Extracellular vesicles carrying cancer specific splice variants

Abstract

The invention relates to a method for detecting a cancer disease, comprising the steps of: - providing a sample that has been obtained from a patient; - isolating, in an isolation step, extracellular vesicles from said sample; - detecting in said extracellular vesicles the presence of a cancer-specific splice variant selected from (i) extra-domain B of fibronectin (EDB-FN); (ii) extra-domain A of fibronectin (EDA-FN); (iii) tenascin C domain A1 (TNC-A1); or detecting in said extracellular vesicles an mRNA encoding said cancer-specific splice variant.

Description

[0001]xr ur rrn nr rn r n,nm r .rr rrn,n wnrnr . rrn rn. r nrr-nnvnn v rrn mrmr n nrnn rn v v n nnrm -, -n -nxr rrmn n nn rnn.rnnvnnrrn vmrn. mnvvrn mrm.nwvnr,mn mr rrrn nnvrmrnnnn mnrn wn n w nnmr n nrnr, r r rnn r mrrwr ,rnr wn r n vrm mrmn-r mn n. r .r mn nmr-rv mrn rr rn, nmrn nm r nmr ,m , n m, ,r nmr nxr r v .mn rv nmn- r r m n vn.n , nmnrmnn nrmrnn ww mv-rr mn rnnvrn rmrrnn nrrn n m, rv , rv nrnrnmn n r rmm mxmr. rm rnr vrr rnvrrm,n, rnnnrrnmm - nm. r -rnrrrnr nnn , r, nr ,rn,n m rmr r nn . rr , -nrnnrn nn vr v rv r -v ., r rmnr nmr nv n, r nr w, .nnrv n nn mr- . , v nn n mrm r nn-mnn n , ,rw mrrm w w v ,- n rrv v,mmr-rrn.rnv m vrn, vr nr.nn rxnr-nmnnnn-- wrnxzr- mmnr- -nn.rnnrnm -mn nn- -,r n- xr r mrx mr,nnn mnxnrrnnmrmr v rn rn , .rvr, r n , ., nnnmrmn rr n ,x r mnmn, m v . , nn nn n v-mnnn rrnrn, . mnn m v rnnvnn rv n rn mrn n,rrv mr mr nn rrm nr . v n - mmn n m rn n, w rrvn n. r n nnm,xm,rnnr rn mmnvrn nvnn vrn nrrnvnn nvnn nrn .nrrrnrmr- nvn r vr m v r-rn, rr r nn,nnrnmr, mrrr nnr m.-mnrnm-nmr-- n n mrnn mr- rv , ,nrn, rr mr . rn nv:nn r m r n nr , m - -rvnn,nnm n,xnr nr vrmrn - vrnnn rxmr r v rnmnmr- xxrr--mn rrnn - n mn rnn - r vrnnnxmrnr v- n m nn nr- xrn, wrn xr--mmnn rrnnn - nn mn n - - rmnn r - vrnnr mrr.r n nr n rmn nn rnr rrr,rmnrrnnnr wn,n wnrnn w n wnvr n nn r wn wn mn n v vr.n vn n nrr rn rrn, rnm “r nr. rmm mvrnn”, “rvn”, “nnn”,n “nn”,n rmrrm,n n-n n , rn,rnn vnn mnnn x v n nmmrrm wnnn wr n mn n xm,nr “mrn”mm,rn mnn,,nm nnn ..m,rnmn.nr mrnmnn,n,.n,r,nnnnn nnnnmrnn, “,mnr nr mr r,nrmm vn r,n r ”nmrrm n ”r mmn “nn r rn “ vnn.” n wrm,rv, nr nrvnn v, n wrm rnnnn nrx r rw,wn rn nm wn r, nrnrvnn v n rn, r rnn nr m, rnxxnmn rn.n mr n n r. rr rrn “ v”r vrmrrrrm.rrxrnn n r vrn r rn m, rn rrrn “ ”n r rrr “ nn,.” nnnnn n m, nrrm “”, “r”n “”n "nr" x r rw. rm wnnr wr wrn rn. , r n w r nrrm" "nrnr"nnnnr "n ",", "rm ,r " n,"n r ""r" "nnn,nn ""r nmn.w ", nwnn:n,,nn,,r r r r n n n nmmnnnrrw,nn. rnnr nn rmr..,nrnvr, mm mnn n mr,rzn n n mr,rnn rn, n r r m r,n,n m m nr,.mrnr ., rnn: rrn, . rnrr rrr, nrnrnr,,n..n .,rr n r , rm n .nn m m . :. x rn n r r n: rm v:r.n 2r4n:. 24:. , .. r rm r rnv n.r rrnrm mnr m r-m, w n xn n r r rrnrn, wr rnrn nrn nnnrnnn-n rn, r-m nn r .rn rnv n,rnxn wn m m vr x ,rnrnn rn, r nn r nrn nrmn n. m vrn,r vrn,n rnnrn w rnnnnrr. rm vrn r m r m rrnrm vrn.nr rmmn, nm r .nr mrnn.nrrnr rmmmnn, rm rr n m. nr rr rrn rnn mnun n rn rrn-mnrvnnrmmnn-rrxrrmr,n.mrdrr .wr rrmn n n rr rrnnr, .. mn . n r wrn rn rnrm mn -r - rmn.n rn w nr nmnr,mn rx nmn rr rr r n r r n r n w:nn ,, ,nnmr,, ,r,nnn,,,,r n ,, n ,, mnn, n , ,, n,, n ,, rnn r,,,rnnn,r,,,nrnnnr,,,n,rnnr,,,rn r,, rmnr n nx r . rm “ n n r mnn n nnrr”n “mnn n ”r nnm rn. nr,mm nm rv nr,rmnnv rv n nn m,rm- rvmrn m.r rrnv nm n nmvnnmnnrm rnrr mn nmmn rnnrr mr. rrnmvnnnnvnnnrmn nr m n rrn m,r r r,rmm.,m ,rm mnmm, m vrr rmnnrrnmnx rn n r n rm v- nn m nr wn nrr w rmn r. r r-9mn m .nu nnnnrnx rn n rr m - m mrrn w n-7m nn nn . rmn - munm r rmr nnrn r rn rrnr,r,n rn, rnxn n -n r rnnnnrn mm , n . rmnn- , nm , nm , m rr rv rm n-r rvrmrm r rn, rr r rn. rrmnr-nn rvrm nn rvrmrn m mn,u rrnm nn rnnmn, n rr, rnnmn , :-, rvrm nnrrnm. . , - :-,rm mn rrm mwnn .rr , mm-r n, n, nn rnn , mn, nwn , nzmnr nz,- n, ,r x r r nn m -. rmn - mur n rn umnz m n . rmn - mu rrnmrmn. mrnm nv mmrn, rmrnrrmnn rrr..nr rrnn vmnn mrn r rnrr..nwnmrmrrnr r n mmn " r.nnr "rrmn"rnr r "rnn"r mn" r w "rm r mn" m n rr nrnvnrm rn.rnnnrmnmn, zn rn mm, ,.. rr, r ,.., r. r nrmnnrr , zn rmr, r,n r rnw. vnn rnr nwnn r rn rn nnvnn rnv rnn ,r m m rrwnnrnrr ,rnrmn nr m rr n rn mmn n m rnr n rnnr n rnrmn,mrn : r m r - -rvnn,nm n nrm n - nnnxn,xr r v rm m vrn rmr rr v rn nr- xxrr--mmnn rrnn - nn mnnn - r nn xr r- vrn, wrn v n m nn nr- xxrr--mmnn rnn - nn m rnn - - rmnn r n - vrnnr mrr.r n nr n nnr mnm.nmrnn,mm-n, vr-n nrm rnrm rnrmn m.nr vrn nrm nrm rnrmnmn mm.nn,rnm-mn v,rnmnn mn-n vr-nn vrn-n vr-n vr.nnrnmmn,mmn,mnn - vr .nrn .nrnmmn,r nn - vrn vrnn - mnn - vrnn -nnvnr’ rvn, - vrn . n. wnn rnr nvn rnr n vnn rx,rnvnnr v mnrnmmn,rn vr nr. rmn,n rrnnr vrnmn rm. vrr wnr, nrmmnrnnn rmn nrmrvr rnnnrnv r nnrnr, v mrnnrnrmnn r n m m,n rr nrnrn mrr m rn n rnv,nvnr rmn, r . n v nn mmrn, rvv n n rrn mn . nnn rn -vrn nr vnrmmn, vrn nn nnnnrn.mnr vmnn., mvn mn nnrn nr- vrn vrn nmr,rn r n v m n n rmnn nvnr wr n n n nr “ n”n. rr,n m v rm mr nr vrn.nvnr n nn vrnn mrmn. , m nnrm nnrr rrn n.n vn vrrnnr n mn n nmrrnvrnrmr-mn vnr,mn nvr vn. rr- vrn mr r . n,r nrrr , rmn nrrnn r , rmn nn. v nr- vrn w r r nn n,r nrnr