woods et al, 2011

27
           D   o   w   n    l   o   a    d   e    d    b   y    C    L    A    R    K    S    O    N    U    N    I    V   o   n    N   o   v   e   m    b   e   r    1    8  ,    2    0    1    1     |    h    t    t   p   :    /    /   p   u    b   s  .   a   c   s  .   o   r   g     P   u    b    l    i   c   a    t    i   o   n    D   a    t   e    (    W   e    b    )   :    N   o   v   e   m    b   e   r    1    7  ,    2    0    1    1     |    d   o    i   :    1    0  .    1    0    2    1    /    b    k      2    0    1    1      1    0    8    3  .   c    h    0    1    2 In Oxidative Stress: Diagnostics, Prevention, and Therapy; Andreescu, S., et al.; ACS Symposium Series; American Chemical Societ y: Washington, DC, 2011.

Upload: lupuloredana

Post on 07-Oct-2015

216 views

Category:

Documents


0 download

TRANSCRIPT

  • &KDSWHU

    %OXH 1DWLYH 3$*( DQG 0DVV 6SHFWURPHWU\ DVDQ $SSURDFK IRU WKH ,QYHVWLJDWLRQ RI 6WDEOH DQG7UDQVLHQW 3URWHLQ3URWHLQ ,QWHUDFWLRQV$OLVD * :RRGV ,]DEHOD 6RNRORZVND 5DPD

  • ERWK WKH SURWHLQ LGHQWLFDWLRQ DQG SURWHLQ TXDQWLWDWLRQ OHYHOVDQG SDUWLFXODUO\ GLIFXOW DW WKH SURWHLQSURWHLQ LQWHUDFWLRQOHYHO +HUH ZH GLVFXVV YDULRXV DSSOLFDWLRQV RI %13$*(LQFOXGLQJ WKH FKDUDFWHUL]DWLRQ RI VWDEOH SURWHLQ FRPSOH[HVWUDQVLHQW SURWHLQSURWHLQ LQWHUDFWLRQV WKH DVVHPEO\ RI SURWHLQVLQWR SRO\PHUV DQG WKH GLVDVVHPEO\ RI SURWHLQ FRPSOH[HV LQWRVXEFRPSOH[HV )XWXUH GLUHFWLRQV IRU %13$*( LQ FRPELQDWLRQZLWK 06 ZLWK D IRFXV RQ R[LGDWLYH VWUHVV DUH DOVR GLVFXVVHG

    ,QWURGXFWLRQ3URWHRPLFV LV WKH VWXG\ RI WKH SURWHRPH 7KH SURWHRPH LV GHQHG DV WKH

    SURWHLQV LQ D SDUWLFXODU FHOO VXEFHOOXODU FRPSDUWPHQW RU RUJDQHOOH WLVVXH RUJDQH[WUDFHOOXODU XLG EORRG XULQH VDOLYD WHDUV RU VSLQDO XLG RU ZKROH RUJDQLVP$ SURWHRPLFV H[SHULPHQW XVXDOO\ LQYROYHV VDPSOH IUDFWLRQDWLRQ GXULQJ ZKLFKSURWHLQV DUH ELRFKHPLFDOO\ VHSDUDWHG XVLQJ WKHLU SK\VLRFKHPLFDO SURSHUWLHV HQ]\PDWLF GLJHVWLRQ RI SURWHLQV LQWR SHSWLGHV DQDO\VLV RI D SHSWLGH PL[WXUHE\ OLTXLG FKURPDWRJUDSK\PDVV VSHFWURPHWU\ OLTXLG FKURPDWRJUDSK\ FRPELQHGZLWK WDQGHP PDVV VSHFWURPHWU\ RU /&0606 D GDWDEDVH VHDUFK DQDO\VLVDQG LQWHUSUHWDWLRQ GDWD YDOLGDWLRQ2[LGDWLYH VWUHVV LV D FHOOXODU UHVSRQVH WKDW LV SURGXFHG DV D UHDFWLRQ WR WKH

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

    Dow

    nloa

    ded

    by C

    LAR

    KSO

    N U

    NIV

    on

    Nov

    embe

    r 18,

    201

    1 | h

    ttp://

    pubs

    .acs

    .org

    P

    ublic

    atio

    n D

    ate

    (Web

    ): N

    ovem

    ber 1

    7, 2

    011

    | doi

    : 10.

    1021

    /bk-

    2011

    -108

    3.ch

    012

    In Oxidative Stress: Diagnostics, Prevention, and Therapy; Andreescu, S., et al.; ACS Symposium Series; American Chemical Society: Washington, DC, 2011.

  • %13$*( DQG &13$*( DV &KRLFHV IRU WKH ,QYHVWLJDWLRQ RI3URWHLQ &RPSOH[HV DQG 3URWHLQ3URWHLQ ,QWHUDFWLRQV

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

    LQYROYHG LQ WKH HOHFWURQ WUDQVSRUW FKDLQ ZKHUH PRVW 526 DUH SURGXFHG DQG ZKHUHD KLJK QXPEHU RI SURWHLQV DUH GDPDJHG E\ R[LGDWLYH VWUHVV 7KHUHIRUH WKH XVHRI %13$*( WR LQYHVWLJDWH WKH R[LGDWLYH VWUHVV DW WKH SURWHLQ FRPSOH[ OHYHO LV RIKLJK LQWHUHVW DQG DGGV D QHZ GLPHQVLRQ WR WKH LQYHVWLJDWLRQ RI R[LGDWLYH VWUHVV DWWKH SURWHLQSURWHLQ LQWHUDFWLRQ OHYHO%13$*( H[SHULPHQWV PD\ SURYLGH LQIRUPDWLRQ DERXW WKH VL]H QXPEHU

    VXEXQLW FRPSRVLWLRQ VWRLFKLRPHWU\ DQG UHODWLYH DEXQGDQFH RI WKHVH SURWHLQFRPSOH[HV 6HSDUDWLRQ RI SURWHLQ FRPSOH[HV IURP WKH %13$*( ODQH LQ D VHFRQGGLPHQVLRQ 6'63$*( PD\ UHYHDO WKH VXEXQLW FRPSRVLWLRQ RI D SDUWLFXODU SURWHLQFRPSOH[ DV ZHOO DV WKH LQWHUDFWLQJ SDUWQHUV RI D SDUWLFXODU SURWHLQ &RPSDUHGZLWK WKH SUHYLRXV PHWKRGV XVHG WR VWXG\ SURWHLQSURWHLQ LQWHUDFWLRQV WKHUH DUHVHYHUDO DGYDQWDJHV RI %13$*( IRU VWXG\LQJ SURWHLQSURWHLQ LQWHUDFWLRQV VHSDUDWLRQ RI WKH SURWHLQ FRPSOH[HV WDNHV SODFH XQGHU QDWLYH FRQGLWLRQV VR HYHQWKH WUDQVLHQW LQWHUDFWLRQV EHWZHHQ SURWHLQV PD\ EH LGHQWLHG VHSDUDWLRQ RISURWHLQ FRPSOH[HV LQ WKH VHFRQG GLPHQVLRQ PD\ UHYHDO WKHLU VXEXQLW FRPSRVLWLRQ WKH PHWKRG PD\ DQDO\]H WKH DVVRFLDWLRQ RI SURWHLQV LQWR SURWHLQ FRPSOH[HVDV D UHVXOW RI D VWLPXOXV %13$*( PD\ FRQUP WKH UHVXOWV REWDLQHG E\LPPXQRSUHFLSLWDWLRQ H[SHULPHQWV )RU H[DPSOH LI SURWHLQV $ % DQG& FRLPPXQRSXULI\ %13$*( ZLOO RIWHQ GLVWLQJXLVK EHWZHHQ $%& $%$& DQG %& FRPSOH[HV )XUWKHUPRUH E\ FRPELQLQJ %13$*( ZLWK PDVVVSHFWURPHWU\ ERWK VWUXFWXUDO DQG IXQFWLRQDO LQIRUPDWLRQ PD\ EH REWDLQHG &13$*( ZDV GHYHORSHG E\ WKH VDPH JURXS WKDW GHYHORSHG %13$*( DQG IXUWKHU PRGLHG E\ RWKHUV &13$*( XVHV H[DFWO\ WKH VDPHSURFHGXUH DV &13$*( H[FHSW WKDW WKH FDWKRGH EXIIHU GRHV QRW FRQWDLQ WKH&RRPDVVLH G\H ,Q DGGLWLRQ WKH VDPSOH EXIIHU GRHV QRW FRQWDLQ WKH &RRPDVVLHG\H HLWKHU 7KHUHIRUH VHSDUDWLRQ RI SURWHLQ FRPSOH[HV LV QRW EDVHG RQ WKH H[WHUQDOFKDUJH LQGXFHG E\ &RRPDVVLH G\H LW LV UDWKHU EDVHG RQ WKH LQWHUQDO FKDUJH RIWKH SURWHLQ FRPSOH[ JLYHQ E\ WKH FKDUJH RI LWV VXEXQLWV $ VFKHPDWLF RI WKHSULQFLSOH RI %13$*( DQG &13$*( DV ZHOO DV WKH GLIIHUHQFHV EHWZHHQ WKHWZR PHWKRGV LV SUHVHQWHG LQ )LJXUH +RZHYHU WKH SULQFLSOH RI VHSDUDWLRQ RI%13$*( DQG &13$*( LV YHU\ GLIIHUHQW HYHQ WKRXJK WKH VHSDUDWLRQ LQ ERWKPHWKRGV WDNHV SODFH XQGHU WKH VDPH QDWLYH FRQGLWLRQV ,I LQ %13$*( VHSDUDWLRQRI WKH SURWHLQ FRPSOH[HV WDNHV SODFH DFFRUGLQJ WR WKHLU 0Z &13$*( GRHV QRW

