Professional Documents
Culture Documents
Effects of N-Glycosylation of The Human TRPA1 On Agonist-Sensitivity PDF
Effects of N-Glycosylation of The Human TRPA1 On Agonist-Sensitivity PDF
BIOSCIENCE REPORTS
ACCEPTED MANUSCRIPT
Ǧ
ͳǦ
Ǧ
Ǧǡ
Ǥ
Ǧ
ǡ
Ǧ
Ǥ
ǡǦ
ǤͳǦ
Ͷ
ͷ͵Ǥǡ
Ǧ
Ͷ
ǦͳǤǡǦ
ͳ
Ǥ
ǡ
Ǧ
ͳǤ
Ǧ
ͳ
Ǥ
7LPRWK\-(JDQ0DULR$$FXxD0DUF\=HQREL:RQJ+DQQV8OULFK=HLOKRIHU'DYLG8UHFK
&DUWLODJH(QJLQHHULQJ 5HJHQHUDWLRQ/DEWKH'HSDUWPHQWRI+HDOWK6FLHQFH 7HFKQRORJ\7KH6ZLVV
GXUHFK#QXPDEFRP
$%675$&7
'HWHUPLQLQJWKHIXQFWLRQDOVLJQLILFDQFHRISRVWWUDQVODWLRQDOPRGLILFDWLRQVDGYDQFHVRXUXQGHUVWDQGLQJRI
PDQ\EURDGO\H[SUHVVHGSURWHLQVDQGSDUWLFXODUO\LRQFKDQQHOV7KHHQ]\PHVWKDWFDWDO\]HWKHVHPRGLIL
FDWLRQV DUH RIWHQ H[SUHVVHG LQ D FHOOW\SH VSHFLILF PDQQHU UHVXOWLQJ LQ FRQVLGHUDEOH VWUXFWXUDO GLYHUVLW\
DPRQJSRVWWUDQVODWLRQDOO\PRGLILHGSURWHLQVWKDWDUHH[SUHVVHGDFURVVDYDULHW\RIFHOOW\SHV753FKDQ
LILHGZLWK1JO\FDQVWKDWFRQWULEXWHWRSURWHLQIXQFWLRQ753$SRVVHVVHVWZRSXWDWLYH1OLQNHGJO\FR
V\ODWLRQVLWHVDW1DQG1WKDWKDYHQRW\HWEHHQVWXGLHGLQGHWDLO+HUHZHVKRZWKDWERWKRIWKHVH
VLWHVFDQEHPRGLILHGZLWKDQ1JO\FDQDQGWKDWWKHJO\FDQDWSRVLWLRQ1PRGXODWHVDJRQLVWVHQVLWLYLW\
RI 753$ in vitro $GGLWLRQDOO\ ZH IRXQG WKDW 1JO\FRV\ODWLRQ DOVR PRGXODWHV FRRSHUDWLYH HIIHFWV RI
WHPSHUDWXUHDQGWKHDJRQLVWFLQQDPDOGHK\GHRQ753$FKDQQHODFWLYDWLRQ&ROOHFWLYHO\WKHVHILQGLQJV
VXJJHVWDG\QDPLFUROHSOD\HGE\WKH1JO\FRV\ODWLRQRIKXPDQ753$7KH\DOVRSURYLGHIXUWKHUHYL
GHQFHRIWKHYHUVDWLOLW\RI1JO\FDQVDQGZLOODVVLVWLQHIIRUWVWRIXOO\XQGHUVWDQGWKHFRPSOH[UHJXODWLRQRI
753$DFWLYLW\
6XPPDU\6WDWHPHQW2XUH[SHULPHQWVFRQILUP1JO\FRV\ODWLRQRIWKHKXPDQFDWLRQFKDQQHO753$
DQGVXJJHVWDUROHRIWKH1JO\FDQDWSRVLWLRQ1LQGHWHUPLQLQJFKDQQHOVHQVLWLYLW\WRYDULRXVDJRQLVWV
)XUWKHUWKHDFWLYLW\PRGXODWLQJHIIHFWVRI753$1JO\FDQVDUHHYLGHQWO\LQIOXHQFHGE\WHPSHUDWXUH
6KRUW7LWOH1JO\FRV\ODWLRQRI753$PRGXODWHVDJRQLVWVHQVLWLYLW\
.H\ZRUGV753FKDQQHOVJO\FRV\ODWLRQJO\FRSURWHLQ753$FLQQDPDOGHK\GHPHQWKRO
FRQFDYDOLQ $ P$E PRQRFORQDO DQWLERG\ ,J* LPPXQRJOREXOLQ * *$3'+ *O\FHUDOGHK\GH
SKRVSKDWH GHK\GURJHQDVH &$ FLQQDPDOGHK\GH $,7& DOO\O LVRWKLRF\DQDWH 5)8 UHODWLYH IOXRUHVFHQFH
XQLWVH*)3HQKDQFHGJUHHQIOXRUHVFHQWSURWHLQ&,FRQILGHQFHLQWHUYDO
,1752'8&7,21
7UDQVLHQW UHFHSWRU SRWHQWLDO 753 FKDQQHOV UHYLHZHG LQ ± DUH D IDPLO\ RI QRQVHOHFWLYH FDWLRQ
SHUPHDEOHLQWHJUDOPHPEUDQHSURWHLQVWKDWFRPPRQO\DFWDVUHFHSWRUVLQYDULRXVVHQVRU\SURFHVVHV)XQF
WLRQDO753$FKDQQHOVDUHFRPSRVHGRIIRXUVXEXQLWVHDFKFRQWDLQLQJVL[WUDQVPHPEUDQHUHJLRQV6
FKDQQHOVDUHPRGXODWHGE\GLYHUVHFKHPLFDODQGSK\VLFDOVWLPXOLDQGVLPXOWDQHRXVH[SRVXUHWRFRPELQD
WLRQVRIWKHVHVWLPXOLRIWHQWULJJHUXQLTXHFKDQQHOUHVSRQVHVUHYHDOLQJFRRSHUDWLYHHIIHFWV,QDGGLWLRQD
VLQJOH753FKDQQHOJHQHFDQJLYHULVHWRPDQ\VWUXFWXUDOO\DQGIXQFWLRQDOO\GLYHUVHFKDQQHOVGXHWRWKH
DELOLW\RIVXEXQLWVWRKHWHURPXOWLPHUL]HDVZHOODVGLVSOD\SRVWFRWUDQVODWLRQDOO\DGGHGPRGLI\LQJJURXSV
VXFKDV1JO\FDQV
753FKDQQHOVDUHKLJKO\FRQVHUYHGDQGVWXGLHVKDYHGHVFULEHGWKH1OLQNHGJO\FRV\ODWLRQRIPDQ\RI
FHVVLQJRIDQROLJRVDFFKDULGHWRVLGHFKDLQVRIOXPHQDO$VQUHVLGXHVWKDWDUHGLVSOD\HGZLWKLQDVSHFLILF
1JO\FDQ FDQ H[KLELW RQH RI WKUHH VWUXFWXUHV KLJKPDQQRVH K\EULG RU FRPSOH[ 7KH UROH RI 1
JO\FDQV LGHQWLILHG LQ VWXGLHV KDV UDQJHG IURP LQYROYHPHQW LQ SURWHLQ IROGLQJ DQG FHOOXODUORFDOL]DWLRQ WR
&6 H[SUHVVLRQ VHQVLWLYLW\ WR DJRQLVWV DFWLYLW\ UHJXODWLRQ DQG WHPSHUDWXUH
VHQVLWLYLW\
WHUPLQXVLVIXUWKHUFKDUDFWHUL]HGE\UHDFWLYH&\VDQG/\VUHVLGXHVZKLFKDFWDVVLWHVIRUUHYHUVLEOHFRYD
5HYHUVLEOHFRYDOHQWELQGLQJRIWKHVHUHVLGXHVLVWKHPHFKDQLVPE\ZKLFKVHYHUDOSXQJHQWFKHPLFDOVVXFK
DWWKHDSSUR[LPDWHSRVLWLRQRIWKHFKDQQHO¶VVXEXQLWSUHVXPDEO\GXHWRWKHGLVSDUDWHZHLJKWVRI³PDWXUH´
DQG³LPPDWXUH´JO\FRSURWHLQ7KLVVWXG\VRXJKWWRFRQILUPWKHSUHVHQFH±DQGFKDUDFWHUL]HWKHUROH±RI
K753$OLQNHGJO\FDQVZLWKWKHDLPRIRIIHULQJYDOXDEOHLQVLJKWVLQWRSURWHLQIXQFWLRQDQGUHJXODWLRQ
2XUSUHVHQWGDWDFRQILUPWKHGLJO\FRV\ODWLRQRI753$DQGLQGLFDWHDIXQFWLRQDOUROHIRUWKHJO\FDQDW
SRVLWLRQ17KHVHILQGLQJVSUHVHQWIXUWKHUHYLGHQFHRIWKHIXQFWLRQDOVLJQLILFDQFHRI1JO\FRV\ODWLRQ
HVSHFLDOO\DVLWUHODWHVWRLRQFKDQQHOV
(;3(5,0(17$/
&ORQLQJ:LOGW\SHDQG0XWDQWK753$)/$*
ZDV VXSSOLHG E\ )LVKHU 6FLHQWLILF $* K753$)/$* F'1$ ZDV DPSOLILHG DQG FORQHG LQWR D
DWLQJ WKH RSHQ UHDGLQJ IUDPH 25) XVLQJ WKH IROORZLQJ SULPHUV IRUZDUG ¶
WLRQVHQFRGHGVLWHGLUHFWHGPXWDJHQHVLVZDVSHUIRUPHGE\RYHUODSSLQJ3&5XVLQJWKHPXWDWLQJSULPHUV
*$7*&&$*77*$&7**$$$*&&$77&&7**¶14IRUZDUG¶&$$&7**&$7&$7&&$**
$$$&7$*7*$7&¶UHYHUVH¶*$7&$&7$*777&&7**$7*$7*&&$*77*¶LQFRPELQDWLRQ
ZDUG ¶$77*77$$&$&$$&&*$7**$7*7&$7*$*$&&¶ UHYHUVH ¶
$77**$7&&7*7$$$77&$**$**$7*7$$$$*&¶
7UDQVLHQW7UDQVIHFWLRQRI+(.FHOOVDQG:HVWHUQ%ORW([SHULPHQWV
FRUGLQJ WR PDQXIDFWXUHU¶V LQVWUXFWLRQV DQG WKH FXOWXUH PHGLXP ZDV H[FKDQJHG a K DIWHU WUDQVIHF
WLRQ&HOOVZHUHDOZD\VKDUYHVWHGKDIWHUWUDQVIHFWLRQ
