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(1)Cover Page. The handle http://hdl.handle.net/1887/19776 holds various files of this Leiden University dissertation. Author: Runtuwene, Vincent Jimmy Title: Functional characterization of protein-tyrosine phosphatases in zebrafish development using image analysis Date: 2012-09-12.

(2) 5 !. Cell Outliner and Cell Roses: New Tools for Automated Cell Membrane Detection and Determination of Cell Polarity Vincent Runtuwene and Jeroen den Hertog. 119.

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(26) ͵‹†‡’‡†‡–‡š’‡”‹‡–•Š‡–‡”‘œ›‰‘—•’–’”ƒΪȀǦϐ‹•Š™‡”‡‹ ”‘••‡†ƒ†–Š‡‹”‘ˆˆ•’”‹‰™ƒ• ‹Œ‡ –‡†™‹–ŠʹͲ’‰ ǦǤŠ‡‡„”›‘•™‡”‡‹ƒ‰‡†ƒ––Š‡ʹǦ͵•‘‹–‡•–ƒ‰‡ƒ†•—„7 -"6&")'#=<$/")0'=4".$3=$-"6&")1()/9$>5"$*"-&#'()/$(2,/"-$+"*"$4*01"--".$3=$!"##$%&'#()"*$,).$ '5"$*"-&#'()/$.,',$+,-$4#0''".$&-()/$!"##$H0-"-9$>5"-"$/*,45-$-50+$'5"$.(-'*(3&'(0)$08$'5"$0*(")7 ','(0)-$08$'5"$1"##-$()$'5"$4*"-02('(1$2"-0."*2$()$*,.(,#$5(-'0/*,2-9$>+0$1(*1#"-$0)$'5"$/*,45-$ ”‡’”‡•‡––Š‡ͷΨȋ‹‡” ‹” Ž‡Ȍƒ†ͳͲΨȋ‘—–‡” ‹” Ž‡Ȍ’‘’—Žƒ–‹‘ˆ”ƒ –‹‘ƒ”‡”•ˆ‘”–Š‡”ƒ†‹ƒŽ 5(-'0/*,29$I('5()$'5"$3,*-$08$'5"$5(-'0/*,2<$,$-"10).$-&3.(;(-(0)$(-$-50+)<$().(1,'()/$'5"$8*"7 6&")1("-$E3=$-&*8,1"$,*",G$,).$()'")-('=$08$"#0)/,'(0)$E,-4"1'$*,'(0$(-$().(1,'".$()$10#0&*$10."$,-$ ().(1,'".$()$'5"$#"/").<$30''02$*(/5'G$08$1"##-$+('5()$'5,'$4,*'(1&#,*$404&#,'(0)9$J'&.")'7'$-','(-7 '(1-$+,-$&-".$'0$1024,*"$'5"$1"##$"#0)/,'(0)$;,#&"-$,).$I,'-0)$K@$-','(-'(1$+,-$&-".$'0$1024,*"$ '5"$0*(")','(0)$.,',$08$'5"$1"##-9. !"#$%##"&'( !"##$%&'#()"*$+,-$."-(/)".$'0$."'"1'$1"##$2"23*,)"-$()$-,24#"-$+('5$5(/5$ ‡ŽŽ†‡•‹–›ǡƒ”‡†™‹–ŠϐŽ—‘”‡• ‡–‡„”ƒ‡ƒ”‡”•ǡ,16&(*".$'5*0&/5$10)7 801,#$(2,/()/9$:'$5,-$3"")$;,#(.,'".$‹˜‹˜‘‹œ‡„”ƒϐ‹•Š‡„”›‘•ˆ‘”–Š‡‡„”ƒ‡ ƒ”‡”•  Ǧǡ ‹ ™Š‹ Š –Š‡ Ǧ–‡”‹ƒŽ Ǧ‘–‹ˆ ‡†‹ƒ–‡• ƒ••‘ ‹ƒ–‹‘ ™‹–Š–Š‡ ‡ŽŽ‡„”ƒ‡ȏʹͳȐǡƒ†‡ ǡ ‘–ƒ‹‹‰–Š‡Ǧ–‡”‹ƒŽ•‡“—‡ ‡ ‘ˆ Ͷ͵ǡ™Š‹ Š‡†‹ƒ–‡•ƒ••‘ ‹ƒ–‹‘™‹–Š–Š‡ ‡ŽŽ‡„”ƒ‡ȏʹͷǡʹ͸ȐǤ