    Dow

    nloa

    ded

    by C

    LAR

    KSO

    N U

    NIV

    on

    Nov

    embe

    r 18,

    201

    1 | h

    ttp://

    pubs

    .acs

    .org

    P

    ublic

    atio

    n D

    ate

    (Web

    ): N

    ovem

    ber 1

    7, 2

    011

    | doi

    : 10.

    1021

    /bk-

    2011

    -108

    3.ch

    012

    In Oxidative Stress: Diagnostics, Prevention, and Therapy; Andreescu, S., et al.; ACS Symposium Series; American Chemical Society: Washington, DC, 2011.

  • VHSDUDWH SURWHLQ FRPSOH[HV DFFRUGLQJ WR WKHLU 0Z $V VXFK WKH DSSOLFDWLRQV RIWKHVH WZR PHWKRGV ZLWK WZR GLIIHUHQW VHSDUDWLRQ SULQFLSOHV GR QRW H[FOXGH HDFKRWKHU EXW UDWKHU FRPSOHPHQW HDFK RWKHU

    )LJXUH %13$*( DQG &13$*( 3ULQFLSOHV VLPLODULWLHV DQG GLIIHUHQFHV%RWK %13$*( DQG &13$*( VHSDUDWH SURWHLQ FRPSOH[HV XQGHU QDWLYH

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

    Dow

    nloa

    ded

    by C

    LAR

    KSO

    N U

    NIV

    on

    Nov

    embe

    r 18,

    201

    1 | h

    ttp://

    pubs

    .acs

    .org

    P

    ublic

    atio

    n D

    ate

    (Web

    ): N

    ovem

    ber 1

    7, 2

    011

    | doi

    : 10.

    1021

    /bk-

    2011

    -108

    3.ch

    012

    In Oxidative Stress: Diagnostics, Prevention, and Therapy; Andreescu, S., et al.; ACS Symposium Series; American Chemical Society: Washington, DC, 2011.

  •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

    %13$*( DV D 7RRO IRU 'HWHUPLQDWLRQ RI WKH 0Z RI 3URWHLQ&RPSOH[HV DQG IRU 'HWHUPLQDWLRQ RI 7UDQVLHQW 3URWHLQ3URWHLQ

    ,QWHUDFWLRQV5HFHSWRU W\URVLQH NLQDVHV 57. DUH UHFHSWRUV IRU D VHULHV RI JURZWK IDFWRUV

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

    Dow

    nloa

    ded

    by C

    LAR

    KSO

    N U

    NIV

    on

    Nov

    embe

    r 18,

    201

    1 | h

    ttp://

    pubs

    .acs

    .org

    P

    ublic

    atio

    n D

    ate

    (Web

    ): N

    ovem

    ber 1

    7, 2

    011

    | doi

    : 10.

    1021

    /bk-

    2011

    -108

    3.ch

    012

    In Oxidative Stress: Diagnostics, Prevention, and Therapy; Andreescu, S., et al.; ACS Symposium Series; American Chemical Society: Washington, DC, 2011.

  •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

    HSKULQ(SK5V PXOWLPHUV WKDW WULJJHU F\WRVNHOHWDO UHPRGHOLQJ 7KLV XQGHUOLHVFHOO DGKHVLRQ DQG PRWLOLW\ LQ OLJDQG DQG UHFHSWRU EHDULQJ FHOOV 7KH UVWFRQVHTXHQFH RI (SK5 DFWLYDWLRQ LV DXWRSKRVSKRU\ODWLRQ RI LWV WKUHH FRQVHUYHGW\URVLQH UHVLGXHV IURP WKH F\WRSODVPLF VLGH RI WKH UHFHSWRU 7KH SKRVSKRU\ODWHGVLWHV SURYLGH GRFNLQJ IRU D QXPEHU RI 6+GRPDLQ FRQWDLQLQJ VLJQDOLQJ DQGDGDSWRU SURWHLQV WKDW PRGXODWH F\WRVNHOHWDO SODVWLFLW\ $ QXPEHU RI SURWHLQV WKDW LQWHUDFW ZLWK SKRVSKRU\ODWHG (SK5V RU PHGLDWH

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

    Dow

    nloa

    ded

    by C

    LAR

    KSO

    N U

    NIV

    on

    Nov

    embe

    r 18,

    201

    1 | h

    ttp://

    pubs

    .acs

    .org

    P

    ublic

    atio

    n D

    ate

    (Web

    ): N

    ovem

    ber 1

    7, 2

    011

    | doi

    : 10.

    1021

    /bk-

    2011

    -108

    3.ch

    012

    In Oxidative Stress: Diagnostics, Prevention, and Therapy; Andreescu, S., et al.; ACS Symposium Series; American Chemical Society: Washington, DC, 2011.

  • )LJXUH $ $ ZRUNRZ IRU WKH ELRFKHPLFDO DQG 06 FKDUDFWHUL]DWLRQ RIHSKULQ VLJQDOLQJ 7KH XQVWLPXODWHG RU VWLPXODWHG FHOOV ZHUH O\VHG DQG WKH O\VDWHVDQGRU WKH S
  • GLUHFWLRQ RI PLJUDWLRQ LQ WKH JHO DQG 0Z PDUNHUV DUH LQGLFDWHG 1RWH WKH KLJK0Z PDUNHUV LQ %13$*( ' IRU SURWHLQ FRPSOH[HV DQG QRUPDO 0Z PDUNHUV LQ6'63$*( ' IRU SURWHLQV ' %13$*( ' WKH H[SHULPHQW LV DV LQ &H[FHSW WKDW WKH JHOV IURP 6'63$*( ' LQVWHDG RI EHLQJ VLOYHU VWDLQHG ZHUHHOHFWUREORWWHG DQG DQDO\]HG IRU WKH SUHVHQFH RI W\URVLQH SKRVSKRU\ODWHG SURWHLQV

    DQG SURWHLQ FRPSOH[HV E\ XVLQJ DQWLSKRVSKRW\URVLQH DQWLERGLHV

    %13$*( DQG 06 DV D 7RRO IRU 'HWHUPLQDWLRQ RI WKH 6XEXQLW&RPSRVLWLRQ RI 6WDEOH +RPR DQG +HWHUR3URWHLQ &RPSOH[HV:H IXUWKHU LQYHVWLJDWHG ZKHWKHU LQWDFW FRQVWLWXWLYHO\ H[SUHVVHG SURWHLQ

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

    Dow

    nloa

    ded

    by C

    LAR

    KSO

    N U

    NIV

    on

    Nov

    embe

    r 18,

    201

    1 | h

    ttp://

    pubs

    .acs

    .org

    P

    ublic

    atio

    n D

    ate

    (Web

    ): N

    ovem

    ber 1

    7, 2

    011

    | doi

    : 10.

