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Cellmap

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Material Information

Title:
Cellmap an automated multielectrode array cell culture analysis system based on electrochemical impedance spectroscopy
Physical Description:
Book
Creator:
Abdur Rahman, Abdur Rub
Publisher:
University of South Florida
Place of Publication:
Tampa, Fla
Publication Date:

Subjects

Subjects / Keywords:
Biosensor
Bioimpedance
Cell-based assays
Whole cell assays
Cell substrate sensing
Dissertations, Academic -- Electrical Engineering -- Doctoral -- USF   ( lcsh )
Genre:
non-fiction   ( marcgt )

Notes

Summary:
ABSTRACT: The objective of this research is to develop fundamental understanding of cell-substrate (CS) and cell-cell (CC) interactions in the culture space for time evolving cell cultures. Space resolved CC and CS interactions are important indicators of cell-density distribution, localized cellular behavior, and multiple cell-layers which are differentiators of normal and abnormal cell behavior. In this research, CS and CC interactions and the variations therein due to a) Cell growth, 2) cell-drug interaction, and 3) effect of Cytotoxin were studied using multielectrode, multi-frequency Electrochemical Impedance Spectroscopy (EIS). Contemporary impedance based methods sense either CC or CS interaction as a space averaged macroscopic quantity. A major contribution of this research is that, both CC and CS interactions are recorded and analyzed with spatio-temporal resolution. This research led to the development of an automated cell culture monitoring system, namely, CellMap.A planar eight electrode sensor was fabricated on a glass substrate and interfaced with a switching circuit. The switching circuit sequentially selects consecutive electrodes upon input of a 5V trigger pulse which is generated by the frequency response analyzer at the end of each frequency scan, thereby facilitating automated switching and recording of multielectrode dataset. Calibration standards and protocols were developed to null the channel parasitics of individual channels. A set of eight impedance measurements for eight electrodes constitutes a "frame". Frames are recorded at regular time intervals over the desired course of time. Impedance mapping of adhesion, spreading, motility and detachment of OvCa429 ovarian cancer cells was performed over a period of 70 hours. The cell-layer resistance, which indicates cell-cell contact, increased as a function of time until confluence, and decreased thereafter due to cell death and detachment.This was also confirmed by optical microscopy observations. Similarly, the cell layer Constant Phase Element (CPE) parameters, which were found to correlate well with cell density distribution, also increased as a function of time until confluence and decreased thereafter. Additionally, the cell-growth mapping revealed that the CellMap system is able to resolve non-uniform cell distributions in the culture space, which may be useful in differentiating between normal and pathological cells.
Thesis:
Dissertation (Ph.D.)--University of South Florida, 2007.
Bibliography:
Includes bibliographical references.
System Details:
Mode of access: World Wide Web.
System Details:
System requirements: World Wide Web browser and PDF reader.
Statement of Responsibility:
by Abdur Rub Abdur Rahman.
General Note:
Title from PDF of title page.
General Note:
Document formatted into pages; contains 138 pages.
General Note:
Includes vita.

Record Information

Source Institution:
University of South Florida Library
Holding Location:
University of South Florida
Rights Management:
All applicable rights reserved by the source institution and holding location.
Resource Identifier:
aleph - 001988961
oclc - 307532795
usfldc doi - E14-SFE0002185
usfldc handle - e14.2185
System ID:
SFS0026503:00001


This item is only available as the following downloads:


