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UBC Theses and Dissertations

Roles of the P2X7 receptor in C6 astroglioma: in vitro and in vivo studies Wei, Wei 2007-12-31

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Ethbr(EttCllr  ll pl d rl t tnfln dd ltdIn  pt llrtrt th 3 KM OxAf hr  37 C prr tpfnthEt t Clrthntrllt d d th S  1 1  ll  1 Eppdf t d th b f 5 n  rttth Et ( Mth  tht trtnt (A  A pl OxA  OxA Aftbnllrntr d pnt drd prr tddtl t trtpl r An Ultrp   pr Ubltrh (GE thr Uppl Sd tthrb  thl  n(thtn vh  Et r vtntn thlfr llth rn b th Et h nd ddt ptrpfd d thrtrnl rtthtrt ntr d prt   pntQntn d bld nn2. S  d n thf thr-d n fl plhpr (b-Eln   199 nd   th  dn Cllrpl tvd rl t tnfln  thd f ll lt On llrnflntthd tr-fr  f  hr tnthfln  th ft Aft thpd  r vnthnflnt nrr IIIplt pttIn  pt llrb th 3 prMOxA f hr  11M G (Sf 15 n prr tpfnthrdvrth  thr t th thr-fr 1dtrtnt pldA d  th OxA ( G thr-fr d Aft  hr  bn (37 C5% COthllrff  nClr  S t d th f th pld 1 M S (1n  rtpAft   Sllrb   (Invr 1 /  S tvlnl ddnll nb thfld  v Clrth   t dt t lvtt  thrn  thd b-n ll  d thI 17b ftr(tn In th r tn  d l  thd rf Ct ptn nthrt thpt rn rrvWth td thnb ll rrt  5 x 5    ptrpfd trl rtrnl rtthnb  llt  l thntr trt lt d prt   pntQntn d bld nn262.8. Stt tprt   dd r   (SEM Stt (p t Stt t-tt    vnfl bStt--Klltl tt  pll(Grhd 3; S  CA2.2. In v2.2. Intrr pltn  C6 llAll  pr rrrd t rdtpr pr bthUAn CEthCttht  thCn Cnl An C MSpr- r(3  Chl v b StCtrth th trl (p jtn  thrhl(7/; dMCbrO d xhrhl(9 /; r InEt O d pl  tp (v Kf Inrt jCA r pltn   llCllrhvtntr d d  S-bf phllt ln (SSd 3   ll  (1x1 5 lljt tthrt   Injtn d f thrt r fltrptr (A1 ; dll (M-3 ; dvtr (-5 fr brA dl  d thp tpthl d t  trltn  C llpfd prld (   In  pt C ll(ACM r dd Cllth nprthlrt nprtEld blt-l llrtn d th ndn C ll( 2.  Cllrljt (l/ntthrt 1lt r tht-ndl jtn r lt  plf  dn 3 nd thndllthdrnAft rthndl th t d thrl tr d th rn tthr 2.2.2. Ith Anrtrdllpf th h ld lfl bpld 1 M phphbf (1 M p7dtxthnrr fr thlpt-f  th fnht d th pl  3 f prtnbrnrth rlfr  pd drd   -7CCn( rt    thtv  thrt thrt d hpp d   prtt ln-fltnrprd f nnthtr  d prl(d M fl nrp bbn S  ntn1% br b (A; S 1 nl  r(Sd   1 f 1 hrStnrth b nht  Cth thflprt tC(Orr fl/h1; SOxfdUK tl frlprn (G rr f tr1 S  tnr nl(rr f nr 1 Chl CA Stnrrd  S th 5% A d b th Alr nj trb I (1; M  Alr 59nj tI (1; M  rtpf hr  thdStnrth   1M St  Sfrt/ p ( St; tt d t  Mntt (rt O Cr thnntrthprttfr thnpr2.2. lnlrn nlnlrn npfd  d prl(d Mn -fltnrb nht  C th xt t prt t(1; Alb rIl n th th t13-l (1; Ab CbrMAtO (1; S  tG(1 S t (1;Ch Stnrth b  xt Alr nj trb I (1; M E O d Alr59nj  tI (1; M  rtpf hr  thd22.2.4. G ntrnIn  pt dr ntrn t rht t CpltnC r(n = 3rjt pth G (5/dlv  St p d d ntr (n = 3rv  vt vp-fr l t t trltnl r d r nrt  d   2.2.2.  l nth pfd tdnt v2.2. Qntn  t vr v rdn fr r rt n(  thtv  n f l br  thlt  th prr I17b (tn Inth lt vth ll t trn  thtfr n d thrt vprd  thpnt tht vtthv thtt hph ltIn vr Intrl l lt  rp ttAInl pt ft th  tAt bn  [C  ] C ll A rrtrp bA (3  prt  r3-1A d thld d rr [C 1(  r /3fl bldrnt ( rp fr n = 3ll[C 1 d bA llh  thpr  thtt OxA (3p(r3-1 n = 3llQntn f thpl  rpfA thn d pr  OxA prt  r3-1C;rprdh b% th pln  OxA (= 5 ptth t  13llf A trtnt d 151 llf A trtnt thpr  OxA31M- CA 1 -----^A 0 M1^  (31-  -2  .0.   M - ^   OxA 0 M A 0 M                         3 (A ^ A + OxAr [C 2lrpd bth t Aprttr  C   rpd bA ( 1(A n =3ll   (   thpr  OxA d hr bA pln (n =3ll(CAl  rpf dft trtntn (= 5ddt ptf  p  32.  rtprd  C6 lld rpr prfltftExpr  d v fltt  -th Cllp tntr (dd d  3 11MA ln (trtntf hr prtrtprt  r3- d   ntr l l b pr   thMC1 d I d th ft G tnt th A rd  pr  d l thfltft(r3- Strt(r3-; fr = ddtpt  th n th thld tldpr  d l thr ft(rtntr fl (b11 MC1 (b5% I(b1% d G(b% SprOxA ft blnthA-d   pr  thprfltft(dn n th pnt  d btG 71111= 7MC1IG==   -tA  110.6E 0.4 0.20CtrA  MC ^ Gr2. Efft A n  C6  ll pr  d prfltft(Aprt-rtf  MC1Id Gf p Ctntr (dd d d A (3 1M f hr 13-t rv  rn dd( Ovl rtfr = ddt pt  3 prn  prt  C6 lld   bAnWtn bltd nh nr tdnhthprn  pr bClld  prtn  thd th ll ptAWtn bl  tn dr thpr  prn  Cll(r3-3Ad ll l fr r brn t r  pntr  rt fth rr bnh n prtrtth d  ndn nf l (r3-3 p lt p A rdlhd prn l(r3-3p rht ph rr  OxA  th A(r3-3l lt p OxA d n t nfprn l(r3-3l rht p Qntn  thrt(fr = 3 ddt pt  prt  r3-3C d t  prn lth A (b3fd rtthtrt ntrd rd hn  thpr  OxA (b% rtA r And n  prn  C6 (AExpr  prn  Cll bWtn blt lll  t t fr r brn t r pntr  (  prtnh nf ntr (plt p A (p rht p A+OxA (l lt pd OxA(l rht p Slb = 1A (Cprn pr  Cllf thdft trtnt(= 3 ddt pt; p5 fA vntr d  f A+OxA vA AK 66 ^  CO ^  A + OxA OxA       C                               < 4^ 4 ^ 400^ -^ v-1 + 44. tn d pftn  C6 llAn hbr(Ett tdpftnl rt(r3- = f  pbn th A (3 ttI5 nd rb (  nbrth t-fd  th trt ntrdpftnd th tntnt  OxA th A tlh d t(b5 tA pln  OxA