(27) ƒ‰‹‰ ‘ˆœ‡„”ƒϐ‹•Š‡„”›‘•™‹–Š—„‹“—‹–‘—•‡š’”‡••‹‘‘ˆ–Š‡•‡ƒ”‡”•›‹‡Ž†‡†‹ƒ‰7 ‡•‘ˆ ‡ŽŽ•Š‡‡–•™‹–ŠŠ‹‰Š†‡•‹–›ƒ†ƒŠ‹‰ŠϐŽ—‘”‡• ‡ ‡‹–‡•‹–›•‹‰ƒŽƒ––Š‡ 1"##$2"23*,)"<$,).$011,-(0),##=$#0+$()'")-('=$;"-(1&#,*$-'*&1'&*"-$()$'5"$1='0-0#9$ ‘‡‹ƒ‰‡•ǡ—•‡†ˆ‘”ƒƒŽ›•‹•ǡ ‘–ƒ‹‡†ϐŽ— –—ƒ–‹‘‘ˆ•‹‰ƒŽ‹–‡•‹–›™‹–Š‹ƒ 2"23*,)"$-"/2")'$0*$,$/#03,#$()'")-('=$/*,.(")'9$>5"$2"'50.$5,-$-50+)$'0$3"$ *03&-'$()$5,).#()/$/*,.&,#$()'")-('=$15,)/"-$,).$5,-$-50+)$'0$3"$,3#"$'0$1027 4")-,'"$80*$-2,##$,3*&4'$15,)/"-$08$()'")-('=<$4*0.&1()/$;"*=$,11&*,'"$*"-&#'-$80*$ '5"-"$2,*?"*-9$ ‡ƒŽ•‘•Š‘™‡†–Š‡‡–Š‘†–‘„‡‡ˆˆ‡ –‹˜‡‹ϐ‹š‡†Ͷ•‘‹–‡•–ƒ‰‡‘—•‡ "23*=0-$ #,3"#".$ +('5$ @$ ,)'(30.("-$ 80*$ A71,.5"*()<$ ."20)-'*,'()/$ ,$ +(."$ 1027 4,'(3(#('=$,).$,44#(1,3(#('=9$!"##$%&'#()"*$1,)$()$4*()1(4#"$3"$&-".$()$0'5"*$20."#$ 0*/,)(-2-$0*$,)=$‹˜‹˜‘$0*$‹˜‹–”‘$20."#$=("#.()/$(2,/"-$."-1*(3()/$.,',$()$@B$ ,**,=$80*2,'$()$+5(15$'5"$1"##$2"23*,)"-$,*"$#,3"#".9$!"##$%&'#()"*$2,=$3"$,44#(7 ƒ„Ž‡ˆ‘”ƒŽ–‡”ƒ–‹˜‡’—”’‘•‡•ƒ•™‡ŽŽǤ ‘™‡˜‡”ǡ‹–‹•‡••‡–‹ƒŽ–‘ ‘ϐ‹”–Šƒ––Š‡ 03C"1'-$08$()'"*"-'$,*"$08$1024,*,3#"$-(D"$E()$4(F"#-G$'0$'5"$1"##-$('$+,-$."-(/)".$'0$ 20."#$-()1"$'5"$)&2"*(1,#$;,#&"-$&-".$()$'5"$,#/0*('52$+"*"$04'(2(D".$80*$'5(-$ 4&*40-"9$ Š‡†‡ϐ‹‹‰’ƒ”ƒ‡–‡”•ˆ‘”–Š‡†‡–‡”‹ƒ–‹‘‘ˆ ‡ŽŽ’‘Žƒ”‹œƒ–‹‘†‡ˆ‡ –• ƒ”‡“—ƒ–‹ϐ‹ ƒ–‹‘•‘ˆ ‡ŽŽ‡Ž‘‰ƒ–‹‘ƒ†‡†‹‘ǦŽƒ–‡”ƒŽ‘”‹‡–ƒ–‹‘Ǥ‘˜‡–‹‘7 ,##=<$'5"-"$,*"$,),#=D".$,).$*"4*"-")'".$-"4,*,'"#=9$:)$.0()/$-0<$'5"*"$(-$,$40'")7 '(,#$#0--$08$()80*2,'(0)$,30&'$'5"$10**"#,'(0)$3"'+"")$'5"$,80*"2")'(0)".$4,*,27 132.