7RKDUYHVWLQWDFWFHOOVZHUHULQVHGLQLFHFROG;3KRVSKDWH%XIIHUHG6DOLQHS+ZLWKRXW&DRU
LFHDJLWDWLQJLQWHUPLWWHQWO\7KHVDPSOHVZHUHWKHQFHQWULIXJHGDWJIRUPLQDW&DQGWKH
ZY *O\FHURO YY P0 ('7$ %URPRSKHQRO %OXH ZY P0 '77 ZDV DGGHG
DQGWKHVDPSOHVZHUHLQFXEDWHGIRUPLQDW57DQGWKHQERLOHGDW&IRUPLQ)RU:%GHWHF
DQG D SHUR[LGDVHFRQMXJDWHG UDEELW DQWLPRXVH ,J* -DFNVRQ ,PPXQR5HVHDUFK &DWDORJXH
ZDVXVHGDVDVHFRQGDU\DQWLERG\/HFWLQPHGLDWHGK753$GHWHFWLRQZDVDOVRSHUIRUPHGDSSO\LQJ
D &RQFDYDOLQ $ &RQ$SHUR[LGDVH FRQMXJDWH 6LJPD$OGULFK &DWDORJXH / WR WUDQVIHU PHP
EUDQHVGLVSOD\LQJLPPRELOL]HGSXULILHGK753$)/$*SXULILFDWLRQPHWKRGGHVFULEHGLQQH[WVHFWLRQ
:% GDWD DQDO\VLV ZDV SHUIRUPHG XVLQJ VLJQDO LQWHQVLWLHV DX QRUPDOL]HG WR WRWDO SURWHLQ WUDQVIHUUHG
ZKLFK ZDV UHYHDOHG E\ VWDLQLQJ WUDQVIHU PHPEUDQHV ZLWK ; $PLGR %ODFN 6WDLQLQJ 6ROXWLRQ 6LJPD
$OGULFKDQGGHVWDLQLQJRYHUQLJKWLQPHWKDQRODQGDFHWLFDFLGLQGG+27RFRPSDUHSURWHLQ
H[SUHVVLRQOHYHOVWKHFRPELQHGQRUPDOL]HGVLJQDOLQWHQVLWLHVRIDOO753$VXEXQLWJO\FRIRUPVGHWHFWHG
DOO753$VXEXQLWJO\FRIRUPVGHWHFWHGLQ:7VDPSOHV
(QGR+DQG31*DVH)'LJHVWLRQRI'HQDWXUHG3URWHLQ
+(.FHOOWUDQVIHFWLRQVZHUHFDUULHGRXWLQIODVNVZLWKDFPJURZWKVXUIDFHDUHD$IWHULQFXEDWLRQ
WKHFHOOVZHUHULQVHGWKUHHWLPHVLQLFHFROG3%6DQGO\VHGLQP//\VLV%XIIHU:LOGW\SH:7RUPX
WDQWK753$)/$*ZDVLPPXQRSUHFLSLWDWHGRXWRIZKROHFHOOO\VDWHVE\LQFXEDWLRQZLWK/0DQ
WL)/$* PDJQHWLF EHDGV 6LJPD$OGULFK URWDWLQJ WKH VDPSOHV RYHUQLJKW DW & $IWHU ULQVLQJ WKH
EHDGV;ZLWKLFHFROG3%6WKHSURWHLQVZHUHHOXWHGIURPWKHEHDGVZLWK/P0*O\FLQH+&O
)URPHDFKHOXDWHWKUHH/VDPSOHVZHUHDOLTXRWHGDQGWKHSURWHLQVDPSOHVZHUHGHQDWXUHGIRUVXE
O\]HGE\:HVWHUQ%ORW
&HOOVXUIDFH%LRWLQ\ODWLRQ
+(.FHOOWUDQVIHFWLRQVZHUHFDUULHGRXWLQIODVNVZLWKDFPJURZWKVXUIDFHDUHD$IWHULQFXEDWLRQ
WKHLQWDFWFHOOVZHUHULQVHGWKUHHWLPHVLQLFHFROG3%6DQGLQFXEDWHGLQPJP/(=/LQN6XOIR1+6
; LQ LFHFROG P0 JO\FLQH±3%6 DQG WKHQ WKUHH DGGLWLRQDO ZDVKHV ZLWK LFHFROG 3%6 ZHUH SHU
IRUPHG7KHFHOOVZHUHO\VHGLQȝ//\VLV%XIIHU$IWHUHTXDOL]LQJWKHSURWHLQFRQWHQWVDOLTXRWVIURP
HDFK O\VDWH VDPSOH ZHUH VHW DVLGH &/ VDPSOHV DQG WKH UHPDLQLQJ O\VDWH ZDV LQFXEDWHG ZLWK /
7KH DJDURVH ZDV SHOOHWHG E\ FHQWULIXJDWLRQ DW J DQG &VXSHUQDWDQWV ZHUH GLVFDUGHG DQG WKH
DJDURVHZDVZDVKHG;WRWDO;LQ:DVK6ROXWLRQ$P07ULV+&OS+P01D&OP0
P0 7ULV+&O S+ 7KH VWUHSWDYLGLQDIILQLW\SUHFLSLWDWH ZDV HOXWHG IURP WKH EHDGV ZLWK /
IRUPLQDW&$OOVDPSOHVZHUHWKHQDQDO\]HGE\:%XVLQJVLJQDOLQWHQVLWLHVQRUPDOL]HGWRWRWDOSUH
FLSLWDWHWUDQVIHUUHGDVGHWHUPLQHGE\GHWHFWLRQZLWKVWUHSWDYLGLQSHUR[LGDVH7RFRQILUPWKHDEVHQFHRI
ERG\WREHGHWHFWHGE\DQ+53FRQMXJDWHGJRDWDQWLUDEELW,J*-DFNVRQ,PPXQR5HVHDUFK&DWDORJXH
0HPEUDQH'HSRODUL]DWLRQ$VVD\VRQWKH)OH[6WDWLRQ
K7UDQVIHFWLRQVDPSOHV/MHW35,0(7UDQVIHFWLRQ%XIIHUDQG/MHW35,0(7UDQVIHF
WLRQ5HDJHQWSHUJSODVPLGZHUHGLOXWHGLQSUHZDUPHGFRPSOHWH'0(0DWDFRQFHQWUDWLRQRIa
JSODVPLGP/7KHROGPHGLXPZDVGLVFDUGHGDQGaQJFPRIWKHDSSURSULDWHSODVPLGZHUHDGGHG
WRHDFKZHOO7KHFHOOVZHUHWKHQLQFXEDWHGIRUK
7KHPHGLXPZDVGLVFDUGHGDQG/;0HPEUDQH3RWHQWLDO$VVD\.LW5(''\HLQ&RPSRQHQW%
FXEDWHGIRUPLQDWHLWKHU&RU&7KHSODWHVZHUHDQDO\]HGLQD)OH[6WDWLRQVFDQQLQJIOXR
URPHWHU 0ROHFXODU 'HYLFHV ,QF DW QP H[FLWDWLRQ DQG QP HPLVVLRQ QP FXWRII ZDYH
OHQJWKV,QLWLDWLRQRIWKHVFDQQLQJSURJUDPUHVXOWHGLQWKHWUDQVIHURI0RIWKH753$DJRQLVW
GDWDZHUHFROOHFWHGXVLQJ6RIW0D[3URY0ROHFXODU'HYLFHV,QFIRUPLQDIWHUDSSOLFDWLRQRI
VHQWHGKHUH1RUPDOL]HGIOXRURPHWULFGDWDDUHSUHVHQWHGDVPD[LPXP5)8$OOGRVHUHVSRQVHFXUYHV
ZHUHILWWHGWRGDWDXSWRWKHPD[LPDOFKDQQHOUHVSRQVHXVLQJ*UDSK3DG3ULVPVRIWZDUH
(OHFWURSK\VLRORJLFDO5HFRUGLQJV
+(.FHOOVZHUHPDLQWDLQHGLQFRPSOHWH'0(0DQGSODWHGRQPPGLVKHVKEHIRUHWUDQVIHF
RISODVPLGHQFRGLQJHQKDQFHGJUHHQIOXRUHVFHQWSURWHLQH*)3XVHGDVDPDUNHURIVXFFHVVIXOWUDQVIHF
WLRQDQGWKHQLQFXEDWHGIRUK2QWKHGD\RIH[SHULPHQWWKHPHGLXPZDVGLVFDUGHGDQGUHSODFHG
ZLWK SUHZDUPHG H[WUDFHOOXODU VROXWLRQ & FRQWDLQLQJ LQ P0 1D&O .&O &D&O
0J&O +(3(6 S+ DQG JOXFRVH :KROHFHOO SDWFKFODPS UHFRUGLQJV ZHUH SHUIRUPHG DW D
KROGLQJ SRWHQWLDO RI P9 XVLQJ DQ (3& DPSOLILHU DQG 3DWFK 0DVWHU Y VRIWZDUH +(.$ (O
0J&O *73 DQG $73 2QO\ H*)3SRVLWLYH FHOOV ZHUH XVHG IRU UHFRUGLQJV :KROHFHOO
753$PHGLDWHG FXUUHQWV ZHUH DFWLYDWHG E\ PLQ DSSOLFDWLRQ RI &$ IHG WKURXJK DQ RXWOHW WXEH
P ,' RI D FXVWRPGHVLJQHG JUDYLW\IHG PLFURSHUIXVLRQ V\VWHP SRVLWLRQHG ±P IURP WKH
UHFRUGHGFHOO7KHWHPSHUDWXUHRIVROXWLRQLQWKHFHOO¶VPLFURHQYLURQPHQWZDVPDLQWDLQHGDW&XVLQJD
+HDW&RROHG7HPSHUDWXUH&RQWURO6\VWHP$XWR0DWH6FLHQWLILF
5(68/76
K753$LV*O\FRV\ODWHGLQYLWUR
7RLQYHVWLJDWH1JO\FRV\ODWLRQDWERWK1DQG1ZHDSSOLHGWKUHHVWUDWHJLHVVLWHGLUHFWHGPXWD
VLGDVH GLJHVWLRQ RI K753$)/$* SXULILHG IURP ZKROHFHOO O\VDWHV &/V DQG OHFWLQ
PHGLDWHGGHWHFWLRQRIJO\FRV\ODWHGK753$)/$*
:%DQDO\VLVRI+(.FHOOVWUDQVLHQWO\H[SUHVVLQJK753$)/$*:7DQG14DQG14PX
