanées sont des tumeurs dermiques et/ou hypodermiques relativement fréquentes. Elles sont dominées par les tumeurs bénignes. A travers une série de 121 cas, nous avons étudié le profil épidémiologique, anatomopathologique et évolutif de ces tumeurs. C'est une étude rétrospective réalisée au service d'anatomie pathologique du CHU Mohammed VI de Marrakech entre 2004 et 2012. Il s'agit de 121 patients. La moyenne d’âge était de 36 ans (1-80ans). Le sex-ratio H/F était de 1,12. La tumeur avait un aspect nodulaire dans 90% des cas. Le membre inférieur était la localisation la plus fréquente (30,5%). L’étude anatomopathologique a porté sur un matériel biopsique dans 100% des cas. Soixante-neuf pour cent de ces tumeurs étaient bénignes. Elles étaient représentées essentiellement par les tumeurs vasculaires, suivies par les tumeurs fibreuses et fibro-histiocytaires. Trente et un pour cent des tumeurs étaient malignes. Il s'agissait essentiellement de tumeurs fibreuses et fibro-histiocytaires, suivies de tumeurs vasculaires. L’étude immunohistochimique était réalisée dans 2cas. Le traitement chirurgical était entrepris dans 73% des cas. L’évolution était précisée dans 19% des cas avec une évolution favorable dans 13% des cas. Un cas de décès et 2 cas de récidive étaient notés. Les tumeurs conjonctives cutanées bénignes sont de bon pronostic, mais posent un problème majeur de nosologie et de classification. D'autre part, la prise en charge diagnostique et thérapeutique ainsi que l’évaluation pronostique des sarcomes cutanés restent difficiles. PMID:25379113

  4. Teaching for Abstraction: A Model

    ERIC Educational Resources Information Center

    White, Paul; Mitchelmore, Michael C.

    2010-01-01

    This article outlines a theoretical model for teaching elementary mathematical concepts that we have developed over the past 10 years. We begin with general ideas about the abstraction process and differentiate between "abstract-general" and "abstract-apart" concepts. A 4-phase model of teaching, called Teaching for Abstraction, is then proposed…

  5. 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....

  6. 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....

  7. 48 CFR 970.3002 - CAS program requirements.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

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

  8. 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....

  9. 48 CFR 970.3002 - CAS program requirements.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

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

  10. 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

  11. 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

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

    PubMed

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

    2015-07-23

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

  13. Automated Supernova Discovery (Abstract)

    NASA Astrophysics Data System (ADS)

    Post, R. S.

    2015-12-01

    (Abstract only) We are developing a system of robotic telescopes for automatic recognition of Supernovas as well as other transient events in collaboration with the Puckett Supernova Search Team. At the SAS2014 meeting, the discovery program, SNARE, was first described. Since then, it has been continuously improved to handle searches under a wide variety of atmospheric conditions. Currently, two telescopes are used to build a reference library while searching for PSN with a partial library. Since data is taken every night without clouds, we must deal with varying atmospheric and high background illumination from the moon. Software is configured to identify a PSN, reshoot for verification with options to change the run plan to acquire photometric or spectrographic data. The telescopes are 24-inch CDK24, with Alta U230 cameras, one in CA and one in NM. Images and run plans are sent between sites so the CA telescope can search while photometry is done in NM. Our goal is to find bright PSNs with magnitude 17.5 or less which is the limit of our planned spectroscopy. We present results from our first automated PSN discoveries and plans for PSN data acquisition.

  14. Stellar Presentations (Abstract)

    NASA Astrophysics Data System (ADS)

    Young, D.

    2015-12-01

    (Abstract only) The AAVSO is in the process of expanding its education, outreach and speakers bureau program. powerpoint presentations prepared for specific target audiences such as AAVSO members, educators, students, the general public, and Science Olympiad teams, coaches, event supervisors, and state directors will be available online for members to use. The presentations range from specific and general content relating to stellar evolution and variable stars to specific activities for a workshop environment. A presentation—even with a general topic—that works for high school students will not work for educators, Science Olympiad teams, or the general public. Each audience is unique and requires a different approach. The current environment necessitates presentations that are captivating for a younger generation that is embedded in a highly visual and sound-bite world of social media, twitter and U-Tube, and mobile devices. For educators, presentations and workshops for themselves and their students must support the Next Generation Science Standards (NGSS), the Common Core Content Standards, and the Science Technology, Engineering and Mathematics (STEM) initiative. Current best practices for developing relevant and engaging powerpoint presentations to deliver information to a variety of targeted audiences will be presented along with several examples.

  15. Covalent Modification of Bacteriophage T4 DNA Inhibits CRISPR-Cas9

    PubMed Central

    Bryson, Alexandra L.; Hwang, Young; Sherrill-Mix, Scott; Wu, Gary D.; Lewis, James D.; Black, Lindsay; Clark, Tyson A.

    2015-01-01

    ABSTRACT The genomic DNAs of tailed bacteriophages are commonly modified by the attachment of chemical groups. Some forms of DNA modification are known to protect phage DNA from cleavage by restriction enzymes, but others are of unknown function. Recently, the CRISPR-Cas nuclease complexes were shown to mediate bacterial adaptive immunity by RNA-guided target recognition, raising the question of whether phage DNA modifications may also block attack by CRISPR-Cas9. We investigated phage T4 as a model system, where cytosine is replaced with glucosyl-hydroxymethylcytosine (glc-HMC). We first quantified the extent and distribution of covalent modifications in T4 DNA by single-molecule DNA sequencing and enzymatic probing. We then designed CRISPR spacer sequences targeting T4 and found that wild-type T4 containing glc-HMC was insensitive to attack by CRISPR-Cas9 but mutants with unmodified cytosine were sensitive. Phage with HMC showed only intermediate sensitivity. While this work was in progress, another group reported examples of heavily engineered CRISRP-Cas9 complexes that could, in fact, overcome the effects of T4 DNA modification, indicating that modifications can inhibit but do not always fully block attack. PMID:26081634

  16. Function of the CRISPR-Cas System of the Human Pathogen Clostridium difficile

    PubMed Central

    Boudry, Pierre; Semenova, Ekaterina; Monot, Marc; Datsenko, Kirill A.; Lopatina, Anna; Sekulovic, Ognjen; Ospina-Bedoya, Maicol; Fortier, Louis-Charles; Severinov, Konstantin; Dupuy, Bruno

    2015-01-01

    ABSTRACT Clostridium difficile is the cause of most frequently occurring nosocomial diarrhea worldwide. As an enteropathogen, C. difficile must be exposed to multiple exogenous genetic elements in bacteriophage-rich gut communities. CRISPR (clustered regularly interspaced short palindromic repeats)-Cas (CRISPR-associated) systems allow bacteria to adapt to foreign genetic invaders. Our recent data revealed active expression and processing of CRISPR RNAs from multiple type I-B CRISPR arrays in C. difficile reference strain 630. Here, we demonstrate active expression of CRISPR arrays in strain R20291, an epidemic C. difficile strain. Through genome sequencing and host range analysis of several new C. difficile phages and plasmid conjugation experiments, we provide evidence of defensive function of the CRISPR-Cas system in both C. difficile strains. We further demonstrate that C. difficile Cas proteins are capable of interference in a heterologous host, Escherichia coli. These data set the stage for mechanistic and physiological analyses of CRISPR-Cas-mediated interactions of important global human pathogen with its genetic parasites. PMID:26330515

  17. 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....

  18. 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....

  19. 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....

  20. 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....

  1. 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...

  2. 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....

  3. 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...

  4. 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....

  5. 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....

  6. 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...

  7. 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....

  8. 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...

  9. 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....

  10. 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....

  11. Abstraction of Drift Seepage

    SciTech Connect

    J.T. Birkholzer

    2004-11-01

    This model report documents the abstraction of drift seepage, conducted to provide seepage-relevant parameters and their probability distributions for use in Total System Performance Assessment for License Application (TSPA-LA). Drift seepage refers to the flow of liquid water into waste emplacement drifts. Water that seeps into drifts may contact waste packages and potentially mobilize radionuclides, and may result in advective transport of radionuclides through breached waste packages [''Risk Information to Support Prioritization of Performance Assessment Models'' (BSC 2003 [DIRS 168796], Section 3.3.2)]. The unsaturated rock layers overlying and hosting the repository form a natural barrier that reduces the amount of water entering emplacement drifts by natural subsurface processes. For example, drift seepage is limited by the capillary barrier forming at the drift crown, which decreases or even eliminates water flow from the unsaturated fractured rock into the drift. During the first few hundred years after waste emplacement, when above-boiling rock temperatures will develop as a result of heat generated by the decay of the radioactive waste, vaporization of percolation water is an additional factor limiting seepage. Estimating the effectiveness of these natural barrier capabilities and predicting the amount of seepage into drifts is an important aspect of assessing the performance of the repository. The TSPA-LA therefore includes a seepage component that calculates the amount of seepage into drifts [''Total System Performance Assessment (TSPA) Model/Analysis for the License Application'' (BSC 2004 [DIRS 168504], Section 6.3.3.1)]. The TSPA-LA calculation is performed with a probabilistic approach that accounts for the spatial and temporal variability and inherent uncertainty of seepage-relevant properties and processes. Results are used for subsequent TSPA-LA components that may handle, for example, waste package corrosion or radionuclide transport.

  12. Comparative Study of the Coverage of Physics Journals by Two Computerized Data Bases-SPIN (Searchable and Physics Information Notes) and CAC (Chemical Abstracts Condensates)

    ERIC Educational Resources Information Center

    Jerome, S.

    1973-01-01

    SPIN coverage is abnormally low, only 70 journals, versus the coverage of information services in other fields. Every journal listed by SPIN is selectively covered by CAS, resulting in a 76 percent overlap. (3 references) (Author/SJ)

  13. 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

  14. 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.

  15. 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.

  16. Lessons Learned on Management of CAS Development.

    ERIC Educational Resources Information Center

    Boyadjieff, Kiril

    1995-01-01

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

  17. 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

  18. 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.

  19. Advance Organizers: Concret Versus Abstract.

    ERIC Educational Resources Information Center

    Corkill, Alice J.; And Others

    1988-01-01

    Two experiments examined the relative effects of concrete and abstract advance organizers on students' memory for subsequent prose. Results of the experiments are discussed in terms of the memorability, familiarity, and visualizability of concrete and abstract verbal materials. (JD)

  20. Accepted scientific research works (abstracts).

    PubMed

    2014-01-01

    These are the 39 accepted abstracts for IAYT's Symposium on Yoga Research (SYR) September 24-24, 2014 at the Kripalu Center for Yoga & Health and published in the Final Program Guide and Abstracts. PMID:25645134

  1. In vivo genome editing using Staphylococcus aureus Cas9

    PubMed Central

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

    2015-01-01

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

  2. Structure and Engineering of Francisella novicida Cas9

    PubMed Central

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

    2016-01-01

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

  3. Structure and Engineering of Francisella novicida Cas9.

    PubMed

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

    2016-02-25

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

  4. Photoactivatable CRISPR-Cas9 for optogenetic genome editing.

    PubMed

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

    2015-07-01

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

  5. 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

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

    PubMed

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

    2012-01-01

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

  7. 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

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

    PubMed

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

    2014-01-01

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

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

    PubMed Central

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

    2015-01-01

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

  10. Expanding the catalog of cas genes with metagenomes.

    PubMed

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

    2014-02-01

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

  11. 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

  12. Efficient Mitochondrial Genome Editing by CRISPR/Cas9

    PubMed Central

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

    2015-01-01

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

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

    PubMed

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

    2016-07-01

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

  14. Mechanical Engineering Department technical abstracts

    SciTech Connect

    Denney, R.M.

    1982-07-01

    The Mechanical Engineering Department publishes listings of technical abstracts twice a year to inform readers of the broad range of technical activities in the Department, and to promote an exchange of ideas. Details of the work covered by an abstract may be obtained by contacting the author(s). Overall information about current activities of each of the Department's seven divisions precedes the technical abstracts.

  15. Recursive Abstractions for Parameterized Systems

    NASA Astrophysics Data System (ADS)

    Jaffar, Joxan; Santosa, Andrew E.

    We consider a language of recursively defined formulas about arrays of variables, suitable for specifying safety properties of parameterized systems. We then present an abstract interpretation framework which translates a paramerized system as a symbolic transition system which propagates such formulas as abstractions of underlying concrete states. The main contribution is a proof method for implications between the formulas, which then provides for an implementation of this abstract interpreter.

  16. Un cas de trichobezoard gastrique

    PubMed Central

    Moujahid, Mountassir; Ziadi, Tarik; Ennafae, Issam; Kechna, Hicham; Ouzzad, Omar; El Kandry, Sifeddine

    2011-01-01

    Le trichobézoard est défini par la présence d'un corps étranger intra gastrique formé par des cheveux ou des fibres textiles. C'est une pathologie rare qui survient habituellement chez des adolescents présentant des troubles psychiques. La symptomatologie clinique est très variée et le diagnostic est souvent suspecté à la radiologie et à l'endoscopie. Le traitement est chirurgical associé à une prise en charge psychologique. Nous rapportons l'observation d'une jeune femme de 26 ans, hospitalisée au service pour une masse épigastrique. La fibroscopie oesogastroduodénale a mis en évidence un trichobézoard confirmé par le scanner abdominal. La patiente a été opérée par une gastrostomie, les suites post opératoires étaient simples. Elle a été confiée par la suite à un psychiatre pour suivi thérapeutique. Le trichobézoard est une pathologie rare qui survient habituellement chez des adolescents présentant des troubles psychiques. PMID:22355429

  17. MUNICIPAL WATER POLLUTION CONTROL ABSTRACTS: NOVEMBER 1976-OCTOBER 1977

    EPA Science Inventory

    The Franklin Institute Research Laboratories, Science Information Services Department, prepared for the Environmental Protection Agency Volume 4 of the Municipal Technology Bulletin, a current-awareness abstracting bulletin covering methods of municipal waste water treatment, pro...

  18. MUNICIPAL WATER POLLUTION CONTROL ABSTRACTS: MAY-OCTOBER 1976

    EPA Science Inventory

    The Franklin Institute Research Laboratories, Science Information Services Department prepared for the Environmental Protection Agency, Volume 4 of a monthly current-awareness abstracting bulletin, Municipal Technology Bulletin, which dealt with methods of municipal waste water t...

  19. MUNICIPAL WATER POLLUTION CONTROL ABSTRACTS: APRIL 1975-MARCH 1976

    EPA Science Inventory

    The Franklin Institute Research Laboratories, Science Information Services Department prepared for the Environmental Protection Agency, Volume 3 of a monthly current-awareness abstracting bulletin, Municipal Technology Bulletin, which dealt with methods of municipal waste water t...

  20. Méthotrexate et psoriasis: à propos de 46 cas

    PubMed Central

    Inani, Kawtar; Meziane, Mariame; Mernissi, Fatimazahra

    2014-01-01

    Le psoriasis est une maladie inflammatoire chronique, son traitement peut être local ou général. Le méthotrexate (MTX) est parmi les traitements systémiques du psoriasis modéré à sévère. Le but de notre étude est d’évaluer la place du MTX dans le traitement du psoriasis dans notre contexte marocain. C'est une étude rétrospective menée au service de dermatologie du CHU HASSAN II FES de 2010 à 2013. 46 patients ont répondus aux critères d'inclusions. Il s'agissait de patients de sexe masculin dans 58,7% des cas, de sujets âgés entre 18 et 45 ans dans 45,7% des cas. Le psoriasis vulgaire était la forme la plus répondue (76,1%), 56,5% avaient une surface corporelle(SC) atteinte comprise entre 25 et 50%, L’évolution était marquée par une rémission complète dans 50% des cas. Le MTX a été utilisé depuis plus de 40 ans dans le traitement du psoriasis modéré à sévère. Dans notre série le recours au MTX était nécessaire et ceci après échec d'autres thérapeutiques. Son efficacité a été constatée chez 50% des patients, avec peu d'effets secondaires. Le MTX est une molécule de référence dans le traitement du psoriasis modéré à sévère, avec un meilleur rapport coût/bénéfice/risque. PMID:25709742

  1. Advances in therapeutic CRISPR/Cas9 genome editing.

    PubMed

    Savić, Nataša; Schwank, Gerald

    2016-02-01

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

  2. Inducible in vivo genome editing with CRISPR/Cas9

    PubMed Central

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

    2015-01-01

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

  3. Abstracts

    NASA Astrophysics Data System (ADS)

    2012-09-01

    Measuring cosmological parameters with GRBs: status and perspectives New interpretation of the Amati relation The SED Machine - a dedicated transient spectrograph PTF10iue - evidence for an internal engine in a unique Type Ic SN Direct evidence for the collapsar model of long gamma-ray bursts On pair instability supernovae and gamma-ray bursts Pan-STARRS1 observations of ultraluminous SNe The influence of rotation on the critical neutrino luminosity in core-collapse supernovae General relativistic magnetospheres of slowly rotating and oscillating neutron stars Host galaxies of short GRBs GRB 100418A: a bridge between GRB-associated hypernovae and SNe Two super-luminous SNe at z ~ 1.5 from the SNLS Prospects for very-high-energy gamma-ray bursts with the Cherenkov Telescope Array The dynamics and radiation of relativistic flows from massive stars The search for light echoes from the supernova explosion of 1181 AD The proto-magnetar model for gamma-ray bursts Stellar black holes at the dawn of the universe MAXI J0158-744: the discovery of a supersoft X-ray transient Wide-band spectra of magnetar burst emission Dust formation and evolution in envelope-stripped core-collapse supernovae The host galaxies of dark gamma-ray bursts Keck observations of 150 GRB host galaxies Search for properties of GRBs at large redshift The early emission from SNe Spectral properties of SN shock breakout MAXI observation of GRBs and short X-ray transients A three-dimensional view of SN 1987A using light echo spectroscopy X-ray study of the southern extension of the SNR Puppis A All-sky survey of short X-ray transients by MAXI GSC Development of the CALET gamma-ray burst monitor (CGBM)

  4. Vague Language in Conference Abstracts

    ERIC Educational Resources Information Center

    Cutting, Joan

    2012-01-01

    This study examined abstracts for a British Association for Applied Linguistics conference and a Sociolinguistics Symposium, to define the genre of conference abstracts in terms of vague language, specifically universal general nouns (e.g. people) and research general nouns (e.g. results), and to discover if the language used reflected the level…

  5. Leadership Abstracts; Volume 4, 1991.

    ERIC Educational Resources Information Center

    Doucette, Don, Ed.

    1991-01-01

    "Leadership Abstracts" is published bimonthly and distributed to the chief executive officer of every two-year college in the United States and Canada. This document consists of the 15 one-page abstracts published in 1991. Addressing a variety of topics of interest to the community college administrators, this volume includes: (1) "Delivering the…

  6. Food Science and Technology Abstracts.

    ERIC Educational Resources Information Center

    Cohen, Elinor; Federman, Joan

    1979-01-01

    Introduces the reader to the Food Science and Technology Abstracts, a data file that covers worldwide literature on human food commodities and aspects of food processing. Topics include scope, subject index, thesaurus, searching online, and abstracts; tables provide a comparison of ORBIT and DIALOG versions of the file. (JD)

  7. Innovation Abstracts, Volume XV, 1993.

    ERIC Educational Resources Information Center

    Roueche, Suanne D., Ed.

    1993-01-01

    This volume of 30 one- to two-page abstracts from 1993 highlights a variety of innovative approaches to teaching and learning in the community college. Topics covered in the abstracts include: (1) role-playing to encourage critical thinking; (2) team learning techniques to cultivate business skills; (3) librarian-instructor partnerships to create…

  8. Student Success with Abstract Art

    ERIC Educational Resources Information Center

    Hamidou, Kristine

    2009-01-01

    An abstract art project can be challenging or not, depending on the objectives the teacher sets up. In this article, the author describes an abstract papier-mache project that is a success for all students, and is a versatile project easily manipulated to suit the classroom of any art teacher.

  9. Abstraction in perceptual symbol systems.

    PubMed Central

    Barsalou, Lawrence W

    2003-01-01

    After reviewing six senses of abstraction, this article focuses on abstractions that take the form of summary representations. Three central properties of these abstractions are established: ( i ) type-token interpretation; (ii) structured representation; and (iii) dynamic realization. Traditional theories of representation handle interpretation and structure well but are not sufficiently dynamical. Conversely, connectionist theories are exquisitely dynamic but have problems with structure. Perceptual symbol systems offer an approach that implements all three properties naturally. Within this framework, a loose collection of property and relation simulators develops to represent abstractions. Type-token interpretation results from binding a property simulator to a region of a perceived or simulated category member. Structured representation results from binding a configuration of property and relation simulators to multiple regions in an integrated manner. Dynamic realization results from applying different subsets of property and relation simulators to category members on different occasions. From this standpoint, there are no permanent or complete abstractions of a category in memory. Instead, abstraction is the skill to construct temporary online interpretations of a category's members. Although an infinite number of abstractions are possible, attractors develop for habitual approaches to interpretation. This approach provides new ways of thinking about abstraction phenomena in categorization, inference, background knowledge and learning. PMID:12903648

  10. La tuberculose extra-ganglionnaire primitive de la sphère ORL: à propos de 15 cas

    PubMed Central

    Touati, Mohamed Mliha; Darouassi, Youssef; Chihani, Mehdi; Lakouichmi, Mohammed; Tourabi, Khalid; Ammar, Haddou; Bouaity, Brahim

    2014-01-01

    Les localisations ORL extra ganglionnaires de la tuberculose sont rares. La symptomatologie clinique ainsi que les examens paracliniques sont souvent trompeurs,posant ainsi le problème de diagnostic différentiel avec la pathologie tumorale. Nous rapportons 15 cas de localisations extra ganglionnaires de tuberculose, colligés au service ORL et CCF de l'Hopital Militaire Avicenne de Marrakech colligés entre 2009 et 2013. L’âge moyen de nos patients est de 33 ans. L’étude topographique a montré 6 cas au niveau du cavum, un cas de miliaire tuberculeuse pharyngée, 4 cas laryngés; 2 localisations auriculaires; 1 parotidienne et 1 localisation sous maxillaire. Le diagnostic était anatomopathologiquedans tous les cas. Tous nos patients ont reçu un traitement antituberculeux avec une bonne évolution. Mots-clés: Tuberculose, amygdale, rhinopharynx, larynx, glandes salivaires,Oreille moyenne. PMID:25815100

  11. Evaporated CaS thin films for AC electroluminescence devices

    NASA Astrophysics Data System (ADS)

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

    1985-08-01

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

  12. Technical abstracts: Mechanical engineering, 1990

    SciTech Connect

    Broesius, J.Y.