rrn rn. mnmrmn, v n- vrn mr nvrmr-rv v. r w r v m vr rv mzn, rrvrm r-vn ,nrnmmn n m vr nnn n n.rnrn r,r v r nnr- vrn n n’nxrmn rvn mzn m , r rrn mrnrnwrr rvn . . nnvn rrmnn nnrrn rn, wnnnm vmr , n r m mn rmmn n rmn vn. r r v,nn rxmn nrr rn nnn n rv r m , n vr ,mrrn,rn mm n rnvrnr-vnrn-v nn. rn rn xr r v ,n w rrrnn , r v, n r m mr rn mrr- v nrnrvr mr m:wr vr n. . •r -m:, r–rnvr v r: rxm mm mm,,r r – m m,r r • r-xm:– m m,r r – m m,r r • -:– mmmm,,r r – m m,r r.rrnr mxm , r m m. • -:– rnmvr v r: rxm m mr r – m m,r r • r-:– m mm,,r r – m m,r r • -xm m m, rxm:– mmmm,r r – m m,r r.rxr- . • r-rn xm:–nr, rrmnvm,rr vr r–: m m,r r • -: m m, r – m m,r r – m m,r r • -xm:– mmmm,, rxm m mr r – m m,r r.r-mnm, rrnv. • -:– m mr,r v r: rxm m m r – m m,r r • r-xm:– m m,,r r – m m,r r • -: m m, r – m m,r r – m m,r r. •rnnxnmrrnmmmr, rmr.nvr v r: r-xm:– m m,r r – m m,r r • -:– mm mm,,r r – m m,r r • r-xm:– mmmm,, rxm m r r– m m,r r.r mnnm, rrnv m. • -:– mrm, vrr: rxm m m, r – m m,r r • r-x:– m m,r r – m m,r r • r-m xm:– mm mmm m,, .r r – m m,r r n rrrn n vv mr: rr--, -,n -nrnvrxr v nn wmr,vn- wn n. v r n n rnn n mnr wr. mmrn n n r n, rnvrnrnnmr-vr mnrrr nn. rn r . mm rv nr n mn n nr-nv vr n n vrn, r r v rn rn nnn n,nrmmn, . •r -m. :., rrnvr v r: • - –.,r r.–.,r r rxm ., • -::..––..,,rrrr..––..,,rrrr rrxm ., .rrn m , rrnvr v r xm .. • • --::..––..,,rr rr..––..,,r: r r rxm ., • -:.–.,r r.– r rxm ., .rxr-rn n .,r r rxm .. • -:.–.r, r,rrnvr v r: • -:.– r.–.,r r rxm ., • - .,r r.–.,r r rxm ., .r-m:n.–m., r,r r.–.,r r rxm .. • - rnvr v r: • -::..––..,,rr rr..––..,,rr rr rrxm ., • -:.–.,r r.–., xm ., .rnnnrrnmmr r r rxm .. • -:.–.,r, rrnvr v r: • -:.– r.–.,r r rxm ., • -:.–..,,rrrr..––..,,rrrr rrxxmm ..., . •r mnn • -m:., r–r.n,vrrr. v–.r,:r r rxm ., • --::..––..,,rrrr..––..,,rrrr rxm ., r m nnn , nmr vrxm,r .. rvmrm n n m rn mr,r r , nn w nm wnr . v rv nnrnn rmrnrnmmnw n nr v. nn-nr. n n,rn v vr mr vn r nwnr rnr rmrnnr vrnnrm nn. rnn, m rv r x n nr r m mrr nrrxmn,r w-n. r mrrn, , -m mnnnnr mnm rnn ,- r r mrrn mrnrnnr mn.rn mnrnnnvv,.nnvr w,rnrnnnnwnwrrrzmnmnn r rrvr ,rnn,rv mmn,nn m r.n nn,n n , nn- mrrvn n r - vwr mn.n r m rr mrvn n,n vr nrmn rn, vrn,r r rnn mnr nn rn nrmznr rrrnn. mn m mmn n rm m , nrn rrnm w n m,rr m- rnn rnn w wnrvr ,x,nr,r nn.mrv n -rrz r vr , mn r mn n r n, r n,n rnrz rn mrrn.rmmn, n n r vnnrnrrv nrmv r-mnr,. vrnn:m-r-,rv-,nrrm-.nnmnn r mrrr n n nn n nr n m n r. r vrrnn mrrnr, rrvnnr nnv mnrrnnnrrrnrmn,n n wr nr nrmzmn mzvrr n.vrmmr-rr mrnr rmmn, n n r vnv . rrn r r rm r nn: nr n . .. nm rrn mrm mnr n rmnnmr n .rr nrmr nmmmn, -rn .n.r rmmn,n mnmnm-mn, -.rn .nr rmmn,n mnn nmnm-mn, -.rn .nr rmmn,n mnrm:rnnm-mn, .xr r v r rrz rmr . z..nz nnn -n , ,,rr r ,rr n n, mnrr-- n n rnmn. nr rnn ,n rv nn ,nr m rm, nn,rnmn rn n, zwn nmn m r .n xr r v vnn . v rm nr rnnr. n . rv n n v zn mr nnr mrmn r z r nr n nnrn nn nnr n. m n n mvmn mr.r rnrrwrnwnn mn mnnnvnr r z mr n v r mv mr r nrrn z r mr r zrm mr n.n r-nn n nnr rnv r nnnnn n mr . n m. wrrrznrnn nnrn,nr r r mnnz, n nnnnr r n n n. m r n mr n nr. nnr nnr,wnw mr-m nn-mnr,nv nrr nr nnrr, nnrr,rnnrnrnrrnr r.n rrnnn. mn nrn rn,nnnmrrrnr rrrn rrrn,nznnnr n vmrrrrr nrr , r. nrzznrnnnnnnrnrnn. rmmmn, xr r v r n n m . ..znrx nn,rmr, . mnr n m n nnn mv r m m. mrn,nrxm’n r mn r .r v wn rnnr, r n, wrrxmw,rnvnnrvmrnrv rnwnn r.nvnnmr r r rnn mmn n rvn mrrmm n nrm n,nrnmmmn,zxnr-nrmr vrnn r nnwmr. w n nnn vrnn,nn mr rn n.nr rmmn, n n n rn n rnn -nm. nxmmrn,n nn r w mr . rm n n rrn, rnrrnnn m , w v rnrmn n rnrn n m. mmn nr r nrnnrr m . m n nzm,rn m ,r v , n n n,mn r. mn rx n r , mrr v z rmn rnn rr n m. n r m nnnrmmrmmmnn,,rnn wnrrn. mr.nr rmmn - vrn w nn wmr. , rn nr- vrn nrnrmnm.n, rn nr- vrn nnr rnv,mr rn mwmr,nrnmr,,rr r nmn mr, r,rnnmn rrn nn n mmr vrm. nn, ,nxr r v wrnrn nrnr,rrm mmrmn, n wr.rn nr vrnnrn n, wrnv n mrnnn. nr- vnrn rnnmmrmr mmn nn,, . m nrmn m v r r . m mr m.nmmmn,nm m.nrnmmmmnn, m mm.nrnmmn,, mm rr mm.nrmmn, m rrrv mm.nnrr,nrmnmnr,,rnnnr mm. nr,rnrmnr,nrnnr, mn nrrnnrnmmn,nrrn nm. r r -mnm, nr rm r m,m rmrnr rmnrn mnr,r, mnnnm,rn m r rrm rnrnnnxrrmn n m r, mnmr.n,rn-nrnnnm rmrmnn,nnnr r - v. rmn. rr ,nnm -.nrn n rnn, w m -n -. m mvmnr n,v- v rr nr xr r r nn, rn mr-rv rvrr vn r.- v r rv n m mz n rrmrn,r rrvnnrm rnvn’nnx,n.r nnnr, rrnn, w rn m rn r n mr v rvn mrr wn rn rn rrn x nv r n w .nrv . . r vr —mmn n rn w rnnv v wnr n— nrnmrrvrnr nrxrnrrn rn -r v nrr vrnnr mrrnvn wnr n . n, rrn mrv n r . rvn nn m rv rn nn n nvnn, xr r v m r r w rr rn nr- nm m n, . m n rm: . m n, .n m n .nn mn m n . mn .n mn rnnn.r- - n r r m mnn rn.rv rwnr nr. m n rnr . -nwn n n n m nrr nrrn xm nr- m n w r r - , nn-m nnnr nm nr,n: nrr:n mr. nnrrm,mn vnmnr mnnmn, - rn,r nr.rnn mnn.r: m nrrvnn nn-m n r w rn, n,nmrnr n r - nrnnm m m nrnn. nr.nrzrm:,nm n nnr n r n rzm,n nrr - rr . m nnmnnr.nrznn,rrn: rrrnmn rn nn- nr w n rn.,n nr n r r - nn-m nnnnr,nnnrr: n m nrmmnnr nr, n rrn r. mrn.n r rxm - nr, nn n r: n m nr nn vr m nnnnnn mr mmn m.v n r nz nrr n mn - m-. rrrn rnnn mrnnmn nrm: - rmn n rmn. .mn, n,n nnr r - nnrnmmn, nr- m n rmn rn m rrzrn, nnr- n mrn.mn wnvnrn mrmvrrrmnn nr m rrmn n,mrnmnrnnn v r, nvn n n rmn m rrnr- m r wr,nrrmn