WKHUWKH753$JO\FRIRUPVH[KLELWLQJWKHORZHVWHOHFWURSKRUHWLFPRELOLW\ORVWFRQVLGHUDEOHVLJQDOLQWHQ
$VQ UHVLGXHV ,W DOVR VHHPV OLNHO\ WKDW WKH IDVWHUPLJUDWLQJ 753$ VXEXQLWV LQ WKH UHVSHFWLYH GRXEOHWV
KDYHEHHQPRGLILHGZLWKDFRUHJO\FDQEHFDXVHXQOLNHWKH14GRXEOHPXWDQWWKH\ZHUHGHWHFWHG
RYHU&RQ$IDLOHGWRGHWHFWDQ\RIWKHVORZHUPLJUDWLQJVXEXQLWVLQWKHGRXEOHWVVXJJHVWLQJWKDWERWK1
JO\FRV\ODWLRQVLWHVXOWLPDWHO\EHDUDFRPSOH[JO\FDQSRVVHVVLQJWKUHHRUPRUHDQWHQQDH
([SUHVVLRQRIK753$LV$IIHFWHGE\14PXWDWLRQ
2YHUWKHFRXUVHRILQGHSHQGHQW+(.WUDQVIHFWLRQVWKHSUHVHQFHRIHLWKHUVLQJOH14PXWDWLRQUH
DOVR LQGLFDWHG WKDW WKH DEVHQFH RI WKH 1 JO\FRV\ODWLRQ VLWH HQKDQFHV WKH JO\FRSURFHVVLQJ RI WKH 1
JO\FDQ DW 1 DV GHWHUPLQHG E\ GLIIHUHQFHV LQ WKH UDWLR RI FRPSOH[FRUHJO\FRV\ODWHG SURWHLQ VLJQDO
,QDGGLWLRQWRWKHGLIIHUHQFHVREVHUYHGLQRYHUDOOSURWHLQH[SUHVVLRQZHGHWHFWHGGLVSDUDWH&6H[SUHV
VLRQ EHWZHHQ PXWDQW DQG :7 K753$ DIWHU LQGHSHQGHQW +(. WUDQVIHFWLRQV )LJXUH () 7KH
ORZHU FHOOVXUIDFH H[SUHVVLRQ RI 14 DQG 14 PXWDQWV UHODWLYH WR :7 14
14ZDVDSSDUHQWO\FRQVLVWHQWZLWKWKHUHGXFWLRQLQWRWDOSURWHLQH[SUHVVLRQGHWHU
PLQHGE\DQDO\VLVRIPHDQUDWLRVRI:7UHODWLYHFHOOVXUIDFHH[SUHVVLRQWR:7UHODWLYHWRWDOK753$H[
'HVSLWHWKH:%UHVXOWVZHREWDLQHGZHGLGQRWREVHUYHVLJQLILFDQWGLIIHUHQFHVEHWZHHQ753$YDUL
DQWVLQPD[LPDOFKDQJHVLQUHODWLYHIOXRUHVFHQFHDQGSHDNDPSOLWXGHVLQRXUIOXRURPHWULFYROWDJHPHDV
PHWULFDVVD\VGLGUHYHDOPHDQPD[LPDOIOXRUHVFHQFHFKDQJHVWKDWZHUHFRQVLVWHQWZLWKUHGXFWLRQVLQFHOO
0HDQZKLOH WKH DEVHQFH RI D QHJDWLYHO\FKDUJHG FDWLRQDWWUDFWLQJ JO\FDQ RQ K753$ 14 PD\ SUR
YLGHDUHDVRQZK\PD[LPDOFHOOUHVSRQVHVFRXOGEHGLPLQLVKHGGHVSLWHVOLJKWO\KLJKHUFHOOVXUIDFHH[SUHV
VLRQ
753$*DWLQJ0HFKDQLFVDUH$OWHUHGE\140XWDWLRQ
:7141414)LJ
XUH%:KLOHWKH(&RI&$GLIIHUHGFRQVLGHUDEO\EHWZHHQYDULDQWVGLIIHUHQFHVLQVDWXUDWLQJFRQ
FHQWUDWLRQVZHUHQRWREYLRXV,QGHHGWKHVWHHSHUVORSHIDFWRUVRIWKH&$GRVHUHVSRQVHFXUYHVSORWWHGIRU
14PXWDQWVUHODWLYHWR:7FRPELQHGZLWKUHGXFHG5)8YDOXHVDWORZHUFRQFHQWUDWLRQVPLJKWVXJ
JHVWWKDWWKHDEVHQFHRIDQ1JO\FDQDW1UHVXOWVLQDOWHUHGDFWLYDWLRQNLQHWLFVUDWKHUWKDQUHGXFHGDJR
QLVWDIILQLW\
,Q WKHRU\ QHLWKHU PXWDWLRQ VKRXOG DOWHU WKH DIILQLW\ RI OLSRSKLOLF VXOIK\GU\ODPLQHUHDFWLYH 753$
WKHLGHDWKDWWKHIXQFWLRQDOGLIIHUHQFHVEHWZHHQ753$YDULDQWVZHUHWKHUHVXOWRIJO\FDQSDUWLFLSDWLRQLQ
L
(DFKIOXRURPHWULFH[SHULPHQWLHLQGHSHQGHQWH[SHULPHQWGHVFULEHGFRQVLVWHGRIWULSOLFDWHUHDGLQJVDWHDFK
DJRQLVWFRQFHQWUDWLRQ
JDWLQJUDWKHUWKDQDJRQLVWELQGLQJ7RWHVWWKLVK\SRWKHVLVDQGWRGHWHUPLQHZKHWKHUWKHDOWHUHGVHQVLWLYLW\
ZHREVHUYHGZDVVSHFLILFWR1WHUPLQDOGRPDLQELQGLQJ753$DJRQLVWVZHSHUIRUPHGDGGLWLRQDOIOXR
WHUDFWLRQ VLWHV DORQJ WKH 6 GRPDLQ )LJXUH &' 7KH VLJQLILFDQW GLVSDULWLHV EHWZHHQ $,7& DQG
PHQWKROGRVHUHVSRQVHFXUYHVZHUHFRQVLVWHQWZLWKWKHYDULDQWVSHFLILFFKDQJHVWR&$VHQVLWLYLW\ZHRE
VHUYHG (& LQ ȝ0 &, $,7& :7 14 14
14 0HQWKRO :7 14 14
14)LJXUH&'
)LQDOO\UHVXOWVIURPHOHFWURSK\VLRORJLFDOH[SHULPHQWVDOVRVXSSRUWWKHDSSDUHQWIXQFWLRQDOVLJQLILFDQFH
UHGXFHGVHQVLWLYLW\WRORZHUFRQFHQWUDWLRQVRI&$DPRQJK753$YDULDQWVEHDULQJWKH14PXWDWLRQ
(& LQ ȝ0 &, :7 14 14
LQDJRQLVWFRQFHQWUDWLRQUHTXLUHGIRUKDOIPD[LPDODFWLYDWLRQRIK753$WKDWLVQRQJO\FRV\ODWHGDWSRVL
WLRQ1
9DULDQWVSHFLILF&RRSHUDWLYH(IIHFWVRI7HPSHUDWXUHDQG&$
$QDSSDUHQWFRRSHUDWLYLW\EHWZHHQFROGWHPSHUDWXUHDQGFKHPLFDODJRQLVWVLQWKHDFWLYDWLRQRI753FKDQ
7530 WKDW LQIOXHQFHG FKDQQHO WHPSHUDWXUH VHQVLWLYLW\ $FFRUGLQJO\ ZH UDQ IOXRURPHWULF DFWLYLW\
H[SHULPHQWVDW&LQSDUDOOHOWRWKRVHFRQGXFWHGDW&WRGHWHUPLQHZKHWKHUVXFKFRRSHUDWLYLW\EH
WZHHQWHPSHUDWXUHDQG&$ZDVHYLGHQWDQGZKHWKHULWFRXOGEHLQIOXHQFHGE\1JO\FRV\ODWLRQ
,QRXUH[SHULPHQWVn LQGHSHQGHQWH[SHULPHQWVZHREVHUYHGDVOLJKWDOEHLWVLJQLILFDQWGHFUHDVHLQ
WKHVHQVLWLYLW\RIK753$)/$*:7DQGWKH14PXWDQWDW&UHODWLYHWRUHFRUGLQJVDW&(&
LQ ȝ0 &, :7 14 )LJXUH $ & ,Q FRQWUDVW WKH DE
VHQFHRIWKH1JO\FRV\ODWLRQVLWHOHGWRHQKDQFHGVHQVLWLYLW\RI753$DWDPELHQWWHPSHUDWXUH(&
ILQGLQJV ZH GLG QRW KRZHYHU REVHUYH DQ\ GLUHFW DJRQLVWLF HIIHFW RI QR[LRXV FROG DV ORZ DV & RQ
K753$)/$* H[SUHVVHG LQ +(. FHOOV GDWD QRW VKRZQ 7KH FROGVHQVLWLYLW\ RI 753$ UHPDLQV
',6&866,21
2XUVWXG\LQWRWKHHIIHFWVRI1JO\FRV\ODWLRQRIK753$SURGXFHGVHYHUDOQHZLQVLJKWV,QDGGLWLRQWR
FRQILUPLQJWKDWERWKFRQVHQVXVVLWHVFDQEHPRGLILHGZLWKDQ1JO\FDQin vitroZHGHWHUPLQHGWKDWWKH
UHPRYDORIWKHVHVLWHVGRHVQRWSUHYHQWIXQFWLRQDODVVHPEO\RUFHOOVXUIDFHWUDIILFNLQJRIWKHSURWHLQ2Q
WKHFRQWUDU\LWDSSHDUVWKDWWKHDEVHQFHRIWKHJO\FDQDWSRVLWLRQ1UHVXOWVLQKLJKHUOHYHOVRISURWHLQ
ORFDOL]HGWRWKHFHOOVXUIDFH$GGLWLRQDOO\ZHLGHQWLILHGZKDWDSSHDUWREHLPSRUWDQWIXQFWLRQDOUROHVRI1
JO\FRV\ODWLRQDWWKH1FRQVHQVXVVLWH6SHFLILFDOO\ZHILUVWREVHUYHGWKDWWKH14PXWDWLRQLQIOX
HQFHVWKHHIILFLHQF\RIJO\FRSURFHVVLQJRIWKH1JO\FDQDW1/DWHUZHREVHUYHGWKDWWKHDEVHQFHRI
WKH 1 JO\FDQ UHGXFHV K753$ VHQVLWLYLW\ WR D YDULHW\ RI DJRQLVWV DQG WKRVH HIIHFWV ZHUH PRUH GUD
