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  • 2005-2009
  • 1985-1989  (127,767)
  • 1820-1829
  • 1987  (127,767)
Material
Years
Year
Language
  • 1
    Book
    Book
    Wien [u.a.] :Springer, ; 1.1977 - 16.2003; damit Ersch. eingest.
    Title: Computing : archives for informatics and numerical computation; Supplementum
    Publisher: Wien [u.a.] :Springer,
    Year of publication: 1977-2003
    Dates of Publication: 1.1977 - 16.2003; damit Ersch. eingest.
    Type of Medium: Book
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  • 2
    Journal/Serial
    Journal/Serial
    New York, NY :ACM, ; 1.1969 - 7.1975/76; N.S. 1.1976 - 21.2001,1; damit Ersch. eingest.
    Title: SIGBIO newsletter /
    Author: Association for Computing Machinery / Special Interest Group on Biomedical Computing
    Publisher: New York, NY :ACM,
    Year of publication: 1969-2001
    Dates of Publication: 1.1969 - 7.1975/76; N.S. 1.1976 - 21.2001,1; damit Ersch. eingest.
    ISSN: 0163-5697
    Type of Medium: Journal/Serial
    Language: Undetermined
    Parallel Title: Internetausg. ---〉:Biomedical computing
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  • 3
    Journal/Serial
    Journal/Serial
    New York, NY :North-Holland, ; 1.1984 - 46.2000
    Title: ¬The¬ journal of logic programming
    Publisher: New York, NY :North-Holland,
    Year of publication: 1984-2000
    Dates of Publication: 1.1984 - 46.2000
    ISSN: 0743-1066
    Type of Medium: Journal/Serial
    Language: Undetermined
    Subsequent Title: Forts. ---〉:¬The¬ journal of logic and algebraic programming
    Parallel Title: Internetausg. ---〉:¬The¬ journal of logic and algebraic programming
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  • 4
    Book
    Book
    New York, NY :ACM, ; Nachgewiesen 2.1971 - 20.1999,4; damit Ersch. eingest.
    Title: Computer personnel : a quarterly publ. of the Special Interest Group on Computer Personnel Research, SIGCPR
    Publisher: New York, NY :ACM,
    Year of publication: 1971-1999
    Dates of Publication: Nachgewiesen 2.1971 - 20.1999,4; damit Ersch. eingest.
    ISSN: 0160-2497
    Type of Medium: Book
    Parallel Title: Internetausg. ---〉:Computer personnel
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  • 5
    Journal/Serial
    Journal/Serial
    Woodbury, NY :AIP, ; 1.1987,1(Nov./Dez.); 2.1988 - 12.1998
    Title: Computers in physics /
    Contributer: American Institute of Physics
    Publisher: Woodbury, NY :AIP,
    Year of publication: 1987-1998
    Dates of Publication: 1.1987,1(Nov./Dez.); 2.1988 - 12.1998
    ISSN: 0894-1866
    Type of Medium: Journal/Serial
    Language: Undetermined
    Subsequent Title: Forts. ---〉:Computing in science & engineering
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  • 6
    Journal/Serial
    Journal/Serial
    New York, NY :ACM, ; 4.1969 - 33.1998,2; damit Ersch. eingest
    Title: SIGNUM newsletter
    Author: Association for Computing Machinery / Special Interest Group on Numerical Mathematics
    Publisher: New York, NY :ACM,
    Year of publication: 1969-1998
    Dates of Publication: 4.1969 - 33.1998,2; damit Ersch. eingest
    ISSN: 0163-5778
    Type of Medium: Journal/Serial
    Former Title: Vorg. ---〉:Association for Computing Machinery / Special Interest Committee on Numerical Mathematics: SICNUM newsletter
    Additional Information: 16,3=3,2 von:Association for Computing Machinery / Technical Committee on Fortran: FORTEC forum
    Parallel Title: Internetausg. ---〉:Numerical mathematics
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  • 7
    Journal/Serial
    Journal/Serial
    Amsterdam [u.a.] :Elsevier [u.a.], ; 9.1985 - 30.1998
    Title: Computer networks and ISDN systems : the international journal of computer and telecommunications networking
    Publisher: Amsterdam [u.a.] :Elsevier [u.a.],
    Year of publication: 1985-1998
    Dates of Publication: 9.1985 - 30.1998
    ISSN: 0169-7552 , 0376-5075
    Type of Medium: Journal/Serial
    Language: Undetermined
    Subsequent Title: Vorg. u. Forts. ---〉:Computer networks
    Note: Computer networks for research in Europe
    Additional Information: In 14,1=15 von:Networkshop: Conference report
    Additional Information: 16,1/2=4; 17,4/5=5 von:European Networkshop: European Networkshop
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  • 8
    Journal/Serial
    Journal/Serial
    San Francisco, Calif. :Miller Freeman, ; 1.1983 - 16.1998,3
    Title: Unix review : the publication for the Unix community
    Publisher: San Francisco, Calif. :Miller Freeman,
    Year of publication: 1983-1998
    Dates of Publication: 1.1983 - 16.1998,3
    ISSN: 0742-3136
    Type of Medium: Journal/Serial
    Language: Undetermined
    Subsequent Title: Forts. ---〉:Unix review's performance computing
    Parallel Title: Internetausg. ---〉:Unix review.com
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  • 9
    Journal/Serial
    Journal/Serial
    Amsterdam :Amsterdam Universities Computing Centre, ; Nr. 1.1984 - 69.1997; damit Ersch. eingest.
    Title: Supercomputer : bimonthly magazine on supercomputing in the Netherlands
    Publisher: Amsterdam :Amsterdam Universities Computing Centre,
    Year of publication: 1984-1997
    Dates of Publication: Nr. 1.1984 - 69.1997; damit Ersch. eingest.
    ISSN: 0168-7875
    Type of Medium: Journal/Serial
    Language: Undetermined
    Note: Teils auch mit Jg.-Zählung
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  • 10
    Journal/Serial
    Journal/Serial
    Thousand Oaks, Calif. :Sage Science Press, ; 1.1987 - 11.1997
    Title: ¬The¬ international journal of supercomputer applications and high performance computing
    Publisher: Thousand Oaks, Calif. :Sage Science Press,
    Year of publication: 1987-1997
    Dates of Publication: 1.1987 - 11.1997
    ISSN: 1078-3482 , 0890-2720
    Type of Medium: Journal/Serial
    Subsequent Title: Forts. ---〉:¬The¬ international journal of high performance computing applications
    Parallel Title: Internetausg. ---〉:¬The¬ international journal of high performance computing applications
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  • 11
    Book
    Book
    Bergheim :DATACOM-Zeitschriften-Verl., | Köln Müller -1993,9 ; 11.1973 - 14.1976; 19.1981 - 20.1982; 1983 - 1994; 32.1995 - 34.1997,10
    Title: Online : erfolgreiches Informationsmanagement, ADI-Nachrichten, ÖVD ; Organ d. ADI - Anwenderverband Deutscher Informationsverarbeiter e.V
    Contributer: Anwenderverband Deutscher Informationsverarbeiter
    Publisher: Bergheim :DATACOM-Zeitschriften-Verl., , Köln Müller -1993,9
    Year of publication: 1973-1997
    Dates of Publication: 11.1973 - 14.1976; 19.1981 - 20.1982; 1983 - 1994; 32.1995 - 34.1997,10
    ISSN: 0340-1545 , 0179-6623 , 0342-9393
    Type of Medium: Book
    Language: Undetermined
    Former Title: Vorg. ---〉:Zeitschrift für Datenverarbeitung
    Subsequent Title: 15.1977 - 18.1980 ---〉:ADL-Verband für Informationsverarbeitung: ADL-Nachrichten, Online
    Subsequent Title: Aufgeg. in ---〉:Information week
    Note: Später ohne Zählung
    Additional Information: Beil. ---〉:Drucker spezial
    Additional Information: Beil. ---〉:Online / special
    Additional Information: Darin ---〉:Anwenderverband Deutscher Informationsverarbeiter: ADI-Nachrichten, ÖVD
    Additional Information: Beil. ---〉:Pro info
    Additional Information: Beil. ---〉:Online-Info
    Additional Information: 1996 darin ---〉:Datacom-Special
    Parallel Title: 19.1981 auch in ---〉:Öffentliche Verwaltung und Datenverarbeitung
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  • 12
    Journal/Serial
    Journal/Serial
    München :Hanser, ; 1.1983 - 14.1996,6
    Title: Unix mail : Europas erster Informationsdienst für Unix-Hersteller und -Anwender
    Publisher: München :Hanser,
    Year of publication: 1983-1996
    Dates of Publication: 1.1983 - 14.1996,6
    ISSN: 0176-8654
    Type of Medium: Journal/Serial
    Language: Undetermined
    Subsequent Title: Forts. ---〉:¬Die¬ blauen Blätter
    Parallel Title: CD-ROM-Ausg. ---〉:Unix mail, die blauen Blätter
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  • 13
    Journal/Serial
    Journal/Serial
    Minneapolis, Minn. :Cray Research, Inc., ; Nachgewiesen 6.1984 - 18.1996,2; damit Ersch. eingest.
    Title: Cray channels : a Cray Research, Inc. publication
    Contributer: Cray Research, Inc. 〈Mendota Heights, Minn.〉
    Publisher: Minneapolis, Minn. :Cray Research, Inc.,
    Year of publication: 1984-1996
    Dates of Publication: Nachgewiesen 6.1984 - 18.1996,2; damit Ersch. eingest.
    Type of Medium: Journal/Serial
    Language: Undetermined
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  • 14
    Title: Nachrichtentechnische Zeitschrift : NTZ ; Zeitschrift für Informationstechnik u. Telekommunikation ; Organ der Nachrichtentechnischen Gesellschaft im VDE
    Contributer: Nachrichtentechnische Gesellschaft / Fachausschuß Informationsverarbeitung
    Publisher: Berlin :VDE-Verl.,
    Year of publication: 1955-1995
    Dates of Publication: 8.1955,10 - 40.1987,2; 40.1987,6 - 48.1995,2
    ISSN: 0027-707X
    Type of Medium: Journal/Serial
    Language: Undetermined
    Former Title: Vorg. ---〉:Fernmeldetechnische Zeitschrift
    Subsequent Title: 40.1987,3-5 u. Forts. ---〉:NTZ
    Note: Mikro-Elektronik
    Additional Information: Beih. ---〉:Nachrichtentechnische Fachberichte
    Additional Information: Index 1/10=11 von:Nachrichtentechnische Fachberichte
    Parallel Title: CD-ROM-Ausg. 1994 - 1995 ---〉:Elektronisches Zeitschriftenarchiv
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  • 15
    Journal/Serial
    Journal/Serial
    Heidelberg :Physica-Verl., ; 16.1972 - 42.1995
    Title: Zeitschrift für Operations-Research : ZOR ; mathematical methods of operations research
    Publisher: Heidelberg :Physica-Verl.,
    Year of publication: 1972-1995
    Dates of Publication: 16.1972 - 42.1995
    ISSN: 0340-9422
    Type of Medium: Journal/Serial
    Language: Undetermined
    Former Title: Vorg. ---〉:Ablauf- und Planungsforschung
    Subsequent Title: Forts. ---〉:Mathematical methods of operations research
    Note: Ser. A, Theorie = H. 1,3,5,7 d. Jg.; Ser. B, Praxis = H. 2,4,6,8 d. Jg. , Deutsche Gesellschaft für Operations-Research
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  • 16
    Journal/Serial
    Journal/Serial
    Oxford [u.a.] :Pergamon Press, ; 1.1960 - 19.1995
    Title: Annual review in automatic programming
    Publisher: Oxford [u.a.] :Pergamon Press,
    Year of publication: 1960-1995
    Dates of Publication: 1.1960 - 19.1995
    ISSN: 0066-4138
    Type of Medium: Journal/Serial
    Language: Undetermined
    Subsequent Title: Forts. ---〉:Annual reviews in control
    Additional Information: 1=3; 2=6, 3=11, 4=12; 5=13,2 von:International tracts in computer science and technology and their application
    Additional Information: 8=7; 9,2/3=8; 10=10; 11=11; 13,1=13; 14,1=15 von:Real time programming
    Additional Information: 12,1-12,2=2 von:Systems analysis and simulation
    Additional Information: 13,2=5 von:Control applications of nonlinear programming and optimization
    Parallel Title: Internetausg. ---〉:Annual reviews in control
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  • 17
    Journal/Serial
    Journal/Serial
    Stuttgart :IBM, ; 22.1972,Apr. - 45.1995 = Nr. 210-323; damit Ersch. eingest.
    Title: IBM-Nachrichten /
    Author: IBM Deutschland GmbH 〈Stuttgart〉
    Publisher: Stuttgart :IBM,
    Year of publication: 1972-1995
    Dates of Publication: 22.1972,Apr. - 45.1995 = Nr. 210-323; damit Ersch. eingest.