Full Text
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Cellmap :
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Text (Electronic dissertation) in PDF format.
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ABSTRACT: The objective of this research is to develop fundamental understanding of cell-substrate (CS) and cell-cell (CC) interactions in the culture space for time evolving cell cultures. Space resolved CC and CS interactions are important indicators of cell-density distribution, localized cellular behavior, and multiple cell-layers which are differentiators of normal and abnormal cell behavior. In this research, CS and CC interactions and the variations therein due to a) Cell growth, 2) cell-drug interaction, and 3) effect of Cytotoxin were studied using multielectrode, multi-frequency Electrochemical Impedance Spectroscopy (EIS). Contemporary impedance based methods sense either CC or CS interaction as a space averaged macroscopic quantity. A major contribution of this research is that, both CC and CS interactions are recorded and analyzed with spatio-temporal resolution. This research led to the development of an automated cell culture monitoring system, namely, CellMap.A planar eight electrode sensor was fabricated on a glass substrate and interfaced with a switching circuit. The switching circuit sequentially selects consecutive electrodes upon input of a 5V trigger pulse which is generated by the frequency response analyzer at the end of each frequency scan, thereby facilitating automated switching and recording of multielectrode dataset. Calibration standards and protocols were developed to null the channel parasitics of individual channels. A set of eight impedance measurements for eight electrodes constitutes a "frame". Frames are recorded at regular time intervals over the desired course of time. Impedance mapping of adhesion, spreading, motility and detachment of OvCa429 ovarian cancer cells was performed over a period of 70 hours. The cell-layer resistance, which indicates cell-cell contact, increased as a function of time until confluence, and decreased thereafter due to cell death and detachment.This was also confirmed by optical microscopy observations. Similarly, the cell layer Constant Phase Element (CPE) parameters, which were found to correlate well with cell density distribution, also increased as a function of time until confluence and decreased thereafter. Additionally, the cell-growth mapping revealed that the CellMap system is able to resolve non-uniform cell distributions in the culture space, which may be useful in differentiating between normal and pathological cells.
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Mode of access: World Wide Web.
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Advisor: Shekhar Bhansali, Ph.D.
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Cell-based assays
Whole cell assays
Cell substrate sensing
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PAGE 1

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PAGE 5

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C nrnr+ rn (rnrrnrr r$rnnn n"rrnnnrn rr' nr"n"& &n"r rrnrr&nr2 "n n'rrnnr rn rr2( 3(n(n nrrnr "n'(n n rrnnr
PAGE 27

Ernn""nn2 r n"rrn '(rn rnrrnrr" nr rnr'(r"rnrn rn"n rnnn rn3nnr n2nn"rn' nrrrnrn nnrn3n""&n" &n3 rrrnrnr'!n%"r &n nnrrr BT-?EC&2 %rnrn3n""nnrn3" 'nrnr rnrrnrr&nr rr rnrnrn'n r rrrnrr2rn& n"':2nr "nr"rnnn rrn"nn nnn&n nn rnnrrn"rn'2r%nrrn2 rn1nn r& &rrn;n2nn' (2r%r1nn rnn"rnrn r
PAGE 28

nnr"nrrn 3"' nrrnrnnn nrnrrr n'nrn3 "nnr"3n rrnrn "nr"2 rnrrnr rnrr'(rrrn& n&"rnn rr@3Ar @2rA'@3 A'rnnr nn2 n"r'$rr"n nr rr2rn%r n"r r'nn"r"n &r rnnn%nr2n'rr"&r rr" "nrr'n 3 rnrnrnrnn "&3 rnrn'nrn rn rrnrnnrn rnr"2 "r"nn"r'(" $r n"rn"nr'(rr2n rn"2r rnrn"r"r''$) //r!r rrrrnr"n "rnn+ rnnnnnnr"rr rnr rH'(rrnn n"rnn2 rrnrrnrnrn rnrrn r'(&rnrnn nnrQ nrn2n"rnnn2r rnnrnnG -?H "rrr2n' n

PAGE 29

n nrnnrnn '2r rnnrnr 2r%n rnrnr'(rn rnr r<=//r2r%n rrrn rnrnrr"nG-8H'n nr& nrrrr n nnrrnn&H&G6?Hr rrG68H&G60H' ,n$" "nnrr3 r3nBC&n n rG6.H'r3r nrrrnr G67H&r"nnn" r rrnr"rBrr nrnr r<6/LC'rr"& n rrrnnrn3'#r<&r Qrrr

PAGE 30

n&r nrBrn 6'0'-C&n rr 3n'(/H rn=&>H' nrr2nrn"nrr rnnrn& nnrnr'(nn r+ 2nrrnrn'n r"rn3 r"rr' '$)$# !/(0r (nnrnnr "nrn&n rSnr%r'A2 r rrn23nn'rn rrrr rrnn&2r"r rn' rnn"&"n rr rn2r'(nrnnn nr "rrr n2&n n"'nn rrr&r
PAGE 31