m n nvnn.n m nrnn v nnrnrm nnn nr- mnrn,rm rnmn, m rn m nrnmnn rmn .rn,r rn r v wmrn,nrnr,nrnrr wn. w nnvvrrnr nrr n n v rn, n,r r nn mnnn nr rw. nnn rrr r n ,n nnr rwnnr. nr. rmn mnm,r nr,nm r -rmmrnnn rn -r m:, m nm w n nnm n wr: mn nrnrv r - rn nr:mr vmrn,rn r n. mnm w m nn n.' nmnn w n w n r,r v rmn, -nrnrv: n. r r rvr nmnn wnmn, n r, m n.mn n w vn mnm rnnrrrrmmnmn,n m n rmn rn, n r nn r wn,n n r,r rn n r rm n r n,n,xm,nmmnmrn. nvvnrr r rn rmrw r r r , w rmmnn vrrnnnrvrv, n nrn-m. nnrnvrrxr,nr nnr,rnn ,n m n nr.nn nrvn rw nr . - n,rr: n m rn n : n w r mnnnr rv r rmn -rn rv:mr n,rn r rmn. nr nr.r n n - n r rrn nr . m n:.nn n rrv r rmn w rn rvn rn mnmn rrvr n r r, - mrn r: n rnmr, nnrn. v rrmn w r n r.mnrr m n n n w vv m n, mnnn vrrnmn w r r n r-n r - xm rxn n r. r , nrnmmnm rx: r mnnn m m n r mnn v nrn n n nr.n ,rvnn vn r,rr n xr rmnn rr rmmn,n nm n nrrn. n n rmn wn n r,r rnmn rn, r nn rzm,rzm,-r n r rm n zm, n, nrn,n nnr m.nnmrmnrrw r r rn nrm rw vrxr ,r rr nrn m nr, rn n , n rvmrrw.rn n wn rnn nmrv m r n w - vnr. nn r -,r mz mnm n m rnr,n::rmrn mnrmnnn - vr rnrn r nmr nr.rzm wr n rnn - rzm r , nmrm rvnrw n. rnr : rrzm,rzm mnnn n mr rrm n. n nmnn wrzmn n n - vr nr,nnn n - - n w. mrrzmnmnn : r mnrzm w xrn .',wnrr- vr mr nr rvrzmn mr.vr nr n w vrv -nn. nr:m mnmn r r nn rn rmn vn r m nrn wnmr r r m nr - rn rnx:nrrnnnrr - v. rn xn vnr rrr-n --vr nr. wnrzm- r,m r n r nrvr nr - n : nwr n r nmnn w rrrrn. n mr rrmn zm r vn nr r m - vnrr rnmr,nmnn, n wrn m . n rmn w n rrnmn rmn rn, n nn rzn,rn n r,r rn n r rm r n mmn,nn,rnn,rnnnrn rnn n, rrrnmn- vmrr rrz m mmn wn nrm rwnnr. n rn nr.rn rrr nn-m nnr n rn rrn n. nnmnrn w n r wnnn n -rzn.rznr:mr m n, nr mr nv rm rnr:mnnnzm-, nv - rn n:r nn nzmrrwnrvv. n nn w vnvr rrnrmrn - v , r rnrn rzn. - nn wn n n m n. ,nnn:n w vvvn r vnrn - v nwnr vnnrnrn m r n rzn. n -rn nr:rvr- v . n w vrr n mrrnrn rzn.rn wn nv n rn - rnn,nrrnn:mr r n m n. rrr vnr-rnrn n r n nr -rn n w v rvnnrr mn m .rn r- v -nrn-: nx-nrn nr r,n r. nnr r rmn - vnrn wnrmn r m n nr r n rr-nrn,nnn v nmn rr rmmn r. n , n rmn rn, n nn rrmn wn r.,r rn n r rm r rn mrw,mmnrn mn,nnnrvvnnr,nvvnnn w "nr " n n. rrn,rr w nr rnvnmnv vm nn.wvr, w n , m n m : nn rrn m n, -rmr mr mr: w rr r rv r .nr nn-mr nnnr w mnr mn,nnnwn -rm nr rw. 'rrrn :mr r, r r m n. m n,nnnr r rmn vr nr w wnr n , r nr,nnr nr. rnrr rmmnm rn,nrnr. n rmn wnn n rmn rn, n nnv nr n r,r rn n r rm r r rn m m nn,nrnrnr m n vr nr,nn m mn n n,r nn m n nrm m ,m,n nrrm. wn r mr mnrw.rn m nzmn , m n rmnr nrn nr. n r - wv rn rvm rr::n n rrv rrmn rvr n v n r mrmnm-m w n rn mmn. - nr. mnnn :n n rrv r rmn wn nn r n.n nr nn mn nzm, m n.vrn nrm m mn rnnnnrrnrmrmmnn w, r-nnrnnnr ,rmrnr rn,rm r nnmn, n, nn, nn n - n rr - rnn,nnrmnn. rmm. rmm nrm rrnzmr m nmrm rn m n n m m .nn - m :nr rrn m . - n rn , m n - wmn v: rrm-m.nmnr rrvvnrzrmrnm n m,rnrmn n mn rnn n - n r: n-nrn nr mn n. mnnnvrm - n r mr nn rmn mrm mn r n.' rv r - n n:nnr n-rmm- v . ,r r nrnn r, nn r rmn - n :nnn- wnrvnv- n r rnrrvnrrnrmn. n n w v n rn rrmn,n mn - nn : r-nrn - n nmn. m - m n,nn r n vrm r n rn r.nn rrm m n, w r n v n n wr r nrnn n rmm- r. rn rrrnn n r nrrr, mnvnn r m rn nr n n n nrmrn : mn n n m r n n .rn n vmn r r rmn nrnrrnmrmmn,rrrnm mmn nn r.nrnmmn, v.,nn rnm r n r m nnnn nr. v n nvrnm - rrn mn mn rnnvmn.nnrrnmn n nnrn- rn mn n mn r r n nnnr . vrnr n mrrm nnn n n nmrrrv n m rvnnrmnrn - vrnnvn n n rn nvnnnrwrnnrmn. wnm: m.: n m nr n nr ,rrwn rr nr -rv,nmrmn: n nrm n - - n vrnnxnr xrr v r vrm mrn nr- xr rm xr--mn rnn - nnmnm rnn - r vrnnn. xrnr v- n m nn nr-. vr mn rn m, wrnrn rrnnr . mmnrn rmn. vr m, wrnv m nn nr-. m n n. nrn nn r nm , wr vrn. m rn . mr rn.nn r nm , wrn v.vn m rn m, wrn n n r. mnrrnv v r . nr n rmr, wnrnrnr nr r vn : m r nn . .. nm rrn mrm mnr rnnmr .r nmr . m rrn nnmr . . m rn nnn rr nnmm, wrn - . - . , wrnn m nn .. m m rrnn nnnn rr nnmm,, w wrrnnn- . - . m nn . m rn nn r nm, wrn - . . m- r.n nn r nm, wrnn m nn. v mr rrnrznnr r nm, wrn xr r . mr rm: ..nz nnn ,,r r nr- n . m rn nnr rr nnr-m, wrn xrn v r m r r .nr n m .zx n, . rmr, . mn n m . vr mr n rnv nn.nn mr nrm, wrn nr- nn vrn,n n r w n. m rn m, w n n. rn rn n nn rnn -.. m. m rrnn nm,m r, wrn n . . m rn nn wrnr nnm, w wrn rn. nr- vrn wm rn mr. r,r r nn w. mnr- rn vnrnn r nnm , wrn rn.xr mr vrn wrnnrnn rn nrnrmr, wnrn vn . rn n,. r m m. nnrnnnr-n r vrnnm, wrn rn n . nrn . mr rn.m, wrn rmn m v r .m m. rn n r nm, wrn m m.. m m rrnn mm,, w wrrnn mm rrrmrvm. . m rn n r nm mm. m , wrn m rm. mrmrrn nnnn nrrrnnnmr, wrn nr nr,rn nr,nnr, mn ,r nr, rnn. m rn m, wrn m. m, nrrn nr rm r rn m mr rr r -mnm,rn m , mnnm, nr, rmrmrnr rmnnr,r. m mnm. rm rn nn r nm, wrnn r v.xr mn. mr vrn n,rrnn r m nnr wn r rm m:r ,r wnrn nr- .. m n, .nn n m n .n m m n . mn .n n . - m mnn ..n - n. r mn, rnn nm, wrrn nr- m n rmn rrz nr- mmnn w,rrrrvrrmn nr . n m nn rrmmnn wrn,: nr r m n, n r rm r n r,r r w nrn n vmrn, rnn .rnmn m n rmn rn, n n r n wnn n r,r rn n r rm m rn, n, n, xm,nmm n .rnrnnm n rmn rn, n n nrmn rn,n w nnn nrnzmr,,rrrzmn,n-rrzm,rmn, .rnmn wrrnnmnn r,rmrn rn, n n r nn rzn,rnrn n r rm n , n,rnn,rnn . rn rmn w n,rrmnr rn, n n nn n n r rm r . n r nn rr. rmn wn n r,mn rn, n n nnv nr mr rnnnn r rm r . - rn n rmn rnnnr m n rmn w rnnn r ,r,rnn n r nnmn, n, nn, nn rm . n - n r nn r mn rn, n n rvrrnvmnr wn rrnnrnrr ,rrnr “ wmrmn. nr rnv m mnr rmr n”,nvnn r mmn nvnn rrn. mmnnvn rnrwn.wnxxmmrn mnr,rrm wnrr n m . vnn .rnwmur-r nm vrn rnnrnn-n n r .. -rv rmnr n.. m rn rv rrmnnv -, -, - nn- nr,r nrn mm n-, m n m n r vrm -.,r-nnn- r , rnv r nn mn n , r v.