    ISSN: 0018-8662
    Type of Medium: Journal/Serial
    Language: Undetermined
    Former Title: Vorg. ---〉:Internationale Büro-Maschinen-Gesellschaft Deutschland 〈Sindelfingen〉: IBM-Nachrichten
    Additional Information: Beil. ---〉:Hollerith-Mitteilungen
    Parallel Title: CD-ROM-Ausg. ---〉:IBM Deutschland GmbH 〈Stuttgart〉: IBM-Nachrichten
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  • 18
    Journal/Serial
    Journal/Serial
    New York, NY :ACM Inc., ; 1.1987/88 - 4.1990/91; 5.1992; 6.1992/93; 7.1994 - 8.1995,2; damit Ersch. eingest.
    Title: LISP pointers /
    Publisher: New York, NY :ACM Inc.,
    Year of publication: 1987-1995
    Dates of Publication: 1.1987/88 - 4.1990/91; 5.1992; 6.1992/93; 7.1994 - 8.1995,2; damit Ersch. eingest.
    ISSN: 1045-3563
    Type of Medium: Journal/Serial
    Note: Special Interest Group on Programming Languages (SIGPLAN)
    Parallel Title: Internetausg. ---〉:Association for Computing Machinery / Special Interest Group on Programming Languages: ACM SIGPLAN LISP pointers
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  • 19
    Journal/Serial
    Journal/Serial
    New York, NY :ACM, ; 2.1971/72 - 16.1985; 19.1988 - 26.1995; damit Ersch. eingest.
    Title: SIGMICRO newsletter : a quarterly publ. of the Special Interest Group on Microprogramming
    Author: Association for Computing Machinery / Special Interest Group on Microprogramming
    Publisher: New York, NY :ACM,
    Year of publication: 1971-1995
    Dates of Publication: 2.1971/72 - 16.1985; 19.1988 - 26.1995; damit Ersch. eingest.
    ISSN: 0163-5751 , 1050-916X
    Type of Medium: Journal/Serial
    Language: Undetermined
    Former Title: Vorg. ---〉:Association for Computing Machinery / Special Interest Committee on Microprocessing: SICMICRO newsletter
    Subsequent Title: 17.1986 - 18.1987 ---〉:Association for Computing Machinery / Special Interest Group on Microprogramming: SIGMICRO TCMICRO newsletter
    Additional Information: Beil. ---〉:Microprogramming bibliography
    Additional Information: 9,4=11; 12,4=14; 13,4=15 von:Micro
    Additional Information: 20,3=22 von:International Workshop on Microprogramming and Microarchitecture: Annual International Workshop on Microprogramming and Microarchitecture
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  • 20
    Title: Data-base 〈New York,NY〉 : db ; a quarterly publication of the Special Interest Group on Business Information Technology of the Association for Computing Machinery
    Contributer: Association for Computing Machinery / Special Interest Group on Business Data Processing , Association for Computing Machinery / Special Interest Group on Business Information Technology
    Publisher: New York, NY,
    Year of publication: 1971-1994
    Dates of Publication: 3.1971 - 25.1994
    ISSN: 0095-0033
    Type of Medium: Book
    Former Title: Vorg. ---〉 SIGBDP news-letter
    Subsequent Title: Forts. ---〉:¬The¬ data-base for advances in information systems
    Additional Information: Einzelne Bd. zugl. Bd. von:Association for Computing Machinery / Special Interest Group on Management of Data: SIGMOD record
    Additional Information: 12,4u.13,1=2 von:Data-base directions
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  • 21
    Journal/Serial
    Journal/Serial
    München :Franzis-Verl., ; 1981 - 1994,6
    Title: MC 〈München〉 : Computerpraxis für technische Anwender
    Publisher: München :Franzis-Verl.,
    Year of publication: 1981-1994
    Dates of Publication: 1981 - 1994,6
    ISSN: 0720-4442 , 0941-777X , 0943-5409
    Type of Medium: Journal/Serial
    Language: Undetermined
    Subsequent Title: Aufgeg. in ---〉:DOS international
    Note: Auch mit fehlerhafter Jg.-Zählung im Impressum
    Additional Information: 1992 Sonderh. ---〉:WINbox
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  • 22
    Title: SIGSMALL - PC notes : a publication of the Special Interest Group on Small and Personal Computing Systems and Applications, Association for Computing Machinery
    Author: Association for Computing Machinery / Special Interest Group on Small and Personal Computing Systems and Applications
    Publisher: New York, NY :ACM,
    Year of publication: 1984-1993
    Dates of Publication: 10.1984,4 - 19.1993/94,2(1993)
    ISSN: 0893-2875
    Type of Medium: Journal/Serial
    Language: Undetermined
    Former Title: Vorg. ---〉:Association for Computing Machinery / Special Interest Group on Small Computing Systems and Applications: SIGSMALL newsletter
    Subsequent Title: Forts. ---〉:Association for Computing Machinery / Special Interest Group on Individual Computing Environments: SIGICE bulletin
    Note: Zählung von "SIGSMALL newsletter" übernommen
    Parallel Title: Internetausg. ---〉:Association for Computing Machinery / Special Interest Group on Small and Personal Computing Systems and Applications: ACM SIGSMALL - PC notes
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  • 23
    Journal/Serial
    Journal/Serial
    London [u.a.], ; 1.1969 - 39.1993
    Title: International journal of man machine studies
    Publisher: London [u.a.],
    Year of publication: 1969-1993
    Dates of Publication: 1.1969 - 39.1993
    ISSN: 0020-7373
    Type of Medium: Journal/Serial
    Language: Undetermined
    Subsequent Title: Forts. ---〉:International journal of human - computer studies
    Note: Index 1/4.1969/72 in: 4.1972
    Additional Information: 10,3=5 von:Man Computer Communications Conference: Proceedings
    Parallel Title: Internetausg. ---〉:International journal of man machine studies
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  • 24
    Title: SIGACT news : publ. by the ACM Special Interest Group on Automata and Computability Theory
    Author: Association for Computing Machinery / Special Interest Group on Automata and Computability Theory
    Contributer: Association for Computing Machinery / Special Interest Committee on Automata and Computability Theory
    Publisher: New York, NY :ACM,
    Year of publication: 1969-1992
    Dates of Publication: 1.1969 - 23.1992,2 = Nr. 1-83
    ISSN: 0163-5700
    Type of Medium: Journal/Serial
    Subsequent Title: Forts. ---〉:Association for Computing Machinery / Special Interest Group on Algorithms and Computation Theory: SIGACT news
    Additional Information: In 12.1980,2 Index 1/12.1969/80 von ---〉:Symposium on Theory of Computing: Conference record of the ... annual ACM Symposium on Theory of Computing
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  • 25
    Journal/Serial
    Journal/Serial
    München :Oldenbourg, ; 28.1986 - 34.1992
    Title: Informationstechnik : it ; Computer, Systeme, Anwendungen
    Publisher: München :Oldenbourg,
    Year of publication: 1986-1992
    Dates of Publication: 28.1986 - 34.1992
    ISSN: 0179-9738 , 0013-5720
    Type of Medium: Journal/Serial
    Language: Undetermined
    Former Title: Vorg. ---〉:Elektronische Rechenanlagen
    Subsequent Title: Forts. ---〉:Informationstechnik und technische Informatik
    Note: it-Seminar
    Additional Information: Beil. ---〉:Euro-KI-Führer
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  • 26
    Title: SIGPLAN notices : a monthly publication of the Special Interest Group on Programming Languages of the Association for Computing Machinery
    Author: Association for Computing Machinery / Special Interest Group on Programming Languages
    Publisher: New York, NY :ACM,
    Year of publication: 1966-1991
    Dates of Publication: 1.1966 - 26.1991,9u.11
    ISSN: 0362-1340
    Type of Medium: Journal/Serial
    Language: Undetermined
    Subsequent Title: 26.1991,10 u. Forts. ---〉:Association for Computing Machinery / Special Interest Group on Programming Languages: ACM SIGPLAN notices
    Additional Information: 6,2=1971 von:Symposium on Data Structures in Programming Languages: Proceedings of a Symposium on Data Structures in Programming Languages
    Additional Information: 17,4=10,2; 22,10=15,5; 24,spec.iss.=17,2; 26,4=19,2 von:Computer architecture news
    Additional Information: 21,10=1986 von:Workshop on Object Oriented Programming: Transcript
    Additional Information: 11,6=10,1 von:Computer graphics
    Additional Information: 16,6=2,1/2 von:Association for Computing Machinery / Special Interest Group on Office Automation: SIGOA newsletter
    Additional Information: 14,8=1979; 17,6=1982; 19,6=1984; 21,7=1986 von:Symposium on Compiler Construction: Proceedings of the SIGPLAN Symposium on Compiler Construction
    Additional Information: 20,7=1985 von:Symposium on Language Issues in Programming Environments: Proceedings of the ACM SIGPLAN ... Symposium on Language Issues in Programming Environments
    Additional Information: 19,5=1; 22,1=2; 24,2=3 von:Software Engineering Symposium on Practical Software Development Environments: Proceedings of the ACM SIGSOFT SIGPLAN Software Engineering Symposium on Practical Software Development Environments
    Additional Information: 22,10=2; 24,spec.iss.=3 von:International Conference on Architectural Support for Programming Languages and Operating Systems: Proceedings
    Additional Information: 23,7=1988; 24,7=1989; 25,6=1990 von:Conference on Programming Language Design and Implementation: Proceedings of the SIGPLAN ... Conference on Programming Language Design and Implementation
    Additional Information: 21,11=1; 22,12=2; 23,11=3; 24,10=4 von:OOPSLA: Conference proceedings
    Additional Information: 22,10=21,4; 24,spec.iss.=23,spec.iss.; 26,4=25,spec.iss. von:Operating systems review
    Additional Information: 17,4=1982 von:Symposium on Architectural Support for Programming Languages and Operating Systems: Proceedings
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  • 27
    Journal/Serial
    Journal/Serial
    Utrecht :VNU Science Press, ; 1.1986 - 6.1991
    Title: Soviet journal of numerical analysis and mathematical modelling
    Publisher: Utrecht :VNU Science Press,
    Year of publication: 1986-1991
    Dates of Publication: 1.1986 - 6.1991
    ISSN: 0169-2895
    Type of Medium: Journal/Serial
    Language: Undetermined
    Subsequent Title: Forts. ---〉:Russian journal of numerical analysis and mathematical modelling
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  • 28
    Title: SIGART newsletter : a bimonthly publ. of the ACM Special Interest Group on Artificial Intelligence
    Author: Association for Computing Machinery / Special Interest Group on Artificial Intelligence
    Publisher: New York, NY :ACM,
    Year of publication: 1969-1989
    Dates of Publication: Nachgewiesen Nr. 15.1969 - 110.1989
    ISSN: 0163-5719
    Type of Medium: Journal/Serial
    Subsequent Title: Forts. ---〉:Association for Computing Machinery / Special Interest Group on Artificial Intelligence: SIGART bulletin
    Additional Information: 73=1; 80=2 von:Workshop on Distributed AI: Report on the Workshop on Distributed AI
    Additional Information: 84=3 von:Workshop on Distributed Artificial Intelligence: Report on the ... Annual Workshop on Distributed Artificial Intelligence
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  • 29
    Journal/Serial
    Journal/Serial
    Braunschweig ; Wiesbaden :Vieweg, ; 13.1971 - 31.1989
    Title: Angewandte Informatik : Applied informatics
    Publisher: Braunschweig ; Wiesbaden :Vieweg,
    Year of publication: 1971-1989
    Dates of Publication: 13.1971 - 31.1989
    ISSN: 0013-5704
    Type of Medium: Journal/Serial
    Language: Undetermined
    Former Title: Vorg. ---〉:Elektronische Datenverarbeitung
    Additional Information: Beil. CAK
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  • 30
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Journal of neurochemistry 49 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Gene cha-1 · unc-17 of the nematode Caenorhabditis elegans is a complex gene, consisting of at least two complementation groups. One part (cha-1 region) of the gene encodes the enzyme choline acetyltransferase (ChAT), but the function of the other part (unc-17 region) is still unclear. We measured the ChAT activity and ACh levels of the cha-1 and unc-17 complex gene mutants. We show here that alterations in ACh levels, rather than the ChAT activity, reflect abnormal phenotypes accompanying cha-1 · unc-17 mutations, that is, the decreased ACh levels in cha-1 mutations and abnormal accumulation in unc-17 mutations. Our results suggest that the unc-17 region may encode functions necessary for storage and/or release of ACh at the presynaptic level.
    Type of Medium: Electronic Resource
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  • 31
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Journal of neurochemistry 49 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Release of norepinephrine from PC12 cells can be stimulated by free Ca2+ in micromolar concentrations after permeabilization with 10 μg/ml of digitonin. This release is time and temperature dependent, half-maximal at 0.3 μM Ca2+, and, after washing out of endogenous ATP, half- maximal at about 0.5 mA/ MgATP when exogenously added. Similar results were obtained with bovine adrenal chromaffin cells using the same protocol. Support for the idea that the mechanism of release from both permeabilized cell types is still exocytosis is demonstrated at the electron microscopic level by immunolabeling chromaffin granule membrane antigens that were introduced into the plasma membrane following stimulation. Electron micrographs furthermore demonstrate that chromaffin granules retain typical dense cores after permeabilization, indicating that leakiness of catecholamines from the granules was not major factor. Pores, formed by digitonin in the plasma membranes, were utilized to introduce antibodies into such exocytosis-competent cells. Anti-actin and anti-chromaffin granule membrane antibodies show a staining pattern similar to conventionally fixed and stained preparations. Our results demonstrate that pores formed by digitonin do not impair the process of exocytosis although they are big enough to allow macromolecules to pass in both directions. The digitonin-permeabilized cell is therefore an ideal in vitro system with which to study the fusion process between chromaffin granules and the plasma membrane.