PDWLFDWSK\VLRORJLFDOWHPSHUDWXUHV
7KH HIIHFWV RI SURWHLQ JO\FRV\ODWLRQ DUH KLJKO\ YDULDEOH %HFDXVH RI WKLV GLYHUVLW\ WKH WDVN RI
FKDUDFWHUL]LQJ1JO\FDQVPXVW±DWOHDVWFXUUHQWO\±EHSHUIRUPHGRQDJO\FRSURWHLQE\JO\FRSURWHLQEDVLV
7KLVDSSHDUVSDUWLFXODUO\WUXHLQWKHFDVHRI753FKDQQHOV)RUH[DPSOH±LQHYLGHQWFRQWUDVWWRK753$
±UHPRYDORIWKH1JO\FDQRQUDW7539DSSHDUVWRHQKDQFHDJRQLVWVHQVLWLYLW\ZLWKUHVHDUFKHUVREVHUY
LQJDaIROGUHGXFWLRQLQWKH(&RIFDSVDLFLQLQQRQJO\FRV\ODWHGPXWDQWV$VVXFKGHWHUPLQLQJ
WKHVSHFLILFIXQFWLRQRI1JO\FRV\ODWLRQIRUHDFK753FKDQQHOSOD\VDFUXFLDOUROHLQH[SDQGLQJRXUXQGHU
VWDQGLQJRIERWKWKLVSURWHLQIDPLO\DQGPRUHEURDGO\WKHVWUXFWXUDODQGIXQFWLRQDOSURWHLQWUDLWVWKDWFDQ
EHFRQIHUUHGE\JO\FDQV6HYHUDOUHVHDUFKHUVKDYHDOUHDG\FRQWULEXWHGKHDYLO\LQWKLVUHJDUG±EXW
PDQ\753FKDQQHOERXQG1JO\FDQVUHPDLQXQVWXGLHG
*LYHQ WKH SRWHQWLDO FOLQLFDO VLJQLILFDQFH RI 753$ ± LQVLJKWV LQWR LWV VWUXFWXUH DQG IXQFWLRQ
FRXOGEHRIFRQVLGHUDEOHYDOXH2XUGDWDVXJJHVWDFULWLFDOUROHRIDQ1JO\FDQLQ753$JDWLQJPHFKDQ
LFVZLWKLWVDEVHQFHUHVXOWLQJLQGLPLQLVKHGFKDQQHOVHQVLWLYLW\WRORZHUFRQFHQWUDWLRQVRIDEURDGDUUD\RI
DJRQLVWV:KLOHWKDWILQGLQJPD\EHRIJUHDWHVWLQWHUHVWZLWKLQWKHFRQWH[WRIRXUUHVHDUFKLWEHDUVPHQ
WLRQLQJWKDWFRQILUPLQJ753$JO\FRV\ODWLRQDQGHOLPLQDWLQJWKHSRVVLELOLW\WKDWLWLVUHTXLUHGIRUHLWKHUD
SURWHLQDVVHPEO\RUEWUDIILFNLQJWRWKHFHOOVXUIDFHLVLQLWVHOIKLJKO\UHOHYDQW7KLVLVHVSHFLDOO\WUXHRI
WKHODWWHUDVWKH1JO\FRV\ODWLRQRI753FKDQQHOVKDVEHHQRFFDVLRQDOO\VKRZQWRKDYHDSURIRXQGLPSDFW
RQWKHOHYHOVRIFHOOVXUIDFHH[SUHVVLRQ
:KLOHWKHH[SUHVVLRQGDWDZHFROOHFWHGUHYHDOHGWKDWDJUHDWHUSURSRUWLRQRIK753$14ZDVOR
FDOL]HGWRWKHFHOOVXUIDFHWKDQWKDWRIK753$:7LWLVLPSRUWDQWWRDOVRFRQVLGHURXUILQGLQJWKDWWKH
UHPRYDORIWKHVPDOOHUJO\FDQDWSRVLWLRQ1UHVXOWHGLQPRUHUREXVWH[SUHVVLRQRIPDWXUHUHODWLYHWR
LPPDWXUHJO\FRSURWHLQ7KLVPLJKWVXJJHVWWKDWWKHHQKDQFHGVXUIDFHH[SUHVVLRQRIWKLV753$PXWDQW
DULVHVGXHWRLWVPRUHHIILFLHQWJO\FRSURFHVVLQJUDWKHUWKDQE\WKHDEVHQFHRIWKH1JO\FDQDW1per se
7KLVZRXOGSRLQWWRDFULWLFDOUROHIRUWKHFRPSOH[1JO\FDQDW1QDPHO\WKDWLWVSUHVHQFHSURPRWHV
RSWLPL]DWLRQRIFHOOVXUIDFHH[SUHVVLRQ7KHH[SUHVVLRQSDWWHUQVRIK753$SURWHLQVEHDULQJWKH14
PXWDWLRQDSSHDUWRVXSSRUWWKLVLGHD%RWKSURWHLQVSRVVHVVLQJWKLVPXWDWLRQH[KLELWHGUHGXFHGWRWDOH[
SUHVVLRQ UHODWLYH WR :7 DQG FRQVHTXHQWO\ UHGXFHG FHOOVXUIDFH H[SUHVVLRQ ,W LV XQFOHDU ZKHWKHU WKHVH
UHGXFWLRQVVLJQLI\OHVVHIILFLHQWSURWHLQSURFHVVLQJRULQFUHDVHGVXVFHSWLELOLW\WRGHJUDGDWLRQ5HJDUGOHVV
LWDSSHDUVWKDWRSWLPDOK753$H[SUHVVLRQSDWWHUQVPD\UHO\RQGLJO\FRV\ODWLRQ
7KHKHLJKWHQHGOHYHORIPDWXUHK753$HOLFLWHGE\WKH14PXWDWLRQFRXOGDULVHIURPVHYHUDOSRV
VLEOHFDXVHV)RUH[DPSOHLWLVEHFRPLQJLQFUHDVLQJO\FOHDUWKDWVHTXHQFHVDGMDFHQWWRWKH1X-67FRQ
VHQVXV VLWH FDQ LQIOXHQFH JO\FRV\ODWLRQ DQGRU JO\FRSURFHVVLQJ ± $GGLWLRQDO HYLGHQFH KLJKOLJKWV
WKHLQIOXHQFHWKDWWKHGLVWDQFHEHWZHHQ1JO\FRV\ODWLRQDFFHSWRUVLWHVFDQKDYHRQWKHHIILFLHQF\RIJO\FR
SURFHVVLQJ:KLOHRQHFRXOGK\SRWKHVL]HWKDWDQXSVWUHDP14PXWDWLRQPLJKWUHVXOWLQXQQDWXUDO
JO\FRSURFHVVLQJ DW SRVLWLRQ 1 SRWHQWLDOO\ UHVXOWLQJ LQ WKHDOWHUHG SURWHLQ IXQFWLRQDOLW\ WKDW FRQWUD
GLFWVRXUILQGLQJWKDWWKH14GRXEOHPXWDQWSURGXFHVVLPLODUDJRQLVWLQGXFHGEHKDYLRUWRWKDWRI
K753$ 14 :H ILQG LW PRUH OLNHO\ WKDW FORVH DSSUR[LPDWLRQ RI WKH JO\FRV\ODWLRQ VLWHV DQGRU WKH
DPLQRDFLGVQHDUE\WKHUHVSHFWLYHVHTXRQVDUHWKHFDXVHRIWKHDSSDUHQWLQWHUSOD\EHWZHHQWKHVHWZRVLWHV
,QOLJKWRIWKHGLVSDUDWHOHYHOVRIFHOOVXUIDFHH[SUHVVLRQZHREVHUYHGLWZDVVXUSULVLQJWKDWWKHPD[L
PDO753$VSHFLILFUHVSRQVHVREVHUYHGLQZKROHFHOOSDWFKFODPSDQGIOXRURPHWULFDVVD\VGLGQRWGLIIHU
VLJQLILFDQWO\EHWZHHQ753$YDULDQWV+RZHYHUPD[LPDOIXQFWLRQDOUHVSRQVHVPD\QRWGLUHFWO\FRUUHODWH
ZLWK753$FHOOVXUIDFHH[SUHVVLRQ$FWLYDWLRQRI753$LQLQWUDFHOOXODURUJDQHOOHVZKRVHH[SUHVVLRQ
LVQRWUHIOHFWHGLQRXUPHPEUDQHIUDFWLRQ:%DQDO\VLV PD\LQIOXHQFHFXUUHQWDQGYROWDJHVHQVLWLYHG\H
UHDGRXWV ,Q DGGLWLRQ LQ WKH SDWFKFODPS H[SHULPHQWV ZH PD\ KDYH KDG D ELDV WRZDUGV VHOHFWLQJ FHOOV
ZLWK D ORZ OHYHO RI 753$ H[SUHVVLRQ DV WKHVH FHOOV DUH OLNHO\ WR DSSHDU ³KHDOWKLHU´ WKDQ FHOOV ZLWK D
KLJKHUH[SUHVVLRQOHYHO8OWLPDWHO\WKHSURGXFWLRQRIVWDEOHFHOOOLQHVZRXOGEHXVHIXOLQUHVROYLQJWKHVH
GLVFUHSDQFLHV
*LYHQWKHFRQVLVWHQF\RIRXUUHVXOWVIURPWKHIXQFWLRQDODQDO\VLVERWKDFURVVGLIIHUHQWDVVD\VDQGEH
WZHHQDJRQLVWVZLWKGLVWLQFWUHDFWLYLW\SURILOHVZHDUHFRQILGHQWLQWKHIXQFWLRQDOVLJQLILFDQFHRIWKHJO\
FDQDWSRVLWLRQ1$VIXUWKHUVXSSRUWWKH(&YDOXHVZHJHQHUDWHGIURPIOXRURPHWULFPHDVXUHPHQWV
RIK753$)/$*:7PHGLDWHGPHPEUDQHGHSRODUL]DWLRQFORVHO\UHVHPEOHWKRVHREVHUYHGE\RWKHUUH
VHDUFKHUV XVLQJ +(. FHOOV LQ VLPLODU DVVD\V IRU DOO WKUHH DJRQLVWV DQG ZH OLNHZLVH RE
VHUYHG,&YDOXHVRIWKH753$VHOHFWLYHDQWDJRQLVW+&WKDWZHUHFRQVLVWHQWZLWKWKHOLWHUDWXUH
GDWDQRWVKRZQ7KHVHILQGLQJVWRRXUWKLQNLQJYDOLGDWHWKHXVHRIWUDQVLHQWO\WUDQVIHFWHG+(.