    1991-03-01

    This document is a compilation of the published, unclassified abstracts produced by mechanical engineers at Lawrence Livermore National Laboratory (LLNL) during the calendar year 1990. Many abstracts summarize work completed and published in report form. These are UCRL-JC series documents, which include the full text of articles to be published in journals and of papers to be presented at meetings, and UCID reports, which are informal documents. Not all UCIDs contain abstracts: short summaries were generated when abstracts were not included. Technical Abstracts also provides descriptions of those documents assigned to the UCRL-MI (miscellaneous) category. These are generally viewgraphs or photographs presented at meetings. An author index is provided at the back of this volume for cross referencing.

  13. Metaphor: Bridging embodiment to abstraction.

    PubMed

    Jamrozik, Anja; McQuire, Marguerite; Cardillo, Eileen R; Chatterjee, Anjan

    2016-08-01

    Embodied cognition accounts posit that concepts are grounded in our sensory and motor systems. An important challenge for these accounts is explaining how abstract concepts, which do not directly call upon sensory or motor information, can be informed by experience. We propose that metaphor is one important vehicle guiding the development and use of abstract concepts. Metaphors allow us to draw on concrete, familiar domains to acquire and reason about abstract concepts. Additionally, repeated metaphoric use drawing on particular aspects of concrete experience can result in the development of new abstract representations. These abstractions, which are derived from embodied experience but lack much of the sensorimotor information associated with it, can then be flexibly applied to understand new situations. PMID:27294425

  14. 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

  15. 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

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

    PubMed

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

    2015-11-26

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

  17. Cas9 as a versatile tool for engineering biology

    PubMed Central

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

    2014-01-01

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

  18. Optical Control of CRISPR/Cas9 Gene Editing

    PubMed Central

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

    2016-01-01

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

  19. NASA Patent Abstracts bibliography: A continuing bibliography. Section 1: Abstracts (supplement 21) Abstracts

    NASA Technical Reports Server (NTRS)

    1982-01-01

    Abstracts are cited for 87 patents and applications introduced into the NASA scientific and technical information system during the period of January 1982 through June 1982. Each entry consists of a citation, an abstract, and in mose cases, a key illustration selected from the patent or patent application.

  20. Les complications tardives de prothèse totale de la hanche: à propos de 42 cas

    PubMed Central

    Azarkane, Mohamed; Boussakri, Hassan; Shimi, Mohamed; Elibrahimi, Abdlehalim; Elmrini, Abdlemeji

    2013-01-01

    L'arthroplastie de la hanche est un moyen fiable dans le traitement des affections de la hanche. En lui rendant sa mobilité sa stabilité et son indolence. Cependant cette chirurgie prothétique expose au risque de la survenue des complications qui peuvent engager le pronostic fonctionnel. Nous avons réalisé une étude rétrospective sur une durée de 8 ans de janvier 2004 au janvier 2012 au service de traumatologie-orthopédie de CHU HASSAN II FEZ. Pendant cette période nous avons opéré 240 patients pour PTH. Après un recul moyen de 5 ans nous avons noté chez 42 (17,4%) patients une complication tardive. Nous noté 13 cas de descellement aseptique soit 5,4%. Cette complication a été survenue dans notre série sur une prothèse cimentée dans 8 cas et non cimentée dans 5 cas. Le traitement que nous avons adopté dans notre série a été une reprise de PTH sans greffe osseuse ni anneau de reconstruction dans 4 cas, reprise avec mise en place d'anneau de Kerboull dans 7 cas et reprise avec greffe osseuse et anneau de kerboull dans 2 cas. Nous avons trouvé 11 cas de sepsis tardive soit 4,6% des cas. Nous avons le diabète comme facteur de risque chez 3 malades. L'agent causal a été staphylococcus épdermidis dans 5 cas, colibacille dans 2 cas et association staphylococcus-BGN dans 1 cas. Les différentes modalités que nous avons utilisé pour traiter l'infection dans notre ont été un lavage simple, système d'irrigation-drainage et réimplantation simple en un seul temps ou en 2 temps avec couverture systématique par une antibiothérapie adaptée selon l'antibiogramme. Nous avons noté également 11 cas de fracture sur PTH intéressant dans tous les cas le fémur, nous avons traité ce type de fracture dans notre série par une tige fémorale prothétique longue dans 4 cas, une plaque vissée cerclée dans 3 cas et cerclage simple dans 4 cas. La consolidation a été obtenue chez 9 patients avec 2 cas de pseudarthrose. Nous avons noté 7 cas de

  1. 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

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

    PubMed

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

    2016-02-01

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

  3. Teaching Abstract Concepts by Metaphor.

    ERIC Educational Resources Information Center

    Sutherland, Judith A.

    2001-01-01

    Defines metaphor and its uses; explains the construction and application of metaphors in nursing education. Describes the transformation of the abstract psychiatric concept of therapeutic milieu into a visual metaphor. (SK)

  4. Deficiencies in structured medical abstracts.

    PubMed

    Froom, P; Froom, J

    1993-07-01

    This study was carried out to determine if the content of structured abstracts conforms with recommendations of the Ad Hoc Working Group for the critical appraisal of the medical literature as adopted by the Annals of Internal Medicine. The study design was a survey. All articles published in Annals of Internal Medicine in 1991, excluding editorials, case-reports, literature reviews, decision analysis, studies in medical education, descriptive studies of clinical and basic phenomena, and papers lacking a structured abstract, were studied. Of a total of 150 articles, 20 were excluded. The abstract and text of each article were assessed for the presence of the following items; patient selection criteria, statements concerning extrapolation of findings, need for further study, and whether or not the information should be used now. Number of refusers, drop outs and reason(s) for drop outs were assessed for intervention and prospective cohort studies only. Deficiencies of assessed items were noted in both abstracts and texts. For abstracts, patient selection criteria, numbers of refusers, number of drop outs and reason(s) for drop outs were reported in 44.6% (58/130), 3.1% (4/130), 16.9% (14/83) and 2.4% (2/83) respectively. These items were reported more frequently in the texts 87.7% (114/130), 9.2% (12/130), 60.2% (50/83) and 37.3% (31/83) respectively (p < 0.05). Statements concerning extrapolation of findings, need for further study and use of information now were also more frequent in texts than abstracts (p < 0.0001). A large number of structured abstracts published in the Annals of Internal Medicine in 1991, lack information recommended by the Ad Hoc Working Group. Our findings should not be extrapolated to other journals requiring structured abstracts. PMID:8326342

  5. Candida albicans Gene Deletion with a Transient CRISPR-Cas9 System

    PubMed Central

    Min, Kyunghun; Ichikawa, Yuichi

    2016-01-01

    ABSTRACT Clustered regularly interspaced short palindromic repeat (CRISPR) and CRISPR-associated gene 9 (CRISPR-Cas9) systems are used for a wide array of genome-editing applications in organisms ranging from fungi to plants and animals. Recently, a CRISPR-Cas9 system has been developed for the diploid fungal pathogen Candida albicans; the system accelerates genetic manipulation dramatically [V. K. Vyas, M. I. Barrasa, and G. R. Fink, Sci Adv 1(3):e1500248, 2015, http://dx.doi.org/10.1126/sciadv.1500248]. We show here that the CRISPR-Cas9 genetic elements can function transiently, without stable integration into the genome, to enable the introduction of a gene deletion construct. We describe a transient CRISPR-Cas9 system for efficient gene deletion in C. albicans. Our observations suggest that there are two mechanisms that lead to homozygous deletions: (i) independent recombination of transforming DNA into each allele and (ii) recombination of transforming DNA into one allele, followed by gene conversion of the second allele. Our approach will streamline gene function analysis in C. albicans, and our results indicate that DNA can function transiently after transformation of this organism. IMPORTANCE The fungus Candida albicans is a major pathogen. Genetic analysis of this organism has revealed determinants of pathogenicity, drug resistance, and other unique biological features, as well as the identities of prospective drug targets. The creation of targeted mutations has been greatly accelerated recently through the implementation of CRISPR genome-editing technology by Vyas et al. [Sci Adv 1(3):e1500248, 2015, http://dx.doi.org/10.1126/sciadv.1500248]. In this study, we find that CRISPR elements can be expressed from genes that are present only transiently, and we develop a transient CRISPR system that further accelerates C. albicans genetic manipulation. PMID:27340698

  6. 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

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

    PubMed

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

    2016-01-01

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

  8. 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…

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

    ERIC Educational Resources Information Center

    Winslow, Carl

    2003-01-01

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

  10. 48 CFR 970.3002-1 - CAS applicability.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

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

  11. 48 CFR 970.3002-1 - CAS applicability.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

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

  12. 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...

  13. 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…

  14. 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...

  15. 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...

  16. 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…

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

    PubMed

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

    2016-04-01

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

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

    PubMed

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

    2016-08-01

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

  19. Recent Progress in CRISPR/Cas9 Technology.

    PubMed

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

    2016-02-20

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

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

    PubMed Central

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

    2015-01-01

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

  1. Conditional Control of CRISPR/Cas9 Function.

    PubMed

    Zhou, Wenyuan; Deiters, Alexander

    2016-04-25

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

  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. Modelling Metamorphism by Abstract Interpretation

    NASA Astrophysics Data System (ADS)

    Dalla Preda, Mila; Giacobazzi, Roberto; Debray, Saumya; Coogan, Kevin; Townsend, Gregg M.

    Metamorphic malware apply semantics-preserving transformations to their own code in order to foil detection systems based on signature matching. In this paper we consider the problem of automatically extract metamorphic signatures from these malware. We introduce a semantics for self-modifying code, later called phase semantics, and prove its correctness by showing that it is an abstract interpretation of the standard trace semantics. Phase semantics precisely models the metamorphic code behavior by providing a set of traces of programs which correspond to the possible evolutions of the metamorphic code during execution. We show that metamorphic signatures can be automatically extracted by abstract interpretation of the phase semantics, and that regular metamorphism can be modelled as finite state automata abstraction of the phase semantics.

  4. Mechanical Engineering Department technical abstracts

    SciTech Connect

    Not Available

    1984-07-01

    The Mechanical Engineering Department publishes abstracts twice a year to inform readers of the broad range of technical activities in the Department, and to promote an exchange of ideas. Details of the work covered by an abstract may be obtained by contacting the author(s). General information about the current role and activities of each of the Department's seven divisions precedes the technical abstracts. Further information about a division's work may be obtained from the division leader, whose name is given at the end of each divisional summary. The Department's seven divisions are as follows: Nuclear Test Engineering Division, Nuclear Explosives Engineering Division, Weapons Engineering Division, Energy Systems Engineering Division, Engineering Sciences Division, Magnetic Fusion Engineering Division and Materials Fabrication Division.

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

    PubMed Central

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

    2015-01-01

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

  6. Abstract communication for coordinated planning

    NASA Technical Reports Server (NTRS)

    Clement, Bradley J.; Durfee, Edmund H.

    2003-01-01

    work offers evidence that distributed planning agents can greatly reduce communication costs by reasoning at abstract levels. While it is intuitive that improved search can reduce communication in such cases, there are other decisions about how to communicate plan information that greatly affect communication costs. This paper identifies cases independent of search where communicating at multiple levels of abstraction can exponentially decrease costs and where it can exponentially add costs. We conclude with a process for determining appropriate levels of communication based on characteristics of the domain.

  7. Complications du diverticule de Meckel (DM) chez l'adulte: à propos de 11 cas au CHU-Yalgado Ouédraogo au Burkina Faso

    PubMed Central

    Ouangré, Edgar; Zida, Maurice; Bazongo, Moussa; Sanou, Adama; Bonkoungou, Gilbert Patindé; Doamba, Rodrigue Namékinsba; Sawadogo, Elie Yamba; Ouédraogo, Sidziguin; Zongo, Nayi; Traore, Si Simon

    2015-01-01

    Le diverticule de Meckel (DM) est la persistance partielle du canal omphalomésentérique. Ses complications sont rares. Le diagnostic est le plus souvent per opératoire. L'objectif a été de décrire les complications du diverticule de Meckel chez l'adulte dans le service de chirurgie générale et digestive du CHU Yalgado Ouédraogo. Il s'est agi d'une étude transversale descriptive sur 10 ans (janvier 2004-décembre 2013) portant sur les dossiers des patients âgés de plus de 15 ans ayant présenté un DM compliqué. Durant la période d’étude, 11 cas ont été colligés. L'incidence annuelle a été de 11 cas. Nous avons noté une prédominance masculine avec un sex-ratio de 4,5. L’âge moyen des patients était de 29,8 ans. Le diagnostic préopératoire a été dans huit cas une occlusion intestinale aiguë, une appendicite aiguë dans deux cas, une péritonite aiguë généralisée dans un cas. Il a été diagnostiqué en peropératoire une occlusion intestinale dans neuf cas; une diverticulite dans un cas et un cas de perforation du DM. Tous les DM avaient été réséqués dont huit résections segmentaires iléales emportant le DM et trois résections cunéiformes. Tous les DM étaient situés à moins d'un mètre de la jonction iléo-caecale. L'histologie réalisée dans deux cas avait conclu à une diverticulite. Les suites opératoires ont été simples dans neuf cas, compliquées dans deux cas dont une éventration et un décès. Les complications du diverticule de Meckel sont rares. Le diagnostic préopératoire est difficile. Le traitement est essentiellement chirurgical. PMID:26958137

  8. Innovation Abstracts, Volume XIX, 1997.

    ERIC Educational Resources Information Center

    Roueche, Suanne D., Ed.

    1997-01-01

    The 52 abstracts in these 29 serial issues describe innovative approaches to teaching and learning in the community college. Sample topics include a checklist for conference presenters, plan to retain students, faculty home page, improvements in writing instruction, cooperative learning, support for high risk students, competitive colleges and the…

  9. Handedness Shapes Children's Abstract Concepts

    ERIC Educational Resources Information Center

    Casasanto, Daniel; Henetz, Tania

    2012-01-01

    Can children's handedness influence how they represent abstract concepts like "kindness" and "intelligence"? Here we show that from an early age, right-handers associate rightward space more strongly with positive ideas and leftward space with negative ideas, but the opposite is true for left-handers. In one experiment, children indicated where on…

  10. Innovation Abstracts, Volume XX, 1998.

    ERIC Educational Resources Information Center

    Roueche, Suanne D., Ed.

    1998-01-01

    The 52 abstracts in these 29 serial issues describe innovative approaches to teaching and learning in the community college. Sample topics include reading motivation, barriers to academic success, the learning environment, writing skills, leadership in the criminal justice profession, role-playing strategies, cooperative education, distance…

  11. Abstract Journal Concept Being Examined

    ERIC Educational Resources Information Center

    Somerville, Brendan F.

    1972-01-01

    In order to control the information explosion, some European chemical groups are studying the idea of abandoning full publication in printed form of all primary journals and, in their place, substituting a new form of abstract journal combined with a microfilm record of full scientific papers. (Author/CP)

  12. Metaphoric Images from Abstract Concepts.

    ERIC Educational Resources Information Center

    Vizmuller-Zocco, Jana

    1992-01-01

    Discusses children's use of metaphors to create meaning, using as an example the pragmatic and "scientific" ways in which preschool children explain thunder and lightning to themselves. Argues that children are being shortchanged by modern scientific notions of abstractness and that they should be encouraged to create their own explanations of…

  13. Abstract Expressionism. Clip and Save.

    ERIC Educational Resources Information Center

    Hubbard, Guy

    2002-01-01

    Provides information on the art movement, Abstract Expressionism, and includes learning activities. Focuses on the artist Jackson Pollock, offering a reproduction of his artwork, "Convergence: Number 10." Includes background information on the life and career of Pollock and a description of the included artwork. (CMK)

  14. ERGONOMICS ABSTRACTS 48347-48982.

    ERIC Educational Resources Information Center

    Ministry of Technology, London (England). Warren Spring Lab.

    IN THIS COLLECTION OF ERGONOMICS ABSTRACTS AND ANNOTATIONS THE FOLLOWING AREAS OF CONCERN ARE REPRESENTED--GENERAL REFERENCES, METHODS, FACILITIES, AND EQUIPMENT RELATING TO ERGONOMICS, SYSTEMS OF MAN AND MACHINES, VISUAL, AUDITORY, AND OTHER SENSORY INPUTS AND PROCESSES (INCLUDING SPEECH AND INTELLIGIBILITY), INPUT CHANNELS, BODY MEASUREMENTS,…

  15. Does "Social Work Abstracts" Work?

    ERIC Educational Resources Information Center

    Holden, Gary; Barker, Kathleen; Covert-Vail, Lucinda; Rosenberg, Gary; Cohen, Stephanie A.

    2008-01-01

    Objective: The current study seeks to provide estimates of the adequacy of journal coverage in the Social Work Abstracts (SWA) database. Method: A total of 23 journals listed in the Journal Citation Reports social work category during the 1997 to 2005 period were selected for study. Issue-level coverage estimates were obtained for SWA and…

  16. The Theatre Audience: An Abstraction.

    ERIC Educational Resources Information Center

    Campbell, Paul Newell

    1981-01-01

    Argues that theater is aimed at and presented to an ideal or abstract audience. Discusses the implications of performing for an actual audience, adaptation to various audiences, and the concept of the audience as an evaluative device. (See CS 705 536.) (JMF)

  17. 1990 National Water Quality Laboratory Services Catalog

    USGS Publications Warehouse

    Pritt, Jeffrey, (Edited By); Jones, Berwyn E.

    1989-01-01

    PREFACE This catalog provides information about analytical services available from the National Water Quality Laboratory (NWQL) to support programs of the Water Resources Division of the U.S. Geological Survey. To assist personnel in the selection of analytical services, the catalog lists cost, sample volume, applicable concentration range, detection level, precision of analysis, and preservation techniques for samples to be submitted for analysis. Prices for services reflect operationa1 costs, the complexity of each analytical procedure, and the costs to ensure analytical quality control. The catalog consists of five parts. Part 1 is a glossary of terminology; Part 2 lists the bottles, containers, solutions, and other materials that are available through the NWQL; Part 3 describes the field processing of samples to be submitted for analysis; Part 4 describes analytical services that are available; and Part 5 contains indices of analytical methodology and Chemical Abstract Services (CAS) numbers. Nomenclature used in the catalog is consistent with WATSTORE and STORET. The user is provided with laboratory codes and schedules that consist of groupings of parameters which are measured together in the NWQL. In cases where more than one analytical range is offered for a single element or compound, different laboratory codes are given. Book 5 of the series 'Techniques of Water Resources Investigations of the U.S. Geological Survey' should be consulted for more information about the analytical procedures included in the tabulations. This catalog supersedes U.S. Geological Survey Open-File Report 86-232 '1986-87-88 National Water Quality Laboratory Services Catalog', October 1985.

  18. CRISPR/Cas9 in Genome Editing and Beyond.

    PubMed

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

    2016-06-01

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

  19. 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

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

    PubMed

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

    2016-07-15

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

  1. Etiologies des hypertensions artérielles endocrines: à propos d'une série de cas

    PubMed Central

    Bouznad, Naima; El Mghari, Ghizlane; El Ansari, Nawal

    2016-01-01

    Les hypertensions artérielles (HTA) d'origine endocrine restent une cause rare d'HTA, sa prévalence globale n'excède pas 4% des hypertendus. L'intérêt de la recherche des HTA endocrines réside dans la gravité de certaines formes parfois mortelles et le caractère potentiellement curable et réversible de ces HTA. Le but du travail est de déterminer le profil clinique, para clinique, étiologique et thérapeutique des HTA secondaires d'origine endocrine chez les patients suivis au service d'endocrinologie au CHU Mohamed IV à Marrakech. Il s'agit d'une étude descriptive prospective s’étalant sur une période de 4 ans incluant 45 patients ayant une HTA endocrinienne. La moyenne d’âge est de 44,89 ans, avec une nette prédominance du sexe féminine (sexe ratio de 0,49). Les étiologies des HTA endocrines étaient dominées par le phéochromocytome (17 cas), l'hypercorticisme (11 cas) et l'acromégalie (8 cas). L'HTA était paroxystique dans 24,4%. Elle était d'emblée sevère classée grade 3 dans 40% des cas. L'HTA a été compliquée de cardiopathie dans 24% des cas et de néphropathie dans 20% des cas. Le traitement curatif a permis une guérison de l'HTA chez 60% (27 cas). Le diagnostic des HTA secondaires endocrines est parfois difficile du fait de l'absence de spécificité clinique. Il n'est pas exceptionnel que l'HTA soit l'unique manifestation de la maladie. Dans notre travail nous notons le caractère paroxystique et sévère de l'HTA. Le caractère éventuellement curable des HTA endocrines, dans plus des deux tiers des cas, fait qu'il est important de la dépister devant toute HTA sévère, résistante au traitement, ou en présence de signes cliniques, biologiques ou radiologiques évocateurs. PMID:27303586

  2. Adaptation in CRISPR-Cas Systems.

    PubMed

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

    2016-03-17

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

  3. International Service Learning: Its Time Has Come.

    ERIC Educational Resources Information Center

    Silcox, Harry C.; Leek, Torun E.