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  • 32
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Incubation of cultured bovine adrenal medullary cells in Na+-free sucrose medium or in Na+-free Cs+ medium enhanced the synthesis of 14C-catecholamines from [14C]tyrosine about two- to threefold or sixfold, respectively. The increment of 14C-catecholamine synthesis produced by Na+-free medium was partially dependent on the presence of Ca2+ in the medium. Dibutyrylcyclic AMP also stimulated the synthesis of 14C-catecholamines in adrenal medullary cells, and the effects of Na+ removal and dibutyryl cyclic AMP (5 mM) on the synthesis were almost additive. The intracellular pH measured by using a weak acid 5, 5-dimethyloxazolidine-2, 4-dione was 7.14 in control cells and when Na+ was replaced by sucrose or Cs+, it shifted down to 6.56 or 5.66, respectively. The fall in intracellular pH and the stimulation of 14C-catecholamine synthesis were similarly dependent on the concentration of Na+ in the medium. The optimal pH of soluble tyrosine hydroxylase was 5.5–6.0 both in control cells and in cells incubated in Na+-free med um. These results suggest that removal of extracellular Na+ increases the synthesis of catecholamines, at least in part, by shifting the intracellular pH toward the optimal pH of tyrosine hydroxylase.
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  • 33
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    Oxford, UK : Blackwell Publishing Ltd
    Journal of neurochemistry 49 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Cells from neural retina from 7-day chick embryos were cultured on polylysine-coated dishes up to 7 days. The small, round-shaped cells at seeding differentiated progressively, and after 4 days in vitro the majority had enlarged bodies and abundant processes. The content of protein and DNA was essentially unchanged during the entire period of culture. The incorporation of radioactivity from [3H]glucosamine into gangliosides declined slightly, reaching about 65% of the initial values at the end of the culture period. The proliferating activity measured by the incorporation of [3H]thymidine into DNA decreased to 10% or less of the initial value after 3 days in vitro. Almost at the same chronological times as in ovo, the synthesis of GD3 and of a ganglioside partially identified as GT3 decreased from 70 and 19% of the total incorporation into gangliosides in the first 20 h of culture to about 7 and 5%, respectively, after 3 days in vitro. Conversely, the synthesis of GDI a increased from about 6% at the beginning to about 70% at the end of the culture times. Immunocytochemical analyses of the expression of gangliotetraosyl gangliosides in cultured cells showed that these gangliosides appeared in the bodies and processes of cells having neuronal morphology; very little immunostaining of the scarce flattened cells, probably Müller cells, was to and. The results indicate that the changes in ganglioside metabolism, which lead to decreased synthesis of gangliosides lacking the galactosyl-N-acetyl-galactosaminyl disaccharide end and to increased synthesis of gangliotetraosyl gangliosides, occur in cells that in culture differentiate into neurons.
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  • 34
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: We observed the presence of the novel pituitary protein „7B2” and its release in the bovine adrenal medulla. The 7B2 concentration (mean ± SEM) in extracts of the bovine adrenal medulla was 952 ± 155 pg/mg tissue (n = 6). 7B2 was distributed in the chromaffin granule fraction prepared from the bovine adrenal medulla and was released by high K+ and/or nicotine from cultured cells of the bovine adrenal medulla. Co-release of 7B2 with catecholamine induced by nicotine from the cultured bovine chromaffin cells was also observed. In an analysis of the bovine adrenal medulla chromaffin granule fraction on gel permeation chromatography, there was a major peak with an apparent molecular weight of 45,000, whereas a major peak with an apparent molecular weight of 20,000 was found in that on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. On reverse-phase HPLC, a major peak with a retention time of 35 min was observed in the bovine chromaffin granule fraction and in the bovine anterior pituitary extract. These findings indicate that 7B2 is a secretory protein in the bovine adrenal medulla. The possibility that 7B2 might be released with catecholamine, possibly in response to stress, warrants investigation.
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  • 35
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    Oxford, UK : Blackwell Publishing Ltd
    Journal of neurochemistry 49 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Two [3H]ketanserin recognition sites are present in the rat striatum. The high-affinity site (Kd, 0.39 nM) is similar to the 5-hydroxytryptamine2 (5-HT2) site previously characterized by various investigators. The low-affinity site (Kd, 21.8 nM) has a unique pharmacologic specificity and is preferentially localized to rat striatum and septum. Conventional 5-HT2 antagonists as well as 5-HT and 5-HT uptake inhibitors are ineffective at inhibiting [3H]ketanserin binding to this low-affinity site. Also, chronic treatment with p-chlorophenylalanine, which depletes brain 5-HT, upregulates only the high-affinity site. Thus, in the striatum and septum, [3H]ketanserin labels a unique recognition site. This site has recently been shown to be associated with dopaminergic nerve endings and may regulate biogenic amine release.
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  • 36
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    Oxford, UK : Blackwell Publishing Ltd
    Journal of neurochemistry 49 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: We have previously shown that, in the myelin-deficient jimpy mutant mouse, 74 nucleotides are absent from the mRNA for proteolipid protein (PLP) as a result of aberrant RNA processing. To define the exact site of the jimpy mutation, we have analyzed the PLP gene obtained from a jimpy mouse genomic library. We find that the nucleotide sequence that is absent from jimpy PLP mRNA is fully preserved in the jimpy PLP gene. The missing segment corresponds to a separate exon, equivalent to exon 5 of the human PLP gene. The nucleotide sequence at the 3’end of intron 4 in the jimpy PLP gene contains a single point mutation. A base change A → G in the 3’acceptor splice site has altered a position that is 100% conserved in all published splice acceptor sequences. We conclude that the primary genetic defect of the jimpy mouse is a single base change in the PLP gene disabling an invariant recognition sequence of RNA splicing.
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  • 37
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    Oxford, UK : Blackwell Publishing Ltd
    Journal of neurochemistry 49 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: The proposed structural protein of peripheral nerve myelin, P0, has been shown to have several covalent modifications. In addition to being glycosylated, sulfated, and acylated, P0 is phosphorylated, with the intracellular site of this latter addition being in question. By employing nerve injury models that exhibit different levels of P0 biosynthesis in the absence and presence of myelin assembly, we have examined the cellular location of P0 phosphorylation. It is demonstrated that there is comparable P0 phosphorylation in both normal and crush-injured adult rat sciatic nerves, although the level of biosynthesis of P0 differs between these myelin maintaining and actively myelinating nerve models, respectively. The glycoprotein does not appear to be phosphorylated readily in the transected adult sciatic nerve, a preparation in which P0 biosynthesis is observed but that lacks myelin membrane. These observations suggest that the modification is not associated with the biosynthesis or maturation of P0 in the endoplasmic reticulum or Golgi, but that it instead occurs after myelin assembly. That P0 phosphorylation occurs in the normal nerve even when translation is inhibited by cycloheximide treatment lends further support to this conclusion. P0 is shown to be phosphorylated on one or more serine residues, with all or most of the phosphate group(s) being labile as evidenced by pulse-chase analysis. Addition of a biologically active phorbol ester, 12-O-tetradecanoylphorbol-13-acetate or 4β-phorbol 12, 13-dibutyrate, substantially increases the extent of [32P]orthophosphate incorporation into the glycoprotein of normal and crushed nerve but not transected nerve. Biologically inactive 4α-phorbol 12, 13-didecanoate has no effect on P0 phosphorylation. Similarly, the addition of the cyclic AMP analog 8-bromo-cyclic AMP causes no appreciable changes in P0 labeling. These findings indicate that the phorbol ester-sensitive enzyme, protein kinase C, may be responsible for the phosphorylation of P0 within the myelin membrane.
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  • 38
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    Oxford, UK : Blackwell Publishing Ltd
    Journal of neurochemistry 49 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Clonal cell line D6P2T, subcloned from an ethylnitrosourea-induced tumor line D6 of the rat peripheral nervous system, has been characterized with particular attention to galactolipid metabolism. Galactosylcerebroside and sulfatide synthesis and expression on the cell surface are highly regulated in D6P2T cells by mechanisms involving serum-and cyclic AMP-mediated pathways. These cells also express 2′,3′-cyclic nucleotide 3′-phosphohydrolase (Wolfgram protein Wla) and laminin. In contrast, myelin basic protein and antigen HNK-1 were not detected. Line D6P2T appears to be a semi-differentiated Schwann cell model, which offers interesting possibilities for studies of galactolipid synthesis, transport, and sorting.
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  • 39
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    Oxford, UK : Blackwell Publishing Ltd
    Journal of neurochemistry 49 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Increased concentrations of the endogenous tryptophan metabolite 3-hydroxykynurenine (3-HK) were measured in the brains of vitamin B6 deficient neonatal rats. Mean concentrations of 3-HK in B6 deficient cerebellum, corpus striatum, frontal cortex, and pons/medulla ranged from 9.7 to 18.6 and 102 to 142 nmol/g of wet tissue at 14 and 18 days of age, respectively. 3-HK was not significantly increased in control neonatal or adult rat brain, vitamin B6 deficient rat brain at 7 days of age, or in brains from adult rats deprived of vitamin B6 for 58 days. The administration of daily intraperitoneal injections of vitamin B6 from the 14th to the 18th day of age decreased the concentration of 3-HK to control levels. 3-HK has been shown by other investigators to produce seizures when injected into the cerebral ventricles of adult rodents. Thus, our studies show the accumulation in brain of a putative endogenous convulsant as the result of a nutritional deficiency.
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  • 40
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    Oxford, UK : Blackwell Publishing Ltd
    Journal of neurochemistry 49 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
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  • 41
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    Oxford, UK : Blackwell Publishing Ltd
    Journal of neurochemistry 49 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: 2-Pyrrolidinone, the lactam of γ-aminobutyric acid (GABA), is identified as the major constituent of total GABA in human CSF. Structural elucidation was done by mass spectrometry. In lumbar CSF of four patients, 2-pyr-rolidinone represented about 54% of GABA found after acid hydrolysis, thus accounting for essentially all of the hitherto unknown GABA fraction in CSF.
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  • 42
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    Oxford, UK : Blackwell Publishing Ltd
    Journal of neurochemistry 49 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: The effects of Ca2+ and Na+ ions on the affinities of rat striatal dopamine D1 and D2 receptors for a wide range of agonists and antagonists were investigated. These experiments were performed at 37°C, since it was found that at this physiological temperature D2 receptor affinities for dopamine and 2-amino-6,7-dihydroxy-1,2,3,4-tetrahy-dronaphthalene were clearly lower than at room tempera-re. No correlation was found between the effects of the cations on the affinities of compounds for D1 and D2 receptors and their agonistic or antagonistic nature. On the other hand, the Hill coefficients of agonists but not of antagonists were consistently and significantly below unity, with either Ca2+ or Na+ ions present and at both receptors. This suggests the existence of yet another type of heterogeneity of D1 and D2 receptor forms, to which agonists but not antagonists are sensitive. It is thus concluded that changes in D1 and D2 receptor affinities induced by cations do not predict the agonistic or antagonistic nature of a compound. However, since dopamine itself was sensitive to Na+ or Ca2+ ions, this mechanism might play a role in the regulation of receptor affinities in synaptic transmission, in addition to that exerted by guanyl nucleotides.
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  • 43
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: The accumulation and utilization of [35S]3′-phos-phoadenosine 5′-phosphosulfate (PAPS) were studied in slices from rat cerebral cortex incubated in the presence of inorganic [35S]sulfate. [35S]PAPS levels were directly evaluated after either isolation by ion-exchange chromatography or quantitative enzymatic transfer of its active [35S]sulfate group to an acceptor phenol under the action of added phenolsulfotransferase activity. [35S]PAPS formation was also indirectly followed by incubating slices in the presence of β-naphthol and measuring the levels of [35S]β-naphthyl sulfate ([35S]β-NS). Whereas [35S]PAPS levels rapidly reached a plateau, [35S]β-NS formation proceeded linearly with time for at least 1h, an observation indicating that the nucleotide was continuously synthesized and utilized for endogenous sulfation reactions. [35S]PAPS formation in ices was completely and rather potently blocked by 2,6-dichloro-4-nitrophenol (IC50= .10 μM), an inhibitor of the PAPS-synthesizing enzyme system in a cytosolic preparation. [35S]PAPS accumulation and [35S]β-NS‘formation were strongly reduced by depolarizing agents such as potassium or veratridine. At millimolar concentrations, various excitatory amino acids (glutamate, aspartate, cysteate, quisqualate, and homocysteate) also elicited similar effects, whereas kainate and N-methyl-D-aspartate were inactive. This suggests that PAPS synthesis is turned off when cerebral cells are strongly depolarized.