    1021

    /bk-

    2011

    -108

    3.ch

    012

    In Oxidative Stress: Diagnostics, Prevention, and Therapy; Andreescu, S., et al.; ACS Symposium Series; American Chemical Society: Washington, DC, 2011.

  • ZDV D VXEXQLW RI WKH N'D KRPRFRPSOH[ FRPSRVHG RI IRXU LGHQWLFDO VXEXQLWVLV VKRZQ LQ )LJXUH

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

    Dow

    nloa

    ded

    by C

    LAR

    KSO

    N U

    NIV

    on

    Nov

    embe

    r 18,

    201

    1 | h

    ttp://

    pubs

    .acs

    .org

    P

    ublic

    atio

    n D

    ate

    (Web

    ): N

    ovem

    ber 1

    7, 2

    011

    | doi

    : 10.

    1021

    /bk-

    2011

    -108

    3.ch

    012

    In Oxidative Stress: Diagnostics, Prevention, and Therapy; Andreescu, S., et al.; ACS Symposium Series; American Chemical Society: Washington, DC, 2011.

  • LV DERXW N'D ,Q RXU H[SHULPHQWV ZH LGHQWLHG WKLV FRPSOH[ LQ WKH VDPH N'D UDQJH VXJJHVWLQJ WKDW %13$*( PD\ SURYLGH LQIRUPDWLRQ DERXW WKH 0Z RID SURWHLQ FRPSOH[

    )LJXUH $QDO\VLV RI VXEFRPSOH[ RI WKH HXNDU\RWLF LQLWLDWLRQ IDFWRU KHWHURSURWHLQ FRPSOH[ E\ %13$*( DQG /&0606 7KH JHO EDQG ZDV

    H[FLVHG DQG GLJHVWHG E\ WU\SVLQ DQG WKH UHVXOWLQJ SHSWLGH PL[WXUH ZDV DQDO\]HG E\/&0606 7KH H[WUDFWHG LRQ FKURPDWRJUDP (,& RI WKH SHDN ZLWK P] RI LGHQWLHG LQ WKH FKURPDWRJUDP D SHDN DW PLQXWHV $ WKDW FRQWDLQHG 06VSHFWUD RI D SHDN ZLWK P] RI % 7KH LVRWRSLF UHVROXWLRQ RI WKLV SHDNLV VKRZQ LQ LQER[ % 7KLV SHDN ZDV VHOHFWHG IRU IUDJPHQWDWLRQ E\ 0606 &DQG SURGXFHG D VHULHV RI E DQG \ IUDJPHQW LRQV 'DWD SURFHVVLQJ DQG GDWDEDVHVHDUFK OHG WR WKH LGHQWLFDWLRQ RI D SHSWLGH ZLWK WKH VHTXHQFH )69/4

  •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

    %13$*( DQG :% DV D 7RRO IRU ,GHQWLFDWLRQ RI 7UDQVLHQW3URWHLQ3URWHLQ ,QWHUDFWLRQV

    ,Q DGGLWLRQ WR WKH GHWHUPLQDWLRQ RI WKH 0Z DQG VXEXQLW FRPSRVLWLRQ RIVWDEOH KRPR DQG KHWHURSURWHLQ FRPSOH[HV %13$*( DQG /&0606 LV DOVRDEOH WR LGHQWLI\ WUDQVLHQW SURWHLQSURWHLQ LQWHUDFWLRQV 7R LGHQWLI\ WKHVH WUDQVLHQWSURWHLQSURWHLQ LQWHUDFWLRQV WKH SURWHLQ FRPSOH[HV IURP WKH XQVWLPXODWHGDQG HSKULQ%)FVWLPXODWHG 1* FHOOV ZHUH LPPXQRDIQLW\ SXULHG XVLQJDQWLSKRVSKRW\URVLQH S

  •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

    )LJXUH 3RVVLEOH WKHRUHWLFDO VFHQDULRV IRU WKH DQDO\VLV RI SURWHLQSURWHLQLQWHUDFWLRQV XVLQJ D S

  • )LJXUH ,GHQWLFDWLRQ RI WUDQVLHQW SURWHLQSURWHLQ LQWHUDFWLRQV LQ SULPDU\QHXURQV E\ %13$*( DQG :% $QDO\VLV RI WKH O\VDWHV RI XQVWLPXODWHG DQG%'1)VWLPXODWHG ZDV SHUIRUPHG XVLQJ SULPDU\ QHXURQDO FHOO FXOWXUHV E\%13$*( 7KH O\VDWHV ZHUH VHSDUDWHG LQ %13$*( ' DQG 6'63$*( 'DQG HOHFWUREORWWHG DQG WKHQ DQDO\]HG IRU WKH SUHVHQFH RI +UV SURWHLQ XVLQJDQWL+UV DQWLERGLHV 7KH LPPXQH UHDFWLRQ ZDV YLVXDOL]HG XVLQJ HQKDQFHG

    FKHPLOXPLQHVFHQW UHDFWLRQ $V REVHUYHG +UV IRUPV D N'D SURWHLQ FRPSOH[XSRQ %'1) VWLPXODWLRQ

    %13$*( DV D 7RRO LQ $QDO\VLV RI 3URWHLQ 6HOI$VVHPEO\ LQWR+LJKHU 0Z 3RO\PHUV

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

    Dow

    nloa

    ded

    by C

    LAR

    KSO

    N U

    NIV

    on

    Nov

    embe

    r 18,

    201

    1 | h

    ttp://

    pubs

    .acs

    .org

    P

    ublic

    atio

    n D

    ate

    (Web

    ): N

    ovem

    ber 1

    7, 2

    011

    | doi

    : 10.

    1021

    /bk-

    2011

    -108

    3.ch

    012

    In Oxidative Stress: Diagnostics, Prevention, and Therapy; Andreescu, S., et al.; ACS Symposium Series; American Chemical Society: Washington, DC, 2011.

  • LQWR KLJKHU RUGHU VWUXFWXUHV 7KHVH QGLQJ ZHUH IXUWKHU FRQUPHG E\ HOHFWURQPLFURVFRS\ ZKHUH WKH IXOO\ DVVHPEOHGSRO\PHUL]HG 9( SURWHLQV ZHUH REVHUYHG DVKLJK 0Z ODPHQWV 7KHUHIRUH %13$*( LV D XVHIXO WRRO IRU WKH DQDO\VLV RI WKHSRO\PHUL]DWLRQ VWDWH RI D SURWHLQ RU PL[WXUH RI SURWHLQV

    %13$*( DV D 7RRO IRU WKH 'HWHUPLQDWLRQ RI WKH 0Z DQG0XOWLPHUL]DWLRQ 6WDWH RI D 6WDEOH 3URWHLQ &RPSOH[ 3URWHFWLYH

    DJDLQVW 2[LGDWLYH 6WUHVV6R IDU %13$*( LV RQH RI WKH IDVWHVW DQG FKHDSHVW ZD\V WR GHWHUPLQH WKH

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

    Dow

    nloa

    ded

    by C

    LAR

    KSO

    N U

    NIV

    on

    Nov

    embe

    r 18,

    201

    1 | h

    ttp://

    pubs

    .acs

    .org

    P

    ublic

    atio

    n D

    ate

    (Web

    ): N

    ovem

    ber 1

    7, 2

    011

    | doi

    : 10.

    1021

    /bk-

    2011

    -108

    3.ch

    012

    In Oxidative Stress: Diagnostics, Prevention, and Therapy; Andreescu, S., et al.; ACS Symposium Series; American Chemical Society: Washington, DC, 2011.