pldpd n t rb rtntrIntt th tn ldtftn  l pp Cllttnn  hh l  A ( 5 M f 7hrh nft tdhll vb(dn n r n  n ldttn  t  nr ph  Cll30 ^24- 2000 00t 00^ zr4. And t hbr= ddt ptf  trtnt; p5 f A vntrd  f A+OxA vA    rt C6  llrll bb C  vt  th rr-d  An  r3-5A (t lt p ltl nn Crv  ntr r-fr ndnv thpr A (31M h bnCll ndlb(r3-5At rht ph llh  thpr  OxA (3;l lt p  b Crv th OxA (r3-5Al rht p Qntn  thrt(fr = ddt pt prt  r3-5d t   CbthptA (b rtthtrt ntrd rd hn thpr  OxA (b91% rtA Intt lCbrhh  ntr  th OxA tth bpr rffnt tpt Cb  llAnh tG l ft blnll bd bA (r3-5C; b7 rtA  trtntth G d n ft Cll nA Untrt A   :^ vltt..., ^ -^ -^" : ^ ^ "  ^  : I ^ A + OxA ^ OxA^ CC 0 42  400. 020000  00  0   400 0Et0200  0  0 .006  K ^ ,,zfr (^ % ^ 4rr A ft C6 rn  thd (Aprtrtf trtntth ntr (p lt p A(p rht p A+OxA (l lt pd OxA (l rhtp Slb = 1 Exptth G  l rt (Qntn f d rn th OxA  thtt frfr ddt ptpfd  trl (CQntn f drn th G  thtt fr fr ddt ptpfd  trl p5 f A vtrt ntr d  fA+OxA  A+G vA 2. In v2. r prr  thr brndt Ct d  tvt 7pr d fn  v  Inl pt nth thtdntprr  t d fr flCjtn tr rtAn r3-l nd t dnt  t (Wdfr (Wt trltnQntn (fr n = 3 p p pr  r3- d thrt   prd  thpnt th pd btht tth  thntr dA2 W 4 Wlt 060400 2002W ^ 4Wr3- r  C(A l ndnt  t  d t Ctrltn l n C t  thntr d( Qntn  t (n = 3 p p42.2. Cln  th C6llIn d tl jt Cll  t    C ll llnprthlrt nprtEld blt((3-l  rl  tnbhth nd nt lln btdAt  t jtnpr  th Cllr dnt   flC jtnn bl n(lt pr3-7Ad nCllrtd bj3-l n((rht p r3-7A A hh dr -ln  d Crv (t pr3-7dth Cllpr  t  Intr rn rd l dn(bt lt pr3-7 bndrn l  trb n p rn(bt rhtr3-744C6-I ^ t   tdrnAr Cln  th C6ll(A l ndnt  C  t Ctrltn (lt pd Cllr td b13-l  (rht p( Cln  (rth C ll(n2. Cln   th lrh d trWth  pr   th l/h(r3- d tr (r3-9(l/h; O rr;tr; Grr CdlO d  rv rt (t pr3- th ltl t  ntr rn(ndpr3-Intr  hh l nf O d(thd pr3-drn -ln  d O  trb n p (  p rn ltrArd ntn prt  r3-9 (t pthll l  rv  ntr rt (nd pr3-9G n  tr rn(thd pr3-9Cln  Gptr th t  thp ptn  thbndrn (bt pr3-9  rv th nr( rrdn nr8. Mrl d n thrt tA hh dr  -ln  l d  7 fnd th trrn r bndrnbh O d    trb np   df tr Ar d  n thrt tGn n  tr rnh ntn hh l Ar -l th  p rn2.4. Adntrn  G r tht ztt G   r th prd tht brnr3-1 rl r nfr t brjt lth l(C  th G (C+ G Adntrn G rdlr thz brn tfd btrlt Cll th ljtnQntn  thprrt(fr n = 3p p prt  r3-1 d tht vr bt  % th G ntrn4A "- ^ AY06 C6 + Glt v  0-(  802dO  40OC6^ C6 + Gr3-1 Adntrn  G r tht z(ASl r nfr t brjt lth l(C th G (C+ G In bh  thlt hphd n rvrrd rv  thht ntrl nd tprrAtr;    ptr( Qntn  t vrt v prd  thpnt thlt vtthv thtt hph n = 3 p pA4. 