(28) ‡–‡”•Ǥ›”‡’”‡•‡–‹‰„‘–Š‘ƒ‡ŽŽ‘•‡ǡ–Š‡™‡ŽŽǦ•–—†‹‡† ‘””‡Žƒ–‹‘„‡–™‡‡ !"#$%&'()"*('+"'%,-()$%,+(,#+%*$",)()$%,+%.+)/"+0*"1%!$)$2+!"1%#"*!+3(1+2'"(*+ .%*+ )/"+ 3$'#)40"+ 1(!0'"15+ 6"+ #"!%,1)*()"#+ )/"+ 2%!7$,"#+ 81"+ %.+ 9"''+ :8)'$,"*+ (,#+9"''+;%1"1+.%*+(,('4<$,-+)/"+!8)(,)+0/",%)40"+%.+”’–’ƒ+$,+)/"+0*"1%!$)$2+!"& •‘†‡”‘ˆœ‡„”ƒϐ‹•Š‡„”›‘•ǤŠ‡ ‡ŽŽ”‘•‡• Ž‡ƒ”Ž›•Š‘™‡†–Š‡ ‘ ‘‹–ƒ–Ž‘•• %.+"'%,-()$%,+(,#+!"#$%&'()"*('+%*$",)()$%,+%.+)/"+2"''1+$,+)/"+!8)(,)5+ !""#$%&'$()*&)+*,()%#-.$() =/"+,"2"11$)4+%.+#")"*!$,$,-+(,#+12%*$,-+$,#$>$#8('+2"''+7"/(>$%8*1+(,#+ 0%'(*$)4+(1+0(*)+%.+(+0%08'()$%,+%*+)$118"+$1+,%)+*"1)*$2)"#+)%+)/"+1)8#4+%.+-(1)*8'(& )$%,5+?,+#">"'%0!",)+('%,"+,8!"*%81+"@(!0'"1+2(,+7"+.%8,#+$,+3/$2/+0%'(*$)4+ ƒ†–‹••—‡‹‰”ƒ–‹‘ƒ”‡‡›ˆƒ –‘”•ǡŽ‹‡‹ƒ‰‹‘‰‡‡•‹•ȏʹ͹Ȑƒ†–Š‡‡•–ƒ„Ž‹•Š& ‡–‘ˆ–Š‡Š‹‰ŠŽ›‘”‰ƒ‹œ‡†•–”— –—”‡‘ˆƒ—†‹–‘”›ƒ†˜‡•–‹„—Žƒ”‡’‹–Š‡Ž‹ƒȏʹͺȐǤ ‘ŽŽ‡ –‹˜‡–‹••—‡‹‰”ƒ–‹‘ƒŽ•‘—†‡”Ž‹‡•™‘—†Š‡ƒŽ‹‰Ǣ–Š‡„ƒ•ƒŽ”ƒ†‘‹& #$>$#8('+!%>"!",)1+%.+)/"+",#%)/"'$('+2"''1+(*"+*"%*$",)"#+)%+2%''"2)$>"'4+0%'(*& $<"#+!$-*()$%,+74+!%#8'(*+2%,)*%'A+$,#"0",#",)+.*%!+-*%3)/+.(2)%*1A+80%,+)/"+ ƒ’’‡ƒ”ƒ ‡‘ˆ ‡ŽŽˆ”‡‡•’ƒ ‡ȏʹͻȐǤ‹‹Žƒ” ‡ŽŽ„‡Šƒ˜‹‘”•Šƒ˜‡ƒŽ•‘„‡‡‘„•‡”˜‡† ‹‡–ƒ•–ƒ–‹ ’”‘ ‡••‡•‹‘—•‡ƒƒ”›–‹••—‡ȏ͵ͲȐǤŠ‡ƒ†˜‡–‘ˆ‘”‡•‘& 0/$1)$2()"#+$!(-$,-+)"2/,%'%-4A+$,2'8#$,-+/(*#3(*"+(,#+(+0'")/%*(+%.+/$-/'4+1",& •‹–‹˜‡ƒ†•’‡ ‹ϐ‹ ϐŽ—‘”‡• ‡–’”‘„‡•ǡƒŽŽ‘™•–Š‡˜‹•—ƒŽ‹œƒ–‹‘‘ˆ–Š‡•‡„‹‘Ž‘‰‹& 2('+0*%2"11"1+$,+8,0*"2"#",)"#+#")($'5+B()(+(,('41$1+%.+)/"1"+/8-"+#()(+1")1+.%*+ !%*0/%'%-$2('+."()8*"1+2(,+7"+-*"()'4+.(2$'$)()"#+74+)/"+81"+%.+('-%*$)/!1+'$C"+9"''+ :8)'$,"*5+ ‡ŽŽ—–Ž‹‡”‹•ƒˆ”‡‡Ž›ƒ˜ƒ‹Žƒ„Ž‡