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  • 44
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    Oxford, UK : Blackwell Publishing Ltd
    Journal of neurochemistry 49 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: Kinetic and equilibrium measurements of [3H]-serotonin (5-hydroxytryptamine) binding to human frontal cortex membranes have been made between 4 and 30°C. The effects of spiperone and ascorbate on binding have also been determined. Under the conditions used, binding was saturable and reversible. Affinity constants derived from kinetic and equilibrium data were comparable. Serotonin binding to several sites had substantial enthalpic as well as entropic components.
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  • 45
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    Oxford, UK : Blackwell Publishing Ltd
    Journal of neurochemistry 49 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: An antiserum was raised against fractions of the Golgi apparatus of neurons from rat brain. Immunoblots of these fractions with the antiserum showed two principal bands of 185 and 150 kilodaltons (kd) in apparent molecular mass. The antiserum reacted with five or six bands of 200, 150, 130, 100–110, 64, and 40 kd in apparent molecular mass in immunoblots of several crude brain membrane fractions. Affinity-purified antibodies from the different gel bands transferred to nitrocellulose paper were used in immunoblot and immunocytochemical studies. Antibodies eluted from the 200-, 150-, 100–110-, and 64-kd bands reacted not only with the corresponding band but also with the other three bands. Antibodies eluted from the 40-kd band stained only the corresponding band. On light and/or electron microscopic immunocytochemistry, the antiserum stained the Golgi apparatus of rat neurons, glia, liver, and kidney tubule cells. Weaker, segmented, and less consistent staining was observed in nuclear envelopes, rough endoplasmic reticulum, and plasma membranes of neurons. Antibodies eluted from the bands at 200, 150, 100–110, and 64 kd stained intermediate cisterns of the Golgi apparatus of neurons. These findings suggest that a group of related polypeptides of brain membranes is preferentially expressed or enriched in the Golgi apparatus of neurons. Polypeptides with apparent molecular masses of 185 and 150 kd probably represent moieties endogenous to membranes of the neuronal Golgi apparatus.
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  • 46
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    Oxford, UK : Blackwell Publishing Ltd
    Journal of neurochemistry 49 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: Application of both phosphorus (31P) and proton (1H) magnetic resonance spectroscopy (MRS) to the study of brain metabolism permits the noninvasive measurement of intracellular pH and brain lactate level. We have used water-suppression 1H MRS with novel lactate-editing techniques, together with 31P MRS, to characterize sequential changes in brain lactate level and pH in vivo over an 8-h period following fluid-percussion brain injury of graded severity in the rat. A transient fall in intracellular pH (from 7.09 ± 0.07 at baséline to 6.88 ± 0.09 at 40 min postinjury) occurred in animals subjected to moderate-(1.5–2.2 atm) and high-(2.5–3.3 atm) but not low-level (0.1–1.2 atm) injury; intracellular pH returned to baseline by 90 min postinjury. Transient elevations in brain lactate level were observed that temporally paralleled and were significantly correlated with the pH changes for all injury levels (r= 0.93, p 〈 0.001). Postinjury alterations in intracellular brain pH and lactate level were identical in magnitude in animals subjected to either moderate or high-level injury. However, animals subjected to moderate injury had a moderate chronic neurological deficit that persisted up to 4 weeks postinjury, whereas animals subjected to a high level of injury showed greater histopathological damage and a more severe chronic neurological deficit. These data suggest that the extent of posttraumatic intracellular cerebral acidosis in our model of experimental head injury is not directly related to the severity of functional neurological deficit.
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  • 47
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: The effects of ganglioside supplementation of culture medium on monoamine oxidase (MAO) type A and B activities in a rat clonal pheochromocytoma cell line, PC12h, were examined. The MAO activity in PC12h cells proved to be mainly due to type A MAO, and type B MAO activity was negligible. After supplementation of the culture medium with ganglioside GM1, the PC 12 cells were found to express type B MAO activity after 4 days of culture, and the amount of type B activity increased with the number of days of culture. After 3 weeks of culture in the presence of GM1, type B activity was about 10% of the total, whereas in control cells type B MAO activity was only about 0.6% of the total. By kinetic analyses of type A and B MAO in PC12h cells after 3 weeks of culture, the increase of type B MAO activity was found to be due to the increase in amount of type B MAO; the Km values were almost the same and only the Vmax values were increased in the cells supplemented with GM1. Among gangliosides tested GM1 was the most effective in causing expression of type B MAO activity, whereas nerve growth factor was not effective. These results suggest that GM1 and other gangliosides may be involved in the expression of type B MAO in nerve cells and in the regulation of levels of the biogenic amines in the brain.
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  • 48
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    Journal of neurochemistry 49 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: Primary cultures of newborn rat cerebrum, which are composed of glial cells (principally astroglia), were used for examining the relationship between dolichol-linked glycoprotein synthesis and DNA synthesis in developing cerebral glia. The cells were synchronized by reducing the content of fetal calf serum in the culture medium from 10 to 0.1% (vol/vol) for 48 h between days 4 and 6 in culture. Reversal of the quiescent state by return of the cultures to 10% serum causes a marked increase in DNA synthesis 12–24 h later. A sharp increase in glycoprotein synthesis (incorporation of [3H]mannose) occurred in the first 12 h after serum repletion, preceding the increase in DNA synthesis. Tunicamycin, an inhibitor of the dolichol-linked pathway to glycoprotein synthesis at the first committed step in oligosaccharide formation, promptly and completely prevented the increase in glycoprotein synthesis and, in addition, the subsequent increase in DNA synthesis. The effects of tunicamycin on glycoprotein and DNA syntheses were reversible, and no comparable effect c n total protein synthesis was observed. When tunicamycin was added only during a temporally circumscribed peiod in G1, i.e., from 3 to 9 h after serum repletion, the increase in DNA synthesis between 12 and 24 h after repletion was still markedly inhibited, i.e., to ∼45% of the value in untreated cultures. The data thus show that there is a requirement for dolichol-linked glycoprotein synthesis for the subsequent occurrence of DNA synthesis and that this requirement is expressed late in the G1 phase of the cell cycle.
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  • 49
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    Journal of neurochemistry 49 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: The ability of cholinergic agonists to activate phospholipase C in bovine adrenal chromaffin cells was examined by assaying the production of inositol phosphates in cells prelabeled with [3H]inositol. We found that both nicotinic and muscarinic agonists increased the accumulation of [3H]inositol phosphates (mainly inositol monophos-phate) and that the effects mediated by the two types of receptors were independent of each other. The production of inositol phosphates by nicotinic stimulation required extracellular Ca2+ and was maximal at 0.2 mMCa2+. Increasing extracellular Ca2+ from 0.22 to 2.2 mM increased the sensitivity of inositol phosphates formation to stimulation by submaximal concentrations of 1,1-dimethyl-4-phenyl-piperazinium iodide (DMPP) but did not enhance the response to muscarine. Elevated K+ also stimulated Ca2+-dependent [3H]inositol phosphate production, presumably by a non-receptor-mediated mechanism. The Ca2+ channel antagonists D600 and nifedipine inhibited the effects of DMPP and elevated K+ to a greater extent than that of muscarine. Ca2+ (0.3–10 μM) directly stimulated the release of inositol phosphates from digitonin-permeabilized cells that had been prelabeled with [3H]inositol. Thus, cholinergic stimulation of bovine adrenal chromaffin cells results in the activation of phospholipase C by distinct muscarinic and nicotinic mechanisms. Nicotinic receptor stimulation and elevated K+ probably increased the accumulation of inositol phosphates through Ca2+ influx and a rise in cytosolic Ca2+. Because Ba2+ caused catechol-amine secretion but did not enhance the formation of inositol phosphates, phospholipase C activation is not required for exocytosis. However, diglyceride and wyo-inositol 1,4,5-trisphosphate produced during cholinergic stimulation of chromaffin cells may modulate secretion and other cellular processes by activating protein kinase C and/or releasing Ca2+ from intracellular stores.
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  • 50
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    Oxford, UK : Blackwell Publishing Ltd
    Journal of neurochemistry 49 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
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  • 51
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    Journal of neurochemistry 49 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: [14C]Acetylcholine (ACh) release and parallel alterations in 45Ca2+ uptake and intrasynaptosomal free Ca2+concentration ([Ca2+]i) were measured in guinea-pig brain cortex synaptosomes. Depolarization by high K+ concentrations caused a rapid transient increase in Ca2+ uptake, terminating within 60 s (rate constant = 0.060 s-1; t1/2= 11.6 s). This resulted in a rapid increase (within 1 s) in [Ca2+]i, which then fell to a maintained but still-elevated plateau level (t1/2 for the decline was 15 s). Peaks of [Ca2+]ishowed a sigmoidal dependence on depolarization, contrasting with the simple linear dependence of plateau levels of [Ca2+]i. The K+-evoked ACh release also had two phases: a fast initial increase (t1/2= 11.3 s), which terminated within 60 s, was followed by a slow additional increase during sustained depolarizations of up to 10 min. Depolarization by veratridine led to a slow gradual increase in Ca2+ uptake (t1/2= 130 s) over a 10-min incubation period, whereas an elevated plateau level of [Ca2+]i was achieved within 2 min (without a rapid peak elevation). The Ca2+-dependent fraction of the veratridine-evoked ACh release correlated with the increase in [Ca2+]i rather than with Ca2+ uptake. Using two different methods of depolarization partially circumvented the time limitations imposed by a buffering Ca2+ indicator and we suggest that, in the main, ACh is released in bursts associated with [Ca2+]i transients.
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  • 52
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: The effects of some anticonvulsant drugs have been investigated on γ-hydroxybutyrate release from rat hippocampal and striatal slices. Sodium valproate and ethosuximide inhibited the depolarization-evoked release of γ-hydroxybutyrate induced by 40 mM K+. The IC50 values for these two drugs are in the concentration range of valproate and ethosuximide that exists in rat brain after administration of anticonvulsant doses to the animals. Trimethadione and pentobarbital are without significant effects. It can be concluded that the inhibition of γ-hydroxybutyrate release, particularly that observed for hippocampus, might explain the protective effect of valproate and ethosuximide on γ-hydroxybutyrate-induced seizures and perhaps on other kinds of epileptoid phenomenon.
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  • 53
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: The distribution of plasma membrane markers, the sodium pump [evaluated as ouabain-sensitive, potassium-stimulated p-nitrophenyl phosphatase (K+-pNPPase)], [3H]saxitoxin binding, and 5′-AMPase, was studied in the subcellular fractions prepared from the ho-mogenates of the longitudinal smooth muscle/myenteric plexus of dog ileum. The K+-pNPPase activity and [3H]-saxitoxin binding were found to be predominantly associated with the synaptosomal fraction as indicated by the high level of these activities in the crude synaptosomal fraction and by the copurification of K+-pNPPase and [3H]saxitoxin binding, but not 5′-AMPase, with several synaptosomal markers during the fractionation of the crude synaptosomal fraction on density gradients. In contrast to the K+-pNPPase activity and [3H]saxitoxin binding, the 5′-AMPase activity was found to be concentrated in the microsomal pellet. Further fractionation of microsomes on a density gradient resulted in copurification of 5′-AMPase, but not K+-pNPPase or [3H]saxitoxin binding, with other smooth muscle plasma membrane-bound enzymes, such as high-affinity Ca2+-ATPase, Mg2+-ATPase, and Ca2+-ATP-ase. It was concluded that in the longitudinal smooth muscle/myenteric plexus, the sodium pump activity is present in higher density in the neuronal plasma membranes whereas 5′-AMPase activity is concentrated in the smooth muscle plasma membranes.
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  • 54
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    Journal of neurochemistry 49 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: Na+-dependent uptake of L-[3H]proline was measured in a crude synaptosomal preparation from the entire rat hippocampal formation or from isolated hippo-campal regions. Among hippocampal regions, Na+-depen-dent proline uptake was significantly greater in areas CA1 and CA2-CA3-CA4 than in the fascia dentata, whereas there was no marked regional difference in the distribution of Na+-dependent γ-[14C]aminobutyric acid ([14C] GABA) uptake. A bilateral kainic acid lesion, which destroyed most of the CA3 hippocampal pyramidal cells, reduced Na+-de-pendent proline uptake by an average of 41% in area CA1 and 52% in area CA2-CA3-CA4, without affecting the Na+-dependent uptake of GABA. In the fascia dentata, neither proline nor GABA uptake was significantly altered. Kinetic studies suggested that hippocampal synaptosomes take up proline by both a high-affinity (KT=6.7 μM) and a low-affinity (KT= 290 μM) Na+-dependent process, whereas L-[14C]glutamate is taken up predominantly by a high-affinity (KT=6.1 μM) process. A bilateral kainic acid lesion reduced the Vmax of high-affinity proline uptake by an average of 72%, the Vmax of low-affinity proline uptake by 44%, and the Vmax of high affinity glutamate Uptake by 43%, without significantly changing the affinity of the transport carriers for substrate. Ipsilateral-commissural projections of CA3 hippocampal pyramidal cells Appear to possess nearly as great a capacity for taking up proline as for taking up glutamate, a probable transmitter of these pathways. Therefore proline may play an important role in transmission at synapses made by the CA3-derived Schaffer collateral, commissural, and ipsilateral associational fibers.