4. In vrO   vr prthf rt f pr bh  dprn l  thpbtrt  CAn th rt th ld A d r trt  [C1h nttprrtnt  llth thttOxAnn rpdt b Cllr dr th r  d pr  prfltfthd n  lld pftn  thll r thrpnd  rdl bIn l  A d r   trl l C fl bldrnphtb l   [C 1flpln  A ptll b n l t bOxAIntt   rv l   [C 1 A pld  C  -fr ln (dn n fd ddntnf A  C h th thdt b Onpb th th t  t prt thtr YtS  lldpr  Yrt(Y Cll(S  d r -rt  pr  thbtY thprt r(dn n rthr A h bnn   t   h d r Yrt(Erb   1993;Wdn   51 hrt th  7pr  llh th C-r d ph ndnd n dtfltrp(   ;    ; Mn   ; Yn  prt dt th tn   7 C   thhd ll pr  v prfltftMC1Id G Alth th-rn  thft Cllf ndn  A n (hr   l d h n  ptn  tth  7tnftd t  dftl tt r pl pr  MC1  lnt  th ll d hftrn (Kln   ;t    Alth thfn rp ftrnlln ll-ddth   tth thllnntrtthl th d vn  brn t(Grb   ;Wt   th Gd Ipt t  tf vl ll(   ; Grb   d ththrt tntprt nvld prr (   ;Grb    Ovlr rtnt th th th Clld h t dnt bnt-fltn d An   ld th fn rphnt  ll n  r-d d pdtn  dldprIn thfr d b Clld dn  ll t vntnt rh rt trl5-r n(E t dnr (n  d  vv  n  C ll(nd    Althpftn  nd   trr  ll dth (l  ;rn   1999 fnd n f ll dth An  Cll nt rh nrltll l hrt th tht tn nrld (ffhl  tn  nrltlll tt thYO-O d thtt  nr; dd t  th-prft thll It  pblth   ll t n(dth d rpld t tn hbn dd tf h fn  C fpl  d lptn th prfltrp Indn ld bn th  d  rtn  fl pth lr tpln  A (1 nd (rn   1999bvthprt rt(   th thtt OxAftbld A-d d t  rt tth   rtdn pt pftn d bAO rd n drth trl ph lntn th th ll fn rp It ld bn thth OxA h thd [C 1d bA d  rn  Cll dn ndprf thprldIntt OxA trtnt (3 AM f hrft blnthA-d   pr  thprfltftMC1Id G A l rt tn th dft pln t  OxA53(dn n tth t p th 7tt n ft vvO rtd dth pr  thprfltft dt bC  -ddt ph l ttn  v n  OxA d n lbl[C ld bArrth th d  ntrtCrp th Ad  Y n bprd  Clld d t fthrl l rp d bA  thpr  thttOxAIn dnA h bn rt   t f vrt bt  1(    Alth r drtdt  Cll   pblth h  fd bprd  thllIn dn ld bn th  thn  Cllrlt d dd  r f  hr  prpl tnthft th ft ll b v  pblth r-frtrtnt d  t thbl br  thll It h bn rt thltr vn ( t9hr h ft ll prt rphd thhn ptn d pr  pY rtC ll(Krz  nl   ptt trnth nrrtlll t dd bpt tlh d dfd ftpr d fn th r d h  ll In dnthprt  lld fr dft tvfr  h  thdd fr nr-d t n bv f l 5thln thC d h pr th  ftn   vr d   v b d bl prt brn t(rth199Ovlr   vr rh fnd C llpr rthdtC   bn d ftn  l pAn  thld th ll