-. w mr r rm rm . wrn nnn w ,vrr nw rmn-nr mnmn n n .n nr,r nrm .n rm,,v mr nvvm wr n n vvn wr n n n vrn m.,w w r r n r rrnvnn n nn n nm , . v nn. rnnrvrrmn- m,n- nmrr- , r rn.nr .-. rm nr - wrnznm mn =n = mn w nn-nrnrn = r n w m ,rn m n =,xr-rn nrn =,-mn rvnr =r,nnnnrrnmmrn=,r mnnmn =.-. r rv - - , - w, - n wr mnr,r wn mn ,rn wn nrn m ,nnnrrnmm xr-rn nr,-mn rnrrv.-.r r ,r mnnm. rrr - wrnznnv nrmn =- mn w- , - , = r n w mn =,rn m n nn-nrnrn n =, -mn mn =,nn =,xr-rn nr mnnmn =.-. nrrnmmrn=,r , - rvr rnv - - r w, - r r n r wn ,xnr-rnwnn nrn n w m ,rn m r mnn r,-mnm,nnnrrnmmr, nrn =m, n.mr r n wrnzn mrm m n =n,-nrnrn = r n w mn = ,rn nrrnmmxrr-n=rn,rn mrnnn =m,n =-mnn nmnn =r,rnn n . wr wn mn n n n rvn..nnz n n-w * <. w, ' m mrn -,-,. v wr. w ** <., *** <., **** <.. n n mnmr nrm mrrn,w mnmnvv n n nnrnvr rnnr r.-.mrn v m n n nnn v r m r nnn rrm rrv nn mr vmn - r nr.-.rr nwnnnn nrn vrm n nvr.-n.nrn rn-r vnn m- rn w nvmrmnmr n n =,,r rn wn =rnn m ,rrv r nvr rvv n,, n =.-. n- nr-rn v m wnvr m w . nrnw =- nrn-v, w n -.nv n m n r nrnmmnm.- ,nr nn m wr ,r r n.rrnmn r r.r r nrn rr nwnm rr n, nr nr n.-. v n -rrn n , nr =nrn n rn n w m.-. n-r n rrrn , n = n w rw - r mnvr vrmrv,v n m , n =,nxr-rn nr n =,rn n -rn v . wn, n = n rnrv, xnn-rn ,.rr mn ..nnz n - n-,-. v wr nnn * <. r, **r <., **r* <n'.rr, *n < ***. . xw. rmn- nrr m rrnr.r.nrrmn-v- .. m r mnvnn w n -rn vnn - n - w wnr mr rrm mr m n n.r rn n rn n - rn-n w vn, r v.. mn m n..rnv -m rnnrm r- wr mrnvnn w vnn - n --rn m r mn mn wnvnn.,r rn w n n - r --m r rnrmv.. n rn rn mnnn rm r mmn mnm n.. --rnrm m m ..n rrrn- vr rm w rrnn n - n --rnnn.. nrn--nxr nn mrn r m r - r rn.rnnnvrxr nr - n -- wnn wrwwn. wrnr wnrnrnn .nnrmnn rn. nr - n -- . mrwn nm rrzrmn.-. rrn mvrnmn rn n mm r nr m , m . r, nm..nrnrrmr rnr n nnrnnz r nr nrnm- rr wn r,nr n mrr nrm r.. mrmr r-r n mr m n rvrm - r mnvm znmnr.- nn n mrr n . mm r . m n.mnnm.r, r,mr nmn n m w nn n -,m-nr wmrmnr n-- m. r r mn mr.rrw nmn nn rrnn . r, nm..w rrnv n nnr rnn n rnnmmrr, . v rnnvwrmr.rn r - . n mrmnnrmrnrmm-n r-nrmrn -n m nrrnrmn rm- r mmr m -- n . wr-n w mmrn- n -v rn n vvnvwr-vvvn rrrnn n m rmmn rn .. nr nr r -, - n r. r, nrmm..- r mm r m n m nnrnnz r nmr -, - rn mnr rm- r mm - nr rr r m n m..m rnrvmnrr-r n mr r rn r - w zmn.rnnmn-nn w n . mm .mmn -, -, n n . r,mr nm. n n rm- r mmr mrrm nrnmr wn n,. mw r rr nnn wm. rnnznnmrmn vrnrnnr n.. w mn =,rn m n =,xr-r = r n mnmn =,nnnrrn n nrn =, n nnr r mmrn=,r mnnmn = v ..rr nrnn . nrn rrn mn m m vr n rn- wnmr wnr ,rwn -nn =,.-.- rrwnn r. rrn rm m n w m v nrmnr nnmr-rv rnm rmr nr, n = rnrvr, n = v .-,mrn mn n rv rmr rrnn rrnr r m rzn nr mnnv nn =. m m mrnn ..,-nnz nn-w w ' ,.rr nr ,w- n’ ,r xnn-rn n * <., rmn rn'rr n- n. v wr. ** <., *** <., **** <.. nwnnnnn mnrnrrmn mn n n - r m,n-r r.-, rv -- rr, - v , r rn-nn vnrrnrv ,rrn nnn m rm m nn n rmnrmm nr n nrrmnrv., rrrv ,.,rrv . ,r rm-n,rr nrrm- - , - n nn wm ,wn m mr nr. rmwn rmm r r, n =..mrm n n r n rr, n = nn n nnnvrrmrmn rnrvn n rr nwnnnnmr vmn v , =.-. v rnr rr r n - rnr nv ,rr n v w, -rr-nrv n.. rv -- , -- nrrm m vnnrnnrnnvrrnn =. ,rn n r nn r rrv.-. v - n n w w r nnvnmrmmnm,r nvn m rnn =,, n =..rrrv n =,,r , nr nn n rn rvnnwnmnrr n n w m .-. n-r nvrrrmvvnn n wnn =rw,n--rnn r mnvr m rn n rr, r v v. w wn.m rm nr n xnn-rn .rr n r mn..nnz n-,-, . v r * <., ** <m.n, *** <r.n', ****rr <.n-. . rwr nrn r n n .-.rr nwnm rnn,r nnnn rnv nnr:mm n w n nw n m n rrr n rnv n = rnrv n rnrvrrrn n , n =r rrrn , n = v rrnvrrv nw n rn m n = rnr = rnrv nrv nw n xr-rn nr nn vr rvv rnv .-. n-rrv mnrn n w n nm w, n =,rrw - mnxr nr , n = rn n m , n =,nxr-rn nr m,, n =,vrn mn w ,,w n - mnr n = n rnrv- =,nxr-rn nr,, mnr ,r,r n =n,vrn- m n w, w nxr-rn nr,, n = n rnr , n =, v - n n w v-nrn m, n = w r n mnr - ,rn m , n =,nxr-rn nr, n =. rrmnn v n . rrn mn ..rr n w .,. <.,rnn'.rr r vn- rrnn,w, m,r w,rnn ,rr.n <,. rw mv -. v wr n * <., ** <.. nrnrnn m-- -r-m.. m xrmn r rnr..rnv r wr-- nmmn r mmn r nnwmr, rrnn rzn vr n n rn rnrmn. - r nmmn r-nr - n w mmmrnn-vn -mv n r rm m- r vvn wr nr,nrrn-n- n .v wr w - vrn vvn mnr w n w r .. m -mn m n m n vr n n rmr w m - --. rn r- nn n-rmz -r..r wn mn rmn .. m -r mnrnmr wnv n wn r-n v-r-n.n r mmnrm w n m m r..r wn mn rn r - nn nrm-z - rmnv nm n rmn ..r r-rrnnn n.mrnrrmnnnnn- n --rnrm m n. r rm rm r - rn m vr --rnrrrm m n rrn mrn rn mnrn .. rnxr nn rrn r -rv m r - n --rn r -nrn mrn rn mnrn . xx w vn.rrmn rnn mn v x ww- - w nnvrxr nr - n --rn wr wn r rrnn, r mnv...nrnnz wrnw-r nn’ r.,.. * n-* <rnv.. wrnnz.-.rnr , n = nrn, n = v mn mn rn nnrmr . wr wn. umrnurn... rrrn w- mvn nrnr+.-r.rr nwn m nn =,nrr nw n -,rnv n -, n = m n.mr n = n rrrn r -mnn mr r.n w rrrmn m rr n w rmn n rn' n mn .. * <., ** < rr n- n. v wr n. m ., *** <., **** <.. rnn n n nr n nr u.-. vr rnrnvr.