(29) ƒ‰‡ ’Ž—‰Ǧ‹ȋŠ––’ǣȀȀ”•„™‡„Ǥ‹ŠǤ‰‘˜Ȁ ‹ŒȀ’Ž—‰‹•ȀȌǡ™Š‹ Š™‘”•™‹–Š

(30) ƒ‰‡ ˜‡”•‹‘ͳǤͶͷŠ‘”Š‹‰Š‡”ǤŠ‡ƒ’’Ž‹ ƒ–‹‘ ‹•˜‡”›•‹’Ž‡ǣ‹•–ƒŽŽƒ•ƒ

(31) ƒ‰‡ ’Ž—‰‹ǡƒ†–Š‡ƒ’’Ž‹ ƒ–‹‘™‹ŽŽƒ’’‡ƒ”‹–Š‡ • ”‘ŽŽǦ†‘™‡—™‹–Š‹–Š‡

(32) ƒ‰‡ ’”‘‰”ƒǤ’’Ž›–‘ƒͺǦ„‹–‹ƒ‰‡ȋ‘”•–ƒ  ‘ˆ‹ƒ‰‡•Ȍ”‡’”‡•‡–‹‰–Š‡ ‡ŽŽ‡„”ƒ‡•ǣ–Š‡‘—–’—– ‘–ƒ‹•–Š‡‘”‹‰‹ƒŽ‹& (-"A+(,+$!(-"+%.+)/"+-","*()"#+!%#"'A+(+0*%D"2)$%,+%.+)/"+#")"2)"#+%7D"2)1+%,)%+ )/"+%*$-$,('+$!(-"+(,#+(+)(7'"+3$)/+)/"+#()(+%.+)/"+(,('4<"#+2"''15+=/"+!")/%#+ "'$!$,()"1+)/"+,""#+.%*+2%,>",)$%,('A+'(7%*&$,)",1$>"A+!(,8('+(,,%)()$%,+%.+)/"+ !"!7*(,"1+(,#+$1+"@)*"!"'4+.(1)A+%7D"2)$>"A+*"'$(7'"A+(,#+*"0*%#82$7'"5+E$,2"+9"''+ :8)'$,"*+.(2$'$)()"1+)/"+2%''"2)$%,+%.+$,.%*!()$%,+*"-(*#$,-+(+2%!!%,'4+81"#+0/"& ,%)40$2+0(*(!")"*+'$C"+2"''+1/(0"A+$)+2(,+7"+(00'$"#+)%+(+3$#"+*(,-"+%.+0/",%)40"1+ ƒ†‹• ‡”–ƒ‹Ž›‘–Ž‹‹–‡†–‘–Š‡œ‡„”ƒϐ‹•Š‘”‘—•‡‘†‡ŽǤ ‡ŽŽ‘•‡•‹•ƒˆ”‡‡Ž›ƒ˜ƒ‹Žƒ„Ž‡’Ž—‰‹ȋ—’‘”‡“—‡•–Ȍˆ‘”ƒ–Žƒ„ǡ™Š‹ Š ƒ 7"+81"#+)%+0'%)+)/"+%8)08)+%.+9"''+:8)'$,"*5+=/"+#()(+-","*()"#+74+9"''+:8)'$,"*A+ ™Š‹ Š‹•‹ƒ‡š ‡Žˆ‘”ƒ–ǡ ƒ„‡‹’‘”–‡†‹–‘ƒ–Žƒ„ƒ†•—„•‡“—‡–Ž›’”‘& 2"11"#5+=/$1+,%>"'+!")/%#+%.+#()(+*"0*"1",)()$%,+,%)+%,'4+2%!7$,"1+)/"+(#>(,& )(-"1+%.+)/"+2%,>",)$%,('+1"0(*()"+*"0*"1",)()$%,+%.+2"''+"'%,-()$%,+(,#+!"#$%& 133. !.