FHOOVLQIOXRURPHWULFDVVD\V
,QRXUUHYLHZRIWKHOLWHUDWXUHZHGLGQRWXQFRYHUPDQ\IXOOGRVHUHVSRQVHFXUYHVRI753$JHQHUDWHG
LQDZKROHFHOOSDWFKFODPSIRUPDWLQKHWHURORJRXVH[SUHVVLRQV\VWHPV+RZHYHU&KXQJHWDOXVHGSUH
FLVHO\ VXFK D IRUPDW WR HYDOXDWH WKH UHVSRQVH RI 753$ WR LWV DJRQLVW HXJHQRO :H IRXQG WKDW
753$H[SUHVVLQJ+(.FHOOVZHUHKLJKO\DPHQDEOHWRHOHFWURSK\VLRORJLFDODQDO\VLVZLWKVHULDOGLOX
WLRQVRI&$DSSOLHGWRVLQJOHFHOOVSURGXFLQJODUJHVORZO\LQDFWLYDWLQJLQZDUGFXUUHQWVWKDWZDQHGRQO\
VORZO\XSRQUHPRYDORIWKHDJRQLVW)LJXUH&7KLVHQDEOHGXVWRFRQILUPWKHHIIHFWVRIWKH14PX
WDWLRQZKLFKRQFHDJDLQGHPRQVWUDWHGDQRWDEOHORVVRIDJRQLVWVHQVLWLYLW\$OWKRXJKWKH(&YDOXHVGH
WHUPLQHGE\ZKROHFHOOSDWFKFODPSDQGIOXRURPHWULFDVVD\VGLIIHUHGFRQVLGHUDEO\ZHFRQVLGHUHGWKLVD
FRQVHTXHQFHRIWKHGLIIHUHQFHVLQKHUHQWWRWKHUHVSHFWLYHDVVD\V,QDGGLWLRQWRGLVFUHSDQFLHVLQDVVD\VHQ
VLWLYLWLHVRWKHUIDFWRUVVXFKDVYROWDJHFRQWURODQGRUWKHWUHDWPHQWRI&$QDwYHFHOOVLQWKH)OH[6WDWLRQ
WRWKHREVHUYHGGLIIHUHQFHV
5HFHQWO\ 'HHULQJ5LFH HW DO SXEOLVKHG UHVXOWV IURP D IXQFWLRQDO DQDO\VLV RI QRQJO\FRV\ODWHG
753$YDULDQWV7KHLUZRUNHQWDLOHGDQDO\]LQJFDOFLXPLQIOX[DIWHUDVLQJOHDSSOLFDWLRQRIDVDWXUDWLQJ
FRQFHQWUDWLRQ RI 753$ DJRQLVWV WR +(. FHOOV WUDQVLHQWO\ H[SUHVVLQJ HLWKHU :7 RU 1$ DQGRU
1$ 753$ YDULDQWV 7R UXOH RXW GLVSDUDWH VXEFHOOXODU SURWHLQ GLVWULEXWLRQ DV D FDXVH RI GLVSDUDWH
FKDQQHO UHVSRQVHV WKH UHVHDUFKHUV HQGHDYRUHG WR DFWLYDWH LQWUDFHOOXODU 753$ LQ LVRODWLRQ E\ DSSO\LQJ
753$DJRQLVWVLQWKHSUHVHQFHRIWKHPHPEUDQHLPSHUPHDEOH753$LQKLELWRUUXWKHQLXPUHG:KLOH
WKH\IRXQGGLPLQLVKHGUHVSRQVHVDPRQJ753$YDULDQWVEHDULQJWKH1$PXWDWLRQWKH\GLGQRW±DV
ZHGLG±LGHQWLI\VLJQLILFDQWGLIIHUHQFHVLQSURWHLQGLVWULEXWLRQ:HEHOLHYHWKLVLVOLNHO\DFRQVHTXHQFHRI
WKHGLIIHUHQWDQDO\WLFDODSSURDFKHVQDPHO\WKHXVHRILPPXQREORWWLQJRILVRODWHGFHOOVXUIDFHSURWHLQVYV
DQLQWUDFHOOXODUDFWLYLW\DVVD\WRGHWHUPLQHGLIIHUHQFHVLQ753$FHOOVXUIDFHH[SUHVVLRQ
7KHLQIOXHQFHRI1JO\FRV\ODWLRQRQWKHDSSDUHQWFRRSHUDWLYHHIIHFWVRIWHPSHUDWXUHDQG&$ZDVDVXU
SULVLQJ ILQGLQJ LQ WKLV VWXG\ ,Q SDUWLFXODU LW LV LQWHUHVWLQJ WKDW WKH SUHVHQFH RI ERWK JO\FRV\ODWLRQ VLWHV
DSSHDUVWRRSWLPL]H753$DJRQLVWVHQVLWLYLW\DWSK\VLRORJLFDODQGDPELHQWWHPSHUDWXUHV3HUKDSVLQWKH
FDVHRI753$GLJO\FRV\ODWLRQVHUYHGDVDNLQGRIHYROXWLRQDU\FDOLEUDWLRQHVSHFLDOO\LQOLJKWRIWKHIDFW
WKDWWKH1JO\FRV\ODWLRQVLWHLVPRUHHYROXWLRQDULO\UHFHQWZLWKFROGEORRGHGDPSKLELDQVDQGUHSWLOHV
ODFNLQJLW,WZRXOGEHLQWHUHVWLQJWRVHHZKHWKHUVLPLODUSDWWHUQVDUHSUHVHQWDPRQJRWKHUWKHUPRVHQVLWLYH
753FKDQQHOV
7KHUH DUH VRPH OLPLWDWLRQV LQKHUHQW WR WKLV DQDO\VLV WKDW EHDU DFNQRZOHGJHPHQW 6SHFLILFDOO\ LW UH
PDLQVXQFOHDUZKHWKHU753$1JO\FRV\ODWLRQRFFXUVLQQDWLYHFHOOOLQHVin vivo+RZHYHUUHVXOWVSXE
OLVKHGE\RWKHUUHVHDUFKJURXSVFRQVLVWHQWO\LQGLFDWHWKDW753FKDQQHOVDUH1JO\FRV\ODWHGin vivo
PXWDWLRQV LQGXFH FRQIRXQGLQJ FKDQJHV WR SURWHLQ VWUXFWXUH WKDW PD\ REVFXUH RWKHU HIIHFWV RI JO\FRV\OD
WLRQ $OVR WKHUH ZHUH PXOWLSOH 753$ VXEXQLW JO\FRIRUPV SUHVHQW LQ VLQJOH FHOOV DV HYLGHQFHG E\ WKH
GRXEOHW EDQGV REVHUYHG LQ VLQJOH VDPSOHV LQ :% DQDO\VLV ,QGLYLGXDO 753$ YDULDQW JO\FRIRUPV PD\
FRQWULEXWH GLVSDUDWHO\ WR IXQFWLRQDO UHDGRXWV DQG WKH H[SHULPHQWV ZH SHUIRUPHG HIIHFWLYHO\ DJJUHJDWH
WKHVHFRQWULEXWLRQV
)LQDOO\753$LVSRWHQWLDWHGDQGLQDFWLYDWHGE\&D,WLVSRVVLEOHWKDW:7DQGPXWDWHG753$
FKDQQHOVDQDO\]HGLQRXUH[SHULPHQWVGLIIHULQWKHLUSHUPHDELOLW\WR&DZKLFKLQWXUQFRXOGDIIHFWFXU
UHQWDPSOLWXGHVWKURXJK&DGHSHQGHQW753$SRWHQWLDWLRQ2QHPLJKWFRQVLGHUVXFKGLVSDULWLHVDV
DQ DOWHUQDWLYH H[SODQDWLRQ IRU WKH REVHUYHG GLVFUHSDQFLHV EHWZHHQ RXU H[SUHVVLRQ DQG IXQFWLRQDO GDWD
+RZHYHULWLVXQFOHDUZKDWHIIHFWVVXFKGLVSDULWLHVZRXOGKDYHLIDQ\RQWKHGRVHUHVSRQVHUHODWLRQVKLSV
EHWZHHQDJRQLVWVDQGWKH753$YDULDQWVXQGHUWKHH[SHULPHQWDOGHVLJQZHDSSOLHG,QRXUH[SHULPHQWV
WKHDEVHQFHRIVLJQLILFDQWGLIIHUHQFHVLQPD[LPDO753$VSHFLILFUHVSRQVHVXSSRUWVWKHVWDWLVWLFDOFRP
SDULVRQ RI QRUPDOL]HG GRVHUHVSRQVH FXUYHV )XUWKHUPRUH ZH GLG QRW REVHUYH D GLVFHUQLEOH SDWWHUQ LQ
753$FKDQQHOIXQFWLRQWKDWZDVVXJJHVWLYHRIWKHSUHVHQFHRIFRQIRXQGLQJVWUXFWXUDOFKDQJHVRUGLVSDU
DWH&DGHSHQGHQWSRWHQWLDWLRQEURXJKWRQE\WKHPXWDWLRQWRHLWKHUVLWH2QWKHFRQWUDU\WKHWZRYDUL
DQWVEHDULQJWKH14PXWDWLRQH[KLELWYHU\GLIIHUHQWH[SUHVVLRQSDWWHUQVEXWSURGXFHVLPLODUPD[LPDO
VLJQDOV DQG GRVHUHVSRQVH FXUYHV ZKHQ VWLPXODWHG E\ GLYHUVH DJRQLVWV DQG XQGHU GLIIHUHQW H[SHULPHQWDO