-r. rv nn n mr v.n n rv rnnn wn m mr rn rrn n wxr-rn m n =.n n rm n w rr m rnnn =r .n n r "mnn" rr r m rnnrn n =. " " rr n r n m nrrn r mrr. . mr m, n nnr rnn rnwn.mr nn nrn rr nr rw r rv nr nn rnrvnrnv wnn n . nnn nrr.n r rvnnnn rr wn rnrnn rn .nr nn rnrvnrnvr xm xmmr-:umur- vrrnr -vrn rnnnnn- w rnnnnn-rrvn-mrnn.nvv nr,mr- mnrrrn,n m mrrrnr wnr nnn mr mnm rnmr, w rmnr nn nrn w nr m rmnnr nn m n n mmnr n rm r n-nr nrrz mrzn mxr.. rmr vnrrmnnrn mm mnm nr mnmnrmnr xr vnr rn rnn mrr,wrn.,rn xr r r v, , .rnmn n rm rn - r . n vrnr rnn v rz r nrn nm., wn rrn, wxr r v-.,n-n vrn--rnrrmrrnmnnnnn rm- rnnn m nr n, nn rm n .. rn mr- vrnnmr- w rnr nrm rnv n -vrn- n . r- r nrm - zn -, -, - w mrr n mr nn vrmnr n m. v , w rmnn zn -, n n r r - n mnn mm r nrrmn- . nrrmrnmn rn mr m n wr n. w n w - n nn- r nr .. nn.nn -rw-mr vnrnrnvnmr-, w w- n mmr -, -n - nrn- nnnn mmrm- rnr n m nmw nrnm.. n nrn n nr r rv n - . -nrm nmnwmr, w m-rv . nn nr, -r -nnnr n rr v nn -vrn nm-rvmr.. rvrr n r nn nr n m w-, wm w nn r n -, -,r-,w w nn-n n r .rz,n wr r . - rnr mrr- z r nn nnnrrmrn - mr n ., r r-r nn mr ..r,n mrrxr n mmn .. , , ,x, xmn :rrmrnrnrn- vrnn rnnrnn- w rm mnvnmrmnvnvn rnnr n rnrm r n,r wn vr rmur-nr v n vrnnz m v r , n nn wmr n =r r r. nrn n mn =,-mnm n ,, mrnnn mmn =n =,nnxrnrnnr,r nr, mnm,n rrr rnnnr n mrn =,mrnr nr, n w ,mr,rnnnr n n .mr nr, v ..wv m ,nxrrn nrx rvr rr r, rmnvwr wm.nnmrv nrnmrnv w.v wnnn - n vrn r nn -rrnnv .-. n vrnnn .r r, rrn vr-nrmn w nvn rrmn, vnnv n rnnr wn m r n nnn nr ,vnrrv -- mrnnm nm.r-rmvrrn, w-rrnrv rrrnnrwnnrnvnn.n n rnnrn , wx . w n n nrr n n nv w n w n.n-nnrn . n rr nrr mrnv rmr n nn-nrnrr rv r nnv nn,rrnnn r rnn .. nn r - , w nrm, wm, vrwr nn r w n rnn nmnrn -rv nv <. n -m nn- m rnr rmn mr mrnr m. rnr.rrrvvr,mrnn rmn nn wnnr m rm. m . n. nn r xnmurn: m m nrmuumururn,w mnmnvv r nrn,nvrn r rmnnnnn m nnnrn, m, w rn nr rnnnr rrmrv nr n rn n n mm rmmnmnrn wr , .n , nrmnn nrrmrrr nrnr n n r . m wr rm nrr mrn wn nv nr nz m r mm n=n rm n mr.nrn.nr mnnnnm, wnrm rrmn m w m-nnvvv n m,n m nn mnn, - n rnnrm- , nnn mrr.-. rr nrrwnn-w n r. n, wnrm .-.n m rrnrvnrnv - vrn rv -.mn n- -nvrnnrrv , w rr n rnrnrnv .,. n-rrn - rrn nn rr w mr vmnrm -r n nn , nrrmrz mn, nnn nr- .,. rr n rrv - r w nnr . n - r mrxnrn .-.n n rvn, wr v mrvnr mrvrm m nrn r.nn w w r rmr rn,n wnn rv nn m , rvr, w rmn nr n mrrr n.-. r v m.rn rmvnn w r vm vnr xrn rrm rrrr nnw n,- rrn n . r rrrn rr-nrmn .. rn rv r - - rrn , rr n nmrrn wr nn .-. nrn n mnxr nnvn r n w mrrn , wn nn wr m n v r n - wr nrvvmr n wwv . n n .n, r n nmnr w w n wrn -.mrrn w mrrn w nr n wxrrn nr .,.. nn .r-.rvr -n --rrn , n r xm: - m r mur rnrm --m n w mr-numrrur- w murrnn n m m r m rnrmn,nn , ,rn, m, wrn nnr r mnm nvv n mr- w r v mr-rv . rn m rrn r m rnrmnmr -m .r, w n nrn-v m,nr w n rrn , w mnr m r n n n w mnrrmn- m.n.m wr rm m mr rvrrnr n r,nrm r m nn r - rm -- . n mnrm nm. n, w wrr n rmr m n. , m- rmr m nrnrm.n-n, n m-rv n nn n - n -- ..rrmr, w rrz rmn - ,vn wxm .m rv w n nn rmv . rn rmnrn..rnr mnrn , vn n nn ,mnn n,rnmnnn n-- , w vrv nwn vrxr n mnrmn mrrrxw.nn,r,rrnrrnrnn n -- rnn rnn ,nm - rn n. , w rrnrmrxr nr nmmn . n n nrmnn n rnvrxr n - wr rn rxnr r mmrx.. w rrn vnn n,rnn nrn - nm - rn .. m ,rvn v n nmr- mr m rrm -- rn rn nr r n mnrnn rnnr. xm w v: vrxrnn nn-mr n nnrmnrrr. nr nr nrmvrn nz rn r r nn vrrmn n wnr. n rn mwr nn n v wrnn v r m nrnn n m n-nr rmr , n r n mrrw n r nnnrm nnn-mrnnnn rn. nx nnrmn wwrmrmr. w rrvn nwrn nnv-n wr mnm, rn nn n-mwnn nnmrmnnrn m wrnnn nn.xn n : • nrmr : • mmnnrmrr nnr • mn mmnmr nnr • mnrn mrv rnnr • mnrm mr n nnr • mn mmm v rn nnr • mnmr v rn nnr nr nr mrv rn nnr • n • m:nmnnr n • -- nmnnrnrnn • mnr r n • n-nr n • -- mnrv m mnm n • m nm n • mnnm n • mnmm n • • mnr nn mmnn r n • rrv nnrr nn xmmrr:nr wr numrnznnmnr -nmnnn mr nrnn r rr..rnnrnz rnnwr n mrznwmnrmrrnnnmrmvn. n n rrrnr,nrnwnmrznxm:mnn mrrnr n rr wnr m.mn wrn r mrrvnn r, w znv m,n v nrrnnrmmnnn.nv,nr, xnvr v. v. r,rnv nvr v v mr rrv n rr n m w n n. m ,nrr m . nn m,xr-rn r. m z n rrr v n wxrn xm: .vmrun vnnnrvrrnrrmvn ,rrvnnrvrrrzr m,r nnn , n m r n rnn m r r n m rn.wvr,rr ,nv,n rv -r nmnr mrrv wr rr rn-nr .nrrnmnr,r. w-n-rn n- n - n nr nr, rn- n-rnxr n mrn , ,n w r, wrn nr, rvn r r nn. mr- vr - mrr vn-nr n, n vn v rn rnnrnn-. n n r xnrrr r r nn rm n mrr,r n rmnr - n r rn mnn,n r m mn mmnnrwmrm w w r n rrn r,wn n n rnr -vv.nrvmn > r vr xrrnmr w n nv - r x.rnrnnr, - mr nn n nrrn mnn.rvr, w r n rv n-m,nnrm. mrv n n .rr,rrn - r rm, wn n r mn, w m,nr r nmnx mnnn n n w rnmr v n rnn m r v n. rv m - r wxnrrmn rm r v wnnvn >. v rrmnnnn m nrnzrmnvvnvrnrrrmrnrn rrnr.v nrr,m mnr n rn n mnrnnrn r w rrrvv n w rv -r n - .vn w n rnrn r ,r mn. m zr rnnr n nvmwn nr mnnxnrnnr .m.rrmrrr,, w wn nn rrmn mnrnrrmn r nr . n nnnvw x,r rnrnnnrn rnvvnrnn n..rrmrn, w rm nr xrnnr mrn m n nv n w w n m,nr mn. nrn n nnmnrn nn, vn nn n n vr , nr nr r rr nr rn mnrn,n rnn nmn n n r,rm r mrn,.mmn rv wmmnr rn- xmnm: r n : r mwn wrmr nrnrmnr n nvr nn rn w nnn nnrvrm rr mm w nrm n r nn. rn n.rnnn rrmnnnn -n n. n mn n rn w m,rn m ,xr-r vr mr,nn m, mnnm,nn n m , r rn mn wr nrm nnnnrnrnrrm.mxrr.mnnvmvnrmmnrr nr wrrrmn rn w rvn r n m - .n n. n mr -r: mmrz rr r nr umurz