(33) !"#$%"!&'%($)#"#(')*&(#&"!+'&",,+&",,(#(')"!&()-'%."#(')&%$/"%,()/&#0$(%&1'%%$!"#(')*& 2'#$)#("!!3&%$4$(!()/&4(#"!&()-'%."#(')5&. !"#$%&'()*(+($,-.  Š‡ ƒ—–Š‘”• ™‘—Ž† Ž‹‡ –‘ –Šƒ ‘‹ƒ ‹ƒŽ‡ ƒ ƒ† ƒ “—‡Ž‹‡ ‡•6 10".2+&-'%&7(),!3&2%'4(,()/&#0$&(."/$+&'-&.'8+$&$.9%3'+&+#"()$,&:(#0&#0$&,(-6 ˆ‡”‡–Ǧ ƒ†Š‡”‹ƒ–‹„‘†‹‡•ƒ† ‡”‘‡ƒ‡”•ƒ† ‡”‘‡—••ƒˆ‘”’Žƒ•6 .(,+& $)1',()/& -8'%$+1$)#& .$.9%")$& ."%7$%+5& ;0(+& :'%7& :"+& +822'%#$,& 93& "& ƒ”‹‡ —”‹‡ ‡•‡ƒ” Š ”ƒ‹‹‰ ‡–™‘” ȋ Ȁ ǦǦʹͲͲ͸ǦͲ͵ͷͺ͵ͲȌǤ ;0$&-8),$%+&0",&)'&%'!$&()&+#8,3&,$+(/)*&,"#"&1'!!$1#(')&"),&")"!3+(+*&,$1(+(')&#'& 289!(+0*&'%&2%$2"%"#(')&'-&#0$&.")8+1%(2#5. 134.

(34) ‡ˆ‡”‡ ‡• ͳǤ‘•œ‘

(35) ǡƒ™ƒ†ƒǡ‘Ž‹ ƒǦ”‡œ‡ŽȋʹͲͲͻȌ‡‰—Žƒ–‹‘‘ˆ ‘˜‡”‰‡ ‡ !"#$%&'%"()*"$+*,%+%"'($#-.)"/$,%.'%0.!'%$/!('.-1!')*"$02$'3%$ –Ȁ’ƒ–Š™ƒ›Ǥ‡‹‡ŽŽ‡˜‹‘ŽʹͲǣͻͺ͸Ǧͻͻ͹Ǥ ʹǤ›‡”•ǡ‡’‹ Šǡ‘Ž‹ ƒǦ”‡œ‡ŽȋʹͲͲʹȌ’ƒ –‹˜‹–›‰”ƒ†‹‡– ”‡‰—Žƒ–‡• ‘˜‡”‰‡–‡š–‡•‹‘†—”‹‰œ‡„”ƒϐ‹•Š‰ƒ•–”—Žƒ–‹‘Ǥ‡˜ ‹‘ŽʹͶ͵ǣͺͳǦͻͺǤ ͵Ǥ›‡”•ǡ‡’‹ Šǡ‘Ž‹ ƒǦ”‡œ‡ŽȋʹͲͲʹȌ‘˜‡”‰‡ ‡ƒ†‡š–‡4 •‹‘‹˜‡”–‡„”ƒ–‡‰ƒ•–”—Žƒ‡ǣ ‡ŽŽ‘˜‡‡–•ƒ ‘”†‹‰–‘‘”‹ •‡ƒ” Š‘ˆ‹†‡–‹–›ǫ”‡†• ‡‡–ͳͺǣͶͶ͹ǦͶͷͷǤ ͶǤ‘’ œ‡™•‹ ǡ‡’‹ Šǡ›‡”•ǡƒŽ‡”ǡ‘”‡•ǡ‡–ƒŽǤȋʹͲͲͳȌŠ‡ œ‡„”ƒϐ‹•Š‰Ž›’‹ ƒ›’‡ ‘–”‘Ž• ‡ŽŽ’‘Žƒ”‹–›†—”‹‰‰ƒ•–”—Žƒ–‹‘ ‘˜‡‡–•‘ˆ ‘˜‡”‰‡–‡š–‡•‹‘Ǥ‡˜‡ŽŽͳǣʹͷͳǦʹ͸ͶǤ ͷǤ ‡••‡ ǡ‘’ œ‡™•‹ ǡ‹‰Šƒǡ‡’‹ Šǡƒ”Ž‘™ ǡ‡–ƒŽǤȋʹͲͲʹȌ‡4 „”ƒϐ‹•Š–”‹Ž‘„‹–‡‹†‡–‹ϐ‹‡•‡™”‘Ž‡•ˆ‘”–”ƒ„‹•—•‹‰ƒ•–”—Žƒ–‹‘ ƒ†‡—”‘ƒŽ‘˜‡‡–•Ǥƒ–‡ŽŽ‹‘ŽͶǣ͸ͳͲǦ͸ͳͷǤ ͸Ǥ‘ Šƒǡ†ƒ• ȋͳͻͻͺȌ”‹‡–‡† ‡ŽŽ†‹˜‹•‹‘•ƒ† ‡ŽŽ—Žƒ”‘”4 ’Š‘‰‡‡•‹•‹–Š‡œ‡„”ƒϐ‹•Š‰ƒ•–”—Žƒƒ†‡—”—Žƒǣƒ–‹‡ǦŽƒ’•‡ ƒƒŽ›•‹•Ǥ‡˜‡Ž‘’‡–ͳʹͷǣͻͺ͵ǦͻͻͶǤ ͹Ǥ‹ ǡ‡’‹ ŠǡŠ‡ǡ‘’ œ‡™•‹ ǡ‹ǡ‡–ƒŽǤȋʹͲͲͷȌ••‡–‹ƒŽ”‘Ž‡• ‘ˆ ȓƒŽ’ŠƒȔͳʹȀͳ͵•‹‰ƒŽ‹‰‹†‹•–‹ – ‡ŽŽ„‡Šƒ˜‹‘”•†”‹˜‹‰œ‡4 „”ƒϐ‹•Š ‘˜‡”‰‡ ‡ƒ†‡š–‡•‹‘‰ƒ•–”—Žƒ–‹‘‘˜‡‡–•Ǥ ‡ŽŽ ‹‘Žͳ͸ͻǣ͹͹͹Ǧ͹ͺ͹Ǥ ͺǤ‡ŽŽ‡” ǡ Š‹†–ǡ‹––„”‘†– ǡ–‡Žœ‡” ȋʹͲͲͺȌ‡ ‘•–”— –‹‘ ‘ˆœ‡„”ƒϐ‹•Š‡ƒ”Ž›‡„”›‘‹ †‡˜‡Ž‘’‡–„›• ƒ‡†Ž‹‰Š–•Š‡‡– ‹ ”‘• ‘’›Ǥ ‹‡ ‡͵ʹʹǣͳͲ͸ͷǦͳͲ͸ͻǤ ͻǤ‡‰ƒ•‘ ȋʹͲͲͻȌ