pr  prfltftd ll n hd ntrtt dnt4.2. In vpr  v  j r thC lld l/h (rr tl l b W fnd phl dn  th ntrn  thrttt lnt  G (Grdlr t zG   th  v d tthflrn (1It  ptd lttt h1v t 1nM  thr (   ( Whdr th  n thbld-brn brr( dn n h   tbpld t(pjtnvn G  bl d  n tfth  thd  n lf   vr    C   ptIn th  vr   hfnd th tn  th ld thll pr  prfltftd ll n h d ntrtt dntS th n lprd bC ll 55vr b    v   d hfn r  rt th dn  thr brnOnptt lb   vrd   v th G hCll n d bA (3 Gt rt z l ptllbhll n  thtnIt  rldr th rprd bt-l  tr rn pr  h d l rt  bh h d r (M   199 df  199 Sh  19; Strt199 Wrzbbrz   199 d  rv rd ll ftrn th t Ar prprtbl tbndrn(dn n Alth ltht tpl t r nr j C fltdthr fn  brn t prlddA nb  bprbl ntrt prnn d  dth fnlvv  r  v  thft  I113d (1 d 1nl t prr d vnd br bbld dt t lIn dn  ptt tnth G   l d tnprnAlth th nrt  t G tll th thpbth thnd h n- ftn  n bd tprnAdtr prt d  t th G ht th bt  bh C lld lvfthvtn  rr tnfthft G ntrn  tdnt d tth r   brn t Cn th rr d fdtnO rh fnd C llpr 7 rth dtC bn d ftn  l pAn  th 7 ld th llpr  prfltftd ll n h d ntrtt dntIn dnpr  v  th Clld t- h/l j ntrtpr  t-brr brnIttl  G  ntrn ht thrt drth th 7 rt   pttdft  Crthr phl dn   v tt dntUnddthr   7  brntld trn tt n tht rth tthrth rr ndtbrrd t tnfthft Gntrn  t dntr  7 t-brbrnd bfth vt bt- ph pr   d fltnll v btdd thl fn   brn t5blhAdhKlr  nd    dUnvtE W  SW rl  199 Enhnt  l th ft rt pr  llbrbt t nr ft ph Cr IIh 33737Adrh Sx CW  S MKh-lt   Ctn 51tn  thtldn prthfn  th rtI 1(1-1Andn CMdd MErnl  rtfn d n nr d (5 7-d  Stn r A  Sl rtl  plSrpl  Mrl xt pt pblr rr ftnCl Cr 9(7llrn thbM  I  nztAGln AltIn0,  C Crl 199 t trll (rtrtrl l d prt 7(15-17 33-37rth 199t brn t d pt nrl th9 C (-d C-1  nl 31 91-1nd tb SG v S W19fftt r l rn tlt S 1(9 37371 lll ACn Mr EGr tln-d rtn d t l 7 (1-133rz Cn E199 Gn d tn ll ll l 5153-1n Stnbhb Svn SClzSAIn CYph 19Extrl A d l n-ltndt  hpl  tl Ad S USA (79-799lEdn  Or  Mtn    Ad lf SExtrl A d tpr d tthr  rt  r ll I 17(7 33Cr Ad SSr GrndGltCSt d /drn  n hpp ltC d M (3939Cr Ar CttM MnGSr GrndGltC Gldrn d  hnrrtn brt tt h Int 33 1-197Cr Ar Sr GrndGltClbpld ll dthft  nrrtn bth rttt b blh Int 33 199-7Chll I S  bl A Mh A rnd EAhr 199Cld fn tn  th rt tt9(1--3CSC tEzz vnE  MtlMrdCStd r  A fr tr  lt Ch 13513ClGhd KhEK-lbMrth l G1997 dtrn  th rthl39 17-13C p  MrU t j  Shn MntA199fftl pr  trnth ft-b1-b d -b3 bbltll trd l I 1(5 11CrAKWdrfM 1999Chdn  tr tlrnd Mprtn  t r