  • N'D GHWHFWHG E\ 1GK( DQG VROXEOH N'D GHWHFWHG E\ 1GK- 1GK. VXEFRPSOH[HV )LJXUH % DQG H[LVWV DV PRQRPHULF DQG GLPHULF IRUPV )LJXUH &

    )LJXUH 6WUXFWXUDO FKDUDFWHUL]DWLRQ RI WKH 1GK FRPSOH[ 7KH WK\ODNRLGPHPEUDQHV ZHUH VHSDUDWHG E\ %13$*( ' DQG 7ULFLQH 3$*( ' DQG WKHQVLOYHU VWDLQHG $ RU HOHFWUREORWWHG DQG DQDO\]HG ZLWK 1GK DQWLERGLHV % & 7KHWK\ODNRLG PHPEUDQHV ZHUH DOVR VHSDUDWHG E\ &13$*( ' DQG 7ULFLQH 3$*(' DQG WKH VLOYHU VWDLQHG ' RU HOHFWUREORWWHG DQG DQDO\]HG ZLWK 1GK DQWLERGLHV( $IWHU VHSDUDWLRQ E\ &13$*( ' WKH SURWHLQ FRPSOH[HV ZHUH IXUWKHU

    Dow

    nloa

    ded

    by C

    LAR

    KSO

    N U

    NIV

    on

    Nov

    embe

    r 18,

    201

    1 | h

    ttp://

    pubs

    .acs

    .org

    P

    ublic

    atio

    n D

    ate

    (Web

    ): N

    ovem

    ber 1

    7, 2

    011

    | doi

    : 10.

    1021

    /bk-

    2011

    -108

    3.ch

    012

    In Oxidative Stress: Diagnostics, Prevention, and Therapy; Andreescu, S., et al.; ACS Symposium Series; American Chemical Society: Washington, DC, 2011.

  • VHSDUDWHG E\ %13$*( ' DQG VWDLQHG E\ &RRPDVVLH ) /RFDWLRQ RI 36 ,$73DVH &\W EI FRPSOH[ DQG 36 ,, LV LQGLFDWHG 7KH 0Z PDUNHUV IRU %13$*(DQG 7ULFLQH 3$*( DUH VKRZQ 7KHUH DUH QR 0Z PDUNHUV IRU &13$*( EHFDXVHWKLV PHWKRG GRHV QRW VHSDUDWH SURWHLQ FRPSOH[HV DFFRUGLQJ WR WKHLU 0Z 7KHGLUHFWLRQ RI PLJUDWLRQ LQ %13$*( 7ULFLQH 3$*( DQG &13$*( LV LQGLFDWHG

    &13$*( DV D 3UH3XULFDWLRQ 7RRO IRU WKH /RFDOL]DWLRQ RID 6WDEOH 3URWHLQ &RPSOH[ 3URWHFWLYH DJDLQVW 2[LGDWLYH 6WUHVV

    LQ %13$*(7KH PDLQ JRDO RI WKLV SURMHFW ZDV WR LVRODWH DQG FKDUDFWHUL]H WKH 1GK FRPSOH[

    +RZHYHU DV REVHUYHG LQ )LJXUH $ WKH 1GK FRPSOH[ LV QRW YLVLEOH LQ WKH %13$*( ' QRU LQ 7ULFLQH3$*( ' 7KHUHIRUH DOWKRXJK %13$*( FDQ EH YHU\XVHIXO LQ WKH IDVW LVRODWLRQ DQG FKDUDFWHUL]DWLRQ RI D SURWHLQ FRPSOH[ LI LQ %13$*( WKH 0Z RI WKH SURWHLQ FRPSOH[ RI LQWHUHVW 1GK FRPSOH[ KDV WKH VDPH0Z DV D GLIIHUHQW FRPSOH[ WKHQ DQ DGGLWLRQDO SXULFDWLRQ SURFHGXUH SULRU WR %13$*( LV UHTXLUHG &13$*( LV VXFK D SURFHGXUH ,W VHSDUDWHV SURWHLQ FRPSOH[HVDFFRUGLQJ WR WKHLU LQWHUQDO FKDUJH )LJXUH VKRZV WKH &13$*( ' DQG 7ULFLQH3$*( ' RI WKH WK\ODNRLG PHPEUDQHV IURP PHVRSK\OO FKORURSODVWV IROORZHGE\ VLOYHU VWDLQLQJ )LJXUH ' RU :% )LJXUH ( XVLQJ DQWLERGLHV DJDLQVW 1GK(PRQLWRUV WKH N'D PHPEUDQH SURWHLQ VXEFRPSOH[ DQG 1GK. PRQLWRUV WKH N'D VROXEOH SURWHLQ VXEFRPSOH[ $V REVHUYHG WKH LQWDFW 1GK FRPSOH[ LVORFDOL]HG LQ WKH VHFRQG PRVW LQWHQVH EDQG RI WKH &13$*( ' )LJXUH (6LQFH WKH :% RI &13$*( ' DQG 7ULFLQH 3$*( ' )LJXUH ( DOORZHG

    XV WR LPPXQRORFDOL]H WKH 1GK FRPSOH[ LQ &13$*( ' WKH VHFRQG PRVWLQWHQVH EDQG ZH FRQWLQXHG RXU SXULFDWLRQ DQG IXUWKHU VHSDUDWHG WKH JHO ODQHIURP &13$*( ' LQ %13$*( ' )LJXUH ) 7KLV VHSDUDWLRQ DOORZHG XVWR EHWWHU ORFDOL]H WKH 1GK FRPSOH[ LQ WKH %13$*( LQIRUPDWLRQ WKDW ZLOO EHXVHG IRU IXUWKHU FKDUDFWHUL]DWLRQ ,Q )LJXUH ) WKH SURWHLQ FRPSOH[HV DUH VHHQDV GDUN VSRWV DQG EDVHG RQ PDVV VSHFWURPHWU\ GDWD QRW VKRZQ WKH\ LQFOXGH3KRWRV\VWHP , 36, N'D VSRWV $73V\QWKDVH FRPSOH[ N'DVSRW &\WRFKURPH EI FRPSOH[ N'D DQG SDUW RI 3KRWRV\VWHP ,, 36,, N'D $V REVHUYHG WKH 1GK FRPSOH[ LV QRW YLVLEOH +RZHYHU VLQFH ZH NQHZ WKDWWKH 1GK FRPSOH[ KDV D 0Z RI N'D DQG LW LV SDUW RI RU FORVH WR WKH VHFRQGEDQG LQ WKH &13$*( WKHQ ZH FRXOG ORFDOL]H LW ZLWKLQ D VTXDUH IRUPHG E\ WKH N'D 36, DQG WKH WZR N'D 36, FRUH ,W VKRXOG EH QRWHG WKDW WKH WZR N'D36, FRPSOH[HV KDYH DQ LGHQWLFDO 0Z EXW RQO\ WKHLU LQWHUQDO FKDUJH LV GLIIHUHQWPRVW OLNHO\ GXH WR SRVWWUDQVODWLRQDO PRGLFDWLRQV VXFK DV SKRVSKRU\ODWLRQV7KHUHIRUH WKH FRPELQDWLRQ RI &13$*( DQG %13$*( LV DEOH WR VHSDUDWH WZRSURWHLQ FRPSOH[HV ZLWK LGHQWLFDO 0Z EXW GLIIHUHQW LQWHUQDO FKDUJH7R LGHQWLI\ WKH H[DFW ORFDWLRQ RI WKH 1GK DQG IRU LWV IXUWKHU FKDUDFWHUL]DWLRQ

    ZH FXW KRUL]RQWDO DQG YHUWLFDO JHO EDQGV IURP WKH DERYH PHQWLRQHG VTXDUH NQRZQWR FRQWDLQ WKH 1GK FRPSOH[ DQG VHSDUDWHG WKHP LQ D 6'63$*( ' IROORZHG E\:% XVLQJ DQWLERGLHV DJDLQVW 1GK VXEXQLWV )LJXUH $ $V REVHUYHG WKH SRVLWLRQRI WKH 1GK FRPSOH[ LQ :% RI WKH 6'63$*( ZDV ORFDOL]HG LQ WKH VHFRQG EDQG LQERWK KRUL]RQWDO DQG YHUWLFDO JHO EDQGV )LJXUH $ 7R FRQUP WKH ORFDOL]DWLRQ RI

    Dow

    nloa

    ded

    by C

    LAR

    KSO

    N U

    NIV

    on

    Nov

    embe

    r 18,

    201

    1 | h

    ttp://

    pubs

    .acs

    .org

    P

    ublic

    atio

    n D

    ate

    (Web

    ): N

    ovem

    ber 1

    7, 2

    011

    | doi

    : 10.