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  • 55
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    Journal of neurochemistry 49 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: The Ca2+/calmodulin (CaM)-dependent protein kinase II system in squid nervous tissue was investigated. The Ca2+/CaM-dependent protein kinase II was found to be very active in the synaptosome preparation from optic lobe, where it was associated with the high-speed particulate fraction. Incubation of the synaptosomal homogenate with calcium, calmodulin, magnesium, and ATP resulted in partial and reversible conversion of the Ca2+/CaM-dependent protein kinase II from its calcium-dependent form to a calcium-independent species. The magnitude of this conversion reaction could be increased by inclusion of the protein phosphatase inhibitor NaF or by substitution of adenosine 5′-O-(3-thiotriphosphate) for ATP. When [γ-32P] ATP was used, proteins of 54 and 58 kilodaltons (kDa) as well as proteins 〉100 kDa were rapidly 32P-labeled in a calcium-dependent manner. Major 125I-CaM binding proteins in the synaptosome membrane fraction were 38 and 54 kDa. The Ca2+/CaM-dependent protein kinase II was purified from the squid synaptosome and was shown to consist of 54-and 58–60-kDa subunits. The purified kinase, like Ca2+/CaM-dependent protein kinase II from rat brain, catalyzed auto-phosphorylation associated with formation of the calcium-independent form. These studies, characterizing the Ca2+/ CaM-dependent protein kinase II in squid neural tissue, are supportive of the putative role of this kinase in regulating calcium-dependent synaptic functions.
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  • 56
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    Journal of neurochemistry 49 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: Expression of the myelin proteolipid protein (PLP) was examined in the nuclei and polysomes of 12–27-day-old quaking, jimpy, and shiverer mouse brains and in 2–27-day-oId normal brains and compared with expression of the myelin basic proteins (MBPs). Northern blots showed the presence of multiple mouse PLP RNAs, the developmental expression of which coincided with my-elination. Two major mouse PLP RNAs, 3.5 and 2.6 kilo-bases in length, were observed in both cytoplasmic polyri-bosomes and nuclei, and, in addition, a larger 4.6-kilobase PLP RNA was observed in nuclei. Quantitative measurements with slot blot analyses showed that the levels of PLP and MBP RNAs peaked simultaneously at 18 days in nuclei but that maximal levels of PLP RNA lagged behind MBP RNA by several days in the polysomes. The developmental expression of both major classes of myelin protein mRNAs was affected in all three mutants. In shiverer brains, the levels of PLP mRNA in polysomes and nuclei were only 30–55% of control levels after 15 days. Thus, the deletion off a portion of the MBP gene appeared to have a major effect on the expression of the PLP gene in this mutant. In jimpy mice, where the mutation has been shown to involve the PLP gene, expression of MBP mRNA was also severely reduced, to 〈25% of control values. In quaking brains, the expression of each gene followed its own developmental course, different from each other and different from the normal mouse. The extent to which the expression of PLP and MBP was affected by the quaking mutation depended on the age at which it was examined.
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  • 57
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    Journal of neurochemistry 49 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: We have measured the free amino acid content of three distinct astroglial cell clones derived from permanent lines obtained after “spontaneous immortalization” of 8-day postnatal mouse cerebellar cultures; these clones show characteristics similar to the Golgi Bergmann glia cells, the fibrous astrocytes, and the velate protoplasmic astrocytes, i.e., the three main types of cerebellar astrocytes. The relative concentrations of amino acids that are thought to act as neurotransmitters were compared in confluent cultures of the different astroglial clones. The most striking result was a high concentration of glycine (20% of free amino acids), even in astroglial cells cultured in a glycine-free medium, a finding suggesting that glycine is synthesized by the astroglial clones. Furthermore, no γ-aminobutyric acid (GABA) was detected. In contrast, a “neuron-like” clone derived from the same cerebellar culture contained GABA, whereas its glycine content was much lower than that of the astroglial clones. The present results, together with our previous finding of glycine synthesis in an astrocytic clone derived from 14-day postnatal mouse cerebella transformed by simian virus 40, indicate that a high glycine content may be characteristic of many cerebellar astroglial types.
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  • 58
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    Journal of neurochemistry 49 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: A variety of neurotransmitters elicit a phos-phoinositide response in the CNS; however, their effects on prostaglandin (PG) formation in the brain are not well characterized. In the present study, we investigated the effect of acetylcholine (ACh) on the synthesis of PCs E and F in slices from various regions of guinea pig brain incubated in glucose-fortified Krebs-Henseleit bicarbonate saline. Slices were prewashed in the presence of 1% albumin to reduce basal PG levels followed by incubation for 30 min at 37°C in the presence or absence of ACh. Under these conditions, 5 mM ACh significantly increased the efflux of PGE and PGF from brain regions enriched in muscarinic cholinergic receptors, i.e., cerebral cortex, temporal cortex, corpus striatum, and hippocampus. Depolarization by 45 mM KCl also significantly enhanced PG synthesis, and the relative magnitude of the effect was similar to that of ACh. The stimulation of PG synthesis by ACh was inhibited by 20 μM atropine, whereas the K+-induced stimulation was not. The effects of potassium and ACh were additive at maximally effective ACh concentrations, an observation that suggests that ACh and K+ increase PG efflux through independent mechanisms. Norepinephrine, histamine, and serotonin, three other neurotransmitters that evoke a phos-phoinositide response in the brain, were ineffective in stimulating PG release from brain cortex slices.
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  • 59
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    Topics: Medicine
    Notes: Abstract: The phosphorylation of tyrosine hydroxylase, purified from rat striatum, was investigated using purified Ca2+/calmodulin (CaM)-dependent protein kinase II. This kinase catalyzed the Ca2+-dependent incorporation of up to 0.8 mol 32PO4/mol tyrosine hydroxylase subunit (62 kilo-daltons). Reverse-phase high-performance liquid chroma-tography mapping of tryptic 32P-peptides established that the Ca2+/CaM-dependent protein kinase II phosphorylated a different serine residue than was phosphorylated by the cyclic AMP-dependent protein kinase. Limited proteolysis sequentially reduced the subunit Mr from 62 to 59 kilodaltons and finally to 57 kilodaltons, resulting in loss of the site phosphorylated by the Ca2+/CaM-dependent protein kinase II, but not the site phosphorylated by the cyclic AMP-dependent protein kinase. Phosphorylation by the Ca2+/ CaM-dependent protein kinase II had little direct effect on the kinetic properties of tyrosine hydroxylase, but did convert it to a form that could be activated twofold by addition of an activator protein. This heat-labile activator protein increased the Vmax without affecting the Km for the pterin cofactor. This effect was specific in that the activator protein was without effect on nonphosphorylated tyrosine hydroxylase or on tyrosine hydroxylase phosphorylated by the cyclic AMP-dependent protein kinase. These results are consistent with the hypothesis that the „Vmax-type” activation of tyrosine hydroxylase observed upon depolarization of neural and adrenal tissues may be mediated by the Ca2+/ CaM-dependent protein kinase II.
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  • 60
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    Journal of neurochemistry 49 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: Receptors for the nerve growth factor protein (NGF) have been isolated from three cell types [embryonic chicken sensory neurons (dorsal root sensory ganglia; DRG), rat pheochromocytoma (PC 12) and human neuroblastoma (LAN-1) cells] and have been shown to be similar with respect to equilibrium dissociation constants. The present results demonstrate that there are multiple molecular weight species for NGF receptors from DRG neurons and PC 12 cells. NGF receptors can be isolated from DRG as four different molecular species of 228, 187, 125, and 112 kilodaltons, and PC 12 cells as three molecular species of 203, 118, and 107 kilodaltons. The NGF receptors isolated from DRG show different pH-binding profiles for high-and low-affinity binding. High-affinity binding displays a bell-shaped pH profile with maximum binding between pH 7.0 and 7.9, whereas low-affinity binding is constant between pH 5.0 and 9.1, with a twofold greater binding at pH 3.6. At 22°C, the association rate constant was found to be 9.5 ± 1.0 ± 106M-1 S-1. Two dissociation rate constants were observed. The fast dissociating receptor has a dissociation rate constant of 3.0 ± 1.5 ± 10-2 S-1, whereas the slow dissociating receptor constant was 2.4 ± 1.0 ± 10-4S-1. The equilibrium dissociation constants calculated from the ratio of dissociation to association rate constants are 2.5 ± 10-11M for the high-affinity receptor (type I) and 3.2 ± 10-9M for the low-affinity receptor (type II). These values are the same as those determined by equilibrium experiments on the isolated receptors. The data are consistent with the hypothesis that there are at least two different receptors for NGF.
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  • 61
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    Topics: Medicine
    Notes: Abstract: The inhibition of adenylate cyclase from rat striatal plasma membranes by guanyl-5′-yl-imidodiphos-phate [Gpp(NH)p] and morphine was compared to determine whether Gpp(NH)p-mediated inhibition accurately reflected hormone-mediated inhibition in this system. Inhibition of adenylate cyclase activity by Gpp(NH)p and morphine was examined with respect to temperature, divalent cation concentration, and the presence of Ca2+/calmodulin (Ca2+/CaM). Gpp(NH)p-mediated inhibition was dependent on the presence of Ca2+/CaM at 24°C; the inhibition was independent of Ca2+/CaM at 18°C; and inhibition could not be detected in the presence, or absence, of Ca2+/ CaM at 30°C. In contrast, naloxone-reversible, morphine-induced inhibition of adenylate cyclase was independent of both temperature and the presence of Ca2+/CaM. Mg2+dose-response curves also reinforced the differences in the Ca2+/CaM requirement for Gpp(NH)p-and morphine-induced inhibition. Because Gpp(NH)p-mediated inhibition was independent of Ca2+/CaM at low basal activities (i.e., 18°C, or below 1 mM Mg2+) and dependent on the presence of Ca2+/CaM at higher basal activities (24°C, or above 1 mM Mg2+), the inhibitory effects of Gpp(NH)p were examined at 1 mM Mg2+ in the presence of 100 nM forskolin. Under these conditions, both Gpp(NH)p-and morphine-induced inhibition of adenylate cyclase were independent of Ca2+/CaM. The results demonstrate that the requirement for Ca2+/CaM to observe Gpp(NH)p-mediated inhibition depends on the basal activity of adenylate cyclase, whereas hormone-mediated inhibition is Ca2+/CaM independent under all conditions. Further, this study shows that, under certain conditions, Gpp(NH)p-mediated inhibition of adenylate cyclase does not mimic the inhibition of adenylate cyclase produced by a complete interaction between the inhibitory hormone receptor, the guanine nucleotide-regulatory component that mediates inhibition, and the catalytic unit of the adenylate cyclase complex in striatal plasma membranes.
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  • 62
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    Journal of neurochemistry 49 (1987), S. 0 
    ISSN: 1471-4159
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    Topics: Medicine
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  • 63
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    Journal of neurochemistry 49 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: Autopsy material from deceased individuals between ages 2 and 90 was used to prepare cerebellum, pons, and other selected regions of the brain, the spinal cord, and peripheral nerves. The concentration of dolichol in these different tissues varied greatly and the increase in concentration during the life span varied between 2.5- and 21-fold. In contrast, dolichyl-phosphate (dolichyl-P) was more evenly distributed in these tissues and its concentration increased to a moderate extent only during childhood. The level of cholesterol displayed smaller regional differences and decreased about 15% between ages 35 and 90. Differences in the total phospholipid content were limited. These results demonstrate enrichment and individual regulation of various lipids in specialized regions of the human brain. The independent regulation of dolichol and dolichyl-P levels in the brain and the possible role of dolichol in the function of the aging nerve cell are also emphasized.
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  • 64
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    Journal of neurochemistry 49 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: A neurotrophic factor that promotes the survival of cholinergic parasympathetic ciliary neurons has been purified ∼20,000-fold from bovine cardiac tissue under nondenaturing conditions using heparin-affinity chromatography. Up to 22 μg of purified factor having a specific activity of 4 × 105 trophic units/mg can be obtained from 250 g of heart muscle. Sodium dodecyl sulfate (SDS)-polyacrylamide gels of the purified material show a broad band that is sometimes resolvable into a closely spaced pair of bands of 22 and 23 kilodaltons. Partially purified factor can be resolved into two peaks of activity (pI 5.6 and 5.0) by highresolution anion-exchange chromatography and chromato focusing, although these procedures have not proved useful as purification methods because of the large losses of activity incurred. It is likely that these two peaks represent the two bands seen on SDS-polyacrylamide gels. The bovine cardiac factor(s) differs from similar factors purified from chick optic tissues and pig brain in that it is irreversibly denatured by SDS.