rmnn n rnnmn rn rnnnnm m n wr r rn nrz wn ,nz .,rn .mr mn wrn ., n mnnr-n rr w n n mr, . n mrvrrmn:rrm mn .r,mmrr vzm w w mr n vnmmr- nm nr wrmn mnn nnnr r-nn -w m, wrmr r n nrm r vmr nrrmn. n,mrvr vnmr w mrmn mm vr’x- mnnzn. rz w mr w r rxmn:r m r,mr r mmr nr vrmxrmr m , w mn r rn r mr wr mrnn. nm.r’r ,m ,n rmr mn mwnn mmrmnnrmr n wnrr :mnrmr r nnr , ,mnrn mrv rn ,mnnnn nnr mrv rn nnr ,mn mmr ,mnrm nnr ,mnmnr mrv rn mr v rn rmr r ,mn mmmr , nnr .mn mnnnm r mmnnr m,nmn n wr m mrvm rn, mmmr,mnr mm mmrnvnrrn , r ,vm.nmnrn v rn ,mnmnr mrv rn mmmr m n mmnnr .mnr , wrm mnnn wr n --n rrm vr nm m-mmrr .mnn wr n n , r, .mnrn ‐ m- n ,mn mrv rn ,mnrm mrv r n w mmmr v rn ,mnmnr mrv r rmr wrrnnn vr w n‐ m-r, . wnmn nn wrr:2m°n nrnn. mnnnr n, ,- ,nnr n, , -, n, , -, mnr nr n-,- mnr nr n-rv mnm n, -- mn mnm, -, - mnm n, , - , mnmn,n,,-, ,, , mnmnrvmnrn,n,-,m-n,r mnnrr n, , nr, - n-, -- w.r rnnv n r m mnmnn. n, m ' m , rm r n n w vv m-r, , ,nr,m,n rm r n, . , --, -- n nnr-m,n wrrwnn mn w vv n, m.n w wrvvrn, - mm rm r wr rn - mm w rmnnrrnm,n. rmn w vv , nnrmn. wr rn mn mrmm rm r n, w m-mn, - rmnnm r n, , nnrmn,. m - . n rmrw r ,rm rn,r ,-- vvn, wrrwnn - mm n mn rm r n mn w vv ,m-mn,.mmrmn nnnn,n-nrr-,2m°rvnrrmnrn.n, . wr r n n nnrmnn mm - , wr rn m2. nnr n, r wr mrv r . nn mm w r mn n rmv . rr w rmv nr n rnr n,nwn rrr n r mn. wr r , r °m. - rnr wx rr - mn r r, nnr wrn r ,nnrr n.m wr r nn r r . n r nr wnrmn . nnrmwn, mr m wnr nn nr, nnr , r mn rmv m. rnn wr vm w- m.m w rr rn mrv . n m rznmm wr w mm rr -°nn . n. wr ,°nrn,r w w ,r mn w , ,. rnn wr rmvn wr rn n . nn rrmnvr,n wrnn , nmrzmxnnmmn w,rrn zx n r znn, . r -nr rn wr rr mrnnr.rmnmun ur mmr nnrnmnnr mrnr- nr wrrv n.n- nv m, . w w . n rm r, wr nr rn -nn, -vrn, ,nn m r ., n r, . nx , rvn r rn, nxr wmn,, rmnnnn wr r-m- wnr mnnvrn w.r wrrm n . n mrr- w rrm n m nn- .r rm- rmrm..rrrn m,n w mr rr-n. wnr wnn nrz n rmnr-nr rv, r, rnrwnrrmn . n w mrrmr mnn r wn nrrrmrnnn nrmn x mn n n vm m w n vrr rn r.m wr nr nnrn wr nrm: :.m mrmn r rm.r rmmn r-n r rrm v - wr rr w mrv, r.n mw w n v w nr . m wr vr wrrzn wz .nn wwr.r rrnmrn rn mr nn w rrzn rn n rmmn r, nv n rrm.r,. mm wr r w r rn n w r,n w n w nx w. r n .r wr rrmnmn r mnn, w, - wrn wn-mnnn . n w wr wn-- m w mn w:, ww wnvrn n.x,°n w nn m-m r n. x - wnn ,n:, wrm-r,n nn w r n r. - w w w nx nvnr. -- r mx wrn r .,r-w wr mxn r. r n. m r,nrrr nm °.rwr, wrm n rn vrnnn°.mrnn. r,w r,w rn wr nx m-r wr r nn n nr .rn nm n wr -n , nm nn nmmn n m wrxmn w r. nv nn mr n rn v n wm mnrnmn rn wrrmm n rm rn. mrn rnr n wn w mnxrrrr'mnrm-- n.rv n wnn n n mr-r wnr, n rnnrnnr n nrn r mnr, rm r n. w n rvrrnr n mmnr ,nn r nrr wr-rrn -n°nrm rm r n, nn n w nmr,r- r.rm mm. m n w rrm rnn r r-,m n wvrnn rnmxnr vrmm , , nrwrrvrr r -, . rm mr,n m rm rn, mn,w nnnrn wnr°rw:nzm vn °° r r mn °,nnnxnn ° rr-m°,nnn °°. rmxrrmr,n n r nrm mxr wrr n vrn n, r n m r wm m . r . w xr w nn-w rr -, n -r. nnrrnr-m,n wmn nnnm r nrr wn rmr: mnv m- n nrn °r n mr °°,w n°° r mnn°°°rnnnnxnn °r mn°, °. mnrn zn, wn ° r n nnrn r n r r n w n n mn- w.rmrnn. w..r wrr,n- n r --: r. : - ’- -’ : ---::’’-- --’ : --:’ ’ : -:- -’ : -:’’-- -’-’ :: n-r w wrrnnznnrnrn,.r n,n r n n nn rmr mun n n ._r:,,nn mrn:,n,- nrm-r- m,n mmr v r v.rr,- rnmr m n n, m m n n w,r , v . rmn wr -r n wnrr rmnnrw nnvrnn, r r mn.nnvnrn w rrmn r n r r rmn. rmn r nnn nn r r, n m mr n, -wnn m-n-rrn w mm, nwv mn wnn r'nrn.r mr-mnrn wrmrrmrnnn m vmn .rn wrrmrr mr- nn, rrwrrvr, rmxrr n -, , n rm nnrmrn, wn m rm rn, . nrn ° r:n nzm vn °r mn,n nzm r °,nnnnxnn °r mn°,w wnrvn °r mn° nn °°. n r n m rm n r-m r nn r n vm . rmxrr m wrnr n-.r-m r n w nrn vrm n m wnrrn,nr-mrn w rrmnn -, , , m x rmx rn, . rmrr-r mxn ,n m rm r mn, m nnn wr:n nzm vn ° mn,wnnzmnnvrnn°°rr mn,nnnnnnx°n.n r°nr mx,r wrr n r-m r , m x rmx- . , rmr-r mx,n n nm-r, vn,nrrnnnrmrnnn°nr wr,:n°°rr m mnnrn nzm nnnxnn,w nzmnvn °r m r r wrnz n r rr- nnnn wr°. r mrm n wmnrnn n:n,n w m rr m = r-. *n wr, wr rrn r rrn vnmrmn , rrn vm n , mr nm.nm w > r n,n rrn vm n . w wrnznr ,rr w wrn n n, rn n nn-m nm.r rmr m n m nn- m n nr,,nvnr rm-rm nrm nrn w-m- -nr,n.vrnr m n rmnvnrn,n rn n nvnr rm w- n. , n wrwmr r, rn- wwn m r r rn nr m- n rnr n, wn,rnr rr . wr n rmvx . nvnr, nnn wn w r xrmn.mrrnnnrn wr n n n n nrm wr nnmnn wrr rn r r r n.rr n nn,n wr nr w r wm r. w-rnnn wn-nnr.n nn -,vn-,r r nnr , wr w n nrnnrnmr- w n .r n, -n w nrm rn nr w nvnr. n,nn vrxnr mn .rw m-r, --n wr n nnrn rmr wn m rnr,n mn r. mn w nr rrn v n nmn rr rn,rn..nr -m rn w rv n. n w rrm n,w vvnw wr n nmnwmr, w nm rm m r.rm r mnmnx. rwr wr rmnwmr n rnr rn mxmmwr rnnn wn , , r ,n ..r rnnn wr nnn mn n ww rn rmv nv wr r r mn wr vrn.. . wrnz nmur-r nm wu nrn m . nr m wnrrrnmnrn nr mrrrr vm. ,xr . m rmnr n, z rm mnr -xr, n,n - rm nr n wr n r .r rn n-n w, n, m wr n w n ,nn -n n- x r-n n- ,n x r-n n--, w r-rm,n-n ,r nrn .n n n mn rr’r x r- n n nvrn, rn nmn nm nr nn wr rn nn nm,rmr nn.nr mrzn r nrn m w nrn nn, vn nn n, rn w rrm m w rnm rn mnr nmrn nm.n nr nn n nnn r.., n znr n, - zn.m w rn n,w-r nn nn- w rm :...rn n rmv v rrnrnn wrnzrrnnnn rn rm r, mm rm.r rr nrnrn mmrn, :,rnn w,rnn-- wmrmnr mn:,,nnn, nn--- mmn m :, n,n- :, m, ,n- :,n m ,n- :, m, m, n ,n-x :, m, nn-rn :, m, . nx,wn n n w n-r -, - n-m r, m-mr, - r nm wr v zn xmn mn- w.r w n , r wmrn rm m m nrm n m r vr .m wnr nzx nrmr mnzn n, - mn mnnn - mmn r.r,n n- wnn ,n nnn w °r-w rvn mn n .nr, n n- n vrn w wrrn wn nn- mn mn r n nn m, w mn . -rnz - -n wr ,n rnnnnn - wr n wn mx m w nn n- n - . - r ..rn nr -.nw nrzn w r- r mn . . r n w-r-rn w rr nw, rm r n w nwzrnnwrrvnnm nnn-n. nrr.nv.nmrmnrnwn.nnm wnm.rrrm w °r. mrmrmr mn , wrm, mn mmr - - mn ,.m, xm mm, mm,rnrn mnw r nmn. . wrn r mnn ,rnrm r w mnrr. r-mn wn nnnrn. rnmnr wrn mrn rnnmznnrr.nrn mn . nvnvrm r,n mrm rm mrz wn wnrwn mz-. mx rmmnnnm wnw w , wn r v ,nrmn w m mnxn n mrmz nnr . . m rmrrm wrnrn rm mzn r n v rnr r r :www...r w n nrm rnn nn n n wrrrm nr vrn r . vrn.,n rvrn.. rv, ., rr , . r ,nn . n ., w r r ,rn rm r , vr r r nnrxrnnnnr, rmnnrnnrn,.n. r n n.rm vnr’n.nn rmn mwrr vrn. wr z r m nr rnmnmm. rrxr nrn mn n n nrnrrv ,nvv wr,m , mr nnx =vr v vn.v −.- vxn wn-rnrmn wn nznnxvn rn nr r- nm-mr n n n rwn nnrr n. n r n w nm w,rrn m r - rvvn n n w nz rr nw xrrn nr.rnrr n n.rn mnnmrrnm rr nnmr- v.. n n nnv.n w n xnrv... nn w wrr rm r n v...n nrmnn. rrmr wn nnn v... nnr- rmnnn -rnnn nrn , m rnn, n z w -, n w rnr rmnmnn r wr. r mr wm- rvr rmnn. mrnrnnmn- r.rv. r-r.n nv w,v ,nnrnnnnrv n,rnrrv. , mvrrvrm vn, , nv w nvr.r.v v ,n m - v wr n <. . nn nnrw., v rrvrmw- n r rnrn:nn n mn n rn nrnrr nr,n.,rn. . . ".mr-xv,m.-n.rnn,.rmrn,. m,. r,.n, - rnr m ." r : ,. . ".m,m.n,.nnrr,r.mmnmrmm,n.zr,.n,. r,.rn. rn r r n m." r-,r.m,,w.r,. ...,.w,., rz,..,nrn,..,nrn n,., …  : n mnmnnrmnr xr r v n.nrnn rxr r ,n :.r. .mnnn,rnmnn n. “.nrn. .” rmrmnnr nrz rnmr : -n v. , : -. w.,nnnnv rrnm,n r n. “ n mnnr-n :. n- n r nn r .” n v., : . rn,r-,rr. nnr nxr r w .n --.- n r n,... :-. : r,.,.nr,.-_.., ": . nn-nrnn vrnn rnrn vrnmrxr:mnn rnrnnnvn n n..-,r.nn,r.rrmnn,v z , .r.'"rr. m :-. : mrnv - rrm,. n.n . "r rm rr : -. rn xr nn ."rn rvrv,.,..n,..vnmv,.. n,..nmm,.rm ,.n.n."n..vz:-. ". r: vr r n -:: :www...nrwrvw n : mn n mn : : r- .. :::www...nrwrvw nnmr= n==""r-nn" nxn" : n mn n : : xnr- ::n =:wwnw.n-.nm.n.vnr _ . ..="nm="n nrn "n_nnm="- -" nn:mn:n:.." n: : mn n : : n vn rn : n nrwn n wnn rn nn rn rv. rn nr ., n wn Heat contro o maasto IDH-moumtaate No cno-ncancer N = N = 11 N tro ener = 1 N = 71 memae 244 2727 Aeae 747 17 24 D MaenanearsD 4± 1 ± 12 ± 11 ± 1 oosass 111 Astrtoma - - - Astrooccttooma, WH 2 - - 2 - Astroctommaa,, W WH - - 21 - oenroomHa, 4 WH 2 - - - - 21 - oenrooma, W -MMTromH - - 2 - Metateoter metaton- 44 - Inncmonectusavtee - - - - -IDH mutat- - W Iteon- 111 - - IDDHH11 R R1122 - - - - 117 - IDH1 R122HW - 7 - IDH2 R17 - 12 - Ben t Nunm - = co 1errsera M Nen m - =nao Ext c 1raa-n7ccrearna me Btars -atanss ene 2 N = 77 N =emaer mae 1 4 4 Ae Dane 1 4 MeaaDrs 4± 1 1± 14 ± 1 ± 14 avoen roronsoms eaxusao 212 - - - ran ma 212 - - - Haemoanaornasoma 212 - - - Poctc astrocttoomma 1 - - - Ptutar aenomaa 412 - - - - - cwannoma 21 - Mennoma 2 - - - Mennoma,, W WHH 12 - - 217 - - Tunscstuoen o, orn - - Aenocarcnoma o te esoaoastrc Breeast AE - - - - 111114 1222 orovrx - - 2 - Enomecta - - 4 Esoaetursum - - - - 1111 11 - nn Ben tunm coerrsera Men mnao Ext craa-nccrearna me Btarsatn N = 1 N = 2 N = 77 ass Tssu N = Hee o orn Kane an nec - - 4 - Bone marro - - - 11 Lver w - - - - 1 - 11 Lun - 1 - Lm - 41 44 Pann - creas - - - - 1 - 11 111 11 - -1 P Preoura - - 2 22 arstate - - 11 11 tomcoma - - 11 11 T Vuroac - - - - 11 - 22 anvcaer o unnownrmar,P - - - - 121 - - utens ecct Postve Neatve EoDBa-sFtoNma A. va-uoe tvt. I to1.Ito re vacuteve re vacuteve EDA-FN 27. .7..27 to..4.7 to. .T.4 1. 1..1.42 to.77.4.to1. .7EDoTmBoN-tFa-nNaEA,t EV1oDsnA o-.7.17411.414E.2+.- 14277. 14..77to71.2. 17..to4.. 7..FN an TN-A1 1 1 mEeBtarsatanssAvuat-uoetevntsIecct Postve Neatve EDDBA--FFNN ..417 22122..47747.2...142t too4.I..4 t re vacuteve re vacuteve too4.2 .TN-A1 .1 14.4 4..1244...1.1472tto1o 717.24..to774..124 1..EDoTmBo-tFanNaE,t EVoDsn o.7441.1E+.7 7.2.1.14.71to.47 .FN an TN-AA-1 1 - 1 1 1 1 1 1 Extra-crana ut-o ens ecct Postve Neatve EcDanBc-FeNrAvauetvt4.2I to4.7Ito re vacuteve re vacuteve EDA-FN ..77 112.. 27..21177.27..27to..47.7..7 to27..211 .. Fn - IDH-m EDBomut -Faantnn NAut-o ens . vauetvt.4I toecct .IP to re voascuttevvee Nree vaacuttevvee EDA-FN .772 2.4 .71..14to..to.7 .TN-A1 .21 1142.. ...7to7.4.to. .Tota EVs ..2E+744..to 42.2.1 to. .EDomnaton o.22.21to 1.77.to. 1.7FN aBn-FN T,N ED-AA-1.1 - 7 1 1 11 7n mtr Be Eaa EDtan Bur -me FnnoNcrsA.777 v 1uat1-uo.e tevn .2ts 2IecctIPreoscttvvee Nreeacttvvee TDNA--FAN1 ..4 1..17 7..71.41.42..27t too..777..44.17 t to vaue vaue o.2 .4...4 ttoo 47.2..444to7..7 ..7EDoTmBo-tFanNaE,t EVoDsnA o-.7.17.-E+1.22.to41. 72 7 t.1 to o77. .FN an TN-A11 1 2MeEnDnB-FoNma A. vuat-uoe tevnts 7. I toecct .IP to re voascuttevvee Nree vaacuttevvee EDA-FN .7744 1. .71..to..7.1to. .TN-A1 . 12. 1.71..to7 1..1to1. 7.T.12E+1...4 to7.74.2.12to1. 7.7omotaann TaENtVosn.2 .7 .21.77..7 1.7F ENDB-FN, ED-AA o-1.1 - 7414 to1 1to1 4oEDBas-FtoNma A.1 v 1ua.t-uoeevntst 1.71I toecct .IP to re voascuttevvee Nree vaacuttevvee EDA-FN .1 1.17 .77.7.12 to..to.7 .TN-A1 .2 1 .411.7.41 to7.4.to4. .Tota EVs1.44E+172..1to2..to.7 .omnaton o.77 4 74.77t 7.72.to4. 