(36) –‘–‘‹ƒ‰‹‰‘ˆ‡„”›‘‰‡‡•‹•™‹–Š ‘ˆ‘ ƒŽ –‹‡ǦŽƒ’•‡‹ ”‘• ‘’›Ǥ‡–Š‘†•‘Ž‹‘ŽͷͶ͸ǣ͵ͳ͹Ǧ͵͵ʹǤ ͳͲǤ‘•ƒŽ‹‰ƒ–‹ǡ ƒ‘‘• ǡ ‡Žƒ•ǡ‘ Š‡ǡ„Š‘Žœ‡”ǡ‡–ƒŽǤȋʹͲͳͲȌ ‹•‘–”‘’‹ Žƒ–‡‹ˆˆ—•‹‘ ‹Ž–‡”‹‰ˆ‘”‡–‡ –‹‘‘ˆ‡ŽŽ‡4 „”ƒ‡•‹͵†‹ ”‘• ‘’›

(37) ƒ‰‡•Ǥ”‘ 

(38) 

(39) –›’‹‘‡†

(40) ƒ‰4 ‹‰ǣͷͺͺǦͷͻͳǤ ͳͳǤ‡‰ƒ•‘ ǡ ”ƒ•‡”ȋʹͲͲ͹Ȍ

(41) ƒ‰‹‰‹•›•–‡•„‹‘Ž‘‰›Ǥ‡ŽŽͳ͵Ͳǣ ͹ͺͶǦ͹ͻͷǤ 135. !.