    1021

    /bk-

    2011

    -108

    3.ch

    012

    In Oxidative Stress: Diagnostics, Prevention, and Therapy; Andreescu, S., et al.; ACS Symposium Series; American Chemical Society: Washington, DC, 2011.

  • WKH 1GK FRPSOH[ ZH VXSHULPSRVHG WKH KRUL]RQWDO DQG YHUWLFDO %13$*( 'JHO EDQGV WKDW ZHUH DQDO\]HG E\ :% DQG VKRZQ LQ )LJXUH $ VLOYHU VWDLQHGWKH %13$*( ' JHO WKDW FRQWDLQHG WKH VTXDUH IRUPHG E\ WKH N'D 36, DQG WKHWZR N'D 36, FRUH )LJXUH % HOHFWUREORWWHG WKH %13$*( ' JHO XQGHUQDWLYH FRQGLWLRQV DQG DQDO\]HG LW ZLWK 1GK DQWLERGLHV )LJXUH % $V REVHUYHGWKH SRVLWLRQ LQ WKH JHO RI WKH 1GK FRPSOH[ ORFDOL]HG E\ :% RI WKH 6'63$*( 'JHOV ZDV FRQUPHG E\ ERWK VLOYHU VWDLQLQJ DQG :% RI WKH %13$*( ' )LJXUH%

    )LJXUH /RFDOL]DWLRQ RI WKH 1GK FRPSOH[ LQ %13$*( 3$*( ' DQG LWVFKDUDFWHUL]DWLRQ E\ 0$/', 06 7KH 1GK FRPSOH[ ZDV VHSDUDWHG E\ &13$*(

    Dow

    nloa

    ded

    by C

    LAR

    KSO

    N U

    NIV

    on

    Nov

    embe

    r 18,

    201

    1 | h

    ttp://

    pubs

    .acs

    .org

    P

    ublic

    atio

    n D

    ate

    (Web

    ): N

    ovem

    ber 1

    7, 2

    011

    | doi

    : 10.

    1021

    /bk-

    2011

    -108

    3.ch

    012

    In Oxidative Stress: Diagnostics, Prevention, and Therapy; Andreescu, S., et al.; ACS Symposium Series; American Chemical Society: Washington, DC, 2011.

  •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

    WKDW LGHQWLHG 1GK+ D VXEXQLW RI WKH 1GK FRPSOH[ LV VKRZQ LQ &

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

    %13$*( DQG 0DVV 6SHFWURPHWU\ DV 0HWKRGV IRU ([DPLQLQJ2[LGDWLYH 6WUHVV

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

    Dow

    nloa

    ded

    by C

    LAR

    KSO

    N U

    NIV

    on

    Nov

    embe

    r 18,

    201

    1 | h

    ttp://

    pubs

    .acs

    .org

    P

    ublic

    atio

    n D

    ate

    (Web

    ): N

    ovem

    ber 1

    7, 2

    011

    | doi

    : 10.

    1021

    /bk-

    2011

    -108

    3.ch

    012

    In Oxidative Stress: Diagnostics, Prevention, and Therapy; Andreescu, S., et al.; ACS Symposium Series; American Chemical Society: Washington, DC, 2011.

  • RI UHSUHVHQWDWLYH ZRUN LQFOXGH EXW DUH QRW OLPLWHG WR ZRUN SHUIRUPHG LQ FHOORUJDQHOOH WLVVXHV RUJDQV RU RUJDQLVPV IURP SODQWV DQLPDOV RU EDFWHULD 7KH PDLQ DSSOLFDWLRQ RI %13$*( HLWKHU DORQH RU LQ FRPELQDWLRQ ZLWK

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

    /LPLWDWLRQV RI %13$*($V GHPRQVWUDWHG KHUH WKURXJK TXLWH D IHZ H[DPSOHV %13$*( KDV D ZLGH

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

    Dow

    nloa

    ded

    by C

    LAR

    KSO

    N U

    NIV

    on

    Nov

    embe

    r 18,

    201

    1 | h

    ttp://

    pubs

    .acs

    .org

    P

    ublic

    atio

    n D

    ate

    (Web

    ): N

    ovem

    ber 1

    7, 2

    011

    | doi

    : 10.

    1021

    /bk-

    2011

    -108

    3.ch

    012

    In Oxidative Stress: Diagnostics, Prevention, and Therapy; Andreescu, S., et al.; ACS Symposium Series; American Chemical Society: Washington, DC, 2011.

  • KDYH LGHQWLFDO RU VLPLODU PROHFXODU PDVV DQG WKH VXEXQLWV RI RQH FRPSOH[ PD\ EHHUURQHRXVO\ DVVLJQHG WR D GLIIHUHQW SURWHLQ FRPSOH[

    &RQFOXGLQJ 5HPDUNV2YHUDOO %13$*( RSHQV QHZ SRVVLELOLWLHV IRU VLPXOWDQHRXVO\ DQDO\]LQJ WKH

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

    $EEUHYLDWLRQV%13$*( %OXH 1DWLYH 3$*( &13$*( &RORUOHVV 1DWLYH 3$*(

    6'63$*( VRGLXP GRGHF\O VXOIDWHSRO\DFU\ODPLGH JHO HOHFWURSKRUHVLV 0ZPROHFXODU ZHLJKW :% :HVWHUQ EORWWLQJ 06 PDVV VSHFWURPHWU\ /&0606OLTXLG FKURPDWRJUDSK\ PDVV VSHFWURPHWU\ 0$/',06 0DWUL[ $VVLVWHG/DVHU 'HVRUSWLRQ ,RQL]DWLRQ 0DVV 6SHFWURPHWU\ P] PDVVFKDUJH &,'FROOLVLRQLQGXFHG GLVVRFLDWLRQ

    $FNQRZOHGJPHQWV7KH DXWKRUV ZRXOG OLNH WR WKDQN /DXUD 0XOGHULJ 7RGG %R\HU 6FRWW 1LFKROV

    1DWKDQ 9DQGHUODQ 7LP 0DVFDO .HLWK &RPSVRQ 0LFKDHO %DOORFK &DWDOLQ'RQHDQX 'DYLG &KDPEHUOLQ &DWK\ %ROODQG -RDQQH %RODQG -DQHW *DOODJHU 6HDQ0F&DUWK\ &ULVWLDQ &RMRFDULX DQG &KDUOHV 3UDJQHOO IURP :DWHUV &RUSRUDWLRQ IRUWKH FROODERUDWLYH GLVFRXQW WKDW DOORZHG XV WR SXUFKDVH D 0LFURPDVV 4XDGUXSROH7LPH RI )OLJKW 472) 0LFUR PDVV VSHFWURPHWHU 7KH DXWKRUV ZRXOG DOVR OLNH WRWKDQN 7KRPDV $ 1HXEHUW IURP WKH 6NLUEDOO ,QVWLWXWH 1HZ

  • 5HIHUHQFHV 6FKDJJHU + 1DWLYH HOHFWURSKRUHVLV IRU LVRODWLRQ RI PLWRFKRQGULDO R[LGDWLYH

    SKRVSKRU\ODWLRQ SURWHLQ FRPSOH[HV 0HWKRGV (Q]\PRO 6FKDJJHU + &UDPHU : $ YRQ -DJRZ * $QDO\VLV RI PROHFXODU PDVVHV

    DQG ROLJRPHULF VWDWHV RI SURWHLQ FRPSOH[HV E\ EOXH QDWLYH HOHFWURSKRUHVLVDQG LVRODWLRQ RI PHPEUDQH SURWHLQ FRPSOH[HV E\ WZRGLPHQVLRQDO QDWLYHHOHFWURSKRUHVLV $QDO %LRFKHP