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  • 65
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    Journal of neurochemistry 49 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: The effects of castration and testosterone replacement on hypothalamic pools of β-endorphin and dynorphin and on the basal and corticotropin-releasing factor (CRF)-stimulated release of these peptides from hypothalamic slices in vitro were studied. The experiments were done in adult male rats. The hypothalamic content of both peptides increased significantly within 1 week of castration, and levels remained elevated for up to 4 weeks. Testosterone treatment, begun at the time of castration, prevented these increases. In addition, testosterone replacement 6 weeks after castration reversed peptide levels to normal. Basal in vitro release rates of β-endorphin and dynorphin were significantly lower from hypothalamic slices derived from 1-week castrated animals than from intact males, and when testosterone was administered in various doses in vivo, basal release rates in vitro increased in a dose-related manner. Hypothalami from rats that had been castrated for 4 weeks, however, showed basal release rates similar to those in tissues from intact controls, a finding indicating that castration initially alters both opioid peptide synthesis and release; later, release is normalized, whereas synthesis remains elevated. CRF was found to stimulate β-endorphin and dynorphin release from hypothalami from intact and from 1- and 4-week-castrated rats, a result indicating that castration does not alter the response of β-endorphin and dynorphin neurons to this stimulus.
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  • 66
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    Journal of neurochemistry 49 (1987), S. 0 
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    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: Rat pheochromocytoma cells (PC 12) permeabilized with staphylococcal α-toxin release [3H]dopamine after addition of micromolar Ca2+. This does not require additional Mg2+-ATP (in contrast to bovine adrenal medullary chromaffin cells). We also observed Ca2+-dependent [3H]-dopamine release from digitonin-permeabilized PC 12 cells. Permeabilization with α-toxin or digitonin and stimulation of the cells were done consecutively to wash out endogenous Mg2+-ATP. During permeabilization, ATP was removed effectively from the cytoplasm by both agents but the cells released [3H]dopamine in response to micromolar Ca2+ alone. Replacement by chloride of glutamate, which could sustain mitochondrial ATP production in permeabilized cells, does not significantly alter catecholamine release induced by Ca2+. However, Mg2+ without ATP augments the Ca2+-induced release. The release was unaltered by thiol-, hydroxyl-, or calmodulin-interfering substances. Thus Mg2+-ATP, calmodulin, or proteins containing -SH or -OH groups are not necessary for exocytosis in permeabilized PC 12 cells.
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  • 67
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    Topics: Medicine
    Notes: Abstract: Intraneural injection of pure glycerol induces Wallerian degeneration with subsequent regeneration. In agreement with other reports, we observed an increase in endoneurial polyunsaturated fatty acids 8 days after the glycerol injection. Levels then fell until day 30. After a period of 5 months, there was an increase in C18:2(n-6) in the intrafascicular tissue, concomitant with a marked fall in this fatty acid in the remaining extrafascicular perineurium. The rise in C18:2(n-6) in endoneurium correlated with infiltration of this tissue by perineurial cells. Interactions between perineurium and endoneurium during nerve regeneration are discussed.
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  • 68
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    Journal of neurochemistry 49 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: In a search for biochemical markers of modified synaptic function following training of day-old chicks on a passive avoidance task, we have assayed two monoclonal antibodies to synaptic vesicle proteins (anti-p65 and anti-SV2) and one raised to postsynaptic densities (411B). We have also measured total acetylcholine (ACh) content. Measurements were made on three forebrain regions known to show metabolic and morphological change consequent on training—the lobus parolfactorius (LPO), paleostriatuni augmentatum (PA), and medial hyperstriatum ventrale (MHV)—in the right and left hemispheres 2 and 24 h after training chicks on a passive avoidance task, in which they learn to avoid pecking a bead coated with methylanthranilate [methylanthranilate-trained (M-trained)]. Control chicks were trained on a water-coated bead [water-trained (W-trained)]. Twenty-four hours after training, 411B levels showed no differences between W-trained and M-trained chicks in any region. M-training reduced the litre of antip65 by 16% in the left PA and 15% in the left MHV and that of anti-SV2 by 19% in the left PA. M-trained chicks showed reduced total ACh content in the LPO by up to 40% and in the PA by up to 48% but had no change in ACh level in the MHV. The decreases in antibody titre were not seen in forebrains analysed 2 h after training, but tendencies toward increases in levels in the right PA and MHV were observed with all three antibodies. Significant differences between right and left hemispheric regions, independent of training, were observed for all the antibodies and for ACh content. In the LPO, antibody litres were some 10% lower in the left compared with the right hemisphere and 34% lower for ACh content, but in the PA, the left hemisphere titre was higher by 14–86%. We discuss these results in the context of morphometric and electrophysiological data oblained from W-trained and M-trained chicks.
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  • 69
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    Journal of neurochemistry 49 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: The ontogenesis of rat forebrain adenosine uptake sites labelled by [3H]nitrobenzylthioinosine ([3H]NBI) was determined and compared to that of rat forebrain adenosine receptors labelled by N6-cyclohexyl[3H]adenosine ([3H]-CHA). [3H]NBI binding is highly invariant with similar levels of [3H]NBI binding sites from embryonic day 19 to day 30 postpartum. Scatchard and Hill analyses reveal the binding of [3H]NBI in 6-day-old tissue to be indistinguishable from such binding in 30-day-old tissue. In contrast, [3H]-CHA binding is highly variant. [3H]CHA binding develops slowly but steadily from about embryonic day 19, with adult binding levels being achieved at around 25 days postpartum. The ontogenetic profile of [3H]CHA appears to coincide with synaptogenesis whereas that of [3H]NBI does not.
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  • 70
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    Journal of neurochemistry 49 (1987), S. 0 
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    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: To study the properties of protein-bound oligosaccharides in neuronally differentiating cells, two model systems were used: murine N1E-115 and N-18 neuroblastoma cells inducible by serum starvation and rat PC12 pheochromocytoma cells inducible by nerve growth factor. Glycopeptides were prepared from cells metabolically labeled with [3H]glucosamine and analyzed by gel nitration. The properties of the high-molecular-weight glycopeptides were studied using enzymatic digestion with neuraminidase and endo-β-galactosidase. In contrast to other cell lines analyzed, the neuroblastoma and pheochromocytoma lines contained predominantly glycopeptides completely cleavable with endo-β-galactosidase, which indicated that they were linear-type poly-N-acetyllactosamine glycans. The proportion of these linear chains in the high-molecular-weight fraction increased during neuronal differentiation in both cell systems. The linear nature of the glycans was also correlated with positive anti-i and negative anti-I reactivity of the cells in immunofluorescence microscopy. Specific cell surface labeling for poly-N-acetyllactosamine glycans and sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed several glycoprotein components, some of which showed changes during neuronal differentiation. The high proportion of linear poly-N-acetyllactosamine chains in these neuronal cell lines and its increase during neuronal differentiation suggests that these glycans may be a characteristic feature of neuronal or neuronally differentiating cells.
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  • 71
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    Journal of neurochemistry 49 (1987), S. 0 
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    Topics: Medicine
    Notes: Abstract: The delivery of proteins to the presynaptic terminals of guinea pig retinal ganglion cells by two of the major components of axonal transport, and the subsequent persistence and turnover of those proteins were examined in this study. Ganglion cell proteins were radiolabeled by intravitreal injection of radiolabeled amino acids and radioactive axonally transported proteins were analyzed in synaptosomes prepared from the superior colliculi. This procedure allowed examination of presynaptic components of ganplion cell synapses without having to compensate for postynaptic or other unidentified contaminants. Each of the two major axonal transport components supplies a large number of proteins to the presynaptic terminal, in relative quantities similar although not identical to those seen in the axon. Proteins conveyed by the fast component of axonal transport reached the terminals by 3 h after intraocular injection, peaked by 24 h, and were largely undetectable by 15 days. Slow component b proteins reached the terminals by 12 days, peaked around 21 days, and persisted up to 63 days in the terminals. Proteins in both components demonstrated differential turnover relative to cotransported proteins once they reached the terminals. Differential turnover may account for change in relative concentration of a particular protein required to meet new functional demands on that protein once it enters the terminal.
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  • 72
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    Topics: Medicine
    Notes: Abstract: We investigated the effect of systemic administration of γ-glutamyl L-3,4-dihydroxyphenylalanine (γ-Glu-DOPA) on catecholamine contents in the brain. γ-Glu-DOPA was transformed to dopamine (DA) in vitro with brain homogenate by the sequential action of γ-glutamyl transpeptidase and aromatic L-amino acid decarboxylase. Intraperitoneal injection of γ-Glu-DOPA to mice increased DA markedly and noradrenaline (NA) moderately in the brain. The increase of endogenous DA was followed by elevation of the main DA metabolites (3,4-dihydroxyphenyl-acetic acid and homovanillic acid). These increases were in a dose-dependent manner. The maximal elevation of DA was observed within 30 min after administration of γ-Glu-DOPA, but a substantial increase of NA was observed 2 h after the administration. These results suggest that γ-Glu-DOPA may be applicable to the treatment of Parkinson's disease.
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  • 73
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    Topics: Medicine
    Notes: Abstract: Measurements of the turnover of dopamine (DA) and DA metabolites have been performed in the striatum and substantia nigra (SN) of the rat. Turnover rates of 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid have been assessed from the disappearance rates after blocking their formation by inhibition of monoamine oxidase by pargyline and of catechol-O-methyltransferase by tropolone. DA turnover has been measured as 3-methoxy-tyramine (3-MT) plus DA accumulation rate after MAO inhibition by pargyline and as accumulation rate of 3,4-dihy-droxyphenylalanine (DOPA) after inhibition of aromatic amino acid decarboxylase by NSD 1015 or NSD 1034. These measures of DA turnover have been compared with α-methyl-p-tyrosine (α-MT)-induced DA disappearance rate. In SN all the different measures of DA turnover are in the same range (55–62 nmol/g protein/h) whereas in striatum DOPA accumulation rate after NSD 1015 and α-MT- induced DA disappearance rate (16–23 nmol/g/h) are much lower than DOPAC disappearance rate after pargyline, 3-MT plus DA accumulation rate after pargyline, and DOPA accumulation rate after NSD 1034 (39–46 nmol/g/h). The data confirm our previous findings indicating that the fractional turnover rate of DA is more rapid in SN than in striatum and that O-methylation of DA is relatively more important in SN. In striatum at least two pools of DA with different turnover rates appear to exist, whereas in SN, DA behaves as if located in a single compartment.
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    Topics: Medicine
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    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Using an affinity-purified monospecific polyclonal antibody against bovine brain synapsin I, the distribution of antigenically related proteins was investigated in the electric organs of the three strongly electric fish Torpedo marmorata, Electrophorus electricus, Malapterurus electricus and in the rat diaphragm. On application of indirect fluorescein isothiocyanate-immunofluorescence and using α-bungarotoxin for identification of synaptic sites, intense and very selective staining of nerve terminals was found in all of these tissues. Immunotransfer blots of tissue homogenates revealed specific bands whose molecular weights are similar to those of synapsin Ia and synapsin Ib. Moreover, synapsin I-like proteins are still attached to the synaptic vesicles that were isolated in isotonic glycine solution from Torpedo electric organ by density gradient centrifugation and chromatography on Sephacryl-1000. Our results suggest that synapsin I-like proteins are also associated with cholinergic synaptic vesicles of electric organs and that the electric organ may be an ideal source for studying further the functional and molecular properties of synapsin I.
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  • 77
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    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: The synthesis of a tritiated derivative of the 5-HT1A photoaffinity probe 8-methoxy-2-[N-n-propyl, N-3-(2-nitro-4-azidophenyl)aminopropyl]aminotetralin ([3H]8-methoxy-3′-NAP-amino-PAT) allowed the use of this probe for attempting the irreversible labeling of specific binding sites in rat brain membranes. Sodium dodecyl-sulfate-polyacrylamide gel electrophoresis of proteins solubilized from hippocampal microsomal membranes that had been incubated with 20 nM [3H]8-methoxy-3′-NAP-amino-PAT under UV light revealed a marked incorporation of 3H label into a 63-kilodalton protein termed PI . As expected of a possible correspondence between P1 and 5-HT1A receptor binding sites, 3H labeling by the photoaffinity probe could be prevented by selective 5-HT1A ligands such as 8-hydroxy-2-(di-n-propylamino)tetralin, ipsapirone, buspirone, and gepirone and by N-ethylmaleimide, but not by the 5-HT2 antagonist ketanserin, noradrenaline-and dopamine-related drugs, monoamine oxidase inhibitors, and chlorimipramine. Furthermore, the regional and subcellular distributions of P1 were identical to those of specific 5-HT1A binding sites. These results indicated that the binding subunit of the 5-HT1A receptor is a 63-kilodalton protein with a functionally important sulfhydryl group(s).
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  • 78
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    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: The distribution of carbon-11-labeled L-deprenyl, an irreversible inhibitor of monoamine oxidase type B (MAO-B), was determined in the baboon brain by positron emission tomography. The irreversible blood-to-brain transfer constant (influx constant, Ki) was measured using a complete metabolite-corrected arterial plasma concentration curve. This influx constant was used as a measure of functional enzyme activity for sequential determinations of MAO-B recovery following a single high dose of unlabeled l-deprenyl. The half-life for turnover of MAO-B was thus determined to be 30 days. Using appropriate irreversible inhibitors, this procedure should be generally useful for determining enzyme turnover rates in any organ in vivo and can be applied to some human studies as well.