DSSURDFKHV
2QHRIWKHFRPPRQIHDWXUHVRI753FKDQQHOVLVWKHLUIXQFWLRQDO DQGVWUXFWXUDOYHUVDWLOLW\7KH\DUH
W\SLFDOO\SRO\PRGDOO\DFWLYDWHGXELTXLWRXVO\H[SUHVVHGDQGFDQERWKKHWHURDQGKRPRPXOWLPHUL]H±
7KHUHYHODWLRQWKDW753$EHKDYLRUFDQEHPRGLILHGE\DOWHUHGJO\FRV\ODWLRQSDWWHUQVSRWHQWLDOO\XQ
ORFNV HYHQ PRUH IXQFWLRQDO GLYHUVLW\ *O\FRV\OWUDQVIHUDVHV DUH GLIIHUHQWLDOO\ H[SUHVVHG GXULQJ GHYHORS
IHUDVHH[SUHVVLRQFRXOGHQVXUHWKHWDUJHWHGSUHVHQFHRIIXQFWLRQDOO\XQLTXH753FKDQQHOVDOORZLQJSUR
WHLQVZLWKLGHQWLFDOSULPDU\VWUXFWXUHVWRDFFRPPRGDWHGLIIHUHQWIXQFWLRQDOUROHV,QWKHFRQWH[WRIWKHU
PRVHQVLWLYH753FKDQQHOVSURFHVVHVVXFKDVJO\FRV\ODWLRQPD\DOVREHDPHDQVWRRSWLPL]HIXQFWLRQRYHU
WKHUDQJHRIWHPSHUDWXUHVWRZKLFKDQH[SUHVVLRQORFXVLVW\SLFDOO\H[SRVHG)RUH[DPSOHVHQVRU\UHFHS
WRUVLQQHUYDWLQJWKHVNLQDUHH[SRVHGWRORZHUWHPSHUDWXUHVWKDQWKDWRIWKHFRUHDQGWKHIXQFWLRQDOGH
PDQGVRIWKHVHUHVSHFWLYHHQYLURQPHQWVFRXOGOLNHZLVHGLIIHU
7KLVVWXG\SURYLGHVQHZLQVLJKWVLQWRWKHFRPSOH[FKDQQHOEHKDYLRUDQGLWVWLJKWO\FRQWUROOHGUHJXOD
WLRQ2XUFRQILUPDWLRQRI753$GLJO\FRV\ODWLRQDQGWKHLQGLFDWLRQVWKDWWKLVPRGLILFDWLRQFDUULHVFRQ
VLGHUDEOHIXQFWLRQDOLPSRUWDQFHIRUWKHSURWHLQHQKDQFHVRXUXQGHUVWDQGLQJRIQRWRQO\753$EHKDYLRU
EXWDOVRRIWKHEURDGHU753FKDQQHOIDPLO\DVZHOODVRIWKHYHUVDWLOLW\RI1JO\FDQV
$&.12:/('*0(176
6SHFLDOWKDQNVDUHRZHGWR6HEDVWLDQ0H\HU7HD*XQGHDQG$OH[DQGUH6LPRQLQIRULQSXWDWYDULRXVFULWL
FDOMXQFWXUHVGXULQJWKLVSURMHFW
'(&/$5$7,2162),17(5(67
7KHDXWKRUVGHFODUHWKDWWKH\KDYHQRFRQIOLFWVRILQWHUHVWZLWKWKHFRQWHQWVRIWKLVDUWLFOH
)81',1*
1XPDE$*DQGWKH,QVWLWXWHRI3KDUPDFRORJ\DQG7R[LFRORJ\DW8=+SURYLGHGWKHIXQGLQJIRUWKLVUH
VHDUFK
$87+25&2175,%87,21
7LPRWK\-(JDQGHVLJQHGWKHVWXG\SHUIRUPHGDOOH[SHULPHQWVDQGZURWHWKHSDSHU0DULR$$FXxDDQG
+DQQV8OULFK=HLOKRIHUKHOSHGGHVLJQWKHHOHFWURSK\VLRORJ\H[SHULPHQWVDQG0DULR$$FXxDDVVLVWHGLQ
WKHSDWFKFODPSH[SHULPHQWV'DYLG8UHFKDQG0DUF\=HQREL:RQJDVVLVWHGLQWKHGHVLJQRIWKHVWXG\
$OODXWKRUVDQDO\]HGWKHVHUHVXOWVDQGUHYLHZHGDQGDSSURYHGWKLVPDQXVFULSWSULRUWRVXEPLVVLRQ
5()(5(1&(6
/DWRUUH5=DHO]HU&%UDXFKL66WUXFWXUHIXQFWLRQDOLQWLPDFLHVRIWUDQVLHQWUHFHSWRUSRWHQWLDO
FKDQQHOV45HY%LRSK\V±
&ODSKDP'(753FKDQQHOVDVFHOOXODUVHQVRUV1DWXUH±
1HOVRQ3/%HFN$&KHQJ+7UDQVLHQWUHFHSWRUSURWHLQVLOOXPLQDWHG&XUUHQWYLHZVRQ753VDQG
GLVHDVH9HW->,QWHUQHW@(OVHYLHU/WG±$YDLODEOHIURP
KWWSG[GRLRUJMWYMO
1LOLXV%2ZVLDQLN*7KHWUDQVLHQWUHFHSWRUSRWHQWLDOIDPLO\RILRQFKDQQHOV*HQRPH%LRO
1LOLXV%2ZVLDQLN*9RHWV73HWHUV-$7UDQVLHQWUHFHSWRUSRWHQWLDOFDWLRQFKDQQHOVLQGLVHDVH
3K\VLRO5HY±
9HQNDWDFKDODP.0RQWHOO&753FKDQQHOV$QQX5HY%LRFKHP±
0RUDQ000F$OH[DQGHU0$%tUy76]DOODVL$7UDQVLHQWUHFHSWRUSRWHQWLDOFKDQQHOVDV
WKHUDSHXWLFWDUJHWV1DW5HY'UXJ'LVFRY>,QWHUQHW@1DWXUH3XEOLVKLQJ*URXS±
$YDLODEOHIURPKWWSG[GRLRUJQUG
&RKHQ'05HJXODWLRQRI753FKDQQHOVE\1OLQNHGJO\FRV\ODWLRQ6HPLQ&HOO'HY%LRO
±
&KDQJ4+RHIV6YDQGHU.HPSD:7RSDOD&1%LQGHOV5-+RHQGHURS-*7KHEHWD
JOXFXURQLGDVHNORWKRK\GURO\]HVDQGDFWLYDWHVWKH7539FKDQQHO6FLHQFH±
'UDJRQL,*XLGD(0F,QW\UH37KHFROGDQGPHQWKROUHFHSWRU7530FRQWDLQVDIXQFWLRQDOO\
LPSRUWDQWGRXEOHF\VWHLQHPRWLI-%LRO&KHP±
(UOHU,$O$QVDU\'00:LVVHQEDFK8:DJQHU7)-)ORFNHU]L91LHPH\HU%D7UDIILFNLQJ
DQGDVVHPEO\RIWKHFROGVHQVLWLYH7530FKDQQHO-%LRO&KHP±
3HUWXVD00DGULG50RUHQLOOD3DODR&%HOPRQWH&9LDQD)1JO\FRV\ODWLRQRI7530LRQ
FKDQQHOVPRGXODWHVWHPSHUDWXUHVHQVLWLYLW\RIFROGWKHUPRUHFHSWRUQHXURQV-%LRO&KHP
±
:LUNQHU.+RJQHVWDG+-DKQHO5+XFKR)DQG,OOHV3&KDUDFWHUL]DWLRQRIUDWWUDQVLHQW
UHFHSWRUSRWHQWLDOYDQLOORLG,UHFHSWRUVODFNLQJWKH1JO\FRV\ODWLRQVLWH11HXURSKDUPDFRO
1HXURWR[LFRORJ\±
:RR6..ZRQ06,YDQRY$*HQJ=*HU]DQLFK96LPDUG-0&RPSOH[1*O\FRV\ODWLRQ
VWDELOL]HVVXUIDFHH[SUHVVLRQRIWUDQVLHQWUHFHSWRUSRWHQWLDOPHODVWDWLQESURWHLQ-%LRO&KHP
±
;X+)X<7LDQ:&RKHQ'0*O\FRV\ODWLRQRIWKHRVPRUHVSRQVLYHWUDQVLHQWUHFHSWRUSRWHQWLDO
FKDQQHO7539RQ$VQLQIOXHQFHVPHPEUDQHWUDIILFNLQJ$P-3K\VLRO5HQDO3K\VLRO
)±
'LHWULFK$0HGHURV<6FKQLW]OHU0(PPHO-.DOZD++RIPDQQ7*XGHUPDQQ71/LQNHG
3URWHLQ*O\FRV\ODWLRQ,VD0DMRU'HWHUPLQDQWIRU%DVDO753&DQG753&&KDQQHO$FWLYLW\-
%LRO&KHP±
6WDQOH\36FKDFKWHU+7DQLJXFKL11*O\FDQV,Q9DUNL$&XPPLQJV5'(VNR-')UHH]H
++6WDQOH\3%HUWR]]L&5HWDOHGLWRUV(VVHQWLDOVRI*O\FRELRORJ\QGHG&ROG6SULQJ
+DUERU1<&ROG6SULQJ+DUERU/DERUDWRU\3UHVV
9DUNLD%LRORJLFDOUROHVRIROLJRVDFFKDULGHVDOORIWKHWKHRULHVDUHFRUUHFW*O\FRELRORJ\
±