7.F END aBn-FN T,N ED-AA-11 - 1 1o7 11 1m EeBtarsatanssnnAvuat-uoe evntstIecct P EDB-FN . 1. 1.477Ire voascuttevvee Nree vaacuttevvee TDNA--FAN1 ..44 1.147 7.1472.21..7E7 .742....77t to.. .1.7toto.4.1t o7.71.7.to .4.to1.4 .to7.14 .o.7 7.oTmotanaEtVosn o.744+1.7 7.2.1.71.47 .F END aBn-FN T,N ED-AA-11 - 1 1 1 1 1 1 Extra-crana ut-o enst ecct Preoscttvvee Nreatve EcancerAvauevt. I to.Ito vaue e vacuteve EDDBA--FFNN ..1 1. 7.227.2.7to..4to.2 .TN-A1 .444 11..142 .127.2.to..44to.12 .Tot2.E+1.1244.7.4 to.74..471to4.21 .7EDomB-FanNaE,tVosn o.2 .47.to .7.to.4 .FN an TN ED-AA-1. - 1 1 11IDH-m Postve Neatve EDoBmutant -FaNAut-o enst . v1a.u2evt7.2I toecct 77.4Ito re vacuteve re vacuteve ETDNA--FAN1 .. 11..172 1..7 17..4.4to...1to1..7 ..44...44ttoo 4...2.1 t too.7 .oTmotanaEtVosn o..2E+7. 2.211.77.. 1.7F END aBn-FN T,N ED-AA-11 - 1 1 1 1 1 1ntr aa Benn Pos E taurmenocrsmA vuat-uoeevntstIecct EDB-FN .74 1.7 2..Ito re vacuttevvee Nree vaacuttevvee TDNA-FN .12 1.2 .7441.14.4...4 to.4t too.7..4..17 t to2. .o.7 .7Tota-A1 .7..7E+.74..to7.417..1 to.7 7.EDomB-FnNaE,t EVosn o.7 1.22.to7.to7. .FN an TND-AA-1. - 1 144 7MeEnDnB-FoNma A.22 v 1ua.7t-u4oeevnst .t2242.7I toecct . .IP to re voascuttevvee Nree vaacuttevvee ETDNA--FAN1 ..24 1..27 7..27.2...24to.7.4.17 to7..222 ....to.71.14..71to. .7Tota EVs .2.2E+.7 .42to1.747.2.1 to.7 1.7 F ENDom aBn-FnN Ta,Nt EoDn-AA o-1. - 1 1 1 1 1 1 n rr ener oasoma, = memae eae eears anan ± oaossosma Base A ut-o enst Postve Neatve Ene vaue vt I ecct I vareuectve vareuectve F E FNDB- .217.4 4..247 to 7..47to72.7 1.7T AN TNDA-- ..72712.11 1.1. to 7 1..4to2. 1.Eo1Vtsa 42.712.1.27E+1 4. 4...24 274 to 7 to 4 477...1 to .4to47.. ..47P Postve Nea E FNDDB- A. vautu-oe vetnst 7.I to ecct 71.4I to v7are .u7ectve va7ret .u7ectveve EFT ANDA-.41171.. 11..74 to 414..7 to4.2 1.TN Eo1Vtsa- . 4.2211..2E+1 4 7 1...2. to 7 77.. .11 to 7.2.21 t too71..4 ..nreoene4.Base Ave vautu-oevetnst I ecct I P varoesuetcvteve N vareeuaetctveve PDvautu-oevetnst I ecct I vareuectve vareuectve n rr ener easac cncara, = memae eae eanears ano ± on-ssmsa ceun cancer, Nan cotsocmaeetrow Pearam EtMean D ReerenceNon-canceronors E FDerane IFDNBrs- NA- ..4 ..41 - 1. .4 -.4 T AN1- - 1.27 .42 -. EDB . .4 - 1. .2 -.77 oastoma E F FDN- NA- . 4.2 - 1.4 4. -.1 TN- . .2 -.4 2.4 - 4. E AD1B- . 2. -. 2. -. Bran metastass E F T FDNA- .4 4.1 - 1.1 4.4 -.47 ANN1- ..4 22..27 - 7.7 2. -. E FDNB- 4.21 .2 - - 1..7 2..427 - - 4.1 Ex ctraan-ccrearnsa E FDNA- . 2. 4.4 T AN1- .1 2.14 - -..1 22..27 - - 4..24 E tantoma FDNB- 4.17 2. -. 4 IDH-nu E T FDA- 2. -.2 ANN1- 22.. 2.1 - 7.1 1.7 -. E FDNB- 1.7 11..2 - - 4.. 21..2 - - 2..1 ntra B tauermnenorcnsma E T FDNA- 2.1 1. -. 1.12 - 2.1 E F ANDN1B-- 1..771 1.7 -. .1 - 2.74 Mennoma E FDNA- 2.1 1..12 - - 1..4 21.7 -.24 T AN1- 2. 1.2 - 4. 1.. - - 22..7 PR PeMeancoesm Dco I Eatrearmsasma asmeasm c Roeeersenmce raansmea coesm FDNB- 1. 4. . - 212. 7.asma Non-canceronors E FDNA- 121.2 .2 4. 4 - 11. T AN1- 1.1 . 2 - 2 -114.1.2 114..7 - - 11127..4 E oma FDNB- 21.11 4.4 - 1 7 oast E T FDA- . 42.2 - 7.21 ANN1- 4414..22 21..2 - - 1.24 4.4 - 4.1 E Bran metastass FDNB- 7 17. 1.2 -1.2 E T FNDNA- .1 4. - 14.1 7.4 -. - 4.4 27. -47.2 .7 - 4.74 E F ADN1B- 44417..2 242..4 - - 1144.7 . -42. Ex ctraan-ccrearnsa E FDNA- 441. 22 2.4 . -. T AN1- .2 24..472 17. - -1.1.1 4..41 - - 42.2 E oma FDNB- 41. 2. - 1. .24. IDH-nutant E T FDA- -.47 ANN1- 414..21 41.21 - 122.1 21. -1.2 E FDNB- 22.7 1247..1 - - 7..2 2127..22 - 47.1 ntra Baernencnma E FDNA- 1. 14.2 - 41. -1.7 tumors T AN1- 1 4 1. - 2. EDB . 17.71 - 4. .1 - 24. Mennoma E F T FDNA-- 4. 14.24 7.1 -24. 22. - 47.2 ANN1- 2241.. 114..24 - -..2147 1214..117 - - 24..4 NTAPeatrearmsar Mtceaensm artcDesm a Rretcereesnmce raansme artcesIm Non-c Broancaesrtomonaors 1.a1sEm+a1 1.1a7sEm+a a 1 -.E+1 21..1.1a 1Esm EE+a ++11 - - EaxntrmetastassTota22..4EE++11 11..EE++11 - -..47EE++11222...7EEE+++111 -IDH-ncuaa tna-ccnretarnsa omaEVs 21..12EE+1 1.2E+1 -.E+1 21...1214EEE+++111 - - ntra Baernencnma+1 1.42E+1 - 4.E+12.4E+1 tumors 1.E+1 1.E+1 -.1E+1 12.1.74EE++11 -Mennoma2.1E+1 2.4E+1 -.71E+112..EE++11 -

Claims

m.n m n,r n nr ,rrwn rr nr - -rvnnmr xrmn: r v n nrm n - nn xr rrm m r v rn nr- vrn xxrm r--mr mnn rrnnnn -- r vrnnnnxmrnr v- n m nn nr- xrn-, wrn xr-mmn rnn - nnnmnrnn -. m rn m, wr- . vr nrn rrnnr . mnmnr rmn. vn m, wrnv m nn nr-. rn n. r mn mrrn nrn. r n m , wrn vr.vn mnrrnvrrm, wrn n n r. m rn m,n v r . vnnr wrn n n r nn:, n rrn r r rm r ..mrmr . n r mn rnnmr .rrrnmnmnr . mrn rn nnmrr. v n m, wrn nr- nn n vnrn,nmrn rn w nn.. m rn m, wrn n nn rnn -.r . mn. m rrnn nnnmrr, wnrnm, wnr . nr- vrn wm n rn . m rn m, wrn r. . m rn nn rwnmrm, n wnrnwmr. xr r v wrnrn nr vrn n, . m mr. nnrnnnr-n r vrnn m, wrnn rn n . rn . mr rn.m, wrn rmn m v r . mrmrrn nnnn nrrrnn m, wrn nr nr,rn nr,nnr, nr,r nr, rnn . m rn m, w mnm. r rn nrrn nr rm . m rmn,mn,rn m , mnnm,rn m . xr r v r n nr wn rmr ,r wnrn nr- . m m n. . mrnnm, wrn m n rm: ..nn n m n .n m mnn ..n m n . - m mnn .r mn, rnn .m, wrn nr- m n rmn rrz nr-n mrmnn. w r rvrrmn nr.n m mr- rn vrnnn r n m, wrn mnn. -, -,n - . m rn nn r n m , wrnmnn . nnnr- vrn -, -,n -

Images