    1997-01-01

    The service-learning movement is widespread in European international schools. The most comprehensive initiative presented at two recent European Council of Independent Schools conferences was the Creativity, Activity, Service (CAS) course that students must pass to graduate from International Baccalaureate schools. Exemplary programs in England…

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

    PubMed

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

    2015-02-01

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

  5. Integrin {alpha}{beta}1, {alpha}{sub v}{beta}, {alpha}{sub 6}{beta} effectors p130Cas, Src and talin regulate carcinoma invasion and chemoresistance

    SciTech Connect

    Sansing, Hope A.; Sarkeshik, Ali; Yates, John R.; Patel, Vyomesh; Gutkind, J. Silvio; Yamada, Kenneth M.; Berrier, Allison L.

    2011-03-11

    Research highlights: {yields} Proteomics of clustered integrin {alpha}{beta}1, {alpha}{sub v}{beta}, {alpha}{sub 6}{beta} receptors in oral carcinoma. {yields} p130Cas, Dek, Src and talin regulate oral carcinoma invasion. {yields} p130Cas, talin, Src and zyxin regulate oral carcinoma resistance to cisplatin. -- Abstract: Ligand engagement by integrins induces receptor clustering and formation of complexes at the integrin cytoplasmic face that controls cell signaling and cytoskeletal dynamics critical for adhesion-dependent processes. This study searches for a subset of integrin effectors that coordinates both tumor cell invasion and resistance to the chemotherapeutic drug cisplatin in oral carcinomas. Candidate integrin effectors were identified in a proteomics screen of proteins recruited to clustered integrin {alpha}{beta}1, {alpha}{sub v}{beta} or {alpha}{sub 6}{beta} receptors in oral carcinomas. Proteins with diverse functions including microtubule and actin binding proteins, and factors involved in trafficking, transcription and translation were identified in oral carcinoma integrin complexes. Knockdown of effectors in the oral carcinoma HN12 cells revealed that p130Cas, Dek, Src and talin were required for invasion through Matrigel. Disruption of talin or p130Cas by RNA interference increased resistance to cisplatin, whereas targeting Dek, Src or zyxin reduced HN12 resistance to cisplatin. Analysis of the spreading of HN12 cells on collagen I and laminin I revealed that a decrease in p130Cas or talin expression inhibited spreading on both matrices. Interestingly, a reduction in zyxin expression enhanced spreading on laminin I and inhibited spreading on collagen I. Reduction of Dek, Src, talin or zyxin expression reduced HN12 proliferation by 30%. Proliferation was not affected by a reduction in p130Cas expression. We conclude that p130Cas, Src and talin function in both oral carcinoma invasion and resistance to cisplatin.

  6. 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...

  7. 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...

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

    Code of Federal Regulations, 2010 CFR

    2010-07-01

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

  9. 41 CFR 102-33.440 - Who must report CAS cost and utilization data?

    Code of Federal Regulations, 2011 CFR

    2011-01-01

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

  10. 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...

  11. Determination of the Physical Dimensions of μ Cas

    NASA Astrophysics Data System (ADS)

    Bach, K.; Kang, W.

    2015-07-01

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

  12. Grossesse intra murale à propos d'un cas

    PubMed Central

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

    2015-01-01

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

  13. Introducing the TI-Nspire CAS Learning Technology

    NASA Astrophysics Data System (ADS)

    Osbourne, Michael

    2008-03-01

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

  14. L’éléphantiasis vulvo-clitoridien: à propos d'un nouveau cas

    PubMed Central

    Sine, Babacar; Bagayogo, Ndeye Aissatou; Thiam, Amath; Sarr, Alioune; Sow, Yaya; Fall, Boubacar; Zakou, Abdou Razak Hamidou; Faye, Samba Thiapato; Diao, Babacar; Fall, Papa Ahmed; Ndoye, Alain khassim; Ba, Mamadou

    2015-01-01

    L’éléphantiasis vulvo-clitoridien d'origine filarienne est une affection très rare. Nous rapportons un nouveau cas chez une femme de 33 ans suivie dans un service de Maladies Infectieuses pour filariose lymphatique. Elle avait une masse vulvo-clitoridienne qui évoluait depuis plus de 10 ans. Une résection clitoridienne et une plastie vulvaire a été réalisée. Les résultats fonctionnels et esthétiques étaient satisfaisants. PMID:26889314

  15. Object Classification via Planar Abstraction

    NASA Astrophysics Data System (ADS)

    Oesau, Sven; Lafarge, Florent; Alliez, Pierre

    2016-06-01

    We present a supervised machine learning approach for classification of objects from sampled point data. The main idea consists in first abstracting the input object into planar parts at several scales, then discriminate between the different classes of objects solely through features derived from these planar shapes. Abstracting into planar shapes provides a means to both reduce the computational complexity and improve robustness to defects inherent to the acquisition process. Measuring statistical properties and relationships between planar shapes offers invariance to scale and orientation. A random forest is then used for solving the multiclass classification problem. We demonstrate the potential of our approach on a set of indoor objects from the Princeton shape benchmark and on objects acquired from indoor scenes and compare the performance of our method with other point-based shape descriptors.

  16. An Abstract Plan Preparation Language

    NASA Technical Reports Server (NTRS)

    Butler, Ricky W.; Munoz, Cesar A.

    2006-01-01

    This paper presents a new planning language that is more abstract than most existing planning languages such as the Planning Domain Definition Language (PDDL) or the New Domain Description Language (NDDL). The goal of this language is to simplify the formal analysis and specification of planning problems that are intended for safety-critical applications such as power management or automated rendezvous in future manned spacecraft. The new language has been named the Abstract Plan Preparation Language (APPL). A translator from APPL to NDDL has been developed in support of the Spacecraft Autonomy for Vehicles and Habitats Project (SAVH) sponsored by the Explorations Technology Development Program, which is seeking to mature autonomy technology for application to the new Crew Exploration Vehicle (CEV) that will replace the Space Shuttle.

  17. Cryogenic foam insulation: Abstracted publications

    NASA Technical Reports Server (NTRS)

    Williamson, F. R.

    1977-01-01

    A group of documents were chosen and abstracted which contain information on the properties of foam materials and on the use of foams as thermal insulation at cryogenic temperatures. The properties include thermal properties, mechanical properties, and compatibility properties with oxygen and other cryogenic fluids. Uses of foams include applications as thermal insulation for spacecraft propellant tanks, and for liquefied natural gas storage tanks and pipelines.

  18. Concrete and abstract Voronoi diagrams

    SciTech Connect

    Klein, R. )

    1989-01-01

    The Voronoi diagram of a set of sites is a partition of the plane into regions, one to each site, such that the region of each site contains all points of the plane that are closer to this site than to the other ones. Such partitions are of great importance to computer science and many other fields. The challenge is to compute Voronoi diagrams quickly. The problem is that their structure depends on the notion of distance and the sort of site. In this book the author proposes a unifying approach by introducing abstract Voronoi diagrams. These are based on the concept of bisecting curves which are required to have some simple properties that are actually possessed by most bisectors of concrete Voronoi diagrams. Abstract Voronoi diagrams can be computed efficiently and there exists a worst-case efficient algorithm of divide-and-conquer type that applies to all abstract Voronoi diagrams satisfying a certain constraint. The author shows that this constraint is fulfilled by the concrete diagrams based no large classes of metrics in the plane.

  19. Section traumatique de l'utérus non gravide chez une accidentée de voie publique: à propos d'un cas

    PubMed Central

    Cédrick, Sangwa Milindi; Marius, Kitembo Feruzi; Mireille, Kakinga Zabibu; Nelly, Mukonda Sompo; Chamy, Cham Lubamba; Numbi, Kabamge; Mutomb, Jean Felix

    2015-01-01

    Dans cet article les auteurs rapportent un rare cas de section traumatique d'un utérus non gravide, chez une accidentée de voie publique reçu dans un tableau d'hemopéritoine aux services des urgences de l'hôpital General Provincial de Reference Jason Sendwe. PMID:26113916

  20. Youth Studies Abstracts. Vol. 4 No. 3.

    ERIC Educational Resources Information Center

    Youth Studies Abstracts, 1985

    1985-01-01

    This volume contains 169 abstracts of documents dealing with youth and educational programs for youth. Included in the volume are 97 abstracts of documents dealing with social and educational developments; 56 abstracts of program reports, reviews, and evaluations; and 16 abstracts of program materials. Abstracts are grouped according to the…

  1. The Structural Biology of CRISPR-Cas Systems

    PubMed Central

    Jiang, Fuguo; Doudna, Jennifer A.

    2015-01-01

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

  2. 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

  3. 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

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

    PubMed

    Waaijers, Selma; Boxem, Mike

    2014-08-01

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

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

    PubMed

    Barrangou, Rodolphe; Marraffini, Luciano A

    2014-04-24

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

  6. 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

  7. 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

  8. Comparison of Cas9 activators in multiple species.

    PubMed

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

    2016-07-01

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

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

    PubMed

    Peng, Rongxue; Lin, Guigao; Li, Jinming

    2016-04-01

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

  10. Target specificity of the CRISPR-Cas9 system

    PubMed Central

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

    2015-01-01

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

  11. Operating System Abstraction Layer (OSAL)

    NASA Technical Reports Server (NTRS)

    Yanchik, Nicholas J.

    2007-01-01

    This viewgraph presentation reviews the concept of the Operating System Abstraction Layer (OSAL) and its benefits. The OSAL is A small layer of software that allows programs to run on many different operating systems and hardware platforms It runs independent of the underlying OS & hardware and it is self-contained. The benefits of OSAL are that it removes dependencies from any one operating system, promotes portable, reusable flight software. It allows for Core Flight software (FSW) to be built for multiple processors and operating systems. The presentation discusses the functionality, the various OSAL releases, and describes the specifications.

  12. IEEE conference record--Abstracts

    SciTech Connect

    Not Available

    1992-01-01

    The following topics were covered in this meeting: basic plasma phenomena and plasma waves; plasma diagnostics; space plasma diagnostics; magnetic fusion; electron, ion and plasma sources; intense electron and ion beams; intense beam microwaves; fast wave M/W devices; microwave plasma interactions; plasma focus; ultrafast Z-pinches; plasma processing; electrical gas discharges; fast opening switches; magnetohydrodynamics; electromagnetic and electrothermal launchers; x-ray lasers; computational plasma science; solid state plasmas and switches; environmental/energy issues in plasma science; vacuum electronics; plasmas for lighting; gaseous electronics; and ball lightning and other spherical plasmas. Separate abstracts were prepared for 278 papers of this conference.

  13. Rational design of a split-Cas9 enzyme complex

    DOE PAGESBeta

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

    2015-02-23

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

  14. 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

  15. CRISPR-Cas9 Based Engineering of Actinomycetal Genomes.

    PubMed

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

    2015-09-18

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

  16. 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

  17. 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

  18. 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.

  19. Greenbook Abstract and Catalog--1.

    ERIC Educational Resources Information Center

    Coole, Walter A.; Reitan, Henry M.

    This catalog is intended to be issued periodically as a means of extending and updating teaching materials which have been previously published through ERIC by these authors. The materials include the Alpha and Gamma levels of the "Greenbook System," pre-professional and entry in-service professional levels of an integrated sequence of five…

  20. Community Service and Continuing Education. Program Abstracts. Second Edition.

    ERIC Educational Resources Information Center

    Miller, Lorna M., Ed.

    This comprehensive volume of innovative continuing education programs contains complete descriptions of some 750 cross-indexed projects in four general areas--human resource development, natural resource development, economic development, and community development--directed at community problems that can benefit from adult education. It brings…

  1. 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

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

    PubMed

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

    2016-01-01

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

  3. Functional Analysis of Porphyromonas gingivalis W83 CRISPR-Cas Systems

    PubMed Central

    Burmistrz, Michał; Dudek, Bartosz; Staniec, Dominika; Rodriguez Martinez, Jose Ignacio; Bochtler, Matthias; Potempa, Jan

    2015-01-01

    ABSTRACT The CRISPR-Cas (clustered regularly interspaced short palindromic repeats/CRISPR-associated genes) system provides prokaryotic cells with an adaptive and heritable immune response to foreign genetic elements, such as viruses, plasmids, and transposons. It is present in the majority of Archaea and almost half of species of Bacteria. Porphyromonas gingivalis is an important human pathogen that has been proven to be an etiological agent of periodontitis and has been linked to systemic conditions, such as rheumatoid arthritis and cardiovascular disease. At least 95% of clinical strains of P. gingivalis carry CRISPR arrays, suggesting that these arrays play an important function in vivo. Here we show that all four CRISPR arrays present in the P. gingivalis W83 genome are transcribed. For one of the arrays, we demonstrate in vivo activity against double-stranded DNA constructs containing protospacer sequences accompanied at the 3′ end by an NGG protospacer-adjacent motif (PAM). Most of the 44 spacers present in the genome of P. gingivalis W83 share no significant similarity with any known sequences, although 4 spacers are similar to sequences from bacteria found in the oral cavity and the gastrointestinal tract. Four spacers match genomic sequences of the host; however, none of these is flanked at its 3′ terminus by the appropriate PAM element. IMPORTANCE The CRISPR-Cas (clustered regularly interspaced short palindromic repeats/CRISPR-associated genes) system is a unique system that provides prokaryotic cells with an adaptive and heritable immunity. In this report, we show that the CRISPR-Cas system of P. gingivalis, an important human pathogen associated with periodontitis and possibly also other conditions, such as rheumatoid arthritis and cardiovascular disease, is active and provides protection from foreign genetic elements. Importantly, the data presented here may be useful for better understanding the communication between cells in larger bacterial

  4. Tuberculose pulmonaire révélée par un purpura thrombopénique chez l'enfant-à propos d'un cas clinique observé au service de pédiatrie des Cliniques Universitaires de Lubumbashi

    PubMed Central

    Lubala, Toni Kasole; Mutombo, Augustin Mulangu; Munkana, Arthur Ndundula; Manika, Michel Muteya

    2012-01-01

    Nous rapportons le cas d'un enfant de 7 ans, de sexe masculin ayant présenté un purpura thrombopénique avec épistaxis, hématémèse, otorragies et pétéchies généralisées. Durant la même hospitalisation, nous avons mis en évidence une tuberculose pulmonaire documentée par la présence de bacilles acido-alcoolo résistants à l'examen des crachats. Nous avons observé une majoration du taux de plaquettes en une semaine de corticothérapie intraveineuse à haute dose, avant l'instauration d'une poly chimiothérapie antituberculeuse. Nous rappelons également la controverse que suscite la prise en charge de cette association rarement rapportée. PMID:23077696

  5. Les tumeurs malignes naso-sinusiennes: à propos de 32 cas et revues de la littérature

    PubMed Central

    Darouassi, Youssef; Touati, Mohamed Mliha; Chihani, Mehdi; El Alami, Jihane; Bouaity, Brahim; Ammar, Haddou

    2015-01-01

    Sous l’appellation tumeurs malignes naso-sinusiennes est regroupé un vaste éventail de tumeurs, aux histologies et localisations variées, mais aux tableaux cliniques souvent similaires. Le diagnostic de ces tumeurs est difficile, nécessitant une approche multidisciplinaire, à savoir oto-rhino-laryngologique, radiologique et anatomopathologique. Notre étude rétrospective concerne 32 cas de tumeurs malignes naso-sinusiennes, colligées au service d’ORL de l’hôpital militaire Avicenne de Marrakech, entre Janvier 2004 et Décembre 2014. L’analyse des données a noté que la fréquence des tumeurs épithéliales (75% des cas) était supérieure à celle des tumeurs non épithéliales (25% des cas), avec en tête de file l’adénocarcinome de l’ethmoïde (31,25%) et le carcinome épidermoïde du sinus maxillaire (18,75%). Ces tumeurs surviennent le plus souvent chez le sujet âgé avec une moyenne d’âge de 52 ans et une répartition équitable entre les deux sexes. Le délai de consultation moyen était de 12 mois avec une symptomatologie dominée par un syndrome nasosinusien (71,8%), associé dans certains cas à des signes ophtalmologiques (12,5%) ou neurologiques (15,6%). Tous nos patients ont bénéficié d’un examen clinique notamment endoscopique, d’une exploration radiologique des tumeurs et de leurs extensions, et d’une confirmation diagnostique par un examen anatomopathologique. Le traitement a consisté en une exérèse chirurgicale de la tumeur dès que cela était possible, soit dans 81,3% des cas (26 patients), généralement complété par un traitement adjuvant radio-chimiothérapique (77%). Le suivi à un an de nos patients a permis de noter une bonne évolution pour 08 d’entre eux (25%), une récidive dans 6 cas (18,75%), le décès de neuf patients (28,1%), et l’absence d’information concernant les autres cas (28,1%). PMID:26985260

  6. Le médulloblastome chez l'adulte: à propos de 13 cas et revue de la literature

    PubMed Central

    Drissi, Jamal; Affane, Mariam; Elomrani, Abdelhamid; Khouchani, Mouna

    2015-01-01

    Le médulloblastome est une tumeur neuro-ectodermique primitive maligne. Il s'agit d'une tumeur rare chez l'adulte, représentant moins de 1% des tumeurs cérébrales. Nous proposons une étude rétrospective réalisée au sein du service d'Oncologie-Radiothérapie du CHU Mohamed VI de Marrakech sur une période de 13 ans. Le but de notre travail est de déterminer le profil épidémioclinique, thérapeutique, évolutif ainsi que les facteurs pronostiques de cette entité pathologique avec une revue de la littérature. Notre série comportait 13 patients, 10 hommes et 3 femmes, l’âge médian a été 20,8 ans. Le tableau clinique a été révélé par un syndrome d'hypertension intracrânienne (100%), associée à un syndrome cérébelleux (84%). La localisation était hémisphérique (31%) et vermio-hémisphérique (54%). 31% des patients ont bénéficié d'une exérèse chirurgicale totale. 85% des cas avaient une variante classique et 15% une variante desmoplasique. 30% des cas avaient été classés à «risque standard» et 70% à «haut risque». La chirurgie avait été complétée d'une radiothérapie de l'ensemble du névraxe selon la technique de «jonctions mobiles» dans tous les cas. Le délai moyen était de 73 jours. Une chimiothérapie adjuvante avait été réalisée chez 9 cas. Avec un recul moyen de 21.3 mois, l’évolution a été marquée par une récidive tumorale (4 cas), une toxicité auditive (6 cas) et des troubles cognitifs chez un cas. La prise en charge du médulloblastome doit être multidisciplinaire associant neurochirurgiens et oncologues radiothérapeutes. Cette collaboration est le seul garant d'une amélioration de son pronostic. PMID:26889307

  7. Les hémorragies post coïtales: à propos de 68 cas et revue de literature

    PubMed Central

    Boukhanni, Lahssen; Dhibou, Hanane; Zilfi, Wafaa; Housseini, Kawtar Iraki; Benkeddour, Yasser Ait; Aboulfalah, Abderrahim; Asmouki, Hamid; Soummani, Abderraouf

    2016-01-01

    L'acte sexuel consenti ou imposé, peut être à l'origine des traumatismes. L'hémorragie post coïtale est un symptôme gynécologique commun. Elle peut révéler de sérieux problèmes. Le but de notre travail est d’étudier le profil épidémiologique, diagnostique et thérapeutique ainsi que les moyens préventifs en cas de déchirure post coïtale. Il s'agit d'une étude prospective, étalée sur deux ans, mené au service de gynécologie obstétrique du CHU Med VI de Marrakech. Nous avons colligé 68 patientes. L’âge moyen est de 27 ans, la majorité des patientes étaient des nullipares soit 89,7% des cas. La moitié des patientes avaient un mariage traditionnel. Le rapport était consentent dans 74% des cas. L'hémorragie génitale a constitue le motif de consultation le plus fréquent soit 98% des cas. Les lésions hyménales ont été retrouvées dans 39% des cas et la lésion siégeait dans le cul de sac postérieur chez 35% des cas. La prise en charge thérapeutique a consisté en une suture chirurgicale chez 97% des cas, associé à une transfusion sanguine chez deux patientes et une abstinence sexuelle pendant minimum deux semaines chez toutes nous patientes. Le contexte social ainsi que le manque d’éducation sexuelle sont souvent incriminé d'où l'intérêt d'une prise en charge psychologique pour prévenir aussi bien le retentissement du traumatisme sur la sexualité que les récidives. PMID:27279958

  8. 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

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

    PubMed

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

    2016-01-01

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

  10. Abstract Expression Grammar Symbolic Regression

    NASA Astrophysics Data System (ADS)

    Korns, Michael F.

    This chapter examines the use of Abstract Expression Grammars to perform the entire Symbolic Regression process without the use of Genetic Programming per se. The techniques explored produce a symbolic regression engine which has absolutely no bloat, which allows total user control of the search space and output formulas, which is faster, and more accurate than the engines produced in our previous papers using Genetic Programming. The genome is an all vector structure with four chromosomes plus additional epigenetic and constraint vectors, allowing total user control of the search space and the final output formulas. A combination of specialized compiler techniques, genetic algorithms, particle swarm, aged layered populations, plus discrete and continuous differential evolution are used to produce an improved symbolic regression sytem. Nine base test cases, from the literature, are used to test the improvement in speed and accuracy. The improved results indicate that these techniques move us a big step closer toward future industrial strength symbolic regression systems.