%DQGHOO06WRU\*0+ZDQJ6:9LVZDQDWK9(LG653HWUXV0-HWDO1R[LRXVFROGLRQ
FKDQQHO753$LVDFWLYDWHGE\SXQJHQWFRPSRXQGVDQGEUDG\NLQLQ1HXURQ±
%ULHUOH\60+XJKHV3D3DJH$-.ZDQ.<0DUWLQ&02¶'RQQHOO7DHWDO7KH,RQ&KDQQHO
753$,V5HTXLUHGIRU1RUPDO0HFKDQRVHQVDWLRQDQG,V0RGXODWHGE\$OJHVLF6WLPXOL
*DVWURHQWHURORJ\>,QWHUQHW@(OVHYLHU,QF±H$YDLODEOHIURP
KWWSG[GRLRUJMJDVWUR
'RHUQHU-)*LVVHOPDQQ*+DWW+:HW]HO&+7UDQVLHQWUHFHSWRUSRWHQWLDOFKDQQHO$LVGLUHFWO\
JDWHGE\FDOFLXPLRQV-%LRO&KHP±
+X+%DQGHOO03HWUXV0-=KX0;=LQFDFWLYDWHVGDPDJHVHQVLQJ753$LRQFKDQQHOV1DW
&KHP%LRO±
.ZDQ.<$OOFKRUQH$-9ROOUDWK0D&KULVWHQVHQ$3=KDQJ'6:RROI&-HWDO753$
&RQWULEXWHVWR&ROG0HFKDQLFDODQG&KHPLFDO1RFLFHSWLRQEXW,V1RW(VVHQWLDOIRU+DLU&HOO
7UDQVGXFWLRQ1HXURQ±
/HH63%XEHU07<DQJ4&HUQH5&RUWpV5<6SURXV'*HWDO7K\PRODQGUHODWHGDON\O
SKHQROVDFWLYDWHWKHK753$FKDQQHO%U-3KDUPDFRO±
0DFSKHUVRQ/-*HLHUVWDQJHU%+9LVZDQDWK9%DQGHOO0(LG65+ZDQJ6HWDO7KH
SXQJHQF\RIJDUOLF$FWLYDWLRQRI753$DQG7539LQUHVSRQVHWRDOOLFLQ&XUU%LRO
±
0DFSKHUVRQ/-'XELQ$((YDQV0-0DUU)6FKXOW]3*&UDYDWW%)HWDO1R[LRXVFRPSRXQGV
DFWLYDWH753$LRQFKDQQHOVWKURXJKFRYDOHQWPRGLILFDWLRQRIF\VWHLQHV1DWXUH
±
6WRU\*03HLHU$05HHYH$-(LG650RVEDFKHU-+ULFLN75HWDO$1.70D753OLNH
FKDQQHOH[SUHVVHGLQQRFLFHSWLYHQHXURQVLVDFWLYDWHGE\FROGWHPSHUDWXUHV&HOO
±
7DNDKDVKL1.XZDNL7.L\RQDND61XPDWD7.R]DL'0L]XQR<HWDO753$XQGHUOLHVD
VHQVLQJPHFKDQLVPIRU21DW&KHP%LRO±
;LDR%'XELQ$(%XUVXOD\D%9LVZDQDWK9-HJOD7-3DWDSRXWLDQ$,GHQWLILFDWLRQRI
WUDQVPHPEUDQHGRPDLQDVDFULWLFDOPROHFXODUGHWHUPLQDQWRIPHQWKROVHQVLWLYLW\LQPDPPDOLDQ
753$FKDQQHOV-1HXURVFL±
=KRQJ-3ROODVWUR)3UHQHQ-=KX=$SSHQGLQR*1LOLXV%/LJXVWLOLGH$QRYHO753$
PRGXODWRU3IOXJHUV$UFK(XU-3K\VLRO±
*UDFKHYD(2,QJROLD17.HOO\<0&RUGHUR0RUDOHV-)+ROORSHWHU*&KHVOHU$7HWDO
0ROHFXODUEDVLVRILQIUDUHGGHWHFWLRQE\VQDNHV1DWXUH±
1LOLXV%3UHQHQ-2ZVLDQLN*,UULWDWLQJFKDQQHOVWKHFDVHRI753$-3K\VLRO3W
±
5*DUULVRQ6/6WXFN\&7KH'\QDPLF753$&KDQQHO$6XLWDEOH3KDUPDFRORJLFDO3DLQ
7DUJHW"&XUU3KDUP%LRWHFKQRO±
6RWRPD\RU0&RUH\'36FKXOWHQ.,QVHDUFKRIWKHKDLUFHOOJDWLQJVSULQJ(ODVWLFSURSHUWLHVRI
DQN\ULQDQGFDGKHULQUHSHDWV6WUXFWXUH±
&RUGHUR0RUDOHV-)*UDFKHYD(2-XOLXV'31$63OXV&\WRSODVPLFDQN\ULQUHSHDWVRIWUDQVLHQW
UHFHSWRUSRWHQWLDO$753$GLFWDWHVHQVLWLYLW\WRWKHUPDODQGFKHPLFDOVWLPXOL3URF1DWO$FDG
6FL(±
&KXQJ*,P67.LP<+-XQJ6-5K\X052K6%$FWLYDWLRQRIWUDQVLHQWUHFHSWRUSRWHQWLDO
DQN\ULQE\HXJHQRO1HXURVFLHQFH>,QWHUQHW@±$YDLODEOHIURP
KWWSZZZVFLHQFHGLUHFWFRPVFLHQFHDUWLFOHSLL6
3DXOVHQ&($UPDFKH-*DR<&KHQJ<-XOLXV'6WUXFWXUHRIWKH753$LRQFKDQQHOVXJJHVWV
UHJXODWRU\PHFKDQLVPV1DWXUH
+LQPDQ$&KXDQJ++%DXWLVWD'0-XOLXV'753FKDQQHODFWLYDWLRQE\UHYHUVLEOHFRYDOHQW
PRGLILFDWLRQ3URF1DWO$FDG6FL86$±
&XPPLQJV5'(W]OHU0($QWLERGLHVDQG/HFWLQVLQ*O\FDQ$QDO\VLV,Q9DUNL$&XPPLQJV
5'(VNR-')UHH]H++6WDQOH\3%HUWR]]L&5HWDOHGLWRUV(VVHQWLDOVRI*O\FRELRORJ\QG
HG&ROG6SULQJ+DUERU1<&ROG6SULQJ+DUERU/DERUDWRU\3UHVV
3HLHU$00RTULFK$+HUJDUGHQ$&5HHYH$-$QGHUVVRQ'D6WRU\*0HWDO$753FKDQQHO
WKDWVHQVHVFROGVWLPXOLDQGPHQWKRO&HOO±
0F.HP\''1HXKDXVVHU:0-XOLXV',GHQWLILFDWLRQRIDFROGUHFHSWRUUHYHDOVDJHQHUDOUROH
IRU753FKDQQHOVLQWKHUPRVHQVDWLRQ1DWXUH±
-RUGW6(%DXWLVWD'0&KXDQJ++0F.HP\''=\JPXQW30+|JHVWlWW('HWDO0XVWDUG
RLOVDQGFDQQDELQRLGVH[FLWHVHQVRU\QHUYHILEUHVWKURXJKWKH753FKDQQHO$1.701DWXUH
±
&KHQ-.DQJ';X-/DNH0+RJDQ-26XQ&HWDO6SHFLHVGLIIHUHQFHVDQGPROHFXODU
GHWHUPLQDQWRI753$FROGVHQVLWLYLW\1DW&RPPXQ>,QWHUQHW@1DWXUH3XEOLVKLQJ*URXS
$YDLODEOHIURP
KWWSZZZSXEPHGFHQWUDOQLKJRYDUWLFOHUHQGHUIFJL"DUWLG WRRO SPFHQWUH] UHQGHUW\SH
DEVWUDFW
=XUERUJ6<XUJLRQDV%-LUD-D&DVSDQL2+HSSHQVWDOO3D'LUHFWDFWLYDWLRQRIWKHLRQFKDQQHO
753$E\&D1DW1HXURVFL±
0RSDUWKL/6XUYHU\6.UHLU06LPRQVHQ&.MHOOERP3+|JHVWlWW('HWDO+XPDQ753$LV
LQWULQVLFDOO\FROGDQGFKHPRVHQVLWLYHZLWKDQGZLWKRXWLWV1WHUPLQDODQN\ULQUHSHDWGRPDLQ3URF
1DWO$FDG6FL>,QWHUQHW@±$YDLODEOHIURP
KWWSZZZSQDVRUJORRNXSGRLSQDV
:RRG+3DLQQHZIDPLOLDOSDLQV\QGURPHFDXVHGE\753$PXWDWLRQ1DW5HY1HXURO
>,QWHUQHW@1DWXUH3XEOLVKLQJ*URXS$YDLODEOHIURP
KWWSG[GRLRUJQUQHXURO
3HWUXV03HLHU$0%DQGHOO0+ZDQJ6:+X\QK72OQH\1HWDO$UROHRI753$LQ
PHFKDQLFDOK\SHUDOJHVLDLVUHYHDOHGE\SKDUPDFRORJLFDOLQKLELWLRQ0RO3DLQ
0F1DPDUD&50DQGHO%UHKP-%DXWLVWD'06LHPHQV-'HUDQLDQ./=KDR0HWDO753$
PHGLDWHVIRUPDOLQLQGXFHGSDLQ3URF1DWO$FDG6FL86$±
/HQQHUW]5&.RVV\UHYD(D6PLWK$.6WXFN\&/753$PHGLDWHVPHFKDQLFDOVHQVLWL]DWLRQLQ
QRFLFHSWRUVGXULQJLQIODPPDWLRQ3/R62QH±