"rn'"r6H' rnnrnr rnQnn2r%n& rnnrrn nrrr2 nrnrr' nn%3 nrn&rnnn rnQr r"nr"2r3nBn""rC< nrn'( nrnrnnr 2rnn rrr<&2r3rnr2r3rn rn'n >H' (nrnrnrrnn"r"nr rr nrn&nn""&rrn& nrn&rnr "rn&2rrrn rrn& n"rnrn&rr&n rn&rr3 rnrn'"rn nnrn r'(&nnrr n2 2rnrnr3rrnr rn
PAGE 32

r C (<< rrnrn rnr< ?H' rnrrr rnrr2& nrn&r&nn' n rrnnrn r%rn B1C2&nn&"rrnr Rrn G>8H'rnr"nnr"" nnr r<<'(nrn nrnr2n rnr';,&r<& rn2r "nr2nn rnrnn rrn nn'(0H' C #rrn nr2nn rnn rrnrn& rr rn'nrn2r rrr &n+ rr nrn&nrr n rnnnn'rr &nnr&

PAGE 33

&nr&n& rrrn2n rnrr'"rrrnnn rrr&rrn nr2r&n"nrrn'r rnnr rnrrrnn&rnrr nnn rnrrG>.H' C (*nrn nnnB''&rnn"C& r n
PAGE 34

rnrrnnrn'rn$ nrnnn nr2nrnnn$n': nn rnr'n$n rnr nUnnrnnnn'9*nrn9$nrnnnnr='?D -nn'( rrnnr Brr 2rr=///rnC2n'( n rnr8rrnnrn rnn7H'"r
PAGE 35

(rnrnnnr n rr',nr nrr rrn&3nr3 n', nrrrnrn3n"" nnnrn3 nn2rrnrn'(r2rrnnn nrn nnrrnr nr2r nrnr'(nrn n2 2nrrn'( 2rn nn"2nnn"rrnrr r'n" nnnrnnr r'(nr 2rn'2n nrn& rnnn'n rnrr nrrrnrnrr 2nr rr"rr nn
PAGE 36

rr2nrn2"2nr nrn nrn3rBCn33 nr''$1 !/r23 4rrr (@2rrnrAn@3 rnrA"n rr"+ nnnr nrnnrnn nnr n"rnn"2" r"& nnG?=H&G?-H'( %nr nnrnrnrn rrr rrnrn"rn&r n"rnJ&n rnrnJrnG?6H'n32rrnrnr rrn r3n'n rrnr2r nrnnrn2r 2n'n rnnrnrrnnr n2n rnn'nrnnrrnrnr "n& rr2nnrrn"=Crrn& -Cnn n&6Cr&>Crnnn?Crr 'nrn& nrnr"rn nB2;+r "n;+C& rn nrr+rn&n &nrrrnn G?>H'"nn rnr rrrnr"G??H'"rn3n"n rn3"rrn nrrG?8H'n nrnr nnr2rr&n &=Cn nn-Crn'nn rn&n%r rnrnr2 nBrrC rnG?0H'#r<&rnrr n2

PAGE 37

r2nnnrrn nn'n r&rrn %nr2nnrn "rn%r"'n rnr&r rrr&2 "2r+n rnr r< rn"n"G?.H'rrr"rn rr' (rnr2rr'#&r rr" n&nnrr nr2n nnnrnnr n&n nrrr'rn&n r+rnrnn nrr"'nr+rn rr nrnrnrnn rn"rnn' n2rrnrrr n& rr+rnnrrr nrn r2nnrn" rn2n rnn')nn n"nr r&nn& 2 nnn'nr&"rn r r2rrnrn r'( r"nrrnn rn&rnrn n';r2"&"nr rnr nnnnr<&2 % n"rnn rr' nrnrrn nnrr nrnnn&2rrn & n&nrrnn"

PAGE 38

'$5 /!r r nn"nrrn nrG$%& '&=776H&H') nr r&r "nnrrn nrrr rrnrr C nnnBC& rn&5V (rnn2r%r"9"n% rn rnrn nnBC" rrrrrnG?738=H' 2rrn n&rnnr-?/W 2r%nr B)CnrnrBC'=W nrn 2nrnnr <"r rrG>/H'( nn "nr"r)'( nr% nrn2n"nnn"n (n3nnnr2r%n)n nnn nnnr& %nr nr'(rr nr+nr&2nr+ nrr rn2nnnn r' #-r2 n' nrrn nr>1;+'3 rnrnr nn $nrnnnr"n '(n