  11. Toward Millimagnitude Photometric Calibration (Abstract)

    NASA Astrophysics Data System (ADS)

    Dose, E.

    2014-12-01

    (Abstract only) Asteroid roation, exoplanet transits, and similar measurements will increasingly call for photometric precisions better than about 10 millimagnitudes, often between nights and ideally between distant observers. The present work applies detailed spectral simulations to test popular photometric calibration practices, and to test new extensions of these practices. Using 107 synthetic spectra of stars of diverse colors, detailed atmospheric transmission spectra computed by solar-energy software, realistic spectra of popular astronomy gear, and the option of three sources of noise added at realistic millimagnitude levels, we find that certain adjustments to current calibration practices can help remove small systematic errors, especially for imperfect filters, high airmasses, and possibly passing thin cirrus clouds.

  12. Experience with abstract notation one

    NASA Technical Reports Server (NTRS)

    Harvey, James D.; Weaver, Alfred C.

    1990-01-01

    The development of computer science has produced a vast number of machine architectures, programming languages, and compiler technologies. The cross product of these three characteristics defines the spectrum of previous and present data representation methodologies. With regard to computer networks, the uniqueness of these methodologies presents an obstacle when disparate host environments are to be interconnected. Interoperability within a heterogeneous network relies upon the establishment of data representation commonality. The International Standards Organization (ISO) is currently developing the abstract syntax notation one standard (ASN.1) and the basic encoding rules standard (BER) that collectively address this problem. When used within the presentation layer of the open systems interconnection reference model, these two standards provide the data representation commonality required to facilitate interoperability. The details of a compiler that was built to automate the use of ASN.1 and BER are described. From this experience, insights into both standards are given and potential problems relating to this development effort are discussed.

  13. Abstraction Planning in Real Time

    NASA Technical Reports Server (NTRS)

    Washington, Richard

    1994-01-01

    When a planning agent works in a complex, real-world domain, it is unable to plan for and store all possible contingencies and problem situations ahead of time. The agent needs to be able to fall back on an ability to construct plans at run time under time constraints. This thesis presents a method for planning at run time that incrementally builds up plans at multiple levels of abstraction. The plans are continually updated by information from the world, allowing the planner to adjust its plan to a changing world during the planning process. All the information is represented over intervals of time, allowing the planner to reason about durations, deadlines, and delays within its plan. In addition to the method, the thesis presents a formal model of the planning process and uses the model to investigate planning strategies. The method has been implemented, and experiments have been run to validate the overall approach and the theoretical model.

  14. Abstraction Planning in Real Time

    NASA Technical Reports Server (NTRS)

    Washington, R.

    1994-01-01

    When a planning agent works in a complex, real-world domain, it is unable to plan for and store all possible contingencies and problem situations ahead of time. This thesis presents a method for planning a run time that incrementally builds up plans at multiple levels of abstraction. The plans are continually updated by information from the world, allowing the planner to adjust its plan to a changing world during the planning process. All the information is represented over intervals of time, allowing the planner to reason about durations, deadlines, and delays within its plan. In addition to the method, the thesis presents a formal model of the planning process and uses the model to investigate planning strategies.

  15. In Situ Resource Utilization (ISRU 3) Technical Interchange Meeting: Abstracts

    NASA Technical Reports Server (NTRS)

    1999-01-01

    This volume contains abstracts that have been accepted for presentation at the In Situ Resource Utilization (ISRU III) Technical Interchange Meeting, February 11-12, 1999, hosted by the Lockheed Martin Astronautics Waterton Facility, Denver, Colorado. Administration and publication support for this meeting were provided by the staff of the Publications and Program Services Department at the Lunar and Planetary Institute.

  16. Fanconi Anemia Gene Editing by the CRISPR/Cas9 System

    PubMed Central

    Osborn, Mark J.; Gabriel, Richard; Webber, Beau R.; DeFeo, Anthony P.; McElroy, Amber N.; Jarjour, Jordan; Starker, Colby G.; Wagner, John E.; Joung, J. Keith; Voytas, Daniel F.; von Kalle, Christof; Schmidt, Manfred; Blazar, Bruce R.

    2015-01-01

    Abstract Genome engineering with designer nucleases is a rapidly progressing field, and the ability to correct human gene mutations in situ is highly desirable. We employed fibroblasts derived from a patient with Fanconi anemia as a model to test the ability of the clustered regularly interspaced short palindromic repeats/Cas9 nuclease system to mediate gene correction. We show that the Cas9 nuclease and nickase each resulted in gene correction, but the nickase, because of its ability to preferentially mediate homology-directed repair, resulted in a higher frequency of corrected clonal isolates. To assess the off-target effects, we used both a predictive software platform to identify intragenic sequences of homology as well as a genome-wide screen utilizing linear amplification-mediated PCR. We observed no off-target activity and show RNA-guided endonuclease candidate sites that do not possess low sequence complexity function in a highly specific manner. Collectively, we provide proof of principle for precision genome editing in Fanconi anemia, a DNA repair-deficient human disorder. PMID:25545896

  17. Tuning CAS Application using AIMS: An Automated Instrumentation and Monitoring System

    NASA Technical Reports Server (NTRS)

    Mehra, P.; Sarukkai, S.; Schmidt, M.; Schulbach, C.; VanVoorst, B.; Yan, Jerry; Woodrow, Thomas (Technical Monitor)

    1994-01-01

    To bring together NASA's scientists and engineers and their counterparts in industry, other government agencies, and academia working in the Computational AeroSciences (CAS) field. This workshop is part of the technology transfer plan of the High Performance Computing and Communications Program (HPCCP). Specific objectives of this Workshop are to: (1) communicate the goals and objectives of HPCCP in the area of CAS; (2) promote and disseminate CAS technology within the appropriate technical communities, including NASA, industry, academia, and other government labs; (3) help promote synergy among CAS scientists; and (4) permit feedback from peer researchers in issues pacing the CAS field in general and the HPCCP CAS program in particular.

  18. Spectroscopic studies of three Cepheids with high positive pulsation period increments: SZ Cas, BY Cas, and RU Sct

    NASA Astrophysics Data System (ADS)

    Usenko, I. A.; Klochkova, V. G.

    2015-07-01

    Three high-resolution spectra have been taken at different times with the 6-m SAO RAS telescope (LYNX and PFES spectrographs) for three Cepheids exhibiting high positive period increments: the small-amplitude (DCEPS) SZ Cas and BY Cas and the classical (DCEP) RU Sct. SZ Cas and RU Sct are members of the Galactic open clusters χ and h Per and Trump 35, respectively. Analysis of the spectra has shown that the interstellar Na I D1 and D2 lines in all objects are considerably stronger than the atmospheric ones and are redshifted in SZ Cas and BY Cas and blushifted in RU Sct. The core of the H α absorption line in BY Cas has an asymmetric knifelike shape, while RU Sct exhibits an intense emission in the blue wing of this line. Such phenomena are observed in long-period Cepheids and bright hypergiants with an extended envelope. In this case, the strong Mg Ib 5183.62 Å and Ba II 5853.67, 6141.713, and 6496.90 Å lines with low χlow in SZ Cas and RU Sct also show characteristic knifelike profiles with an asymmetry in the red region, while the Ba II 4934.095 Å line shows similar profiles in the blue one. The absorption lines of neutral atoms and singly ionized metals with different lowerlevel excitation potentials exhibit different degrees of asymmetry: from a pronounced one with secondary components in BY Cas (similar to those in the small-amplitude Cepheid BG Cru pulsating in the first overtone and having an envelope) to its insignificance or virtual absence in SZ Cas and RU Sct. Analysis of the secular changes in mean T eff determined from photometric color indices and spectra over the last 55 years for these stars has revealed periodic fluctuations of 200 K for SZ Cas and BY Cas and 500 K for RU Sct. For SZ Cas and RU Sct, T eff determined in some years from some color indices show much lower values, which together with the temperature fluctuations can be associated with mass loss and dust formation. Based on these facts, we hypothesize the existence of

  19. An abstract approach to music.

    SciTech Connect

    Kaper, H. G.; Tipei, S.

    1999-04-19

    In this article we have outlined a formal framework for an abstract approach to music and music composition. The model is formulated in terms of objects that have attributes, obey relationships, and are subject to certain well-defined operations. The motivation for this approach uses traditional terms and concepts of music theory, but the approach itself is formal and uses the language of mathematics. The universal object is an audio wave; partials, sounds, and compositions are special objects, which are placed in a hierarchical order based on time scales. The objects have both static and dynamic attributes. When we realize a composition, we assign values to each of its attributes: a (scalar) value to a static attribute, an envelope and a size to a dynamic attribute. A composition is then a trajectory in the space of aural events, and the complex audio wave is its formal representation. Sounds are fibers in the space of aural events, from which the composer weaves the trajectory of a composition. Each sound object in turn is made up of partials, which are the elementary building blocks of any music composition. The partials evolve on the fastest time scale in the hierarchy of partials, sounds, and compositions. The ideas outlined in this article are being implemented in a digital instrument for additive sound synthesis and in software for music composition. A demonstration of some preliminary results has been submitted by the authors for presentation at the conference.

  20. Ozone Conference II: Abstract Proceedings

    SciTech Connect

    1999-11-01

    Ozone Conference II: Pre- and Post-Harvest Applications Two Years After Gras, was held September 27-28, 1999 in Tulare, California. This conference, sponsored by EPRI's Agricultural Technology Alliance and Southern California Edison's AgTAC facility, was coordinated and organized by the on-site ATA-AgTAC Regional Center. Approximately 175 people attended the day-and-a-half conference at AgTAC. During the Conference twenty-two presentations were given on ozone food processing and agricultural applications. Included in the presentations were topics on: (1) Ozone fumigation; (2) Ozone generation techniques; (3) System and design applications; (4) Prewater treatment requirements; (5) Poultry water reuse; (6) Soil treatments with ozone gas; and (7) Post-harvest aqueous and gaseous ozone research results. A live videoconference between Tulare and Washington, D.C. was held to discuss the regulators' view from inside the beltway. Attendees participated in two Roundtable Question and Answer sessions and visited fifteen exhibits and demonstrations. The attendees included university and governmental researchers, regulators, consultants and industry experts, technology developers and providers, and corporate and individual end-users. This report is comprised of the Abstracts of each presentation, biographical sketches for each speaker and a registration/attendees list.

  1. 1986 annual information meeting. Abstracts

    SciTech Connect

    Not Available

    1986-01-01

    Abstracts are presented for the following papers: Geohydrological Research at the Y-12 Plant (C.S. Haase); Ecological Impacts of Waste Disposal Operations in Bear Creek Valley Near the Y-12 Plant (J.M. Loar); Finite Element Simulation of Subsurface Contaminant Transport: Logistic Difficulties in Handling Large Field Problems (G.T. Yeh); Dynamic Compaction of a Radioactive Waste Burial Trench (B.P. Spalding); Comparative Evaluation of Potential Sites for a High-Level Radioactive Waste Repository (E.D. Smith); Changing Priorities in Environmental Assessment and Environmental Compliance (R.M. Reed); Ecology, Ecotoxicology, and Ecological Risk Assessment (L.W. Barnthouse); Theory and Practice in Uncertainty Analysis from Ten Years of Practice (R.H. Gardner); Modeling Landscape Effects of Forest Decline (V.H. Dale); Soil Nitrogen and the Global Carbon Cycle (W.M. Post); Maximizing Wood Energy Production in Short-Rotation Plantations: Effect of Initial Spacing and Rotation Length (L.L. Wright); and Ecological Communities and Processes in Woodland Streams Exhibit Both Direct and Indirect Effects of Acidification (J.W. Elwood).

  2. Attracting Girls into Physics (abstract)

    NASA Astrophysics Data System (ADS)

    Gadalla, Afaf

    2009-04-01

    A recent international study of women in physics showed that enrollment in physics and science is declining for both males and females and that women are severely underrepresented in careers requiring a strong physics background. The gender gap begins early in the pipeline, from the first grade. Girls are treated differently than boys at home and in society in ways that often hinder their chances for success. They have fewer freedoms, are discouraged from accessing resources or being adventurous, have far less exposure to problem solving, and are not encouraged to choose their lives. In order to motivate more girl students to study physics in the Assiut governorate of Egypt, the Assiut Alliance for the Women and Assiut Education District collaborated in renovating the education of physics in middle and secondary school classrooms. A program that helps in increasing the number of girls in science and physics has been designed in which informal groupings are organized at middle and secondary schools to involve girls in the training and experiences needed to attract and encourage girls to learn physics. During implementation of the program at some schools, girls, because they had not been trained in problem-solving as boys, appeared not to be as facile in abstracting the ideas of physics, and that was the primary reason for girls dropping out of science and physics. This could be overcome by holding a topical physics and technology summer school under the supervision of the Assiut Alliance for the Women.

  3. Repurposing CRISPR/Cas9 for in situ functional assays

    PubMed Central

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

    2013-01-01

    RNAi combined with next-generation sequencing has proven to be a powerful and cost-effective genetic screening platform in mammalian cells. Still, this technology has its limitations and is incompatible with in situ mutagenesis screens on a genome-wide scale. Using p53 as a proof-of-principle target, we readapted the CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR associated 9) genome-editing system to demonstrate the feasibility of this methodology for targeted gene disruption positive selection assays. By using novel “all-in-one” lentiviral and retroviral delivery vectors heterologously expressing both a codon-optimized Cas9 and its synthetic guide RNA (sgRNA), we show robust selection for the CRISPR-modified Trp53 locus following drug treatment. Furthermore, by linking Cas9 expression to GFP fluorescence, we use an “all-in-one” system to track disrupted Trp53 in chemoresistant lymphomas in the Eμ-myc mouse model. Deep sequencing analysis of the tumor-derived endogenous Cas9-modified Trp53 locus revealed a wide spectrum of mutants that were enriched with seemingly limited off-target effects. Taken together, these results establish Cas9 genome editing as a powerful and practical approach for positive in situ genetic screens. PMID:24298059

  4. Measurement of Flux Density of Cas A at Low Frequencies

    NASA Astrophysics Data System (ADS)

    Patil, Ajinkya; Fisher, R.

    2012-01-01

    Cas A is used as a flux calibrator throughout the radio spectrum. Therefore it is important to know the spectral and secular variations in its flux density. Earlier observations by Scott et. al. (1969) and Baars et. al. (1972) suggested a secular decrease in flux density of Cas A at a rate of about 1% per year at all frequencies. However later observations by Erickson & Perley (1975) and Read (1977) indicated anomalously high flux from Cas A at 38 MHz. Also, these observations suggested that the original idea of faster decay of the flux density rate at low frequencies may be in error or that something more complex than simple decay is affecting the flux density at low frequencies. The source changes at 38 MHz still remains a mystery. We intend to present the results of follow up observations made from 1995 to 1998 with a three element interferometer in Green Bank operating in frequency range 30 to 120 MHz. We will discuss the problems at such low frequencies due to large beamwidth and unstable ionosphere. We will also discuss the strategies we have used so far to to find the flux density of Cas A by calculating the ratio of flux density of Cas A to that of Cyg A, assuming flux density of Cyg A to be constant. Above mentioned work was performed in summer student program sponsored by National Radio Astronomy Observatory.

  5. Displacement of p130Cas from focal adhesions links actomyosin contraction to cell migration.

    PubMed

    Machiyama, Hiroaki; Hirata, Hiroaki; Loh, Xia Kun; Kanchi, Madhu Mathi; Fujita, Hideaki; Tan, Song Hui; Kawauchi, Keiko; Sawada, Yasuhiro

    2014-08-15

    Cell adhesion complexes provide platforms where cell-generated forces are transmitted to the extracellular matrix (ECM). Tyrosine phosphorylation of focal adhesion proteins is crucial for cells to communicate with the extracellular environment. However, the mechanisms that transmit actin cytoskeletal motion to the extracellular environment to drive cell migration are poorly understood. We find that the movement of p130Cas (Cas, also known as BCAR1), a mechanosensor at focal adhesions, correlates with actin retrograde flow and depends upon actomyosin contraction and phosphorylation of the Cas substrate domain (CasSD). This indicates that CasSD phosphorylation underpins the physical link between Cas and the actin cytoskeleton. Fluorescence recovery after photobleaching (FRAP) experiments reveal that CasSD phosphorylation, as opposed to the association of Cas with Src, facilitates Cas displacement from adhesion complexes in migrating cells. Furthermore, the stabilization of Src-Cas binding and inhibition of myosin II, both of which sustain CasSD phosphorylation but mitigate Cas displacement from adhesion sites, retard cell migration. These results indicate that Cas promotes cell migration by linking actomyosin contractions to the adhesion complexes through a dynamic interaction with Src as well as through the phosphorylation-dependent association with the actin cytoskeleton. PMID:24928898

  6. Les Brulures Electriques par Haut Voltage - A Propos de 10 Cas

    PubMed Central

    Belmir, R.; Fejjal, N.; El Omari, M.; El Mazouz, S.; Gharib, N.; Abassi, A.; Belmahi, A.

    2008-01-01

    Summary Les accidents électriques par haute tension (AEHT) provoquent des brûlures profondes par effet Joule le long des axes vasculo-nerveux entre les points d'entrée et de sortie, qui sont le siège de lésions délabrantes. Les Auteurs rapportent une série de dix cas d'AEHT admis au service de chirurgie réparatrice et de brûlés de l'Hôpital Ibn Sina de Rabat à travers laquelle ils étudient les caractéristiques épidémiologiques, cliniques et thérapeutiques. Tous les patients étaient des adultes de sexe masculin dont l'âge moyen était de 31 ans. Dans 70% des cas, ces brûlures étaient secondaires à un contact avec les distributeurs d'électricité avec une surface brûlée inférieure à 20%. Le traitement des lésions électrothermiques a nécessité des interventions itératives avec amputation des segments de membres nécrosés dans 70% des cas, dont les suites étaient marquées par des séquelles fonctionnelles invalidantes. La prévention des AEHT, en particulier pour les accidents du travail au sein des professions exposées, reste fondamentale. PMID:21991124

  7. La pneumonie tuberculeuse: une nouvelle série de 27 cas

    PubMed Central

    Bakouh, Ouiam; Aniked, Sarra; Bourkadi, Jamaleddine

    2014-01-01

    Dans le but d’étudier les aspects cliniques, radiologiques et évolutifs de la pneumonie tuberculeuse (PT) au Maroc, pays à forte prévalence de tuberculose, une étude rétrospective s’étalant de Janvier au Septembre 2013 a été menée au service de phtisiologie femme de l'hôpital Moulay Youssef de Rabat. 27 cas de PT ont été diagnostiqués, dont 2 VIH séropositives. La fièvre et l'altération de l’état général étaient rapportées chez toutes les patientes, précédant les signes respiratoires comme La toux et la dyspnée. L'hémoptysie est rapportée chez 7 cas. Diagnostiquées souvent tardivement, du fait de la non spécificité de ses signes, 14 pneumonies sur 27 étaient excavées. Avec prédominance des lésions au lobe supérieur droit. Le traitement antituberculeux était efficace dans la majorité des cas. On a déploré 2 décès. La décision de mise en route du traitement antituberculeux même en l'absence de certitude bactériologique doit être prise dans un délai raisonnable de 15 jours vue la gravité du tableau et les séquelles persistantes. PMID:25745530

  8. Cancer du sein de l'homme: à propos de 6 cas

    PubMed Central

    Laabadi, Kamilia; Jayi, Sofia; Alaoui, Fatimazohra Fdili; Bouguern, Hakima; Chaara, Hikmat; Melhouf, My Abdelilah; Hassani, Karim Ibn Majdoub; Laalim, Said Ait; Anoun, Hicham; Toughrai, Imane; Mazaz, Khalid

    2013-01-01

    Le but de ce travail était d'analyser les caractéristiques cliniques, histologiques, thérapeutiques et pronostiques du cancer du sein chez l'homme. Il s'agissait d'une étude rétrospective portant sur six patients colligés au service de gynécologie obstétrique II, CHU Hassan II durant la période 2009-2012. L’âge moyen de nos patients est de 65.3 ans. Il s'agit dans 83.3% des cas, d'une tumeur rétroaréolaire dont la taille moyenne est de 44.16 mm. Nous avons retrouvé 4 (66.7%) T4, 1 (16.7%) T3 et dans un cas, une tumeur inclassable. Le type histologique le plus représenté est le carcinome canalaire infiltrant (66.7%). Le taux d'envahissement ganglionnaire axillaire est de 66.7%. L'hormonodépendance de ces tumeurs est prouvée dans 100% des cas. La survie à cinq ans est en cours d’évaluation. L'envahissement ganglionnaire, l'invasion du derme, le stade clinique TNM sont des facteurs qui influencent significativement la survenue de métastases. Aucun de ces facteurs de risque n'est apparu significatif en termes de survie globale. Le cancer du sein chez l'homme est une maladie rare (environ 1% des cancers du sein) au pronostic sombre. Le diagnostic est le plus souvent tardif et les lésions sont traitées à des stades avancés. PMID:24711870

  9. GENOME EDITING IN HUMAN CELLS USING CRISPR/CAS NUCLEASES

    PubMed Central

    Wyvekens, Nicolas; Tsai, Shengdar; Joung, J. Keith

    2016-01-01

    The clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas) system has been broadly adopted for highly efficient genome editing in a variety of model organisms and human cell types. Unlike previous genome editing technologies such as Zinc Finger Nucleases (ZFNs) and Transcription Activator-Like Effector Nucleases (TALENs), the CRISPR/Cas technology does not require complex protein engineering and can be utilized by any researcher proficient in basic molecular biology and cell culture techniques. Here we describe protocols for design and cloning of vectors expressing single or multiplex gRNAs, for transient transfection of human cell lines, and for quantitation of mutation frequencies by T7 Endonuclease I assay. These protocols also include guidance for using two improvements that increase the specificity of CRISPR/Cas nucleases: truncated gRNAs and dimeric RNA-guided FokI nucleases. PMID:26423589

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

    PubMed Central

    Torres-Ruiz, Raul; Rodriguez-Perales, Sandra

    2015-01-01

    The cancer-modelling field is now experiencing a conversion with the recent emergence of the RNA-programmable CRISPR-Cas9 system, a flexible methodology to produce essentially any desired modification in the genome. Cancer is a multistep process that involves many genetic mutations and other genome rearrangements. Despite their importance, it is difficult to recapitulate the degree of genetic complexity found in patient tumors. The CRISPR-Cas9 system for genome editing has been proven as a robust technology that makes it possible to generate cellular and animal models that recapitulate those cooperative alterations rapidly and at low cost. In this review, we will discuss the innovative applications of the CRISPR-Cas9 system to generate new models, providing a new way to interrogate the development and progression of cancers. PMID:26389881

  11. DNA fragment editing of genomes by CRISPR/Cas9.

    PubMed

    Jinhuan, Li; Jia, Shou; Qiang, Wu

    2015-10-01

    The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated nuclease 9 (Cas9) system from bacteria and archaea emerged recently as a new powerful technology of genome editing in virtually any organism. Due to its simplicity and cost effectiveness, a revolutionary change of genetics has occurred. Here, we summarize the recent development of DNA fragment editing methods by CRISPR/Cas9 and describe targeted DNA fragment deletions, inversions, duplications, insertions, and translocations. The efficient method of DNA fragment editing provides a powerful tool for studying gene function, regulatory elements, tissue development, and disease progression. Finally, we discuss the prospects of CRISPR/Cas9 system and the potential applications of other types of CRISPR system. PMID:26496751

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

    PubMed

    Torres-Ruiz, Raul; Rodriguez-Perales, Sandra

    2015-01-01

    The cancer-modelling field is now experiencing a conversion with the recent emergence of the RNA-programmable CRISPR-Cas9 system, a flexible methodology to produce essentially any desired modification in the genome. Cancer is a multistep process that involves many genetic mutations and other genome rearrangements. Despite their importance, it is difficult to recapitulate the degree of genetic complexity found in patient tumors. The CRISPR-Cas9 system for genome editing has been proven as a robust technology that makes it possible to generate cellular and animal models that recapitulate those cooperative alterations rapidly and at low cost. In this review, we will discuss the innovative applications of the CRISPR-Cas9 system to generate new models, providing a new way to interrogate the development and progression of cancers. PMID:26389881

  13. Abstraction and reformulation in artificial intelligence.

    PubMed Central

    Holte, Robert C.; Choueiry, Berthe Y.