brn l d h ll ll lvr Gl (3 1-1Cz  nhz WIln  r fftl ft Y1 d Y1nltrt E1/Edphph 3-nld ll prn  r-dr dnv  Cll  l 1(3 7-5Cz   S r A- phphCn d  hn   Cllr thr tdt nltrt Y(1d Y(1 tt 513(3 179-1nlr Sl KAnlGGrn  GdrAhUCrn n KrtCtthnltrt  ntnltlprn- d l-ttn tptl bdtfbtl l I 1(1-1 rl  1995pt trnr nfltn d ll dthI d 111 5-560 SAndn CMKnECCh YCh YSnn rt-dt r  tfr tr (13-13 S rt td l tC fnGl 57 737Erb    SCIW GA rt trl l Ar5 555Erb   K Sl rn W GA1993nnpr d phffl  U prt  tlAd S U S A 9( 1-153EChnEbl Adj   AtrESt-rYn nC-zr trrtn  thtrtlrn9 nd bI1 d ph  llv-pp 7(3351535155Ebl  MSt-rYMdl 199 In vrpr  MMd MM9   llflptfltdt h h A13(1 -9b-Eln AS AAbr MMSrtz M199Invvt  d- ft r brn trrp t j  vrlnCl Invt 97 (1 1-171rr Chzz lzSl SMClGl G rl  1997A-dt t ll ll hl3915-13rr Chzz lzSn S rl  1997brdn tln-1 br fr ll ll th btl dExp M 1(3 579-5rr Mr MKndl WlbSSzOhf K1999pt ln d tn   th dt r t d nr tn ll dth tt 7(1 71-75rr zzCAdfElCtAIdz Mnth Erl   rtpl  I1 prnd rI 177 37-33rr llbMChzz lzSrdCtn  rl 199 M ll llprpl rpt btrlAI 151531-1539lpAffSvM 199 Extrl A  lphtnpblr ft fn  tl Ad S U S A (-71rrt    Mtln   dhl th ft d d prn  ll ll  131-93-9  Gr Slh  Kr UAllr CIll  rtpr  tr  thnl r 1(3 1-9  Gth AGr Sdt  n Shdt b-hrt  Snd  Ill   rt pr t  thrr rt  r h Exp  (7 9 Wrh Str WG 199 A bdt nprn rt lth II l-tA nth tt1--Whl-SlGnth EKl KExpr  Y1Y Y d Yrt bt  thr r Invt Ophthl S(1337Gtt CEW 1997ndK pt tt thrt  h lph l 1(13-1Gdr  S GYGz W GAMh   rt-dt E1/phphn  htrll Al Cl l C571-GMYSt  1997tft p phphn drn-d d lt hpp nr h (-G SGtASh SK199 Intln-dt trthprn  h bltlt ll lUMGh 71(517-15Grb M Sth KrtzbGWMrl  brn t Gl (9Grb    Sl t C  pt d f th bltth d vnCl  3137-GSlt  Srrt KKSh A Unll S rd 62Cl A tr SW An Ar tGln pphth thA-bdt l fn  thh  rt Ch9 (3317-315 S rrAKtnn  199 t thpfn pt  Wll ll 75(1 7-1lft  Krh  W  -Kl Urn Anln EStb  199Grth hn  nltlh h  ll lblhftnl 39(1 1-1lft  th  Wtnn  Yn nln E Stb 199Grth 15(135-1Crn I W rn  tr KC Cdtrt  pr  h blt  ld 3 tdrnAh 1(171-17dInMInAjKK SInKtYExtrl A trrt nr ft-phr fr r lh 75(3 9-97  W  Krr Clh S n nd prnphphpr  trh l l  rdh (tn CCz SKSdf Wnn 1997Intln 1prd  h  vd  ll prn dt vr Antr 17(5A 37-3Ill  Aldrbr M phl  rt thntr nvE l3(1 5-17InKMr tn bp d pGl (151InKjKM KhYKzSIll  K S199A n  C+ -ddt pln r fr lt ll 1(7 13-131vt nn  llAS   Inhn   A-dttrt  rt nt btht nntrn plnf   tnM htr1(3 5-7Mnz A rth Sprt Alnt lG ltbl