(42) ͳʹǤ —””—•ǡƒ‹˜ƒǡƒ–ƒƒ„‡ǡ†‡”•‘ ǡ ‘‡•ǡ‡–ƒŽǤȋʹͲͳͲȌ‡–‡ ! "#$%&$'&%()*$%&+(+,*-%(.&#%&(/(0"*$%&+#0*$.0$12&#+-3(.&).#%3&-&.(*#-/& ‡—”ƒŽ‡–™‘”ƒ” Š‹–‡ –—”‡Ǥ‡†

(43) ƒ‰‡ƒŽͳͶǣ͹͹ͲǦ͹ͺ͵Ǥ ͳ͵ǤǤǤ—‡‰‘Ǧ”‘œǡǤƒ•–” ϐ‹ǡǤƒ˜›Ǥ ƒ—”‡ǡǤ‘„ƒ”†‘ǡǡǤ‘—”‰‹‡ ƒȋʹͲͲͺȌƒ˜‘”‘‘‹†‹ƒ‰”ƒ‘†‡Ž ‡ŽŽ‰‡‘‡–”‹‡•‹‡ƒ”Ž›•‡ƒ —” Š‹‡„”›‘‰‡‡•‹•ǫ

(44) 

(45) ǣͷͲͶǦͷͲ͹Ǥ ͳͶǤ‡•–‡”ϐ‹‡Ž†ȋͳͻͻͷȌŠ‡œ‡„”ƒϐ‹•Š„‘‘ǣ‹˜‡”•‹–›‘ˆ”‡‰‘’”‡••ǡ—‰‡‡ǡ 4*(3$%5 ͳͷǤƒ™•‘ȋͳͻͻͻȌ ƒ–‡ƒ’’‹‰–Š‡‘—•‡‡„”›‘Ǥ

(46) – ‡˜‹‘ŽͶ͵ǣ͹͹͵Ǧ ͹͹ͷǤ ͳ͸Ǥƒ™Ž‡› ȋʹͲͲ͸Ȍ ƒ†„‘‘‘ˆ„‹‘Ž‘‰‹ ƒŽ ‘ˆ‘ ƒŽ‹ ”‘• ‘’›Ǥ ͳ͹Ǥƒ˜‹‡•ȋͳͻͻͲȌƒ Š‹‡˜‹•‹‘ǣ–Š‡‘”›ǡƒŽ‰‘”‹–Š•ƒ†’”ƒ –‹ ƒŽ‹–‹‡•ǣ ƒ! 6(+#0&1*(..5 ͳͺǤ‡”•‡ Ǥ›ƒ‹ Š”‡•Š‘Ž†‹‰‘ˆ ”‡›Ǧ‡˜‡Ž