    6FKDJJHU + YRQ -DJRZ * %OXH QDWLYH HOHFWURSKRUHVLV IRU LVRODWLRQ RIPHPEUDQH SURWHLQ FRPSOH[HV LQ HQ]\PDWLFDOO\ DFWLYH IRUP $QDO %LRFKHP

    'DULH & & %LQLRVVHN 0 / :LQWHU 9 0XWVFKOHU % +DHKQHO :,VRODWLRQ DQG VWUXFWXUDO FKDUDFWHUL]DWLRQ RI WKH 1GK FRPSOH[ IURP PHVRSK\OODQG EXQGOH VKHDWK FKORURSODVWV RI =HD PD\V )(%6 -

    (XEHO + %UDXQ + 3 0LOODU $ + %OXHQDWLYH 3$*( LQ SODQWV $ WRRO LQDQDO\VLV RI SURWHLQSURWHLQ LQWHUDFWLRQV 3ODQW 0HWKRGV

    YDQ /LV 5 $WWHLD $ 0HQGR]D+HUQDQGH] * *RQ]DOH]+DOSKHQ ',GHQWLFDWLRQ RI QRYHOPLWRFKRQGULDO SURWHLQ FRPSRQHQWV RI&KODP\GRPRQDVUHLQKDUGWLL $ SURWHRPLF DSSURDFK 3ODQW 3K\VLRO

    3ORVFKHU 0 *UDQYRJO % =RU\DQ 0 5HLVLQJHU 9 (LFKDFNHU / $0DVV VSHFWURPHWULF FKDUDFWHUL]DWLRQ RI PHPEUDQH LQWHJUDO ORZ PROHFXODUZHLJKW SURWHLQV IURP SKRWRV\VWHP ,, LQ EDUOH\ HWLRSODVWV 3URWHRPLFV

    +HLQHPH\HU - (XEHO + :HKPKRQHU ' -DQVFK / %UDXQ + 33URWHRPLF DSSURDFK WR FKDUDFWHUL]H WKH VXSUDPROHFXODU RUJDQL]DWLRQ RISKRWRV\VWHPV LQ KLJKHU SODQWV 3K\WRFKHPLVWU\

    6LOYHVWUL ( &LRI ) *OLQQL ' &HFFDUHOOL 0 /RPEDUGL $ GH /DQJH 3&KDPEHU\ $ 6HYHULQR 9 /DQQL $ *RJOLD ) 0RUHQR 0 3DWKZD\VDIIHFWHG E\ GLLRGROWK\URQLQH LQ OLYHU RI KLJK IDWIHG UDWV HYLGHQFH IURPWZRGLPHQVLRQDO HOHFWURSKRUHVLV EOXHQDWLYH 3$*( DQG PDVV VSHFWURPHWU\0RO %LRV\VW

    $QGULQJD . . %DLOH\ 6 0 'HWHFWLRQ RI SURWHLQ WKLROV LQ PLWRFKRQGULDOR[LGDWLYH SKRVSKRU\ODWLRQ FRPSOH[HV DQG DVVRFLDWHG SURWHLQV 0HWKRGV(Q]\PRO

    $QGULQJD . . .LQJ $ / (FFOHVWRQ + % 0DQWHQD 6 ./DQGDU $ -KDOD 1 & 'LFNLQVRQ ' $ 6TXDGULWR * / %DLOH\ 6 0$QDO\VLV RI WKH OLYHU PLWRFKRQGULDO SURWHRPH LQ UHVSRQVH WR HWKDQRO DQG6DGHQRV\OPHWKLRQLQH WUHDWPHQWV 1RYHO PROHFXODU WDUJHWV RI GLVHDVH DQGKHSDWRSURWHFWLRQ $P - 3K\VLRO *DVWURLQWHVW /LYHU 3K\VLRO *

    5HLVLQJHU 9 (LFKDFNHU / $ 2UJDQHOOH SURWHRPLFV UHGXFWLRQ RI VDPSOHFRPSOH[LW\ E\ HQ]\PDWLF LQJHO VHOHFWLRQ RI QDWLYH SURWHLQV 0HWKRGV 0RO%LRO

    .OHSVFK 0 6FKOHJHO 6 :LFNVWURP ' )ULVR * YDQ :LMN . -3HUVVRQ - 2 GH *LHU - : :DJQHU 6 ,PPRELOL]DWLRQ RI WKH UVWGLPHQVLRQ LQ ' EOXH QDWLYH6'63$*( DOORZV WKH UHODWLYH TXDQWLFDWLRQRI PHPEUDQH SURWHRPHV 0HWKRGV

    Dow

    nloa

    ded

    by C

    LAR

    KSO

    N U

    NIV

    on

    Nov

    embe

    r 18,

    201

    1 | h

    ttp://

    pubs

    .acs

    .org

    P

    ublic

    atio

    n D

    ate

    (Web

    ): N

    ovem

    ber 1

    7, 2

    011

    | doi

    : 10.

    1021

    /bk-

    2011

    -108

    3.ch

    012

    In Oxidative Stress: Diagnostics, Prevention, and Therapy; Andreescu, S., et al.; ACS Symposium Series; American Chemical Society: Washington, DC, 2011.

  • 5HLVLQJHU 9 (LFKDFNHU / $ 6ROXELOL]DWLRQ RI PHPEUDQH SURWHLQFRPSOH[HV IRU EOXH QDWLYH 3$*( - 3URWHRPLFV

    %DLOH\ 6 0 $QGULQJD . . /DQGDU $ 'DUOH\8VPDU 9 0 3URWHRPLFDSSURDFKHV WR LGHQWLI\ DQG FKDUDFWHUL]H DOWHUDWLRQV WR WKH PLWRFKRQGULDOSURWHRPH LQ DOFRKROLF OLYHU GLVHDVH 0HWKRGV 0RO %LRO

    5HLVLQJHU 9 (LFKDFNHU / $ +RZ WR DQDO\]H SURWHLQ FRPSOH[HV E\ ' EOXHQDWLYH 6'63$*( 3URWHRPLFV 6XSSOHPHQW

    $LYDOLRWLV 0 .DUDV 0 7VLRWLV * $Q DOWHUQDWLYH VWUDWHJ\ IRU WKHPHPEUDQH SURWHRPH DQDO\VLV RI WKH JUHHQ VXOIXU EDFWHULXP &KORURELXPWHSLGXP XVLQJ EOXH QDWLYH 3$*( DQG ' 3$*( RQ SXULHG PHPEUDQHV -3URWHRPH 5HV

    %DEXVLDN 0 0DQ 3 3HWUDN - 9\RUDO ' 1DWLYH SURWHRPLF DQDO\VLV RISURWHLQ FRPSOH[HV LQ PXULQH LQWHVWLQDO EUXVK ERUGHU PHPEUDQHV 3URWHRPLFV

    $LYDOLRWLV 0 .DUDV 0 7VLRWLV * +LJK WKURXJKSXW WZRGLPHQVLRQDO EOXHQDWLYH HOHFWURSKRUHVLV $ WRRO IRU IXQFWLRQDO SURWHRPLFV RI F\WRSODVPDWLFSURWHLQ FRPSOH[HV IURP &KORURELXP WHSLGXP 3KRWRV\QWK 5HV

    'L 3DQFUD]LR ) %LVHWWR ( $OYHUGL 9 0DYHOOL , (VSRVLWR * /LSSH *'LIIHUHQWLDO VWHDG\VWDWH W\URVLQH SKRVSKRU\ODWLRQ RI WZR ROLJRPHULF IRUPVRI PLWRFKRQGULDO ))$73V\QWKDVH $ VWUXFWXUDO SURWHRPLF DQDO\VLV3URWHRPLFV