A-t r  rt M l 51 -hn G Spn WdrfM SCn C Extrlnlt dftllrtln-lb prhtrplnf fltnpr (13r  Splh  zES1999Invvt   ptd thnrph [3ptfl  bt-th dhl  r hpp n 3(1-1-19Kln  v j   W MC1 pr  C-1trll plnf hftrn tt v nl 5(1 1-1Klh C W r thnvphd nd nIntnn An f  CrKlh Oh  AzzA199 Gl InKtnn  n( lOxfd Unt Oxfdpp 113KrtzbGW199 Mrlnr f ph t thCd 19(3131Krz Sl  Cz  r r Exprd fn tn  Y1 d Y1nltrt ltr-dr  Cll  (19719Kh -fn ft A/ rt tn t nr ft-phr  l trh 93 5-535fntAd  AlzKGMCr-S 1999Expr  vl dhl th ft (G d pl-d thft rt-b(G-b  h  M  13(1-177-1 G Cpbl l S  W ttn  b d rt  h f l 1(1137-11nd C Gr CWn AGC Orl S r  Mt-t/trl -r nn1/h U d l frl prn pr dr thprn d n  C ll  1 (19-193364 M Sh Sz dvn AGnlnKr thp rt  fltnInfl(nd 5b-r  K  SE199 S hthn t nr ft-phfr nrll ll th lI 159-SYWM ChCh ASYn S 1997Mttrtt prn-1 pr d hftrn   Ah (rl93(5 515Mt Ant Gnbn Sh  Mrl 1993tn tln-1 bl  rp tb  rf n-l -1rtnn (1 53-Mn rdt  C+ bn dfn rp lft l l A (33-35Mn Ch Wr K SUrtbnl-dt  trtn  l fr Alzhr d pnt  (3 5Mn Wr Ch Upr pr p7rt  Alzhr d d d-bpttrtl d  ptjt r hpph Exp  (11117MKQn MMMYhMdY rtpr d fn  h-t lt r r l tt 153-15M  b K Strt W 199   ll lltpt r Gl 175-79Mrrn A Stl Spl  SSbCGnz GtttAMInd rt   ll l ttrl A tnl 71(135-1MrOlrGnn  Stl W SWM Mr EdMInz GtttAM In vbltth  rbt r  dC Cr ;  MMn Szz ppM rl  1993Ox AAnrblh  thhprt Ch611 99-MphGtt MAAdt KSYWd C 1995Expr  tln-11 d d bbltll tt 193 153-15nAnEMKYYr 1995Mrtlrn d th tlrn  h  (3r SE YSCn C tIlbtrtlh rt pr d fn  h tr(5-ltCKhf  Ktnn  1997Epl th ft  tft f ll ll vr  f EGrt prE  9(11rbWnh G-t f  199 Chtn dpblfn  5trhphrt t r l111(3 9-95rth M phl rt l v (11-OhtYMMS MExpr  dlnr nth d prtn  nr d btrhphlt r l tt 3(1 77Ol AMKKz Ct Entr Uld C 199nth ft-b1-b d -b3  bhbltll l Idtn  l d dft f  l ltl Ch 7( 1919l CM CAr thv 1 rt prr h  fS (9 9-11l Crdl MzSM CA1999S t rprr th tr th  h  ll l brth 1rt  Cr 79(3n Wj  rx Artrh  W  Ynhf Mr -lrt tn  trnttrtt prn-1 pr vt-t prn n  (17135-7166rvhrthnSGrC btS bt  t  dt pprtn  prl d -r  tr d  Alzhr d Ch15 133-13317t Md SGrd S7rt fr n lvftn  l p  n  t tt 513(3 3-nfld W19 Mrl d thpr ph   Ath17-97t MKrz An UArSEKInnn SWl MMttrtt prn-1 ll ftrn dv  Ann  53 3-39ffhl Chzz lzS rl   rt nthth  h nrltllthr bddt  Cr  (9-91 A K Oln Ml S Mrtt IExpr  fn-1 rt rlGl 37(3 7dr  l G rn CClGrth Srt A1997pA rt Cld pr  h  Ch 9 55rtA19 Srlf  l tn r prpAr Cdl t  1-df WStrp S W199 rn d tn l/h  h brn t Ah 93 3M