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  • 79
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    Journal of neurochemistry 49 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
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  • 80
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: The metabolism of 3-fluoro-3-deoxy-D-glucose (3-FDG) in rat brain in vivo was investigated noninvasively using 19F nuclear magnetic resonance (NMR) Spectroscopy. Following an intravenous infusion of 3-FDG, 400 mg/kg, four resonances assigned to the α and β anomers of 3-FDG, 3-fluoro-3-deoxy-D-sorbitol, and 3-fluoro-3-deoxy-D-fructose were clearly resolved in brain, a result indicating that 3-FDG is metabolized primarily into the aldose reductase sorbitol (ARS) pathway. An orally administered aldose reductase inhibitor, sorbinil, caused reduction of the flux of 3-FDG into the ARS, an observation suggesting that the method can be applied in quantitative studies of ARS path way activities. Studies of 24-h urine specimens showed that in addition to the two metabolites observed in brain, F-was excreted into the urine. 3-FDG appears to be a suitable metabolic probe for assessing glucose metabolism in the ARS pathway by in vivo 19F NMR Spectroscopy.
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  • 81
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: The Ca2+-binding proteins parvalbumin (Mr= 12K) and calbindin D28K [previously designated vitamin D-dependent Ca2+-binding protein (Mr= 28K)] are neuronal markers, but their functional roles in mammalian brain are unknown. The expression of these two proteins was studied by immunocytochemical methods in serum-free cultures of hypothalamic cells from 16-day-old fetal mice. Parvalbumin is first detected in all immature neurons, but during differentiation, the number of parvalbumin-immunoreactive neurons greatly declines to a level reminiscent of that observed in vivo, where only a subpopulation of neurons stains for parvalbumin. In contrast, calbindin D28K was expressed throughout the period investigated only in a distinct subpopulation of neurons. Depolarization of fully differentiated hypothalamic neurons in culture resulted in a dramatic decrease of parvalbumin immunoreactivity but not of calbindin D28K immunoreactivity. The parvalbumin staining was restored on repolarization. Because the anti-parvalbumin serum seems to recognize only the metal-bound form of parvalbumin, the loss of immunoreactivity may signal a release of Ca2+ from intracellular parvalbumin during depolarization of the cells. We suggest that parvalbumin might be involved in Ca2+-dependent processes associated with neurotransmitter release.
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  • 82
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: The main electric organ of Electrophorus electricus is particularly rich in thiamine triphosphate, which represents 87% of the total thiamine content in this tissue. The thiamine pyrophosphate concentration, however, is very low in the eel electric organ and skeletal muscle as compared with other eel or rat tissues. Furthermore, electroplax membranes contain a whole set of enzymes responsible for the dephosphorylation of thiamine tri-, pyro-, and monophos-phate. Thiamine triphosphatase has a pH optimum of 6.8 and is dependent on Mg2+. The real substrate of the enzyme is probably a 1:1 complex of Mg2+ and thiamine triphosphate. Thiamine pyrophosphatase is activated by Ca2+. The apparent Km for thiamine triphosphate and Vmax are found to be, respectively, 1.76 mM and 5.95 nmol/mg of protein/min. Thiamine triphosphatase activity is inhibited at physiological K+ concentrations (up to 90 mM) and increasing Na+ concentrations (50% inhibition at 300 mM). ZnCl2 (10 mM) inhibits 90% of the enzyme activity. ATP and ITP are also strongly inhibitory. No significant effect of neurotoxins is seen. Membrane-associated thiamine triphosphatase is affected differently by proteolytic enzymes and is partially inactivated by pretreatment with phospholipase C and neuraminidase. The physiological significance of thiamine triphosphatase is discussed in relation to a specific role of thiamine in the nervous system.
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  • 83
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    Journal of neurochemistry 49 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: The requirement for the sterol biosynthetic pathway for the occurrence of DNA synthesis in glial cells and, in particular, the relative roles of cholesterol and of mevalonate have been studied. Primary cultures of developing glial cells were synchronized by reducing the content of fetal calf serum (FCS) in the culture medium from 10% to 0.1% (vol/ vol) for 48 h between days 4 and 6 in culture. Reversal of the resulting quiescent state by the return of the cultures to 10% serum caused after 24 h a marked increase in DNA synthesis, and this increase was prevented by the simultaneous addition of mevinolin, a specific inhibitor of the sterol biosynthetic pathway at the 3-hydroxy-3-methylglutaryl coenzyme A reductase step, at the time of serum repletion. A dose-dependent reversal of the mevinolin inhibition of DNA synthesis occurred with simultaneous addition of mevalonate to the culture medium. The induction of DNA synthesis by serum repletion, its inhibition by mevinolin, and the reversal of the inhibition by mevalonate were unaffected by a 95% reduction in exogenous cholesterol produced by utilization of lipoprotein-poor serum (LPPS) rather than FCS. Similarly, return of quiescent cultures to 10% LPPS containing mevinolin and sufficient low-density lipoprotein (LDL) to raise the cholesterol concentration 80-fold failed to restore DNA synthesis. In addition, reversal of the mevinolin inhibition of DNA synthesis by mevalonate occurred despite the continuous presence of mevinolin if mevalonate was added as late as 12 h after serum repletion, but not if added after 16 h or more. This temporal aspect of the requirement for mevalonate was unaffected by the presence of a 90-fold excess of cholesterol, provided as LDL. Thus, these data establish a requirement for mevalonate or for a presumably nonsterol derivative of mevalonate for the occurrence of DNA synthesis in synchronized glial primary cultures, and demonstrate that this requirement is expressed at a specific time in the cell cycle.
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  • 84
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: For clarification of the beneficial effects of the extract of Ginkgo biloba (EGB) on triethyltin (TET) toxicity in rats, the phosphodiesterase (PDE) activities of the cerebral tissue were measured under in vitro and ex vivo conditions. Under in vitro conditions, low concentrations of EGB (0.25–4.0 mg/L) activated the enzyme, whereas after higher concentrations (5–250 mg/L), dose-dependent inhibition of the enzyme activity was observed. In the lower concentration range, the extract also partially restored the high-affinity PDE activity (measured with 0.25 μM cyclic AMP) of the particulate fraction of the brain inhibited by TET in vitro. In contrast, the inhibitory influence of TET on the low-affinity PDE activity (measured with 50 μM cyclic AMP) of the particulate fraction was enhanced by the extract. Although treatment with a single large dose of EGB lowered the particulate PDE activities of the brain of normal rats, no effects of the extract could be detected in animals after repeated daily administrations of EGB during a 4-day period. Curative treatment of the TET-intoxicated rats with EGB during a 7-day period accelerated the recovery of the edematous state of the white matter caused by the intoxication and also normalized the lowered PDE activity of the particulate fraction of the edematous brain tissue. Furthermore, when preventively administered, EGB counteracted both the edema formation and the fall in PDE activity observed with treatment by TET alone. These observations strongly suggest that some beneficial effects of EGB might be due to its modulating influences on cellular cyclic AMP levels via activation of membrane-bound PDE.
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  • 85
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    Journal of neurochemistry 49 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: : The neurotoxic compound 1-[methyl-3H]-4-phenylpyridinium ([3H]MPP+) was actively taken up by human, rabbit, and guinea pig platelets incubated in plasma. In human platelets, the apparent Km of this uptake (22.6 μM) was 50 times higher than that for serotonin [5-hydroxytryptamine (5-HT)]. The uptake of [3H]MPP+ by human platelets was inhibited by selective 5-HT uptake blockers [cianopramine, (−)-paroxetine, and clomipramine], by metabolic inhibitors (KCN and ouabain), and by drugs that interfere with amine storage in the 5-HT organelles (reserpine, mepacrine, and Ro 4–1284). Impairment of the trans-membrane proton gradient by ionophores (monensin and nigericin) induced a marked release of radioactivity from platelets preincubated with [3H]MPP+. Fractionation of homogenates of rabbit platelets preincubated with [3H]MPP+ showed that the drug was concentrated to a great extent in the 5-HT organelle fraction. MPP+ competitively inhibited [14CJ5-HT uptake by human platelets and reduced the endogenous 5-HT content of human, rabbit, and guinea pig platelets. These investigations show that MPP+ is transported into the platelets via the 5-HT carrier and is accumulated predominantly in the subcellular organelles that store 5-HT and other monoamines. It is suggested that an accumulation of MPP+ in amine storage vesicles of neurons may be involved in the effects of the drug in the CNS, e.g., by protecting other subcellular compartments from exposure to high concentrations of MPP+, by sustaining a gradual release of the toxin, or both.
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  • 86
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    Journal of neurochemistry 49 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: : The effects of chronic ethanol treatment on the membrane order of synaptosomes from the cerebral cortex, striatum, cerebellum, brainstem, and hippocampus of rats were determined by measuring the fluorescence polarization of diphenylhexatriene (DPH) that had been incorporated into the synaptosomal membranes. Fischer-344 rats either were fed a nutritionally complete ethanol-containing liquid diet for 5 months or pair-fed with a diet that contained sucrose substituted isocalorically for ethanol. Polarization values for synaptosomes from all the brain regions studied were similar except for those from cerebral cortical synaptosomal membranes, which were significantly less ordered. Ethanol in vitro (30–500 mM) decreased the polarization values in synaptosomes from sucrose-control rats for all brain regions, although the sensitivity of cerebellar synaptosomes to the membrane disordering effects of ethanol in vitro was significantly greater than that of synaptosomes from other brain regions. Chronic ethanol treatment did not alter baseline polarization for any brain region. Cerebellar and brainstem synaptosomes from the ethanol-fed rats were significantly less susceptible to the membrane disordering effects of ethanol in vitro compared to their sucrose controls, suggesting that chronic ethanol administration results in tolerance to ethanol's membrane effects. Striatal synaptosomes exhibited intermediate tolerance, whereas the sensitivities of cortical and hippocampal synaptosomes to membrane disordering by ethanol in vitro were not significantly affected by the chronic ethanol treatment. These results suggest that synaptosomal membranes have different membrane order requirements depending on the brain region from which they are prepared. Variations in brain regional neuronal membrane sensitivity to ethanol and differential tolerance development may contribute to some of the acute and chronic behavioral effects of ethanol.
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  • 87
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: Investigations into the effects of culturing bovine adrenal chromaffin cells in the presence (72 h) of dibutyryl cyclic AMP, forskolin, and reserpine on the level and release of [Met]enkephalyl-Arg6-Phe7 immunoreactivity, noradrenaline, and adrenaline are reported. The assay for [Met]enkephalyl-Arg6-Phe7 immunoreactivity recognises both peptide B, the 31-amino acid carboxy-terminal segment of proenkephalin, and its heptapeptide fragment, [Met]enkephalyl-Arg6-Phe7. Treatments that elevate cyclic AMP increase the amount of peptide immunoreactivity in these cells; this is predominantly peptide B-like immunoreactivity in both control cells and cyclic AMP-elevated cells. Treatment with reserpine gives no change in total immunoreactivity levels, but does result in increased accumulation of the heptapeptide [Met]enkephalyl-Arg6-Phe7 at the expense of immunoreactivity that elutes with its immediate precursor, peptide B. Cyclic AMP treatment causes either no change or a decrease in levels of accumulated noradrena-line and adrenaline. However, the release of [Met]enkephalin-Arg6-Phe7 immunoreactivity, noradrenaline, and adrenaline is increased by 72-h pretreatment with forskolin or dibutyryl cyclic AMP, whether release is stimulated by nicotine or elevated potassium. In each case the molecular form of [Met]enkephalyl-Arg6-Phe7 immunoreactivity that is released approximately reflects the cell content. Pretreatment with reserpine has no effect on the total [Met]enkephalyl-Arg6-Phe7 immunoreactivity released, but does result in an increased release of the heptapeptide and a decrease in release of peptide B-like immunoreactivity. The studies suggest that the levels of [Met]enkephalyl-Arg6-Phe7 and peptide B available for release are controlled both at the level of proenkephalin synthesis and at the level of doublebasic residue proteolysis.
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  • 88
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    Journal of neurochemistry 49 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: : Guinea-pig synaptosomes possess two functional pools of 4-aminobutyrate (GABA). One is rapidly labelled by added [I4C]GABA, is steadily released in a Ca2+-independent manner when the Na+ electrochemical potential across the plasma membrane is collapsed, and is depleted by the GABA analogue 2,4-diaminobutyrate (DABA), all of which is consistent with a cytosolic location. A second, noncytosolic compartment only slowly equilibrates with exogenous [14C]GABA, is not depleted by DABA, but can release 350 pmol of endogenous GABA/mg of protein (8% of the total intrasynaptosomal GABA) within 15 s of depolarization in the presence of Ca2+. Ca2+-independent release occurs by thermodynamic reversal of the plasma membrane uptake pathway following artifactually prolonged depolarization, whereas Ca2+-dependent release is consistent with physiological exocytosis from vesicular stores.