    &DPDFKR&DUYDMDO 0 0 :ROOVFKHLG % $HEHUVROG 5 6WHLPOH 96FKDPHO : : 7ZRGLPHQVLRQDO %OXH QDWLYH6'6 JHO HOHFWURSKRUHVLVRI PXOWLSURWHLQ FRPSOH[HV IURP ZKROH FHOOXODU O\VDWHV $ SURWHRPLFVDSSURDFK 0RO &HOO 3URWHRPLFV

    0F'RQDOG 7 * 9DQ (\N - ( 0LWRFKRQGULDO SURWHRPLFV 8QGHUFRYHU LQWKH OLSLG ELOD\HU %DVLF 5HV &DUGLRO

    'DULH & & -DQVVHQ : * /LWVFKHU ( 6 :DVVDUPDQ 3 0 3XULHGWURXW HJJ YLWHOOLQH HQYHORSH SURWHLQV 9(EHWD DQG 9(JDPPD SRO\PHUL]H LQWRKRPRPHULF EULOV IURP GLPHUV LQ YLWUR %LRFKLP %LRSK\V $FWD

    /LWVFKHU ( 6 -DQVVHQ:* 'DULH & & :DVVDUPDQ 30 3XULHGPRXVHHJJ ]RQD SHOOXFLGD JO\FRSURWHLQV SRO\PHUL]H LQWR KRPRPHULF EULOV XQGHUQRQGHQDWXULQJ FRQGLWLRQV - &HOO 3K\VLRO

    'DULH & & 6KHWW\ 9 6SHOOPDQ ' 6 =KDQJ * ;X & &DUGDVLV + /%ODLV 6 )HQ\R ' 1HXEHUW 7 $ %OXH 1DWLYH 3$*( DQG 0DVV 6SHFWURPHWU\$QDO\VLV RI WKH (SKULQ 6WLPXODWLRQ 'HSHQGHQW 3URWHLQ3URWHLQ ,QWHUDFWLRQVLQ 1*(SK% &HOOV 6SULQJHU9HUODJ 'VVHOGRUI *HUPDQ\

    'DULH & & 'HLQKDUGW . =KDQJ * &DUGDVLV + 6 &KDR 0 91HXEHUW 7 $ ,GHQWLI\LQJ WUDQVLHQW SURWHLQSURWHLQ LQWHUDFWLRQV LQ (SK%VLJQDOLQJ E\ %OXH 1DWLYH 3$*( DQG PDVV VSHFWURPHWU\ 3URWHRPLFV

    5HLIVFKQHLGHU 1 + *RWR 6 1DNDPRWR + 7DNDKDVKL 5 6XJDZD 0'HQFKHU 1 $ .UDXVH ) 'HQLQJ WKH PLWRFKRQGULDO SURWHRPHV IURP YHUDW RUJDQV LQ D SK\VLRORJLFDOO\ VLJQLFDQW FRQWH[W XVLQJ ' EOXHQDWLYH6'63$*( - 3URWHRPH 5HV

    Dow

    nloa

    ded

    by C

    LAR

    KSO

    N U

    NIV

    on

    Nov

    embe

    r 18,

    201

    1 | h

    ttp://

    pubs

    .acs

    .org

    P

    ublic

    atio

    n D

    ate

    (Web

    ): N

    ovem

    ber 1

    7, 2

    011

    | doi

    : 10.

    1021

    /bk-

    2011

    -108

    3.ch

    012

    In Oxidative Stress: Diagnostics, Prevention, and Therapy; Andreescu, S., et al.; ACS Symposium Series; American Chemical Society: Washington, DC, 2011.

  • :RRGV $ * *XWKULH . 0 .XUODZDOOD 0 $ *DOO & 0'HDIIHUHQWDWLRQLQGXFHG LQFUHDVHV LQ KLSSRFDPSDO LQVXOLQOLNH JURZWKIDFWRU PHVVHQJHU 51$ H[SUHVVLRQ DUH VHYHUHO\ DWWHQXDWHG LQ PLGGOH DJHGDQG DJHG UDWV 1HXURVFLHQFH

    :RRGV $ * 3RXOVHQ ) 5 *DOO & 0 'H[DPHWKDVRQH VHOHFWLYHO\VXSSUHVVHV PLFURJOLDO WURSKLF UHVSRQVHV WR KLSSRFDPSDO GHDIIHUHQWDWLRQ1HXURVFLHQFH

    3LWXOHVFX 0 ( $GDPV 5 + (SKHSKULQ PROHFXOHV$ KXE IRU VLJQDOLQJDQG HQGRF\WRVLV *HQHV 'HY

    +ROODQG 6 - *DOH 1 : *LVK * ' 5RWK 5 $ 6RQJ\DQJ =&DQWOH\ / & +HQNHPH\HU 0

  • 0LDR + %XUQHWW ( .LQFK 0 6LPRQ ( :DQJ % $FWLYDWLRQ RI(SK$ NLQDVH VXSSUHVVHV LQWHJULQ IXQFWLRQ DQG FDXVHV IRFDODGKHVLRQNLQDVHGHSKRVSKRU\ODWLRQ 1DW &HOO %LRO

    9HDULQJ & - /DFNPDQQ 0 (SK UHFHSWRU VLJQDOOLQJ GLPHULVDWLRQ MXVW LVQWHQRXJK *URZWK )DFWRUV

    1RUHQ 1 . 3DVTXDOH ( % (SK UHFHSWRUHSKULQ ELGLUHFWLRQDO VLJQDOV WKDWWDUJHW 5DV DQG 5KR SURWHLQV &HOO 6LJQDO

    (OOLV & .DVPL ) *DQMX 3 :DOOV ( 3DQD\RWRX * 5HLWK $ ' $MX[WDPHPEUDQH DXWRSKRVSKRU\ODWLRQ VLWH LQ WKH (SK IDPLO\ UHFHSWRU W\URVLQHNLQDVH 6HN PHGLDWHV KLJK DIQLW\ LQWHUDFWLRQ ZLWK SI\Q 2QFRJHQH

    3DQGH\ $ $QGHUVHQ - 6 0DQQ 0 8VH RI PDVV VSHFWURPHWU\ WR VWXG\VLJQDOLQJ SDWKZD\V 6FL 67.( SO

    3DQGH\ $ 'XDQ + 'L[LW 9 0 &KDUDFWHUL]DWLRQ RI D QRYHO 6UFOLNHDGDSWHU SURWHLQ WKDW DVVRFLDWHV ZLWK WKH (FN UHFHSWRU W\URVLQH NLQDVH - %LRO&KHP

    3DQGH\ $ /D]DU ' ) 6DOWLHO $ 5 'L[LW 9 0 $FWLYDWLRQ RI WKH (FNUHFHSWRU SURWHLQ W\URVLQH NLQDVH VWLPXODWHV SKRVSKDWLG\OLQRVLWRO NLQDVHDFWLYLW\ - %LRO &KHP

    3DQGH\ $ 3RGWHOHMQLNRY $ 9 %ODJRHY % %XVWHOR ; 5 0DQQ 0/RGLVK + ) $QDO\VLV RI UHFHSWRU VLJQDOLQJ SDWKZD\V E\ PDVV VSHFWURPHWU\LGHQWLFDWLRQ RI YDY DV D VXEVWUDWH RI WKH HSLGHUPDO DQG SODWHOHWGHULYHGJURZWK IDFWRU UHFHSWRUV 3URF 1DWO $FDG 6FL 8 6 $

    6WHLQ ( &HUUHWWL ' 3 'DQLHO 7 2 /LJDQG DFWLYDWLRQ RI (/. UHFHSWRUW\URVLQH NLQDVH SURPRWHV LWV DVVRFLDWLRQ ZLWK *UE DQG *UE LQ YDVFXODUHQGRWKHOLDO FHOOV - %LRO &KHP

    6WHLQ ( +X\QK'R 8 /DQH $ $ &HUUHWWL ' 3 'DQLHO 7 2 1FNUHFUXLWPHQW WR (SK UHFHSWRU (SK%(/. FRXSOHV OLJDQG DFWLYDWLRQ WR F-XQNLQDVH - %LRO &KHP