.UHPH\HU%/RSHUD)&R[--0RPLQ$5XJLHUR)0DUVK6HWDO$*DLQRI)XQFWLRQ0XWDWLRQ
LQ753$&DXVHV)DPLOLDO(SLVRGLF3DLQ6\QGURPH1HXURQ±
(LG65&URZQ('0RRUH(//LDQJ+D&KRRQJ.&'LPD6HWDO+&D753$
VHOHFWLYHDQWDJRQLVWDWWHQXDWHVLQIODPPDWRU\DQGQHXURSDWK\LQGXFHGPHFKDQLFDO
K\SHUVHQVLWLYLW\0RO3DLQ
&KHQ--RVKL6.'LGRPHQLFR63HUQHU5-0LNXVD-3*DXYLQ'0HWDO6HOHFWLYHEORFNDGHRI
753$FKDQQHODWWHQXDWHVSDWKRORJLFDOSDLQZLWKRXWDOWHULQJQR[LRXVFROGVHQVDWLRQRUERG\
WHPSHUDWXUHUHJXODWLRQ3DLQ>,QWHUQHW@,QWHUQDWLRQDO$VVRFLDWLRQIRUWKH6WXG\RI3DLQ
±$YDLODEOHIURPKWWSG[GRLRUJMSDLQ
%DXWLVWD'0-RUGW6(1LNDL77VXUXGD355HDG$-3REOHWH-HWDO753$0HGLDWHVWKH
,QIODPPDWRU\$FWLRQVRI(QYLURQPHQWDO,UULWDQWVDQG3URDOJHVLF$JHQWV&HOO±
%Dxy3ROR0%DOGLQ)7DPERUHUR60DUWL5HQRP0D0LQJDUUR,NJO\FRV\ODWLRQHIILFLHQF\
LVGHWHUPLQHGE\WKHGLVWDQFHWRWKH&WHUPLQXVDQGWKHDPLQRDFLGSUHFHGLQJDQ$VQ6HU7KU
VHTXRQ3URWHLQ6FL>,QWHUQHW@±$YDLODEOHIURP
KWWSGRLZLOH\FRPSUR
%HQ'RU6(VWHUPDQ15XELQ(6KDURQ1%LDVHVDQGFRPSOH[SDWWHUQVLQWKHUHVLGXHVIODQNLQJ
SURWHLQ1JO\FRV\ODWLRQVLWHV*O\FRELRORJ\>,QWHUQHW@±$YDLODEOHIURP
KWWSZZZQFELQOPQLKJRYSXEPHG
3HWUHVFX$-0LODF$/3HWUHVFX60'ZHN5$:RUPDOG056WDWLVWLFDODQDO\VLVRIWKHSURWHLQ
HQYLURQPHQWRI1JO\FRV\ODWLRQVLWHVLPSOLFDWLRQVIRURFFXSDQF\VWUXFWXUHDQGIROGLQJ
*O\FRELRORJ\>,QWHUQHW@±$YDLODEOHIURP
KWWSZZZJO\FRERXSMRXUQDOVRUJFJLGRLJO\FREFZK
6KULPDO6*LOPRUH5*O\FRV\ODWLRQRIFORVHO\VSDFHGDFFHSWRUVLWHVLQKXPDQJO\FRSURWHLQV-
&HOO6FL3W±
'HHULQJULFH&(6KDSLUR'5RPHUR(*6WRFNPDQQ&%HYDQV763KDQ40HWDO$FWLYDWLRQRI
7UDQVLHQW5HFHSWRU3RWHQWLDO$QN\ULQE\,QVROXEOH3DUWLFXODWH0DWHULDODQG$VVRFLDWLRQZLWK
$VWKPD±
1LIRUDWRV:=KDQJ;/DNH05:DOWHU.D1HHODQGV7+RO]PDQ7)HWDO$FWLYDWLRQRI
753$&KDQQHOVE\WKH)DWW\$FLG$PLGH+\GURODVH,QKLELWRUȳ&DUEDPR\OELSKHQ\O\O0RO
3KDUPDFRO±
:DQJ<<&KDQJ5%:DWHUV+10F.HP\''/LPDQ(57KHQRFLFHSWRULRQFKDQQHO753$
LVSRWHQWLDWHGDQGLQDFWLYDWHGE\SHUPHDWLQJFDOFLXPLRQV-%LRO&KHP±
-&3DXOVRQ.-&0LQLUHYLHZ*O\FRV\OWUDQVIHUDVHV-%LRO&KHP±
%ORP16LFKHULW]3RQWpQ7*XSWD5*DPPHOWRIW6%UXQDN63UHGLFWLRQRISRVWWUDQVODWLRQDO
JO\FRV\ODWLRQDQGSKRVSKRU\ODWLRQRISURWHLQVIURPWKHDPLQRDFLGVHTXHQFH3URWHRPLFV
±
.OHHQH5%HUJHU(*7KHPROHFXODUDQGFHOOELRORJ\RIJO\FRV\OWUDQVIHUDVHV%LRFKLP%LRSK\V
$FWD±
)LJXUH 6WUXFWXUHRI753$DQGRULHQWDWLRQRILWV1JO\FRV\ODWLRQVLWH
$*UDSKLFDOUHSUHVHQWDWLRQRIWKHVWUXFWXUHRIDQK753$VXEXQLWLQFOXGLQJWKH1JO\FRV\ODWLRQVLWHV
DORQJWKH(GRPDLQWKH1WHUPLQDODQN\ULQUHSHDWGRPDLQWKHORFDWLRQVRIUHDFWLYH&\VILYHDQG/\V
RQHUHVLGXHVDQGWKHORFDWLRQRIWKHWZRUHVLGXHVRQ70FULWLFDOWRPHQWKRODFWLYDWLRQ6 7
%6HTXHQFHDOLJQPHQWRIWKH(GRPDLQRI753$UHWULHYHGIURPWKH1&%,GDWDEDVHDFURVVYDULRXV
PDPPDOLDQVSHFLHVUHYHDOLQJWKHFRQVHUYDWLRQRIERWKJO\FRV\ODWLRQVLWHV5HVLGXHVKLJKOLJKWHGLQ\HO
ORZLQGLFDWHQRQLGHQWLW\ZLWKKXPDQ753$
)LJXUH (IIHFWVRI1JO\FRV\ODWLRQRQ753$H[SUHVVLRQ
$ 0PHGLDWHG GHWHFWLRQ RI K753$)/$* :7 DQG PXWDQWV KDUYHVWHG IURP &/V RI WUDQVLHQWO\
WUDQVIHFWHG+(.FHOOV%5DWLRRIFRPSOH[WRFRUHJO\FRV\ODWHGK753$DQDO\]HGE\D6WXGHQW¶Vt
WHVWn LQGHSHQGHQWWUDQVIHFWLRQV
SGDWDDUHH[SUHVVHGDVPHDQ6(0&0PHGLDWHG
GHWHFWLRQDIWHUJO\FRVLGDVH31*DVH)RU(QGR+RUPRFNGLJHVWLRQRI0SXULILHGK753$)/$*:7
14DQG14PXWDQWVUHSUHVHQWDWLYHRILQGHSHQGHQWH[SHULPHQWV'&RQ$PHGLDWHGGHWHFWLRQ
RI 0DIILQLW\SUHFLSLWDWHG K753$)/$* JO\FRIRUPV UHSUHVHQWDWLYH RI LQGHSHQGHQW H[SHULPHQWV
( 0PHGLDWHG GHWHFWLRQ RI &6 K753$)/$* LVRODWHG E\ ELRWLQ\ODWLRQ RI LQWDFW FHOOV IROORZHG E\
VWUHSWDYLGLQDIILQLW\SUHFLSLWDWLRQ)5HODWLYHQRUPDOL]HGWRWRWDOSURWHLQFRQWHQW&/RUWRWRWDOVWUHS
WDYLGLQDIILQLW\SUHFLSLWDWH&6H[SUHVVLRQRIPXWDQWYV:7K753$)/$*DQDO\]HGE\D6WXGHQW¶Vt
WHVWn LQGHSHQGHQWWUDQVIHFWLRQV
S
S
S
SGDWDDUHH[SUHVVHG
DVPHDQ6(0
)LJXUH (IIHFWVRI1JO\FRV\ODWLRQRQ753$DJRQLVWVHQVLWLYLW\
'RVHUHVSRQVH FXUYHV GHSLFWLQJ PHPEUDQH GHSRODUL]DWLRQ RI +(. FHOOV H[SUHVVLQJ K753$)/$*
JO\FRIRUPV DV PHDVXUHG RQ D )OH[6WDWLRQ GDWD DUH H[SUHVVHG DV PHDQ 6(0 $ 5HVSRQVH RI
K753$)/$* WR LQGLFDWHG FRQFHQWUDWLRQV RI &$ % 1RUPDOL]HG PD[LPXP 5)8 UHVSRQVH RI
K753$)/$*WRLQGLFDWHGFRQFHQWUDWLRQVRI&$&1RUPDOL]HGUHVSRQVHRIK753$)/$*WRLQGL
FDWHGFRQFHQWUDWLRQVRIPHQWKRO'1RUPDOL]HGUHVSRQVHRIK753$)/$*WRLQGLFDWHGFRQFHQWUDWLRQV
RI$,7&
)LJXUH 5REXVWDFWLYLW\RIERWK1JO\FRV\ODWHGDQGQRQ1JO\FRV\ODWHG753$
)LJXUH (OHFWURSK\VLRORJLFDODVVD\VFRQILUPUHGXFHGVHQVLWLYLW\RI14PXWDQWV
)LJXUH &RRSHUDWLYHHIIHFWVRIWHPSHUDWXUHDQG&$RQ753$DFWLYLW\