Cr-S   ErMMC M19Mlr ll ftrd A- pr  l trAnnt    Ah (rl73 1-M  ErMMC nl 19It  nlr ll ftr lnt  Ah (rl73 9-7M n CdEll Cpt  ErMMM19nlr ll ftr th rv  hh-trAh (rl71-193Ch Mn Cb nlpl ptrtnt hft  t th fltnd6d nr l  t  d  nt d 1 (15-1K  ChS tKACh MCMn KSUtrltn  h nr  llprtdn rt nr r d  nt dl 11 -77Mn Ml OS hn bl3-nrd bld-brn brr ln d bptjt rhppExp  1955557MOnMIz YhSW KnKKM199Indtn  vl dhl th ft bt nr ft phh  ll lr  S1 Ch 1( S Cz  z KKKKzr  rl bt  G prn-l nltrt Y(1d Y( vv  l l  Cll  l 13393-SjYnMMMUKS1 MndMn zKh MK  nflttI1 bpr t th  lrnbdtn   ft v  t-r ttnI 1(1 9-5Sz  rl  Kd  A-ddt I1 br fr ll ll I 1(9 39-9Snd  fn AzzM nt 199C dnr l  lnt l npl77(5-3ShMChCCS SMK 19 Ittn  hftr  brn t Crn th p (9-1Splh  r MGzEStt ft  tlln-1 btA d r fr r hpp 151(1-33-39Splh  zESW K    thnvl 73-3Strt W 199Cl nrp  brn t h Appll (5-S S n nACK 1999A- C+ fl d68phpht r brn-d ttrll l A- dt thr  rt h 73(1 333Sdtt CStnbrKGb Mdj  SC199 Mrhd   55(3 Sprt Adr  KhErth l G 199 trt f trl A td  rt S (5735-73S dIIdKK SInKtYtn d r nrrtt nr ft b rt-t l (1 1-7SASMY M 1993nthft-bpprtn d prn  l  vr I151(3155dMSYSSSKOht A Ab 1993Cld trp thh ntr nv ttrrlntrn  t nr ft phf thtrtnt  lnt Cr I Ih 31-9rr GEEn t MM1999t l rtb Spt th rd tpblptn  Ch 153-59 Gdn CKGl GY1995 dhl th ft h  ll ld n bEG Gd b (5 3rd CMtlMA dt l b trd ll ll dn bI- I 1(1-91nE rrrA ff-MztMG Sr E n MChrEArnd M Ev th A pt  thphhlppd tp lflrrnthdWtn bl  Ep (Spl5 7-9rn CCh  rth Sprt A1997Efft dt tnprd ld thr  rthl3915-1rn CMKz Arth Sprt A1999 K ll ldtd p llprnthr  rtl519  335-36rn CMKz Adr rth Sprt A1999brdn  nr rt n t  (315-3nt SnzMCGrAvG 1999dft nrrtt bA  r ll 519( 3 7-13ltCCttM Ar ht  SpdM1999rtft d  rt prlt C nrh l39 1335-13ltCMl 199 St  pt dprt t lt rl nr  (1-193r EShv Kr YCnEnhrh  IrYnlCAAptI1  rr f t vd tl Ad S USA 1(5 -5Wn SCz SGrf Mn G S KSn  dfW Knn 1999Mrll/hpr  tln 1h bltInt Cr (1 11WYn  dS tz 199 Arthr Ar rrtn f ll vrn tl Ad SU S A 93( 1-1WtStn d  Mrl fn  brn t  (3 7-5Wh rnG  rtd nrdl S (1-7Wrzbbrz  KhnIME199 tn  ll llh tr (prrt l h 31-Wdn SSUnn KExtd phl pr  rYd r Yrtd thr ntttrl  d  n l 1 (7 1177-11WtAWt  Wr Ml  Stl  rtntrhl prtn bll ll tl Ad S U S A1(9 339YnYr  rrbr  Chll I  AtrCnAnd  CO C d -nd  tll llh  ltll d trh l l rdC   1   MSIndtn  hth ft (  rll llbprtld E((3 3-395nCMtCltCMrrndG Efft  pt tt d rrt r lt nr n1--35

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