    2003-01-01

    This paper contributes in two ways to the aims of this special issue on abstraction. The first is to show that there are compelling reasons motivating the use of abstraction in the purely computational realm of artificial intelligence. The second is to contribute to the overall discussion of the nature of abstraction by providing examples of the abstraction processes currently used in artificial intelligence. Although each type of abstraction is specific to a somewhat narrow context, it is hoped that collectively they illustrate the richness and variety of abstraction in its fullest sense. PMID:12903653

  14. Annotating user-defined abstractions for optimization

    SciTech Connect

    Quinlan, D; Schordan, M; Vuduc, R; Yi, Q

    2005-12-05

    This paper discusses the features of an annotation language that we believe to be essential for optimizing user-defined abstractions. These features should capture semantics of function, data, and object-oriented abstractions, express abstraction equivalence (e.g., a class represents an array abstraction), and permit extension of traditional compiler optimizations to user-defined abstractions. Our future work will include developing a comprehensive annotation language for describing the semantics of general object-oriented abstractions, as well as automatically verifying and inferring the annotated semantics.

  15. Applications of CRISPR-Cas systems in neuroscience

    PubMed Central

    Heidenreich, Matthias; Zhang, Feng

    2016-01-01

    Genome editing tools, and in particular those based on CRISPR-Cas systems, are accelerating the pace of biological research and enabling targeted genetic interrogation in virtually any organism and cell type. These tools have opened the door to the development of new model systems for studying the complexity of the nervous system, including animal and stem cell-derived in vitro models. Precise and efficient gene editing using CRISPR-Cas systems has the potential to advance both basic and translational neuroscience research. PMID:26656253

  16. High-throughput functional genomics using CRISPR-Cas9

    PubMed Central

    Shalem, Ophir; Sanjana, Neville E.; Zhang, Feng

    2015-01-01

    Forward genetic screens are powerful tools for the discovery and functional annotation of genetic elements. Recently, the RNA-guided CRISPR (clustered regularly interspaced short palindromic repeat)-associated Cas9 nuclease has been combined with genome-scale guide RNA libraries for unbiased, phenotypic screening. In this Review, we describe recent advances using Cas9 for genome-scale screens, including knockout approaches that inactivate genomic loci and strategies that modulate transcriptional activity. We discuss practical aspects of screen design, provide comparisons with RNA interference (RNAi) screening, and outline future applications and challenges. PMID:25854182

  17. 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

  18. 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

  19. V845 Cas - ein RRab-Stern mit Blazhko-Effekt

    NASA Astrophysics Data System (ADS)

    Maintz, Gisela

    2015-02-01

    CCD observations of V845 Cas (RA= 23 26 15.02, DE= +57 23 55.5 (2000)) were obtained at my private observatory. For V845 Cas 13 maxima were gained, showing a light curve, that varies from epoch to epoch. The irregularity of the lightcurves of V845 Cas is due to the Blazhko effect. Revised elements are given as: V845 Cas Max = 2456221.3450 + 0.570845 * E.

  20. Functional annotation of native enhancers with a Cas9-histone demethylase fusion.

    PubMed

    Kearns, Nicola A; Pham, Hannah; Tabak, Barbara; Genga, Ryan M; Silverstein, Noah J; Garber, Manuel; Maehr, René

    2015-05-01

    Understanding of mammalian enhancers is limited by the lack of a technology to rapidly and thoroughly test the cell type-specific function. Here, we use a nuclease-deficient Cas9 (dCas9)-histone demethylase fusion to functionally characterize previously described and new enhancer elements for their roles in the embryonic stem cell state. Further, we distinguish the mechanism of action of dCas9-LSD1 at enhancers from previous dCas9-effectors. PMID:25775043

  1. 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

  2. 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

  3. The Cas6e ribonuclease is not required for interference and adaptation by the E. coli type I-E CRISPR-Cas system

    PubMed Central

    Semenova, Ekaterina; Kuznedelov, Konstantin; Datsenko, Kirill A.; Boudry, Pierre M.; Savitskaya, Ekaterina E.; Medvedeva, Sofia; Beloglazova, Natalia; Logacheva, Maria; Yakunin, Alexander F.; Severinov, Konstantin

    2015-01-01

    CRISPR-Cas are small RNA-based adaptive prokaryotic immunity systems protecting cells from foreign DNA or RNA. Type I CRISPR-Cas systems are composed of a multiprotein complex (Cascade) that, when bound to CRISPR RNA (crRNA), can recognize double-stranded DNA targets and recruit the Cas3 nuclease to destroy target-containing DNA. In the Escherichia coli type I-E CRISPR-Cas system, crRNAs are generated upon transcription of CRISPR arrays consisting of multiple palindromic repeats and intervening spacers through the function of Cas6e endoribonuclease, which cleaves at specific positions of repeat sequences of the CRISPR array transcript. Cas6e is also a component of Cascade. Here, we show that when mature unit-sized crRNAs are provided in a Cas6e-independent manner by transcription termination, the CRISPR-Cas system can function without Cas6e. The results should allow facile interrogation of various targets by type I-E CRISPR-Cas system in E. coli using unit-sized crRNAs generated by transcription. PMID:26013814

  4. OIL POLLUTION ABSTRACTS. VOLUME 6, NUMBER 1

    EPA Science Inventory

    Oil Pollution Abstracts (formerly entitled Oil Pollution Reports) is a quarterly compilation of abstracts of current oil pollution related literature and research projects. Comprehensive coverage of oil pollution and its prevention and control is provided, with emphasis on the aq...

  5. An algorithm for generating abstract syntax trees

    NASA Technical Reports Server (NTRS)

    Noonan, R. E.

    1985-01-01

    The notion of an abstract syntax is discussed. An algorithm is presented for automatically deriving an abstract syntax directly from a BNF grammar. The implementation of this algorithm and its application to the grammar for Modula are discussed.

  6. 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

  7. Ultra-superovulation for the CRISPR-Cas9-mediated production of gene-knockout, single-amino-acid-substituted, and floxed mice

    PubMed Central

    Nakagawa, Yoshiko; Nishimichi, Norihisa; Yokosaki, Yasuyuki; Yanaka, Noriyuki; Takeo, Toru; Nakagata, Naomi; Yamamoto, Takashi

    2016-01-01

    ABSTRACT Current advances in producing genetically modified mice using genome-editing technologies have indicated the need for improvement of limiting factors including zygote collection for microinjection and their cryopreservation. Recently, we developed a novel superovulation technique using inhibin antiserum and equine chorionic gonadotropin to promote follicle growth. This method enabled the increased production of fertilized oocytes via in vitro fertilization compared with the conventional superovulation method. Here, we verify that the ultra-superovulation technique can be used for the efficient generation of clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 9 (Cas9)-mediated knockout mice by microinjection of plasmid vector or ribonucleoprotein into zygotes. We also investigated whether single-amino-acid-substituted mice and conditional knockout mice could be generated. Founder mice bearing base substitutions were generated more efficiently by co-microinjection of Cas9 protein, a guide RNA and single-stranded oligodeoxynucleotide (ssODN) than by plasmid microinjection with ssODN. The conditional allele was successfully introduced by the one-step insertion of an ssODN designed to carry an exon flanked by two loxP sequences and homology arms using a double-cut CRISPR-Cas9 strategy. Our study presents a useful method for the CRISPR-Cas9-based generation of genetically modified mice from the viewpoints of animal welfare and work efficiency. PMID:27387532

  8. Les tumeurs annexielles cutanées: étude anatomopathologique à propos de 96 cas

    PubMed Central

    El Ochi, Mohamed Réda; Boudhas, Adil; Allaoui, Mohammed; Rharrassi, Issam; Chahdi, Hafsa; Bouzidi, Abderrahman Al; Oukabli, Mohammed

    2015-01-01

    Les tumeurs annexielles cutanées sont des tumeurs primitives cutanées à la fois rares et hétérogènes. Elles sont le plus souvent bénignes et rarement malignes. Elles sont dominées du point de vu morphologique par leur polymorphisme lésionnel. Le but de cette étude est de relever le profil épidémiologique et les différents aspects anatomopathologiques de ce groupe de tumeurs dans une cohorte de population marocaine et de les comparer avec les données de la littérature. Il s'agit d'une étude rétrospective de 96 cas de tumeurs annexielles cutanées colligées au service d'anatomie et de cytologie pathologique de l'Hôpital Militaire d'Instruction Mohammed V de rabat durant une période de 6 ans, de Janvier 2009 à Décembre 2014. Le pic de fréquence est situé entre 31 et 40 ans. L’âge moyen est de 36 ans avec une prédominance masculine (63,5%). Le siège de prédilection est la région de la tête et cou (47,9%). Les tumeurs bénignes (97,9%) sont plus fréquentes que les tumeurs malignes. La différenciation est folliculaire dans 51% des cas, eccrine/apocrine dans 44,8% des cas et sébacée dans 4,2% des cas. Le type histologique le plus fréquent est le pilomatrixome (33,4%) suivi par l'hidradénome (12,5%) et le spiradénome eccrine (11,5%). Les tumeurs annexielles cutanées sont rares et très variées. Le profil épidémiologique et les aspects anatomopathologiques qui ressortent sont globalement superposables à ceux rapportés dans la littérature. Elles sont majoritairement bénignes, à prédominance masculine et dominées par le pilomatrixome et l'hidradénome nodulaire. Les tumeurs malignes sont rares, agressives et surviennent à un âge plus avancé. PMID:26185579

  9. Le cavernome porte chez l'enfant: a propos de 11 cas

    PubMed Central

    Ilham, Tadmori; Mounia, Lakhdar Idrissi; Moustapha, Hida

    2014-01-01

    Le cavernome portal est la conséquence d'une occlusion chronique, du système porte extra-hépatique formé d'un réseau de veines dont le calibre est augmenté et au sein desquelles chemine un sang portal hépatopéte. Chez l'enfant, est une cause majeure d'hypertension portale dite «pré ou infra-hépatique» ou encore «extra-hépatique». Onze cas de cavernome porte parmi 78 cas d'hypertension portale ont été colligés au service sur une période allant du Janvier 2003 au Septembre 2012. L’âge de nos patients variait entre 2 et 15 ans et le sexe ratio est de 1,75. Tous nos patients étaient admis au stade d'hypertension portale avec la splénomégalie SMG (100% des cas); hémorragies digestives (63%); ascite (36%); la circulation veineuse collatérale CVC et l'hépatomégalie HMG (27%). L'exploration endoscopique a objectivé la présence de varices ‘sophagiennes dans tous les cas avec une gastropathie hypertensive dans 27% et des varices ectopiques chez 36%. Les perturbations biologiques étaient dominées par la pancytopénie. Le bilan de thrombophilie était demandé pour tous les malades mais réalisé mais n'est réalisé que chez trois revenus normaux chez deux et a objectivé une baisse de protéine S chez le troisième. L’échographie abdominale était le moyen de diagnostic positif et l’écho-doppler a confirmé l'HTP chez tous nos patients. Aucun de nos malades n'a pu être opéré pour le moment. La ligature a été réalisée chez 54,5%. Dix patients ont nécessité une transfusion sanguine. L’évolution globale de nos patients est favorable. La durée moyenne d’évolution du cavernome porte chez nos patients est de quatre ans. PMID:25870732

  10. La maladie de Wilson chez l'enfant: à propos de 20 cas

    PubMed Central

    Idrissi, Mounia Lakhdar; Babakhoya, Abdeladim; Khabbache, Kawtar; Souilmi, Fatimzohra; Benmiloud, Sara; Abourrazak, Sanae; Chaouki, Sanae; Atmani, Samir; Bouharrou, Abdelhak; Hida, Moustapha

    2013-01-01

    La maladie de Wilson ou dégénérescence hépato-lenticulaire est une affection génétique autosomique récessive caractérisée par une accumulation toxique de cuivre dans l'organisme, essentiellement dans le foie, le système nerveux central et la cornée. L'objectif de ce travail était de soulever les difficultés diagnostiques et thérapeutiques dans la prise en charge de la maladie de Wilson dans notre contexte. Nous avons réalisé une étude rétrospective portant sur 20 cas de maladie de Wilson colligés au sein du service de pédiatrie du CHU HASSAN II de Fès sur une période de 7 ans et demi. Il s'agit de 13 garçons et 7 filles dont l’âge moyen est de 9 ans avec des extrêmes allant de 5 à 13 ans. La consanguinité est retrouvée chez 13 malades. Sur le plan clinique, l'ictère est noté dans 13 cas, un syndrome oedémateux est retrouvé dans 13 cas aussi et un syndrome hémorragique dans 6 cas. Les signes neurologiques sont présents dans 7cas. Trois enfants étaient asymptomatiques diagnostiqués à l'occasion d'un dépistage. Sur le plan biologique les signes d'insuffisance hépatocellulaire sont retrouvés chez 17 malades avec une cytolyse dans 8 cas. Une anémie hémolytique est retrouvée chez 8 malades (soit 40%). La céruléoplasminémie est abaissée chez 17 malades, la cuprurie réalisée chez 19 malades s'est révélée augmentée chez 17 soit 89,4%. L'anneau de Kayser- Fleischer est retrouvé chez 14 patients. L’échographie abdominale a montré des signes d'hypertension portale (HTP) sur foie de cirrhose chez 16 malades soit 80%. La D pénicillamine est instaurée chez 17 patients et trois sont mis sous sulfate de zinc. Trois malades ont bénéficié de la vitamine B6. L’évolution est favorable chez 11 malades avec un recul moyen de 3 ans. Nous déplorons 4 décès chez des malades ayant consulté au stade de cirrhose décompensé. Le pronostic de la maladie de Wilson dépend de la précocité du traitement. Le dépistage chez

  11. 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…

  12. After 16 Years of Publishing Standards, Do CAS Standards Make a Difference?

    ERIC Educational Resources Information Center

    Arminio, Jan; Gochenauer, Patty

    2004-01-01

    Using members of professional associations who are a part of the Council for the Advancement of Standards in Higher Education (CAS) consortia as a sample, this study investigated who uses CAS Standards, how and why they are used, and whether CAS Standards are associated with enhanced student learning. Using a quantitative analysis, this study…

  13. Creating Genome Modifications in C. elegans Using the CRISPR/Cas9 System.

    PubMed

    Calarco, John A; Friedland, Ari E

    2015-01-01

    The clustered, regularly interspaced, short, palindromic repeat (CRISPR)-associated (CAS) nuclease Cas9 has been used in many organisms to generate specific mutations and transgene insertions. Here we describe a method using the S. pyogenes Cas9 in C. elegans that provides a convenient and effective approach for making heritable changes to the worm genome. PMID:26423968

  14. 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...

  15. 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...

  16. 48 CFR 9904.412-60.1 - Illustrations-CAS Pension Harmonization Rule.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 48 Federal Acquisition Regulations System 7 2013-10-01 2012-10-01 true Illustrations-CAS Pension... AND COST ACCOUNTING STANDARDS COST ACCOUNTING STANDARDS 9904.412-60.1 Illustrations—CAS Pension... cost on or after the Applicability Date of the CAS Harmonization Rule. The illustrations present...

  17. 48 CFR 9904.412-60.1 - Illustrations-CAS Pension Harmonization Rule.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 48 Federal Acquisition Regulations System 7 2014-10-01 2014-10-01 false Illustrations-CAS Pension... AND COST ACCOUNTING STANDARDS COST ACCOUNTING STANDARDS 9904.412-60.1 Illustrations—CAS Pension... cost on or after the Applicability Date of the CAS Harmonization Rule. The illustrations present...

  18. 48 CFR 9904.412-60.1 - Illustrations-CAS Pension Harmonization Rule.

    Code of Federal Regulations, 2012 CFR

    2012-10-01

    ... 48 Federal Acquisition Regulations System 7 2012-10-01 2012-10-01 false Illustrations-CAS Pension... AND COST ACCOUNTING STANDARDS COST ACCOUNTING STANDARDS 9904.412-60.1 Illustrations—CAS Pension... cost on or after the Applicability Date of the CAS Harmonization Rule. The illustrations present...

  19. 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...

  20. 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…

  1. 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...

  2. 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...

  3. 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…

  4. 48 CFR 9904.412-64.1 - Transition Method for the CAS Pension Harmonization Rule.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... CAS Pension Harmonization Rule. 9904.412-64.1 Section 9904.412-64.1 Federal Acquisition Regulations... Transition Method for the CAS Pension Harmonization Rule. Contractors or subcontractors that become subject... through 7 Notes (Note 1) Actuarial Accrued Liability $2,470,500 $14,225,000 2 CAS Actuarial Value...

  5. Guide RNAs: A Glimpse at the Sequences that Drive CRISPR-Cas Systems.

    PubMed

    Briner, Alexandra E; Barrangou, Rodolphe

    2016-01-01

    CRISPR-Cas systems provide adaptive immunity in bacteria and archaea. Although there are two main classes of CRISPR-Cas systems defined by gene content, interfering RNA biogenesis, and effector proteins, Type II systems have recently been exploited on a broad scale to develop next-generation genetic engineering and genome-editing tools. Conveniently, Type II systems are streamlined and rely on a single protein, Cas9, and a guide RNA molecule, comprised of a CRISPR RNA (crRNA) and trans-acting CRISPR RNA (tracrRNA), to achieve effective and programmable nucleic acid targeting and cleavage. Currently, most commercially available Cas9-based genome-editing tools use the CRISPR-Cas system from Streptococcus pyogenes (SpyCas9), although many orthogonal Type II systems are available for diverse and multiplexable genome engineering applications. Here, we discuss the biological significance of Type II CRISPR-Cas elements, including the tracrRNA, crRNA, Cas9, and protospacer-adjacent motif (PAM), and look at the native function of these elements to understand how they can be engineered, enhanced, and optimized for genome editing applications. Additionally, we discuss the basis for orthogonal Cas9 and guide RNA systems that would allow researchers to concurrently use multiple Cas9-based systems for different purposes. Understanding the native function of endogenous Type II CRISPR-Cas systems can lead to new Cas9 tool development to expand the genetic manipulation toolbox. PMID:27371605

  6. 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

  7. 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

  8. 2013 SYR Accepted Poster Abstracts.