PAGE 39

n n22nrG8-38 >H':&r n9rG8-Hr "nnnn' rnnn rrrnrn nrnrnrnrn2rnn' (rnr r"3r+rn rnr rnr3n3nrn' #r<&n#6& nnn' #-rnr'rG ?=H' >1;+&nnnn nrnr nn'nrr 2nn rrnn '"rnr nnrn2& n2r%r )nG8?H&,rG88Hn9rG8-H'

PAGE 40

#6rr+n rnn' rr'r'nnrr+rnr& r "nrnr'(r "nnr rrnr&rnr r&n nrnrnrrrn"n &2 rnrrrnr nG80H C 3rnr&nr!n"&-//6 r%rG?=H&rn>/r nrr +rnrr rrnrn n';rrnrn2 nr rnr rrn'( "rrnr2nn# >'r%rr nrrr+r nn' ("r2=//E&2r" rnn&2 r<-?ErrG80H'5 rn3n"r2 r"nr'
PAGE 41

#>rrrnrn rnnnr 'rG?=H' C n3nnrnrrn+ rn&n rn!n"&-//>' ;nG8.H"rrn r "r'(n"2rn" r&Cn r"2")2 nCnr <)2rrn';r2"&nr nr rnrn2r%n nrr' rr"&2nr+r n$nrn nnn'5nG87H"2"rr" r$2r%rn r"rr" r2rnrn';nr nr3nrnnr n"2 r"nrr3rrn nn2nr rn"rrn'(r"n & nrrn#?';r2"&n nQn2 2%'n"&nnr rn+ rrn>r&rn n+rn" r6/LPrrn2r rr'-n

PAGE 42

r r2nr22r r"n rnr' rn2 "rn /'=D/'. /'/? rn2 "rn /'=D/'. /'/? #?rrnrr rnrn'r G88H' C 3rnnnB(3FC& rn n&nr 3(rnnnB(3FC& n rn&rnrn" rnrrn nrnnrrrr nrr3 '(3F2r%n nr nrrn&n&rn nrnrnr'#r 3&r2nnn" &nr3rnnn2 rr3nnn nr'(n rrrn" nnn nrnnrnr nn"'( rnnrr"nrrnrnn "&2n

PAGE 43

rn"r%nr2nn<&B nr nr+rnrrnCG0 /H' (rnnrrnrn 3 rnnnB(3FC'r3 rr< n2rnrr rn rrnrnr%rn nrnn rnrnr' rrr "nrrr3n nr"'( nr2rnr r
PAGE 44

n< (B;rC / /'? = ='? -'? / /'/'> /'8 /'. ='= #rnn r3,3,n n< (B;rC / /'? = ='? -'? / /'/'> /'8 /'. ='= #rnn r3,3,n #0:r(3F'rG80H (rn&nnrnnr nn rn
PAGE 45

nrr"nnrr3 r3 nrnPnrr"nrrn rnrn' CE:;n&)r rnnnB) C& r&#, ) 2rn(nnr nB(C n'(("rnr nnr r'(nr r"3" r'(n '(nr2 rrrrrrr&2n rn r'rnr( rn r2nn#.' #.(n:r2E: n' ) %nr&2 (rn 2'(rrn r nrn&nnrr r3-n nr2nrrnnrnrrrr r

PAGE 46

nnB;(C->n782r' 3rnr& nnr2 rnrn' C1F&<&n#nr&
PAGE 47

#71Fr rr' C( (nrnr" "rn +rnnrn"r nnn '(r n nnn& 9((6/& r" 2nr3r3nrnr&2n n(, rnr"r& n nr"rnr'(% r =Crnrn-Cn n"+rn& n6Cr-7 '(rn2 n"rrr n33nrnrnr nnrr2n rn'

PAGE 48

(=rrnrrnrn" rn3 nrn' "J r n +rn n n n rr 6 "+rn FnrnrVnr 5nnrnrnr 1Fnrnrnrnr E:Fnrnrnrnr (3Fnrnrnrnr r%rnrnrnr (=+rnrn3 3 n'nrrnrn <&nrn3 rrnrnrn&n nn r3n3+ rn"nrn rn'(rnrn r"n nr"n nrrr nrn3nrn