(47) ƒ‰‡•Ǣͳͻͺ͸Ǥ ͳͻǤ‹„Žƒ ȋͳͻͺ͸Ȍ‹–”‘†— –‹‘–‘‹‰‹–ƒŽ

(48) ƒ‰‡”‘ ‡••‹‰ǣ”‡–‹ ‡Ǧ ƒŽŽǤ ʹͲǤƒ—˜‘Žƒ ǡ‹‡–ƒ•‹‡ȋʹͲͲͲȌ†ƒ’–‹˜‡‘ —‡–

(49) ƒ‰‡‹ƒ”‹œƒ–‹‘Ǥƒ–! –‡”‡ ‘‰‹–‹‘͵͵ǣʹʹͷǦʹ͵͸Ǥ ʹͳǤŠ‘›ǡŠ‹—ǡ‹ŽŽ‡––‹ ǡ ‡‘–‹•–‘˜ǡ‘”‹‘–‘ǡ‡–ƒŽǤȋͳͻͻͻȌ†‘‡! „”ƒ‡–”ƒˆϐ‹ ‹‰‘ˆ”ƒ•ǣ–Š‡‘–‹ˆ–ƒ”‰‡–•’”‘–‡‹•–‘–Š‡ƒ†. ‘Ž‰‹Ǥ‡ŽŽͻͺǣ͸ͻǦͺͲǤ ʹʹǤ—”‡Šƒ‡” ȋͳͻͻͳȌ‘”‘‘‹‹ƒ‰”ƒ•Ǧ—”˜‡›‘ˆƒ —†ƒ‡–ƒŽ ‡‘! ‡–”‹ ƒ–ƒ–”— –—”‡Ǥ‘’—–‹‰—”˜‡›•ʹ͵ǣ͵ͶͷǦͶͲͷǤ ʹ͵Ǥ˜ƒ‡‡Ž‡ǡ—–—™‡‡ǡ˜‡”˜‘‘”†‡ ǡ†‡ ‡”–‘‰ ȋʹͲͳͲȌƒŽ’Šƒ ƒ†‡’•‹Ž‘•‹‰ƒŽ‹‰˜‹ƒ ›Ȁ‡•ƒ†Š‘‹•‡••‡–‹ƒŽˆ‘”œ‡„”ƒϐ‹•Š ‘˜‡”‰‡ ‡ƒ†‡š–‡•‹‘ ‡ŽŽ‘˜‡‡–•†—”‹‰‰ƒ•–”—Žƒ–‹‘Ǥ‡˜‹‘Ž ͵ͶͲǣ͸ʹ͸Ǧ͸͵ͻǤ ʹͶǤŽƒ†‡ ȋͳͻ͸ͺȌ‡–‡‘”‘Ž‘‰›ƒ†ƒ–‘‹ ‡‡”‰›ǣ–‘‹ ‡”‰›‘! ‹••‹‘ǤʹͺǦ͵Ͳ’Ǥ ʹͷǤ ”‡ƒ˜‡• ǡ”‡• ‘–– ǡ —ƒ–ƒǡ —ƒ–ƒǡƒŽƒ—ǡ‡–ƒŽǤȋʹͲͲͻȌŠ‡Š›! †”‘’Š‘„‹  ›•–‡‹‡Ǧ”‹ Š†‘ƒ‹‘ˆʹͷ ‘—’Ž‡•™‹–Š†‘™•–”‡ƒ *(.#6)(.&"$&+(6#-"(&+(+,*-%(&#%"(*-0"#$%.&-%6&*(0$3%#"#$%&,2&7889& ’ƒŽ‹–‘›Ž–”ƒ•ˆ‡”ƒ•‡•Ǥ‘Ž‹‘Ž‡ŽŽʹͲǣͳͺͶͷǦͳͺͷͶǤ 136.

(50) ʹ͸Ǥ ”‡ƒ˜‡• ǡƒŽƒ—ǡ —ƒ–ƒǡ —ƒ–ƒǡŠƒ„‡”Žƒ‹ ȋʹͲͲͺȌƒŽ‹–‘›! "#$%&'(#')(*+*,-#'+(%'$+-#.$%&'/(&0($1+('+2-&3-&$+.$%4+(.1#3+-&'+( ›•–‡‹‡Ǧ•–”‹‰’”‘–‡‹Ǥ ‹‘ŽŠ‡ʹͺ͵ǣʹͷͲͳͶǦʹͷͲʹ͸Ǥ ʹ͹Ǥ‹”‘‡ǡ‹ǡ ”‹‡•„ƒ Š ǡŠƒ‰ǡŽ—•ƒ”•‹ǡ‡–ƒŽǤȋʹͲͲͺȌ”‘Ž‡ˆ‘” ’Žƒƒ” ‡ŽŽ’‘Žƒ”‹–›•‹‰ƒŽ‹‰‹ƒ‰‹‘‰‡‡•‹•Ǥ‰‹‘‰‡‡•‹•ͳͳǣ͵Ͷ͹Ǧ͵͸ͲǤ ʹͺǤ‡ŽŽ›ǡŠ‡ȋʹͲͲ͹ȌŠƒ’‹‰–Š‡ƒƒŽ‹ƒƒ—†‹–‘”›•‡•‘”›‘”‰ƒ„› –Š‡’Žƒƒ” ‡ŽŽ’‘Žƒ”‹–›’ƒ–Š™ƒ›Ǥ

(51) – ‡˜‹‘Žͷͳǣͷ͵ͷǦͷͶ͹Ǥ ʹͻǤ‹–‘”‹‘ǡ‡›‡”ȋʹͲͲͺȌ‘†—Žƒ” ‘–”‘Ž‘ˆ‡†‘–Š‡Ž‹ƒŽ•Š‡‡–‹‰”ƒ–‹‘Ǥ. ‡‡•‡˜ʹʹǣ͵ʹ͸ͺǦ͵ʹͺͳǤ ͵ͲǤ› ‘ˆˆ ǡƒ‰ǡ‹ǡ‹ ǡ ‘•™ƒ‹ǡ‡–ƒŽǤȋʹͲͲ͹Ȍ‹”‡ –˜‹•—ƒŽ‹œƒ–‹‘ &0(*#.-&31#5+!#//%/$+)($2*&-(.+""(%'$-#4#/#$%&'(%'(*#**#-6($2*&-/7( ƒ ‡”‡•͸͹ǣʹ͸ͶͻǦʹ͸ͷ͸Ǥ. 137. !.

(52) 138.

(53)

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