    PubMed

    2013-01-01

    SYR 2013 Accepted Poster abstracts: 1. Benefits of Yoga as a Wellness Practice in a Veterans Affairs (VA) Health Care Setting: If You Build It, Will They Come? 2. Yoga-based Psychotherapy Group With Urban Youth Exposed to Trauma. 3. Embodied Health: The Effects of a Mind�Body Course for Medical Students. 4. Interoceptive Awareness and Vegetable Intake After a Yoga and Stress Management Intervention. 5. Yoga Reduces Performance Anxiety in Adolescent Musicians. 6. Designing and Implementing a Therapeutic Yoga Program for Older Women With Knee Osteoarthritis. 7. Yoga and Life Skills Eating Disorder Prevention Among 5th Grade Females: A Controlled Trial. 8. A Randomized, Controlled Trial Comparing the Impact of Yoga and Physical Education on the Emotional and Behavioral Functioning of Middle School Children. 9. Feasibility of a Multisite, Community based Randomized Study of Yoga and Wellness Education for Women With Breast Cancer Undergoing Chemotherapy. 10. A Delphi Study for the Development of Protocol Guidelines for Yoga Interventions in Mental Health. 11. Impact Investigation of Breathwalk Daily Practice: Canada�India Collaborative Study. 12. Yoga Improves Distress, Fatigue, and Insomnia in Older Veteran Cancer Survivors: Results of a Pilot Study. 13. Assessment of Kundalini Mantra and Meditation as an Adjunctive Treatment With Mental Health Consumers. 14. Kundalini Yoga Therapy Versus Cognitive Behavior Therapy for Generalized Anxiety Disorder and Co-Occurring Mood Disorder. 15. Baseline Differences in Women Versus Men Initiating Yoga Programs to Aid Smoking Cessation: Quitting in Balance Versus QuitStrong. 16. Pranayam Practice: Impact on Focus and Everyday Life of Work and Relationships. 17. Participation in a Tailored Yoga Program is Associated With Improved Physical Health in Persons With Arthritis. 18. Effects of Yoga on Blood Pressure: Systematic Review and Meta-analysis. 19. A Quasi-experimental Trial of a Yoga based Intervention to Reduce Stress and

  9. Observational Activities at Manipur University, India (Abstract)

    NASA Astrophysics Data System (ADS)

    Singh, K. Y.; Meitei, I. A.; Singh, S. A.; Singh, R. B.

    2015-06-01

    (Abstract only) We have innovatively designed and constructed three observatories each costing a few hundred USD for housing three small Schmidt-Cassegrain type telescopes namely, Celestron CGE925, Celestron CGE1400, Meade 12-inch LX200GPS. These observatories are completely different in design and are found to be perfectly usable for doing serious work on astronomical observation and measurements. The observatory with the Celestron CGE1400 telescope has been inducted, since January 2012, as one of the observatories of the international “Orion Project” headquartered at Phoenix, Arizona, which is dedicated for photometric and spectroscopic observations of five bright variable stars of the Orion constellation namely, Betelgeuse (alpha Ori), Rigel (beta Ori), Mintaka (delta Ori), Alnilam (epsilon Ori) and Alnitak (zeta Ori). Using this observatory, we have been producing BVRI photometric data for the five stars of the Orion project. The other observatory with the Meade 12-inch LX200GPS telescope is being inducted into service for CCD photometric study of SU UMa stars in connection with implementation of a project funded by Indian Space Research Organization (ISRO). In the present paper, we would like to describe our self-built observatories, our observational facilities, the BVRI photometric data that we acquired for the Orion project, and our future plan for observation of variable stars of interest.

  10. At the HeART of Abstraction

    ERIC Educational Resources Information Center

    Berdit, Nancy

    2006-01-01

    Abstraction has long been a concept difficult to define for students. Students often feel the pressure of making their artwork "look real" and frustration can often lead to burnout in the classroom. In this article, the author describes how her lesson on abstraction has alleviated much of that pressure as students created an abstract acrylic…

  11. 37 CFR 1.438 - The abstract.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 37 Patents, Trademarks, and Copyrights 1 2010-07-01 2010-07-01 false The abstract. 1.438 Section 1... COMMERCE GENERAL RULES OF PRACTICE IN PATENT CASES International Processing Provisions The International Application § 1.438 The abstract. (a) Requirements as to the content and form of the abstract are set forth...

  12. 37 CFR 1.438 - The abstract.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 37 Patents, Trademarks, and Copyrights 1 2011-07-01 2011-07-01 false The abstract. 1.438 Section 1... COMMERCE GENERAL RULES OF PRACTICE IN PATENT CASES International Processing Provisions The International Application § 1.438 The abstract. (a) Requirements as to the content and form of the abstract are set forth...

  13. 37 CFR 1.438 - The abstract.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... 37 Patents, Trademarks, and Copyrights 1 2013-07-01 2013-07-01 false The abstract. 1.438 Section 1... COMMERCE GENERAL RULES OF PRACTICE IN PATENT CASES International Processing Provisions The International Application § 1.438 The abstract. (a) Requirements as to the content and form of the abstract are set forth...

  14. 37 CFR 1.438 - The abstract.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 37 Patents, Trademarks, and Copyrights 1 2014-07-01 2014-07-01 false The abstract. 1.438 Section 1... COMMERCE GENERAL RULES OF PRACTICE IN PATENT CASES International Processing Provisions The International Application § 1.438 The abstract. (a) Requirements as to the content and form of the abstract are set forth...

  15. 37 CFR 1.438 - The abstract.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... 37 Patents, Trademarks, and Copyrights 1 2012-07-01 2012-07-01 false The abstract. 1.438 Section 1... COMMERCE GENERAL RULES OF PRACTICE IN PATENT CASES International Processing Provisions The International Application § 1.438 The abstract. (a) Requirements as to the content and form of the abstract are set forth...

  16. Writing a Structured Abstract for the Thesis

    ERIC Educational Resources Information Center

    Hartley, James

    2010-01-01

    This article presents the author's suggestions on how to improve thesis abstracts. The author describes two books on writing abstracts: (1) "Creating Effective Conference Abstracts and Posters in Biomedicine: 500 tips for Success" (Fraser, Fuller & Hutber, 2009), a compendium of clear advice--a must book to have in one's hand as one prepares a…

  17. 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

  18. Les fistules œsotrachéales congénitales isolées à propos de 2 cas

    PubMed Central

    El Biache, Imad; Lechqar, Maryem; Rami, Mohammed; Bouabdallah, Youssef

    2014-01-01

    Les auteurs rapportent 2 cas de fistules oesotrachéales isolées sans atrésie de l'oesophage, colligés au service de chirurgie pédiatrique au CHU Hassan II de Fès au Maroc entre 2008 et 2013. Il s'agit d'une anomalie rare représentée par un fin canal ascendant entre l'oesophage et la face postérieure de la trachée, à la hauteur du défilé cervico-thoracique. Elle se manifeste cliniquement par une symptomatologie respiratoire parfois digestive. Le diagnostic a été confirmé par le transit oesophagien dans les 2 cas et a permis aussi de déterminer le siège de la fistule. Le traitement était chirurgical, il a permis de supprimer la communication anormale entre l'oesophage et la trachée par un abord cervical avec interposition musculaire dans les 2 cas. Les suites post-opératoires et l’évolution à long terme étaient simples. Le but de ce travail est d'exposer les différents moyens diagnostique et thérapeutique. PMID:25328600

  19. Research & writing basics: elements of the abstract.

    PubMed

    Krasner, D; Van Rijswijk, L

    1995-04-01

    Writing an abstract is a challenging skill that requires precision and care. Criteria for well-formulated abstracts and abstract guidelines for 2 types of articles (empirical studies and reviews or theoretical articles) as well as a description of the content of a structured abstract are presented. Details were gleaned from a review of the literature including the American Medical Association Manual of Style, Eighth Edition and the Publication Manual of the American Psychological Association, Fourth Edition. A good abstract is like a crystal: it is a clear, sharp synthesis that elucidates meaning for the reader. PMID:7546111

  20. 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…

  1. 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…

  2. Research Needed on the Use of CAS Standards and Guidelines.

    ERIC Educational Resources Information Center

    Creamer, Don G.

    2003-01-01

    This article suggests research projects that would extend the knowledge base about the use of Council for the Advancement of Standards in Higher Education (CAS) standards and guidelines in useful ways. Included are five research questions and specific research methodologies to guide researchers. (Contains 20 references.) (Author)

  3. 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…

  4. 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…

  5. Targeted mutagenesis in chicken using CRISPR/Cas9 system.

    PubMed

    Oishi, Isao; Yoshii, Kyoko; Miyahara, Daichi; Kagami, Hiroshi; Tagami, Takahiro

    2016-01-01

    The CRISPR/Cas9 system is a simple and powerful tool for genome editing in various organisms including livestock animals. However, the system has not been applied to poultry because of the difficulty in accessing their zygotes. Here we report the implementation of CRISPR/Cas9-mediated gene targeting in chickens. Two egg white genes, ovalbumin and ovomucoid, were efficiently (>90%) mutagenized in cultured chicken primordial germ cells (PGCs) by transfection of circular plasmids encoding Cas9, a single guide RNA, and a gene encoding drug resistance, followed by transient antibiotic selection. We transplanted CRISPR-induced mutant-ovomucoid PGCs into recipient chicken embryos and established three germline chimeric roosters (G0). All of the roosters had donor-derived mutant-ovomucoid spermatozoa, and the two with a high transmission rate of donor-derived gametes produced heterozygous mutant ovomucoid chickens as about half of their donor-derived offspring in the next generation (G1). Furthermore, we generated ovomucoid homozygous mutant offspring (G2) by crossing the G1 mutant chickens. Taken together, these results demonstrate that the CRISPR/Cas9 system is a simple and effective gene-targeting method in chickens. PMID:27050479

  6. 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…

  7. CRISPR/Cas9-targeted mutagenesis in Caenorhabditis elegans.

    PubMed

    Waaijers, Selma; Portegijs, Vincent; Kerver, Jana; Lemmens, Bennie B L G; Tijsterman, Marcel; van den Heuvel, Sander; Boxem, Mike

    2013-11-01

    The generation of genetic mutants in Caenorhabditis elegans has long relied on the selection of mutations in large-scale screens. Directed mutagenesis of specific loci in the genome would greatly speed up analysis of gene function. Here, we adapt the CRISPR/Cas9 system to generate mutations at specific sites in the C. elegans genome. PMID:23979586

  8. CRISPR/Cas9 advances engineering of microbial cell factories.

    PubMed

    Jakočiūnas, Tadas; Jensen, Michael K; Keasling, Jay D

    2016-03-01

    One of the key drivers for successful metabolic engineering in microbes is the efficacy by which genomes can be edited. As such there are many methods to choose from when aiming to modify genomes, especially those of model organisms like yeast and bacteria. In recent years, clustered regularly interspaced palindromic repeats (CRISPR) and its associated proteins (Cas) have become the method of choice for precision genome engineering in many organisms due to their orthogonality, versatility and efficacy. Here we review the strategies adopted for implementation of RNA-guided CRISPR/Cas9 genome editing with special emphasis on their application for metabolic engineering of yeast and bacteria. Also, examples of how nuclease-deficient Cas9 has been applied for RNA-guided transcriptional regulation of target genes will be reviewed, as well as tools available for computer-aided design of guide-RNAs will be highlighted. Finally, this review will provide a perspective on the immediate challenges and opportunities foreseen by the use of CRISPR/Cas9 genome engineering and regulation in the context of metabolic engineering. PMID:26707540

  9. CRISPR-Cas9 genome editing in Drosophila

    PubMed Central

    Gratz, Scott J.; Rubinstein, C. Dustin; Harrison, Melissa M.; Wildonger, Jill; O’Connor-Giles, Kate M.

    2015-01-01

    The CRISPR-Cas9 system has transformed genome engineering of model organisms from possible to practical. CRISPR-Cas9 can be readily programmed to generate sequence-specific double-strand breaks that disrupt targeted loci when repaired by error-prone non-homologous end joining or to catalyze precise genome modification through homology-directed repair (HDR). Here we describe a streamlined approach for rapid and highly efficient engineering of the Drosophila genome via CRISPR-Cas9-mediated HDR. In this approach, transgenic flies expressing Cas9 are injected with plasmids to express guide RNAs (gRNAs) and positively marked donor templates. We detail target site selection; gRNA plasmid generation; donor template design and construction; and the generation, identification and molecular confirmation of engineered lines. We also present alternative approaches and highlight key considerations for experimental design. The approach outlined here can be used to rapidly and reliably generate a variety of engineered modifications, including genomic deletions and replacements, precise sequence edits, and incorporation of protein tags. PMID:26131852

  10. Indispensable Manual Calculation Skills in a CAS Environment.

    ERIC Educational Resources Information Center

    Herget, Wilfried; Heugl, Helmut; Kutzler, Bernhard; Lehmann, Eberhard

    Which manual calculation skills are still needed when students use graphic/symbolic calculators or computers with computer algebra systems (CAS)? What should students be able to do manually, i.e. just using paper and pencil? This text is the outcome of a two-day discussion on these questions, held by the four authors. Our answers and proposals are…

  11. Some Reflections on CAS Assisted Proofs of Theorems

    ERIC Educational Resources Information Center

    Dana-Picard, Thierry

    2005-01-01

    A mathematician's work consists of proving theorems, calculating, and making mathematics understandable. An assistant for all three components is a Computer Algebra System. We describe and discuss various CAS-assisted processes for proving theorems, and discuss the constraints which can appear regarding efficiency, confidence in the result and…

  12. Technological Discourse on CAS-Based Operative Knowledge

    ERIC Educational Resources Information Center

    Mann, Giora; Dana-Picard, Thierry; Zehavi, Nurit

    2007-01-01

    This article begins with a comparison of two groups of teachers, working on the same tasks in Analytic Geometry. One group has only basic experience in CAS-assisted problem solving, and the other group has extensive experience. The comparison is discussed in terms of the interplay between reflection, operative knowledge and execution. The findings…

  13. Duchenne muscular dystrophy: CRISPR/Cas9 treatment.

    PubMed

    Mendell, Jerry R; Rodino-Klapac, Louise R

    2016-05-01

    A novel approach to gene correction by genome editing shows great promise as a treatment for Duchenne muscular dystrophy (DMD). CRISPR/Cas9 delivered by adeno-associated virus to a mouse model for DMD demonstrated improvement in function and histology. PMID:26926391

  14. 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…

  15. 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

  16. Maximizing mutagenesis with solubilized CRISPR-Cas9 ribonucleoprotein complexes.

    PubMed

    Burger, Alexa; Lindsay, Helen; Felker, Anastasia; Hess, Christopher; Anders, Carolin; Chiavacci, Elena; Zaugg, Jonas; Weber, Lukas M; Catena, Raul; Jinek, Martin; Robinson, Mark D; Mosimann, Christian

    2016-06-01

    CRISPR-Cas9 enables efficient sequence-specific mutagenesis for creating somatic or germline mutants of model organisms. Key constraints in vivo remain the expression and delivery of active Cas9-sgRNA ribonucleoprotein complexes (RNPs) with minimal toxicity, variable mutagenesis efficiencies depending on targeting sequence, and high mutation mosaicism. Here, we apply in vitro assembled, fluorescent Cas9-sgRNA RNPs in solubilizing salt solution to achieve maximal mutagenesis efficiency in zebrafish embryos. MiSeq-based sequence analysis of targeted loci in individual embryos using CrispRVariants, a customized software tool for mutagenesis quantification and visualization, reveals efficient bi-allelic mutagenesis that reaches saturation at several tested gene loci. Such virtually complete mutagenesis exposes loss-of-function phenotypes for candidate genes in somatic mutant embryos for subsequent generation of stable germline mutants. We further show that targeting of non-coding elements in gene regulatory regions using saturating mutagenesis uncovers functional control elements in transgenic reporters and endogenous genes in injected embryos. Our results establish that optimally solubilized, in vitro assembled fluorescent Cas9-sgRNA RNPs provide a reproducible reagent for direct and scalable loss-of-function studies and applications beyond zebrafish experiments that require maximal DNA cutting efficiency in vivo. PMID:27130213

  17. Genome Editing with CRISPR-Cas9: Can It Get Any Better?

    PubMed

    Haeussler, Maximilian; Concordet, Jean-Paul

    2016-05-20

    The CRISPR-Cas revolution is taking place in virtually all fields of life sciences. Harnessing DNA cleavage with the CRISPR-Cas9 system of Streptococcus pyogenes has proven to be extraordinarily simple and efficient, relying only on the design of a synthetic single guide RNA (sgRNA) and its co-expression with Cas9. Here, we review the progress in the design of sgRNA from the original dual RNA guide for S. pyogenes and Staphylococcus aureus Cas9 (SpCas9 and SaCas9). New assays for genome-wide identification of off-targets have provided important insights into the issue of cleavage specificity in vivo. At the same time, the on-target activity of thousands of guides has been determined. These data have led to numerous online tools that facilitate the selection of guide RNAs in target sequences. It appears that for most basic research applications, cleavage activity can be maximized and off-targets minimized by carefully choosing guide RNAs based on computational predictions. Moreover, recent studies of Cas proteins have further improved the flexibility and precision of the CRISPR-Cas toolkit for genome editing. Inspired by the crystal structure of the complex of sgRNA-SpCas9 bound to target DNA, several variants of SpCas9 have recently been engineered, either with novel protospacer adjacent motifs (PAMs) or with drastically reduced off-targets. Novel Cas9 and Cas9-like proteins called Cpf1 have also been characterized from other bacteria and will benefit from the insights obtained from SpCas9. Genome editing with CRISPR-Cas9 may also progress with better understanding and control of cellular DNA repair pathways activated after Cas9-induced DNA cleavage. PMID:27210042

  18. CRISPR-Cas and Restriction-Modification Act Additively against Conjugative Antibiotic Resistance Plasmid Transfer in Enterococcus faecalis

    PubMed Central

    Price, Valerie J.; Huo, Wenwen; Sharifi, Ardalan

    2016-01-01

    ABSTRACT Enterococcus faecalis is an opportunistic pathogen and a leading cause of nosocomial infections. Conjugative pheromone-responsive plasmids are narrow-host-range mobile genetic elements (MGEs) that are rapid disseminators of antibiotic resistance in the faecalis species. Clustered regularly interspaced short palindromic repeat (CRISPR)-Cas and restriction-modification confer acquired and innate immunity, respectively, against MGE acquisition in bacteria. Most multidrug-resistant E. faecalis isolates lack CRISPR-Cas and possess an orphan locus lacking cas genes, CRISPR2, that is of unknown function. Little is known about restriction-modification defense in E. faecalis. Here, we explore the hypothesis that multidrug-resistant E. faecalis strains are immunocompromised. We assessed MGE acquisition by E. faecalis T11, a strain closely related to the multidrug-resistant hospital isolate V583 but which lacks the ~620 kb of horizontally acquired genome content that characterizes V583. T11 possesses the E. faecalis CRISPR3-cas locus and a predicted restriction-modification system, neither of which occurs in V583. We demonstrate that CRISPR-Cas and restriction-modification together confer a 4-log reduction in acquisition of the pheromone-responsive plasmid pAM714 in biofilm matings. Additionally, we show that the orphan CRISPR2 locus is functional for genome defense against another pheromone-responsive plasmid, pCF10, only in the presence of cas9 derived from the E. faecalis CRISPR1-cas locus, which most multidrug-resistant E. faecalis isolates lack. Overall, our work demonstrated that the loss of only two loci led to a dramatic reduction in genome defense against a clinically relevant MGE, highlighting the critical importance of the E. faecalis accessory genome in modulating horizontal gene transfer. Our results rationalize the development of antimicrobial strategies that capitalize upon the immunocompromised status of multidrug-resistant E

  19. Production of genome-edited pluripotent stem cells and mice by CRISPR/Cas [Review].

    PubMed

    Horii, Takuro; Hatada, Izuho

    2016-03-31

    Clustered regularly at interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) nucleases, so-called CRISPR/Cas, was recently developed as an epoch-making genome engineering technology. This system only requires Cas9 nuclease and single-guide RNA complementary to a target locus. CRISPR/Cas enables the generation of knockout cells and animals in a single step. This system can also be used to generate multiple mutations and knockin in a single step, which is not possible using other methods. In this review, we provide an overview of genome editing by CRISPR/Cas in pluripotent stem cells and mice. PMID:26743444

  20. 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

  1. 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

  2. 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

  3. 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.

  4. Structural Basis for the Altered PAM Specificities of Engineered CRISPR-Cas9.

    PubMed

    Hirano, Seiichi; Nishimasu, Hiroshi; Ishitani, Ryuichiro; Nureki, Osamu

    2016-03-17

    The RNA-guided endonuclease Cas9 cleaves double-stranded DNA targets bearing a PAM (protospacer adjacent motif) and complementarity to the guide RNA. A recent study showed that, whereas wild-type Streptococcus pyogenes Cas9 (SpCas9) recognizes the 5'-NGG-3' PAM, the engineered VQR, EQR, and VRER SpCas9 variants recognize the 5'-NGA-3', 5'-NGAG-3', and 5'-NGCG-3' PAMs, respectively, thus expanding the targetable sequences in Cas9-mediated genome editing applications. Here, we present the high-resolution crystal structures of the three SpCas9 variants in complexes with a single-guide RNA and its altered PAM-containing, partially double-stranded DNA targets. A structural comparison of the three SpCas9 variants with wild-type SpCas9 revealed that the multiple mutations synergistically induce an unexpected displacement in the phosphodiester backbone of the PAM duplex, thereby allowing the SpCas9 variants to directly recognize the altered PAM nucleotides. Our findings explain the altered PAM specificities of the SpCas9 variants and establish a framework for further rational engineering of CRISPR-Cas9. PMID:26990991

  5. Chemical and Biophysical Modulation of Cas9 for Tunable Genome Engineering.

    PubMed

    Nuñez, James K; Harrington, Lucas B; Doudna, Jennifer A

    2016-03-18

    The application of the CRISPR-Cas9 system for genome engineering has revolutionized the ability to interrogate genomes of mammalian cells. Programming the Cas9 endonuclease to induce DNA breaks at specified sites is achieved by simply modifying the sequence of its cognate guide RNA. Although Cas9-mediated genome editing has been shown to be highly specific, cleavage events at off-target sites have also been reported. Minimizing, and eventually abolishing, unwanted off-target cleavage remains a major goal of the CRISPR-Cas9 technology before its implementation for therapeutic use. Recent efforts have turned to chemical biology and biophysical approaches to engineer inducible genome editing systems for controlling Cas9 activity at the transcriptional and protein levels. Here, we review recent advancements to modulate Cas9-mediated genome editing by engineering split-Cas9 constructs, inteins, small molecules, protein-based dimerizing domains, and light-inducible systems. PMID:26857072

  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. Analysis of complex networks using aggressive abstraction.