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  • 89
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    Journal of neurochemistry 49 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: : A depletion of not only myo-inositol (MI) but also taurine and other amino acids was observed in the sciatic nerve of a galactosemic rat. Treatment of the galactosemic rats with sorbinil, an aldose reductase (AR) inhibitor, was found to block galactitol formation and protect against the loss of MI, taurine, and other amino acids. Incubation studies of sciatic nerve have revealed that [3H]MI and [3H]-taurine were actively taken up and concentrated. Incubation of the nerve in a high-galactose medium showed a decrease in the accumulation of [3H]MI and [3H]taurine whereas the galactitol level increased. Time-course studies have shown that the galactitol level reached a plateau before a substantial decrease in the accumulation of [3H]MI and [3H]taurine occurred. The addition of AR inhibitors in the galactose medium significantly protected against the loss in the capacity of the nerve to accumulate [3H]MI and [3H]taurine. Hypertonicity of the galactose medium also seemed to have a protective effect similar to that of AR inhibitors.
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  • 90
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: Extracellular (EC) adenosine, hypoxanthine, xanthine, and inosine concentrations were monitored in vivo in the striatum during steady state, 15 min of complete brain ischemia, and 4 h of reflow and compared with purine and nucleotide levels in the tissue. Ischemia was induced by three-vessel occlusion combined with hypotension (50 mm Hg) in male Sprague-Dawley rats. EC purines were sampled by microdialysis, and tissue adenine nudeotides and purine catabolites were extracted from the in situ frozen brain at the end of the experiment. ATP, ADP, and AMP were analyzed with enzymatic fluorometric techniques, and adenosine, hypoxanthine, xanthine, and inosine with a modified HPLC system. Ischemia depleted tissue ATP, whereas AMP, adenosine, hypoxanthine, and inosine accumulated. In parallel, adenosine, hypoxanthine, and inosine levels increased in the EC compartment. Adenosine reached an EC concentration of 40 μM after 15 min of ischemia. Levels of tissue nucleotides and purines normalized on reflow. However, xanthine levels increased transiently (sevenfold). In the EC compartment, adenosine, inosine, and hypoxanthine contents normalized slowly on reflow, whereas the xanthine content increased. The high EC levels of adenosine during ischemia may turn off spontaneous neuronal firing, counteract excitotoxicity, and inhibit ischemic calcium uptake, thereby exerting neuroprotective effects.
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  • 91
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    Journal of neurochemistry 49 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: CNS receptors for thyrotropin-releasing hormone (TRH) and its analogs are likely to mediate the experimentally and clinically observed net excitatory effect of these peptides on lower motor neurons. Previous findings suggest that several types of TRH receptors with distinct TRH analog specificities may be present in rat CNS. In particular, based on competition isotherm assays with unlabeled analog γ-butyrolactone-γ-carbonyl-L-histidyl-L-proli neamide (DN-1417), Funatsu et al. claim the existence of a limbic forebrain site that binds this peptide and TRH with high affinity but that does not bind [3-methyl-histidyl2]-TRH (MeTRH). Using saturation and competition isotherm experiments, we have examined the binding of [3H]TRH and [3H]DN-1417 in three regions of rat CNS: pyriform cortex/amygdala, limbic forebrain, and lumbosacral spinal cord. In all three regions, saturation assays with [3H]TRH (0.4–100 μM) resolved only a single, saturable receptor with high affinity (KD= 12–14 nM) for TRH; in no case could more than one saturable site be identified. When [3H]DN-1417 was substituted as the assay ligand, no high-affinity binding component for this analog could be detected in the three regions. Competition curves for the binding of unlabeled DN-1417 to limbic forebrain and lumbosacral spinal cord ([3H]TRH as assay ligand) were monophasic (not biphasic like those of Funatsu et al.) and indicative of low-affinity binding of DN-1417 in these regions (Ki values = 2–3 μM; in agreement with values obtained in similar assays with [3H]MeTRH). We found that bacitracin in the assay buffer was useful in preventing the deamidation of [3H]TRH otherwise seen during incubation with limbic forebrain membranes. Our data do not support the existence of TRH receptor subtypes in the CNS. Rather, they favor the view that the neurotransmitter-like effects of high-and low-affinity TRH agonists are mediated by a single type of TRH receptor.
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  • 92
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    Journal of neurochemistry 49 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: Abnormal biogenic amine biosynthesis has been observed in humans and animals with endogenous and exogenous disturbances in folate metabolism. In an attempt to study this interaction biochemically, rats were depleted or repleted with folate for 10 weeks. Folate levels in depleted animals in serum and CSF correlated with stores in liver and brain, respectively. In depleted or repleted animals, there was no significant effect on biogenic amine metabolism in the CNS, as determined by quantitation of biogenic amines in brain and their respective metabolites in brain and CSF. These results are contrary to studies by other investigators. We suspect, however, that specific genetic defects in folate metabolism do result in impaired biogenic amine metabolism and probably at the level of disturbed biopterin cofactor functions.
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  • 93
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    Journal of neurochemistry 48 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: Conditions have been developed for an L-[3H]glu-tamate binding assay in which 85-95% of the specific binding is to a site that corresponds to the N-methyl-D-aspartate subclass of acidic amino acid receptors. Incubation of synaptic plasma membranes with L-[3H]glutamate in 50 mM Tris/acetate, pH 7.4, for 2-20 min at 2°C results in binding with pharmacological characteristics of the electrophysio-logically defined N-methyl-D-aspartate receptor. The fraction of glutamate binding to this subclass of receptors, relative to the total, decreases with both increased time and temperature. This binding is reversible, is concentrated in the synaptic plasma membrane fraction, has a pH optimum of 7.0-7.4, and is linear with respect to tissue protein concentration. The binding is unaffected by 1 mM concentrations of the anions sulfate, chloride, bromide, thiocyanate, phosphate, acetate, nitrate, or carbonate and the monovalent cations potassium or ammonium. However sodium and the divalent cations copper, cobalt, zinc, cadmium, and manganese decrease binding to this N-methyl-D-aspartate site.
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  • 94
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    Journal of neurochemistry 48 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: The concentrations of the acidic dopamine (DA) catabolites homovanillic acid (HVA) and 3,4-dihydroxy-phenylacetic acid (DOPAC) measured in human CSF are supposed to reflect the “turnover” of DA in the brain. The notion of “turnover” is, however, not synonymous with impulse nerve activity in the dopaminergic systems. Significant amounts of DOPAC and HVA could, indeed, be demonstrated in brain structures wherein dopaminergic inner-vation has not been documented. It must also be noted that DA is not only a neurotransmitter itself, but also a precursor of norepinephrine and epinephrine. Furthermore, in lumbar CSF, levels of biogenic amine catabolites partially reflect metabolism in the spinal cord and may have limited relevance to neurotransmission in the brain. To elucidate these points further, we determined the concentrations of DOPAC and HVA in 22 areas of six human brains and eight levels of six human spinal cords. The data were correlated with the concentration of DA. Quantitative determinations were done using HPLC with electrochemical detection, after solvent and ion-pair extraction. In this study, significant amounts of both DOPAC and HVA were demonstrated in brain structures not previously associated with dopaminergic innervation. The relatively lower DA concentration in these structures suggests that in these regions, the DOPAC and HVA concentrations are unrelated to dopaminergic neurotransmission. The possible role of capillary walls and glial cells in the catabolism of DA must be further evaluated. The demonstration of DOPAC and HVA in the spinal cord is another argument against the hypothesis that CSF levels of HVA and DOPAC reflect closely the activity of the dopaminergic systems in the brain.
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  • 95
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    Journal of neurochemistry 49 (1987), S. 0 
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    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
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  • 96
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    Topics: Medicine
    Notes: Abstract: To investigate aspects of the biochemical nature of membrane-bound dopamine D1 receptors, rat striatal homogenates were pretreated with heavy metal cations and some other chemical agents, and their effects on D1 receptors were subsequently determined using a standard [3H]CR)-(+)-8-chloro-2,3,4,5-tetrahydro-3-methyl-5-phenyl-1-N-3-benzazepine ([3H]SCH 23390) binding assay. Incubation of striatal membranes with as little as 1 μM Hg2+, 10 μM Cu2+, and 10 μM Cd2+ completely prevented specific [3H]SCH 23390 binding. The effect of Cu2+, 1.5 μM was noncompetitive in nature, whereas 3-5 μM Cu2+ afforded mixed-type inhibition. The inhibitory effect of Cu2+ was fully reversed by dithiothreitol (0.1-1 mM). Cu2+ (2 μM) did not affect the affinity of m-flupenthixol or clo-zapine for remaining [3H]SCH 23390 sites. A second series of cations, Co2+ (30 μM), Ni2+ (30 μM), Mn2+ (1 mM), Ca2+ (25 mM), and Ba2+ (20 mM), inhibited specific [3H]SCH 23390 binding by 50% at the concentrations indicated. The thiol alkylating reagent N-ethylmaleimide (NEM) (0.2 mM) reduced specific binding by 70%. The effect of NEM was completely prevented by coincubation with a D1 receptor saturating concentration of SCH 23390 (20 nM) or dopamine (10 μM). The results indicated that the dopamine D1 receptor is a thiol protein and that a thiol group is essential for the ligand binding.
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  • 97
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    Journal of neurochemistry 48 (1987), S. 0 
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    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: The sensitivity of choline transport to inhibition by ethylcholine mustard aziridinium (ECMA) was studied in several tissues. Choline transport was found to be inhibited irreversibly by ECMA in guinea pig and rat synapto-somes but not inhibited in erythrocytes or kidney slices. If this finding can be extended to other tissues ECMA sensitivity may provide a simple criterion for identifying the choline carrier associated with cholinergic tissue.
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  • 98
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    Journal of neurochemistry 48 (1987), S. 0 
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    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: Traumatic spinal cord injury has recently been shown to cause a rapid increase in free fatty acids (FFAs) and lipid degradation in cats. The present studies report a more delayed, time-dependent increase in FFAs and a concomitant decrease in phospholipids following traumatic spinal injury in rats. The largest percentage increases were found for polyunsaturated fatty acids, particularly arachi-donic acid. Associated with these changes were a reduction in the activity of Na+,K+-ATPase and development of spinal cord edema. These findings support the hypothesis that traumatic spinal cord injury leads to delayed, as well as early, hydrolysis of membrane phospholipids, resulting in the liberation of FFAs. Such changes may contribute to secondary spinal cord injury either through direct effects on membranes or through the actions of secondary metabolic products such as the eicosanoids. The latter may cause tissue injury by contributing to the reduction in spinal cord blood flow or through inflammatory responses that follow trauma.
    Type of Medium: Electronic Resource
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  • 99
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Journal of neurochemistry 48 (1987), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: The oligodendroglial enzyme, 2′,3′-cyclic nucleo-tide 3′-phosphohydrolase (CNP), is a valuable marker for expression of oligodendroglial differentiation in glial primary cultures, and the inducibility of this enzyme by dibu-tyryl-3′,5′-cyclic AMP (dBcAMP) appears to be limited to immature or developing oligodendroglia. To investigate the relationship between the induction of CNP and the sterol biosynthetic pathway, primary cultures of glia dissociated from the brains of newborn rats were maintained in 10% fetal calf serum (FCS) and exposed to 1 mM dBcAMP on day 7 in culture. Cultures so treated for either 48 h or 72 h demonstrated a three- to fourfold induction of CNP specific activity. The magnitude of this induction was not affected when the cholesterol content of the culture medium was reduced by 〉95% by placing the cultures in 10% lipoprotein-poor serum rather than 10% FCS during the exposure to dBcAMP. Mevinolin (10 μM), a specific inhibitor of 3-hy-droxy-3-methylglutaryl coenzyme A reductase, the rate-limiting enzyme of the sterol biosynthetic pathway, completely inhibited the induction of CNP by dBcAMP, while not affecting either the accumulation of cellular protein per flask or rate of protein synthesis. Simultaneous addition of mevalonate (20 mM) prevented the inhibition of the induction of CNP by mevinolin. However, simultaneous addition of low-density lipoprotein sufficient to increase the cholesterol content of the medium 80-fold failed to correct mevi-nolin's inhibition of the induction of CNP. Thus, these results demonstrate that the presence of mevalonate is obligatory for the induction of CNP in primary cultures of developing glia, and suggest that a critical nonsterol derivative of mevalonate is required for this expression of oligodendroglial differentiation.
    Type of Medium: Electronic Resource
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  • 100
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: Activities of six lysosomal enzymes in the cerebellum of jaundiced homozygous (jj) Gunn rats were examined from 5 to 20 days of life and compared with those in heterozygotes (j+). Significantly higher enzyme activities were first detected at 8 days. The jj/j+ activity ratios of all enzymes peaked at 15 days. The ratios of β-glycerophospha-tase, β-mannosidase, and acid lipase were only 1.3–1.7, whereas those of arylsulfatase and cathepsin were 2.0 and 3.1, respectively. The most striking increase in activity was observed with β-glucuronidase, the ratio of which was 8.4. These results indicate a selective increase in activities of certain lysosomal enzymes in the hypoplastic cerebellum of jj rats.
    Type of Medium: Electronic Resource
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