    6WHLQ ( /DQH $ $ &HUUHWWL ' 3 6FKRHFNOPDQQ + 2 6FKURII $ '9DQ (WWHQ 5 / 'DQLHO 7 2 (SK UHFHSWRUV GLVFULPLQDWH VSHFLF OLJDQGROLJRPHUV WR GHWHUPLQH DOWHUQDWLYH VLJQDOLQJ FRPSOH[HV DWWDFKPHQW DQGDVVHPEO\ UHVSRQVHV *HQHV 'HY

    =KDQJ * )HQ\R ' 1HXEHUW 7 $ 6FUHHQLQJ IRU (SK% VLJQDOLQJ HIIHFWRUVXVLQJ 6,/$& ZLWK D OLQHDU LRQ WUDSRUELWUDS PDVV VSHFWURPHWHU - 3URWHRPH5HV

    =KDQJ * 6SHOOPDQ ' 6 6NROQLN (

  • %DXHU ' ( +DW]LYDVVLOLRX * =KDR ) $QGUHDGLV & 7KRPSVRQ &% $73 FLWUDWH O\DVH LV DQ LPSRUWDQW FRPSRQHQW RI FHOO JURZWK DQGWUDQVIRUPDWLRQ 2QFRJHQH

    0F1DXJKW . 6 2ODQRZ & : +DOOLZHOO % ,VDFVRQ 2 -HQQHU 3)DLOXUH RI WKH XELTXLWLQSURWHDVRPH V\VWHP LQ 3DUNLQVRQV GLVHDVH 1DW 5HY1HXURVFL

    %URZQ 0 * 0RQDFR - - %LRFKHPLFDO SXULFDWLRQ RI GLVWLQFW SURWHDVRPHVXEVHWV (Q]\PH 3URWHLQ

    +HQGLO . % :HOLQGHU . * 3HGHUVHQ ' 8HUNYLW] : .ULVWHQVHQ 36XEXQLW VWRLFKLRPHWU\ RI KXPDQ SURWHDVRPHV (Q]\PH 3URWHLQ

    6HHOLJ $ %RHV % .ORHW]HO 3 0 &KDUDFWHUL]DWLRQ RI PRXVH SURWHDVRPHVXEXQLW 0& DQG LGHQWLFDWLRQ RI SURWHDVRPH VXEW\SHV ZLWK GLIIHUHQWFOHDYDJH FKDUDFWHULVWLFV 3URWHDVRPH VXEXQLWV SURWHDVRPH VXESRSXODWLRQV(Q]\PH 3URWHLQ

    +HQGLO . % .ULVWHQVHQ 3 8HUNYLW] : +XPDQ SURWHDVRPHV DQDO\VHGZLWK PRQRFORQDO DQWLERGLHV %LRFKHP - 3DUW

    $NDLVKL 7 6KLRPL 7 6DZDGD +

  • 'RXGQD - $ 5RELQVRQ & 9 0DVV VSHFWURPHWU\ UHYHDOV PRGXODULW\ DQG DFRPSOHWH VXEXQLW LQWHUDFWLRQ PDS RI WKH HXNDU\RWLF WUDQVODWLRQ IDFWRU H,)3URF 1DWO $FDG 6FL 8 6 $

    /X / .RPDGD 0 .LWDPXUD 1 +XPDQ +UV D W\URVLQH NLQDVH VXEVWUDWHLQ JURZWK IDFWRUVWLPXODWHG FHOOV F'1$ FORQLQJ DQG PDSSLQJ RI WKH JHQH WRFKURPRVRPH *HQH

    .RED\DVKL + 7DQDND 1 $VDR + 0LXUD 6 .\XXPD 0 6HPXUD .,VKLL 1 6XJDPXUD . +UV D PDPPDOLDQ PDVWHU PROHFXOH LQ YHVLFXODUWUDQVSRUW DQG SURWHLQ VRUWLQJ VXSSUHVVHV WKH GHJUDGDWLRQ RI (6&57 SURWHLQVVLJQDO WUDQVGXFLQJ DGDSWRU PROHFXOH DQG - %LRO &KHP

    6SHOOPDQ ' 6 'HLQKDUGW . 'DULH & & &KDR 0 9 1HXEHUW 7$ 6WDEOH LVRWRSLF ODEHOLQJ E\ DPLQR DFLGV LQ FXOWXUHG SULPDU\QHXURQV $SSOLFDWLRQ WR EUDLQGHULYHG QHXURWURSKLF IDFWRUGHSHQGHQWSKRVSKRW\URVLQHDVVRFLDWHG VLJQDOLQJ 0RO &HOO 3URWHRPLFV

    'DULH & & %LQLRVVHN 0 / *DZLQRZLF] 0 $ 0LOJURP

  • -RHW 7 &RXUQDF / 3HOWLHU * +DYDX[ 0 &\FOLF HOHFWURQ RZ DURXQGSKRWRV\VWHP , LQ & SODQWV ,Q YLYR FRQWURO E\ WKH UHGR[ VWDWH RIFKORURSODVWV DQG LQYROYHPHQW RI WKH 1$'+GHK\GURJHQDVH FRPSOH[ 3ODQW3K\VLRO

    &DVDQR / 0 =DSDWD - 0 0DUWLQ 0 6DEDWHU % &KORURUHVSLUDWLRQ DQGSRLVLQJ RI F\FOLF HOHFWURQ WUDQVSRUW 3ODVWRTXLQRQH DV HOHFWURQ WUDQVSRUWHUEHWZHHQ WK\ODNRLG 1$'+ GHK\GURJHQDVH DQG SHUR[LGDVH - %LRO &KHP

    6D]DQRY / $ %XUURZV 3 $ 1L[RQ 3 - 7KH FKORURSODVW 1GK FRPSOH[PHGLDWHV WKH GDUN UHGXFWLRQ RI WKH SODVWRTXLQRQH SRRO LQ UHVSRQVH WR KHDWVWUHVV LQ WREDFFR OHDYHV )(%6 /HWW

    &DWDOD 5 6DEDWHU % *XHUD $ ([SUHVVLRQ RI WKH SODVWLG QGK) JHQHSURGXFW LQ SKRWRV\QWKHWLF DQG QRQSKRWRV\QWKHWLF WLVVXHV RI GHYHORSLQJEDUOH\ VHHGOLQJV 3ODQW &HOO 3K\VLRO

    %XUURZV 3 $ 6D]DQRY / $ 6YDE = 0DOLJD 3 1L[RQ 3 -,GHQWLFDWLRQ RI D IXQFWLRQDO UHVSLUDWRU\ FRPSOH[ LQ FKORURSODVWV WKURXJKDQDO\VLV RI WREDFFR PXWDQWV FRQWDLQLQJ GLVUXSWHG SODVWLG QGK JHQHV (0%2-

    6D]DQRY / $ %XUURZV 3 $ 1L[RQ 3 - 7KH SODVWLG QGK JHQHV FRGH IRU DQ1$'+VSHFLF GHK\GURJHQDVH ,VRODWLRQ RI D FRPSOH[ , DQDORJXH IURP SHDWK\ODNRLG PHPEUDQHV 3URF 1DWO $FDG 6FL 8 6 $

    *XHGHQH\ * &RUQHLOOH 6 &XLQH 6 3HOWLHU * (YLGHQFH IRU DQDVVRFLDWLRQ RI QGK % QGK - JHQH SURGXFWV DQG IHUUHGR[LQ1$'3UHGXFWDVHDV FRPSRQHQWV RI D FKORURSODVWLF 1$'3+ GHK\GURJHQDVH FRPSOH[ )(%6/HWW

    /DGLJ 5 6RPPHU 0 6 +DKQ $ /HLVHJDQJ 0 6 3DSDVRWLULRX '* ,EUDKLP 0 (ONHKDO 5 .DUDV 0 =LFNHUPDQQ 9 *XWHQVRKQ 0%UDQGW 8 .ORVJHQ 5 % 6FKOHLII ( $ KLJKGHQLWLRQ QDWLYHSRO\DFU\ODPLGH JHO HOHFWURSKRUHVLV V\VWHP IRU WKH DQDO\VLV RI PHPEUDQHFRPSOH[HV 3ODQW -

    3HQJ