    SciTech Connect

    Colbaugh, Richard; Glass, Kristin.; Willard, Gerald

    2008-10-01

    This paper presents a new methodology for analyzing complex networks in which the network of interest is first abstracted to a much simpler (but equivalent) representation, the required analysis is performed using the abstraction, and analytic conclusions are then mapped back to the original network and interpreted there. We begin by identifying a broad and important class of complex networks which admit abstractions that are simultaneously dramatically simplifying and property preserving - we call these aggressive abstractions -- and which can therefore be analyzed using the proposed approach. We then introduce and develop two forms of aggressive abstraction: 1.) finite state abstraction, in which dynamical networks with uncountable state spaces are modeled using finite state systems, and 2.) onedimensional abstraction, whereby high dimensional network dynamics are captured in a meaningful way using a single scalar variable. In each case, the property preserving nature of the abstraction process is rigorously established and efficient algorithms are presented for computing the abstraction. The considerable potential of the proposed approach to complex networks analysis is illustrated through case studies involving vulnerability analysis of technological networks and predictive analysis for social processes.

  8. An Evaluation of References to Indexes and Abstracts in Ulrich's 17th Edition.

    ERIC Educational Resources Information Center

    Wyndham, Diana

    1980-01-01

    Ulrich's performance in listing references to indexes was tested by comparing its statement of where a journal is indexed with the list of the indexed journals in the indexing/abstracting services themselves. (Author)

  9. Prefrontal cortex organization: dissociating effects of temporal abstraction, relational abstraction, and integration with FMRI.

    PubMed

    Nee, Derek Evan; Jahn, Andrew; Brown, Joshua W

    2014-09-01

    The functions of the prefrontal cortex (PFC) underlie higher-level cognition. Varying proposals suggest that the PFC is organized along a rostral-caudal gradient of abstraction with more abstract representations/processes associated with more rostral areas. However, the operational definition of abstraction is unclear. Here, we contrasted 2 prominent theories of abstraction--temporal and relational--using fMRI. We further examined whether integrating abstract rules--a function common to each theory--recruited the PFC independently of other abstraction effects. While robust effects of relational abstraction were present in the PFC, temporal abstraction effects were absent. Instead, we found activations specific to the integration of relational rules in areas previously shown to be associated with temporal abstraction. We suggest that previous effects of temporal abstraction were due to confounds with integration demands. We propose an integration framework to understand the functions of the PFC that resolves discrepancies in prior data. PMID:23563962

  10. 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

  11. Developing Creativity and Abstraction in Representing Data

    ERIC Educational Resources Information Center

    South, Andy

    2012-01-01

    Creating charts and graphs is all about visual abstraction: the process of representing aspects of data with imagery that can be interpreted by the reader. Children may need help making the link between the "real" and the image. This abstraction can be achieved using symbols, size, colour and position. Where the representation is close to what…

  12. Content Differences for Abstract and Concrete Concepts

    ERIC Educational Resources Information Center

    Wiemer-Hastings, Katja Katja; Xu, Xu

    2005-01-01

    Concept properties are an integral part of theories of conceptual representation and processing. To date, little is known about conceptual properties of abstract concepts, such as idea. This experiment systematically compared the content of 18 abstract and 18 concrete concepts, using a feature generation task. Thirty-one participants listed…

  13. Annual Abstract Series of Educational Material.

    ERIC Educational Resources Information Center

    Shelesnyak, M. C., Ed.

    1980-01-01

    This is the fifth annual collection of abstracts of educational materials presented by the Educational Materials Review Board of the American Physiological Society under the direction of the Education Committee. The collection includes abstracts of articles, papers, textbooks, books, handbooks, and symposia which are valuable in teaching…

  14. Abstracting in the Context of Spontaneous Learning

    ERIC Educational Resources Information Center

    Williams, Gaye

    2007-01-01

    There is evidence that spontaneous learning leads to relational understanding and high positive affect. To study spontaneous abstracting, a model was constructed by combining the RBC model of abstraction with Krutetskii's mental activities. Using video-stimulated interviews, the model was then used to analyze the behavior of two Year 8 students…

  15. National Workplace Literacy Program. 1993 Abstracts.

    ERIC Educational Resources Information Center

    Office of Vocational and Adult Education (ED), Washington, DC. National Workplace Literacy Program.

    This publication presents the abstracts of the 57 National Workplace Literacy Program 1993 projects. Each abstract provides the following information: project title; award number; project director; awardee; address; telephone and fax numbers; funds by fiscal year (federal and nonfederal); award period; federal project officer; objectives;…

  16. A Hybrid Method for Abstracting Newspaper Articles.

    ERIC Educational Resources Information Center

    Liu, James; Wu, Yan; Zhou, Lina

    1999-01-01

    Introduces a hybrid method for abstracting Chinese text that integrates the statistical approach with language understandings, incorporating some linguistics heuristics and segmentation into the abstracting process. Initial responses from application to Chinese newspaper articles show that the method contributes much to the flexibility and…

  17. Abstractions of Awareness: Aware of What?

    NASA Astrophysics Data System (ADS)

    Metaxas, Georgios; Markopoulos, Panos

    This chapter presents FN-AAR, an abstract model of awareness systems. The purpose of the model is to capture in a concise and abstract form essential aspects of awareness systems, many of which have been discussed in design essays or in the context of evaluating specific design solutions.

  18. Foundations of the Bandera Abstraction Tools

    NASA Technical Reports Server (NTRS)

    Hatcliff, John; Dwyer, Matthew B.; Pasareanu, Corina S.; Robby

    2003-01-01

    Current research is demonstrating that model-checking and other forms of automated finite-state verification can be effective for checking properties of software systems. Due to the exponential costs associated with model-checking, multiple forms of abstraction are often necessary to obtain system models that are tractable for automated checking. The Bandera Tool Set provides multiple forms of automated support for compiling concurrent Java software systems to models that can be supplied to several different model-checking tools. In this paper, we describe the foundations of Bandera's data abstraction mechanism which is used to reduce the cardinality (and the program's state-space) of data domains in software to be model-checked. From a technical standpoint, the form of data abstraction used in Bandera is simple, and it is based on classical presentations of abstract interpretation. We describe the mechanisms that Bandera provides for declaring abstractions, for attaching abstractions to programs, and for generating abstracted programs and properties. The contributions of this work are the design and implementation of various forms of tool support required for effective application of data abstraction to software components written in a programming language like Java which has a rich set of linguistic features.

  19. Interpreting Abstract Interpretations in Membership Equational Logic

    NASA Technical Reports Server (NTRS)

    Fischer, Bernd; Rosu, Grigore

    2001-01-01

    We present a logical framework in which abstract interpretations can be naturally specified and then verified. Our approach is based on membership equational logic which extends equational logics by membership axioms, asserting that a term has a certain sort. We represent an abstract interpretation as a membership equational logic specification, usually as an overloaded order-sorted signature with membership axioms. It turns out that, for any term, its least sort over this specification corresponds to its most concrete abstract value. Maude implements membership equational logic and provides mechanisms to calculate the least sort of a term efficiently. We first show how Maude can be used to get prototyping of abstract interpretations "for free." Building on the meta-logic facilities of Maude, we further develop a tool that automatically checks and abstract interpretation against a set of user-defined properties. This can be used to select an appropriate abstract interpretation, to characterize the specified loss of information during abstraction, and to compare different abstractions with each other.

  20. Tour the Galaxy of the Abstract.

    ERIC Educational Resources Information Center

    Kennedy, Patricia

    2003-01-01

    Describes an abstract art unit in which students in an introductory art course created abstract art inspired by the work of M. C. Escher. Explains that some students are unsure of their drawing ability. States this unit helps them overcome their fears. (CMK)

  1. Some Call It Stone: Teaching Abstract Sculpture

    ERIC Educational Resources Information Center

    Asher, Rikki

    2004-01-01

    Abstract visual art is not for everybody. Some people find it threatening, uncomfortable, and often, inaccessible. Understandably, this can result in a lack of attention paid to nonrepresentational works of art in the visual arts curriculum. This article describes an experiential, hands-on, field trip that sought to demystify abstract sculpture,…

  2. Third LDEF Post-Retrieval Symposium Abstracts

    NASA Technical Reports Server (NTRS)

    Levine, Arlene S. (Compiler)

    1993-01-01

    This volume is a compilation of abstracts submitted to the Third Long Duration Exposure Facility (LDEF) Post-Retrieval Symposium. The abstracts represent the data analysis of the 57 experiments flown on the LDEF. The experiments include materials, coatings, thermal systems, power and propulsion, science (cosmic ray, interstellar gas, heavy ions, micrometeoroid, etc.), electronics, optics, and life science.

  3. Romanian Scientific Abstracts, Volume 10 Number 3.

    ERIC Educational Resources Information Center

    Caracas, Angela, Ed.

    The material included in the monthly issues of the "Romanian Scientific Abstracts" as bibliographic references or abstracts is arranged according to discipline and main numerical sequence. The December issue includes a subject index for the material included throughout the year. It also indicates, in an appended table, the numerical symbol of…

  4. Romanian Scientific Abstracts, Volume 10 Number 1.

    ERIC Educational Resources Information Center

    Caracas, Angela, Ed.

    The material included in the monthly issues of the "Romanian Scientific Abstracts" as bibliographic references or abstracts is arranged according to discipline and main problems and is provided with an index and with key-words. The entries are recorded in numerical sequence. The December issue includes a subject index for the material included…

  5. Romanian Scientific Abstracts, Volume 10 Number 6.

    ERIC Educational Resources Information Center

    Caracas, Angela, Ed.

    The material included in the monthly issues of the "Romanian Scientific Abstracts" as bibliographic references or abstracts is arranged according to discipline and main problems and is provided with an index and with key-words. The entries are recorded in numerical sequence. The December issue includes a subject index for the material included…

  6. Romanian Scientific Abstracts, Volume 10 Number 4.

    ERIC Educational Resources Information Center

    Caracas, Angela, Ed.

    The material included in the monthly issues of the "Romanian Scientific Abstracts" as bibliographic references or abstracts is arranged according to discipline and main problems and is provided with an index and with key-words. The entries are recorded in numeric sequence. The December issue includes a subject index for the material included…

  7. Romanian Scientific Abstracts, Volume 10 Number 5.

    ERIC Educational Resources Information Center

    Caracas, Angela, Ed.

    The material included in the monthly issues of the "Romanian Scientific Abstracts" as bibliographic references or abstracts is arranged according to discipline and main problems and is provided with an index and with key-words. The entries are recorded in numeric sequence. The December issue includes a subject index for the material included…

  8. Romanian Scientific Abstracts, Volume 9 Number 11.

    ERIC Educational Resources Information Center

    Caracas, Angela, Ed.

    The material included in the monthly issues of the "Romanian Scientific Abstracts" as bibliographic references or abstracts is arranged according to discipline and main problems and is provided with an index and with key-words. The entries are recorded in numerical sequence. The December issue includes a subject index for the material included…

  9. Romanian Scientific Abstracts, Volume 9 Number 12.

    ERIC Educational Resources Information Center

    Caracas, Angela, Ed.

    The material included in the monthly issues of the "Romanian Scientific Abstracts" as bibliographic references or abstracts is arranged according to discipline and main problems and is provided with an index and with key-words. T8e entries are recorded in numerical sequence. The December issue includes a subject index for the material included…

  10. Romanian Scientific Abstracts, Volume 10 Number 2.

    ERIC Educational Resources Information Center

    Caracas, Angela, Ed.

    The material included in the monthly issues of the "Romanian Scientific Abstracts" as bibliographic references or abstracts is arranged according to discipline and main problems and is provided with an index and with key-words. T entries are recorded in numerical sequence. The December issue includes a subject index for the material included…

  11. Youth Studies Abstracts. Vol. 4 No. 1.

    ERIC Educational Resources Information Center

    Youth Studies Abstracts, 1985

    1985-01-01

    This volume contains abstracts of 76 projects (most of which were conducted in Australia and New Zealand) concerned with programs for youth and with social and educational developments affecting youth. The abstracts are arranged in the following two categories: (1) Social and Educational Developments: Policy, Analysis, Research; and (2) Programs:…

  12. Interactional Metadiscourse in Research Article Abstracts

    ERIC Educational Resources Information Center

    Gillaerts, Paul; Van de Velde, Freek

    2010-01-01

    This paper deals with interpersonality in research article abstracts analysed in terms of interactional metadiscourse. The evolution in the distribution of three prominent interactional markers comprised in Hyland's (2005a) model, viz. hedges, boosters and attitude markers, is investigated in three decades of abstract writing in the field of…

  13. Writing Abstracts for Free-Text Searching.

    ERIC Educational Resources Information Center

    Fidel, Raya

    1986-01-01

    This study surveyed abstracting policies and guidelines used by producers of bibliographic databases that aim to enhance free-text retrieval. Results indicate editors consider content of abstracts and their language as primary factors in retrieval enhancement. Most recommend that concepts and form be coordinated with controlled vocabulary…

  14. A Microfilm Index to "Chemical Abstracts"

    ERIC Educational Resources Information Center

    Robinson, F.

    1973-01-01

    To improve access to the recent Chemical Abstracts,'' a cumulative quarterly index, based on the keyword phrases, has been produced in microfilm form. The index is available soon after the end of each quarter. Abstract titles are included in the index, thus increasing its value as a working tool. (4 references) (Author/SJ)

  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. 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

  18. Nucleosomes impede Cas9 access to DNA in vivo and in vitro.

    PubMed

    Horlbeck, Max A; Witkowsky, Lea B; Guglielmi, Benjamin; Replogle, Joseph M; Gilbert, Luke A; Villalta, Jacqueline E; Torigoe, Sharon E; Tjian, Robert; Weissman, Jonathan S

    2016-01-01

    The prokaryotic CRISPR (clustered regularly interspaced palindromic repeats)-associated protein, Cas9, has been widely adopted as a tool for editing, imaging, and regulating eukaryotic genomes. However, our understanding of how to select single-guide RNAs (sgRNAs) that mediate efficient Cas9 activity is incomplete, as we lack insight into how chromatin impacts Cas9 targeting. To address this gap, we analyzed large-scale genetic screens performed in human cell lines using either nuclease-active or nuclease-dead Cas9 (dCas9). We observed that highly active sgRNAs for Cas9 and dCas9 were found almost exclusively in regions of low nucleosome occupancy. In vitro experiments demonstrated that nucleosomes in fact directly impede Cas9 binding and cleavage, while chromatin remodeling can restore Cas9 access. Our results reveal a critical role of eukaryotic chromatin in dictating the targeting specificity of this transplanted bacterial enzyme, and provide rules for selecting Cas9 target sites distinct from and complementary to those based on sequence properties. PMID:26987018

  19. 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

  20. 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

  1. 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

  2. Force Sensing by Extension of the Src Family Kinase Substrate, p130Cas

    PubMed Central

    Sawada, Yasuhiro; Tamada, Masako; Dubin-Thaler, Benjamin J.; Cherniavskaya, Oksana; Sakai, Ryuichi; Tanaka, Sakae; Sheetz, Michael P.

    2009-01-01

    SUMMARY Physical force is implicated in many cell functions. However, the molecular mechanisms of force sensing are poorly understood. Here, we show that mechanical extension of p130Cas (Cas) in vitro and in vivo causes phosphorylation by Src family kinases with no apparent change in Src kinase activity and that Cas phosphorylation is involved in stretch-dependent activation of the small GTPase, Rap1. In vitro, we mechanically extended bacterially expressed Cas substrate domain (CasSD) and found a remarkable enhancement of phosphorylation by specific kinases. Using an antibody that recognized extended CasSD in vitro, Cas extension in intact cells was observed in the peripheral regions of spreading cells where higher traction forces are expected and phosphorylated Cas was detected, suggesting that the in vitro extension and phosphorylation of CasSD is relevant to physiological force transduction. Thus, Cas acts as a primary force-sensor through extension of the substrate domain, which primes it for phosphorylation. PMID:17129785

  3. Highly efficient targeted chromosome deletions using CRISPR/Cas9.

    PubMed

    He, Zuyong; Proudfoot, Chris; Mileham, Alan J; McLaren, David G; Whitelaw, C Bruce A; Lillico, Simon G

    2015-05-01

    The CRISPR/Cas9 system has emerged as an intriguing new technology for genome engineering. It utilizes the bacterial endonuclease Cas9 which, when delivered to eukaryotic cells in conjunction with a user-specified small guide RNA (gRNA), cleaves the chromosomal DNA at the target site. Here we show that concurrent delivery of gRNAs designed to target two different sites in a human chromosome introduce DNA double-strand breaks in the chromosome and give rise to targeted deletions of the intervening genomic segment. Predetermined genomic DNA segments ranging from several-hundred base pairs to 1 Mbp can be precisely deleted at frequencies of 1-10%, with no apparent correlation between the size of the deleted fragment and the deletion frequency. The high efficiency of this technique holds promise for large genomic deletions that could be useful in generation of cell and animal models with engineered chromosomes. PMID:25362885

  4. Energy biotechnology in the CRISPR-Cas9 era.

    PubMed

    Estrela, Raissa; Cate, Jamie Harrison Doudna

    2016-04-01

    The production of bioenergy from plant biomass previously relied on using microorganisms that rapidly and efficiently convert simple sugars into fuels and chemicals. However, to exploit the far more abundant carbon fixed in plant cell walls, future industrial production hosts will need to be engineered to leverage the most efficient biochemical pathways and most robust traits that can be found in nature. The CRISPR-Cas9 genome editing technology now enables writing the genome at will, which will allow biotechnology to become an 'information science.' This review covers recent advances in using CRISPR-Cas9 to engineer the genomes of a wide variety of organisms that could be use in the industrial production of biofuels and renewable chemicals. PMID:26874259

  5. Application of CRISPR/Cas9 Technology to HBV

    PubMed Central

    Lin, Guigao; Zhang, Kuo; Li, Jinming

    2015-01-01

    More than 240 million people around the world are chronically infected with hepatitis B virus (HBV). Nucleos(t)ide analogs and interferon are the only two families of drugs to treat HBV currently. However, none of these anti-virals directly target the stable nuclear covalently closed circular DNA (cccDNA), which acts as a transcription template for viral mRNA and pre-genomic RNA synthesis and secures virus persistence. Thus, the fact that only a small number of patients treated achieve sustained viral response (SVR) or cure, highlights the need for new therapies against HBV. The clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 gene editing system can specifically target the conserved regions of the HBV genome. This results in robust viral suppression and provides a promising tool for eradicating the virus. In this review, we discuss the function and application of the CRISPR/Cas9 system as a novel therapy for HBV. PMID:26540039

  6. Chromosome engineering in zygotes with CRISPR/Cas9

    PubMed Central

    Boroviak, Katharina; Doe, Brendan; Banerjee, Ruby; Yang, Fengtang

    2016-01-01

    SUMMARY Deletions, duplications, and inversions of large genomic regions covering several genes are an important class of disease causing variants in humans. Modeling these structural variants in mice requires multistep processes in ES cells, which has limited their availability. Mutant mice containing small insertions, deletions, and single nucleotide polymorphisms can be reliably generated using CRISPR/Cas9 directly in mouse zygotes. Large structural variants can be generated using CRISPR/Cas9 in ES cells, but it has not been possible to generate these directly in zygotes. We now demonstrate the direct generation of deletions, duplications and inversions of up to one million base pairs by zygote injection. genesis 54:78–85, 2016. © 2016 The Authors. genesis Published by Wiley Periodicals, Inc. PMID:26742453

  7. Next Generation Prokaryotic Engineering: The CRISPR-Cas Toolkit.

    PubMed

    Mougiakos, Ioannis; Bosma, Elleke F; de Vos, Willem M; van Kranenburg, Richard; van der Oost, John

    2016-07-01

    The increasing demand for environmentally friendly production processes of green chemicals and fuels has stimulated research in microbial metabolic engineering. CRISPR-Cas-based tools for genome editing and expression control have enabled fast, easy, and accurate strain development for established production platform organisms, such as Escherichia coli and Saccharomyces cerevisiae. However, the growing interest in alternative production hosts, for which genome editing options are generally limited, requires further developing such engineering tools. In this review, we discuss established and emerging CRISPR-Cas-based tools for genome editing and transcription control of model and non-model prokaryotes, and we analyse the possibilities for further improvement and expansion of these tools for next generation prokaryotic engineering. PMID:26944793

  8. 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.

  9. 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.

  10. Dissociation of CAS phase in the uppermost lower mantle

    NASA Astrophysics Data System (ADS)

    Ishibashi, Kazufusa; Hirose, Kei; Sata, Nagayoshi; Ohishi, Yasuo

    2008-05-01

    The high-pressure stability limit of calcium aluminosilicate (CAS) phase has been examined in its end-member CaAl4Si2O11 composition at 18 39 GPa and 1,670 2,300 K in a laser-heated diamond-anvil cell (LHDAC). The in-situ synchrotron X-ray diffraction measurements revealed that the CAS phase decomposes into three-phase assemblage of cubic Al-bearing CaSiO3 perovskite, Al2O3 corundum, and SiO2 stishovite above 30 GPa and 2,000 K with a positive pressure temperature slope. Present results have important implications for the subsolidus mineral assemblage of subducted sediment and the melting phase relation of basalt in the lower mantle.

  11. Plasmocytome costal solitaire: à propos d'un cas

    PubMed Central

    Razafimanjato, Narindra Njarasoa Mihaja; Ravoatrarilandy, Manjakaniaina; Rakotoarisoa, Andriamihaja Jean Claude; Hasiniatsy, Rodrigue; Hunald, Allen Francis; Rakototiana, Auberlin Felantsoa; Rafaramino, Florine; Rakotovao, Hanitrala Jean Louis

    2014-01-01

    Les auteurs rapportent un cas de plasmocytome solitaire particulière par leur localisation costale. Le diagnostic est basé sur la mise en évidence d'une tumeur localisée, constituée de cellules plasmocytaires monoclonales cytologiquement identiques à celles du myélome multiple, en l'absence d'autres signes en faveur d'une forme disséminée. Nous rapportons un cas de plasmocytome solitaire à localisation costale et nous discutons les aspects diagnostiques et thérapeutiques de cette affection potentiellement menacée dans son évolution par la transformation en myélome multiple. PMID:25419306

  12. Hématome sous capsulaire de foie compliquant une pré-éclampsie: à propos de 6 cas

    PubMed Central

    Mamouni, Nisrine; Derkaoui, Ali; Bougern, Hakima; Bouchikhi, Chehrazad; Chaara, Hikmat; Banani, Abdelaziz; Abdelilah, Melhouf Moulay

    2011-01-01

    L'hématome sous capsulaire du foie (HSCF) est une complication rare mais gravissime de la grossesse. Devant une symptomatologie clinique souvent non spécifique et un tableau biologique retardé, son diagnostic est basé essentiellement sur les moyens de l'imagerie (échographie, TDM, IRM). Son traitement est fonction de l'intégrité ou non de la capsule de Glisson. Nous rapportons les observations de 6 patientes, à travers une étude rétrospective s’étalant sur la période du Janvier 2005 à Octobre 2008, incluant tous les cas de preeclampsie colligés au service de gynécologie obstétrique du CHU Hassan II. Durant la période d’étude, L'incidence de l'hématome sous capsulaire de foie chez les patientes préeclamptiques admises durant la période d’étude est de 1,49 %. Aucune des patientes n'a benificié d'un suivi prénatal au sein de notre formation. La moyenne d’âge des patientes est de 37,6 ans avec des extrêmes allant de 33 à 45 ans. La gestité moyenne était de 4,8 avec une parité moyenne de 4,5.l'hematome sous capsulaire est survenu en post partum chez tous nos cas avec un délai moyen de 4 jours et des extrêmes allant de J0 et J10 du post partum .Toutes les patientes ont présenté un HELLP syndrome concomitant à la survenue de cette complication gravissime.Le diagnostic positif s'est basé sur les données échographiques dans 5 cas (hemoperitoine –HSCF).l’équipe a opté pour une abstention thérapeutique avec surveillance armée chez 2 cas et l'exploration chirurgicale a été indiquée chez quatre patientes en instabilité hemodynamique.Nous avons déploré deux cas de décès maternel. PMID:22145072

  13. 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

  14. Motions of alloying additions in the CAS steelmaking operations

    NASA Astrophysics Data System (ADS)

    Mazumdar, D.; Guthrie, R. I. L.

    1993-08-01

    Water model studies in a pilot scale ladle ( D = 1.12 m and L = 0.93 m) were carried out to investigate the subsurface motion of both buoyant and sinking additions during the CAS (com-position adjustment by sealed argon bubbling systems) alloy addition procedure in steelmaking. This technique involves placing a refractory baffle around a rising gas/liquid plume within a stirred ladle of steel. Alloy additions are then made by projecting them into the slag-free region of steel within the baffled region. It was found that such particles while moving through the upwelling two-phase plume region can experience a significant reduction in drag forces, causing buoyant particles to penetrate more deeply than anticipated for a homogeneous fluid. Therefore, considering reduced drag on particles penetrating the upwelling gas liquid plume region, predictions were made for the trajectories of spherical-shaped particles using Newton’s law of motion. Predictions were in very reasonable agreement with those measured. Incorporating the velocity fields in industrial size vessels already reported by the present authors, trajectories of spherical-shaped additions (diameter ˜ 80 mm) in a 150-ton ladle during CAS operations were then predicted. The industrial implications of such trajectories, together with the alloy’s dissolution and dispersion behavior, were also analyzed. Finally, advantages of the CAS alloy addition procedure over conventional methods, in terms of the recovery rates of buoyant additions, are discussed.

  15. 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

  16. CRISPR-Cas systems for genome editing, regulation and targeting

    PubMed Central

    Sander, Jeffry D.; Joung, J. Keith

    2014-01-01

    Targeted genome editing using engineered nucleases has rapidly transformed from a niche technology to a mainstream method used by many biological researchers. This widespread adoption has been largely fueled by the emergence of the clustered regularly interspaced short palindromic repeat (CRISPR) technology, an important new platform for generating RNA-guided nucleases (RGNs), such as Cas9, with customizable specificities. RGN-mediated genome editing is facile, rapid and has enabled the efficient modification of endogenous genes in a wide variety of biomedically important cell types and novel organisms that have traditionally been challenging to manipulate genetically. Furthermore, a modified version of the CRISPR-Cas9 system has been developed to recruit heterologous domains that can regulate endogenous gene expression or label specific genomic loci in living cells. Although the genome-wide specificities of CRISPR-Cas9 systems remain to be fully defined, the capabilities of these systems to perform targeted, highly efficient alterations of genome sequence and gene expression will undoubtedly transform biological research and spur the development of novel molecular therapeutics for human disease. PMID:24584096

  17. p130Cas Scaffolds the Signalosome To Direct Adaptor-Effector Cross Talk during Kaposi's Sarcoma-Associated Herpesvirus Trafficking in Human Microvascular Dermal Endothelial Cells

    PubMed Central

    Bandyopadhyay, Chirosree; Veettil, Mohanan Valiya; Dutta, Sujoy

    2014-01-01

    ABSTRACT Kaposi's sarcoma-associated herpesvirus (KSHV) interacts with cell surface receptors, such as heparan sulfate, integrins (α3β1, αVβ3, and αVβ5), and EphrinA2 (EphA2), and activates focal adhesion kinase (FAK), Src, phosphoinositol 3-kinase (PI3-K), c-Cbl, and RhoA GTPase signal molecules early during lipid raft (LR)-dependent productive macropinocytic entry into human dermal microvascular endothelial cells. Our recent studies have identified CIB1 as a signal amplifier facilitating EphA2 phosphorylation and subsequent cytoskeletal cross talk during KSHV macropinocytosis. Although CIB1 lacks an enzymatic activity and traditional adaptor domain or known interacting sequence, it associated with the KSHV entry signal complex and the CIB1-KSHV association was sustained over 30 min postinfection. To identify factors scaffolding the EphA2-CIB1 signal axis, the role of major cellular scaffold protein p130Cas (Crk-associated substrate of Src) was investigated. Inhibitor and small interfering RNA (siRNA) studies demonstrated that KSHV induced p130Cas in an EphA2-, CIB1-, and Src-dependent manner. p130Cas and Crk were associated with KSHV, LRs, EphA2, and CIB1 early during infection. Live-cell microscopy and biochemical studies demonstrated that p130Cas knockdown did not affect KSHV entry but significantly reduced productive nuclear trafficking of viral DNA and routed KSHV to lysosomal degradation. p130Cas aided in scaffolding adaptor Crk to downstream guanine nucleotide exchange factor phospho-C3G possibly to coordinate GTPase signaling during KSHV trafficking. Collectively, these studies demonstrate that p130Cas acts as a bridging molecule between the KSHV-induced entry signal complex and the downstream trafficking signalosome in endothelial cells and suggest that simultaneous targeting of KSHV entry receptors with p130Cas would be an attractive potential avenue for therapeutic intervention in KSHV infection. IMPORTANCE Eukaryotic cell adaptor molecules

  18. Abstract spatial reasoning as an autistic strength.

    PubMed

    Stevenson, Jennifer L; Gernsbacher, Morton Ann

    2013-01-01

    Autistic individuals typically excel on spatial tests that measure abstract reasoning, such as the Block Design subtest on intelligence test batteries and the Raven's Progressive Matrices nonverbal test of intelligence. Such well-replicated findings suggest that abstract spatial processing is a relative and perhaps absolute strength of autistic individuals. However, previous studies have not systematically varied reasoning level--concrete vs. abstract--and test domain--spatial vs. numerical vs. verbal, which the current study did. Autistic participants (N = 72) and non-autistic participants (N = 72) completed a battery of 12 tests that varied by reasoning level (concrete vs. abstract) and domain (spatial vs. numerical vs. verbal). Autistic participants outperformed non-autistic participants on abstract spatial tests. Non-autistic participants did not outperform autistic participants on any of the three domains (spatial, numerical, and verbal) or at either of the two reasoning levels (concrete and abstract), suggesting similarity in abilities between autistic and non-autistic individuals, with abstract spatial reasoning as an autistic strength. PMID:23533615

  19. L'otospongiose: étude rétrospective à propos de 36 cas

    PubMed Central

    Bouaity, Brahim; Chihani, Mehdi; Touati, Mohammed; Darouassi, Youssef; Nadour, Karim; Ammar, Haddou

    2014-01-01

    L'objectif de l’étude est de rapporter notre expérience concernant la prise en charge de l'otospongiose et comparer nos résultats avec ceux de la littérature. Nous rapportons une étude rétrospective concernant 36 cas d'otospongiose colligés au service d'Oto-rhino-laryngologie de l'hôpital militaire Avicenne de Marrakech, entre Janvier 2009 et Décembre 2012. L’âge moyen des patients était de 38 ans avec une nette prédominance masculine (77%). Notre étude a révélé un retard diagnostique (stades audiométriques II et III). Ce diagnostic est le plus souvent simple, très fortement suspecté dès la première consultation devant l'existence d'une surdité de transmission ou mixte à tympan normal qui constitue le signe de découverte le plus prépondérant et constant dans notre série (100%), accompagnée parfois d'acouphènes (41%). L'existence d'antécédents familiaux de la maladie a été retenue chez 33,33%. L'audiométrie tonale liminaire et la tympanométrie ont été réalisées chez tous nos patients (100%) évoquant le diagnostic dans la majorité des cas. La tomodensitométrie, non obligatoire, reste indispensable pour le diagnostic différentiel et surtout permet de prévoir les difficultés opératoires ainsi que les éventuelles associations pathologiques. Tous les patients ont subit une intervention chirurgicale comportant soit une platinectomie partielle ou totale(83%) ou une platinotomie. L’évolution et les résultats ont été généralement satisfaisants, par ailleurs nous avons constaté la survenue d'un cas de fistule périlymphatique, la persistance des acouphènes chez six patients et une surdité de transmission chez trois patients. PMID:25426200

  20. Writing, reviewing, and presenting an abstract.

    PubMed

    Strauss, R G

    1991-01-01

    Abstracts afford an opportunity to report data at professional meetings and, when published, in the literature. Accordingly, they should be prepared with great care. When writing an abstract, anticipate questions the reviewer will ask when judging it and provide complete answers. The presentation of an abstract should follow similar thought processes. State why a problem or question is important, how you addressed it, what you found, and how your findings can be applied to the issue at hand. Slides and text should provide coordinated visual and auditory input, respectively, to ensure complete comprehension. PMID:1816248

  1. 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

  2. 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

  3. Transcriptional Regulation with CRISPR/Cas9 Effectors in Mammalian Cells.

    PubMed

    Pham, Hannah; Kearns, Nicola A; Maehr, René

    2016-01-01

    CRISPR/Cas9-based regulation of gene expression provides the scientific community with a new high-throughput tool to dissect the role of genes in molecular processes and cellular functions. Single-guide RNAs allow for recruitment of a nuclease-dead Cas9 protein and transcriptional Cas9-effector fusion proteins to specific genomic loci, thereby modulating gene expression. We describe the application of a CRISPR-Cas9 effector system from Streptococcus pyogenes for transcriptional regulation in mammalian cells resulting in activation or repression of transcription. We present methods for appropriate target site selection, sgRNA design, and delivery of dCas9 and dCas9-effector system components into cells through lentiviral transgenesis to modulate transcription. PMID:26463376

  4. 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

  5. Long-term monitoring of orbital modulation and secondary-star irradiation in Nova Cas 1995 (V723 Cas)

    NASA Astrophysics Data System (ADS)

    Ochner, P.; Moschini, F.; Munari, U.; Frigo, A.

    2015-11-01

    We present optical spectroscopy collected at seven epochs and BVRCIC photometry obtained at 1227 epochs of nova V723 Cas, covering the time interval between 2007 and 2015. The mean magnitude during this period, stable at ˜3 mag brighter than in quiescence, and the continuous presence of strong [Fe X] and other high-ionization emission lines, indicates that the nuclear burning at the surface of the white dwarf is continuing 20 years past the initial outburst. The light curve shows a large amplitude (2 mag) orbital modulation, which is governed by the visibility of the irradiated side of the secondary star. Our observations do not confirm the reported increase with time of the orbital period of V723 Cas, a period of P=16.638 383 ± 0.000 025 h satisfying equally well all available observations in all bands. Our observations also do not confirm the presence of an additional periodicity around P=15.2397 h from which V723 Cas was classified as an intermediate-polar system.

  6. Reliability Abstracts and Technical Reviews January - December 1970. Volume 10, Nos. 1-12; R70-14805 - R70-15438

    NASA Technical Reports Server (NTRS)

    1970-01-01

    Reliability Abstracts and Technical Reviews is an abstract and critical analysis service covering published and report literature on reliability. The service is designed to provide information on theory and practice of reliability as applied to aerospace and an objective appraisal of the quality, significance, and applicability of the literature abstracted.

  7. Introduction of p130cas signaling complex formation upon integrin-mediated cell adhesion: a role for Src family kinases.

    PubMed Central

    Vuori, K; Hirai, H; Aizawa, S; Ruoslahti, E

    1996-01-01

    Integrin-mediated cell adhesion triggers intracellular signaling cascades, including tyrosine phosphorylation of intracellular proteins. Among these are the focal adhesion proteins p130cas (Cas) and focal adhesion kinase (FAK). Here we identify the kinase(s) mediating integrin-induced Cas phosphorylation and characterize protein-protein interactions mediated by phosphorylated Cas. We found that expression of a constitutively active FAK in fibroblasts results in a consecutive tyrosine phosphorylation of Cas. This effect required the autophosphorylation site of FAK, which is a binding site for Src family kinases. Integrin-mediated phosphorylation of Cas was not, however, compromised in fibroblasts lacking FAK. In contrast, adhesion-induced tyrosine phosphorylation of Cas was reduced in cells lacking Src, whereas enhanced phosphorylation of Cas was observed Csk- cells, in which Src kinases are activated. These results suggest that Src kinases are responsible for the integrin-mediated tyrosine phosphorylation of Cas. FAK seems not to be necessary for phosphorylation of Cas, but when autophosphorylated, FAK may recruit Src family kinases to phosphorylate Cas. Cas was found to form complexes with Src homology 2 (SH2) domain-containing signaling molecules, such as the SH2/SH3 adapter protein Crk, following integrin-induced tyrosine phosphorylation. Guanine nucleotide exchange factors C3G and Sos were found in the Cas-Crk complex upon integrin ligand binding. These observations suggest that Cas serves as a docking protein and may transduce signals to downstream signaling pathways following integrin-mediated cell adhesion. PMID:8649368

  8. Pulmonary toxicology of respirable particles. [Lead abstract

    SciTech Connect

    Sanders, C.L.; Cross, F.T.; Dagle, G.E.; Mahaffey, J.A.

    1980-09-01

    Separate abstracts were prepared for the 44 papers presented in these proceedings. The last paper (Stannard) in the proceedings is an historical review of the field of inhalation toxicology and is not included in the analytics. (DS)

  9. OIL POLLUTION ABSTRACTS. VOLUME 6, NUMBER 2

    EPA Science Inventory

    Oil Pollution Abstracts (formerly entitled Oil Pollution Reports) is a quarterly compilation of current literature and research project summaries. Comprehensive coverage of oil pollution and its prevention and control is provided, with emphasis on the aquatic environment. This is...

  10. Program Aims at Improving Abstract Reasoning

    ERIC Educational Resources Information Center

    Chemical and Engineering News, 1977

    1977-01-01

    Describes a program being conducted within the chemistry department of Xavier University, New Orleans, Louisiana, to improve the abstract reasoning abilities of freshmen science majors. The project is based upon the philosophy developed by Jean Piaget. (SL)

  11. Introducing Abstraction to Junior High Students.

    ERIC Educational Resources Information Center

    Costanzo, Nancy

    1981-01-01

    Suggests a way to introduce abstract art to junior high school students who, more than students of any other age, emphasize realism both in their artwork and in their appreciation of works of art. (Author/SJL)

  12. Masking failures of multidimensional sensors (extended abstract)

    NASA Technical Reports Server (NTRS)

    Chew, Paul; Marzullo, Keith

    1990-01-01

    When a computer monitors a physical process, the computer uses sensors to determine the values of the physical variables that represent the state of the process. A sensor can sometimes fail, however, and in the worst case report a value completely unrelated to the true physical value. The work described is motivated by a methodology for transforming a process control program that can not tolerate sensor failure into one that can. In this methodology, a reliable abstract sensor is created by combining information from several real sensors that measure the same physical value. To be useful, an abstract sensor must deliver reasonably accurate information at reasonable computational cost. Sensors are considered that deliver multidimensional values (e.g., location or velocity in three dimensions, or both temperature and pressure). Geometric techniques are used to derive upper bounds on abstract sensor accuracy and to develop efficient algorithms for implementing abstract sensors.

  13. Surveying for Historical Supernovae Light Echoes in the Milky Way Field (Abstract)

    NASA Astrophysics Data System (ADS)

    Welch, D. L.

    2014-12-01

    (Abstract only) Very luminous, transient events can produce detectable “light echoes”—light scattered by interstellar dust which can arrive much later than the direct light from an outburst. In the last 1,000 years, there have been half a dozen supernovae in the Milky Way which are capable of producing detectable light echoes. Light echo systems have already been found for Tycho (SN 1572) and Cas A. The three-dimensional distribution of light echoes provides one of the few menas for an astronomical source to be inspected from more than one viewpoint. Indications of the degree of asymmetry of supernovae are extremely valuable for understanding the details of the event itself. Amateurs are well-equipped to find the brighter light echoes and in this work I will provide practical guidance on how such surveys may be accomplished and the various science opportunities they provide.

  14. 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

  15. 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

  16. 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

  17. Prediction and Validation of Native and Engineered Cas9 Guide Sequences.

    PubMed

    Briner, Alexandra E; Henriksen, Emily D; Barrangou, Rodolphe

    2016-01-01

    Cas9-based technologies rely on native elements of Type II CRISPR-Cas bacterial immune systems, including the trans-activating CRISPR RNA (tracrRNA), CRISPR RNA (crRNA), Cas9 protein, and protospacer-adjacent motif (PAM). The tracrRNA and crRNA form an RNA duplex that guides the Cas9 endonuclease to complementary nucleic acid sequences. Mechanistically, Cas9 initiates interactions by binding to the target PAM sequence and interrogating the target DNA in a 3'-to-5' manner. Complementarity between the guide RNA and the target DNA is key. In natural systems, precise cleavage occurs when the target DNA sequence contains a PAM flanking a sequence homologous to the crRNA spacer sequence. Currently, the majority of commercial Cas9-based genome-editing tools are derived from the Type II CRISPR-Cas system of Streptococcus pyogenes However, a diverse set of Type II CRISPR-Cas systems exist in nature that are potentially valuable for genome engineering applications. Exploitation of these systems requires prediction and validation of both native and engineered dual and single guide RNAs to drive Cas9 functionality. Here, we discuss how to identify the elements of these immune systems to develop next-generation Cas9-based genome-editing tools. We first discuss how to predict tracrRNA sequences and suggest a method for designing single guide RNAs containing only critical structural modules. We then outline how to predict the PAM sequence, which is crucial for determining potential targets for Cas9. Finally, validation of the system elements through transcriptome analysis and interference assays is essential for developing next-generation Cas9-based genome-editing tools. PMID:27371591

  18. Modified RNAs in CRISPR/Cas9: An Old Trick Works Again.

    PubMed

    Latorre, Alfonso; Latorre, Ana; Somoza, Álvaro

    2016-03-01

    Old tricks, new dog: CRISPR/Cas9 is a powerful tool for gene editing that requires an endonuclease (Cas9) and RNA strands. It has been shown that chemical modification of the RNA structures, an approach that has been used to improve the efficiency of RNA interference, can also be applied to enhance the activity of CRISPR/Cas9 and reduce its off-target effects. PMID:26880106

  19. 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

  20. Earth Sciences Division collected abstracts: 1980

    SciTech Connect

    Henry, A.L.; Hornady, B.F.

    1981-10-15

    This report is a compilation of abstracts of papers, reports, and talks presented during 1980 at national and international meetings by members of the Earth Sciences Division, Lawrence Livermore National Laboratory. The arrangement is alphabetical (by author). For a given report, a bibliographic reference appears under the name of each coauthor, but the abstract itself is given only under the name of the first author (indicated in capital letters) or the first Earth Sciences Division author.