PAGE 49

n ) !r )$# !n&rrr nrrBC n'nnnrn nrnr< nr
PAGE 50

n&rnnn' (nn nrnrnnnr rnrr r'nnrn +rn& rrrn&nrnrG80H'( "nrn rnr&&nrn3n""n2 rr" 2nr3rnn2n" rnG06H'n &nnnn"n n&n& "rr&r"%nr2 nn nn"'n&rr+ rnn2n-&6 r>nrnnnn nnn rnn'#rn& nnr nrnrr nBrrCn rnrn nBr< nnrC'nnrnr rnnr %nr2rrn&rn n" rnrnrrr rrnr"rn nr< n&nr"n'nr rrnr rnrrn rrrnn rn"rrn&2rrnn "n nnr3Brr n
PAGE 51

nr%nr2nr "r" n&nnrnrn nr '(r2r'n& r 2rrrnnrn %r<2 2rn'rnnr %r<& rrrn'( n"r' rnrrrr &r nnrrnn"r #rnrn3nrr & nnnnrnr nr nr"r3 rn n'#rnrnn n "r&E(X(J#X%(Jr-?E-?Y& nr2n nrnnrn rr nrn
PAGE 52

r )$) 0r )$)$# 6/%rrr(2rrrrrn& nnrn3' nrnr r n'nrnnrnr "nQ2& r#r' n rnrnr%nn"r 2nrr' ;r2"rrnnrrnnr2 nnrn' rnrn3r' rrnr rnnnrn3r&n nnn rnnnrn 'rnnrn3 rr2nrrn %';nr "rrn2"nn rnrnr ')$)$' &r2nrnr r+ rB C'nnnrr n2n rnrn&"rn rrrrr& 2rnrrn rnr rrnr'( n"nr rn#=/'(rrnrrr "'(r rr&nnrnnr"n rnr rr'( r r&;r+rn'(nr r rnrnnn;r+ nB; C'( rrnr nr"rnrrr n: ;r+ nB:; C'(r"rn rnrn

PAGE 53

r'rrnnrrn& r"rn nnrnrn !""#$% &"$ #%' ( !""#$% &"$ #%' ( rrn r"nr r" rn ; :; !""#$% &"$ #%' ( !""#$% &"$ #%' ( rrn r"nr r" rn ; :; #=/rnrnn"n rr'; nn;r+ n&:; :;r+ n' (r%nrnrn rrnnr rrn'$rr"rrr r3r nrnr'(n rr 2rn=J" %nr2n n&2rrn'( nnnrn rnnrr'#== rr"rnn nnrnrrrnn rn'

PAGE 54

) )) ) ) )) ) )n )n)n )n ) )) ) ) )) ) ) )) ) )n )n)n )n ) )) ) !# !# !# !#*+* *+**+* *+* )(,!-. )(,!-. )(,!-. )(,!-. )(,!-.n )(,!-.n )(,!-.n )(,!-.n )(,!-. )(,!-. )(,!-. )(,!-. (,!-.) (,!-.) (,!-.) (,!-.) nn ErnEr /'/=&,I6/n /'/?&,I8n /'=&,I6n =&,I/'6n /'/'> /'8 /'. = /'> /'8 /'. = Z=/3. /'/ / ) )) ) ) )) ) )n )n)n )n ) )) ) ) )) ) ) )) ) )n )n)n )n ) )) ) !# !# !# !#*+* *+**+* *+* )(,!-. )(,!-. )(,!-. )(,!-. )(,!-.n )(,!-.n )(,!-.n )(,!-.n )(,!-. )(,!-. )(,!-. )(,!-. (,!-.) (,!-.) (,!-.) (,!-.) nn ErnEr /'/=&,I6/n /'/?&,I8n /'=&,I6n =&,I/'6n /'/'> /'8 /'. = /'> /'8 /'. = Z=/3. /'/ / #==nnrnrr rnnrn')$)$) r% r7%8nrQnn[I =J$N\NP2[ n&\I-NNPnn&2 nn +&nn'nr nn n3rn&nrnr r =JBC=nn&=JBCnnn&%nr2nr22 nn'(r2rn&rnnr" rnrr ""r&rr
PAGE 55

rnrr'rrr 3n& 2rrnrn rn' )nnnrrr& nr n 8 =/ <=/> / = 6 > ? 8 0 7 =/ <=/> nBCn nB3nCn nI/'? nI/'8 nI/'0 nI/'. nI/'7 #=-(r
PAGE 56

rn3 '#=-r< nnrrrnrnr rn &r rnrn3"'(rr n rnrrnn nB C&2r r n n nn''''''''''''''''''''''''''''''''''''''''''''''''''' ''''''''''''''''''''''''B-'=C 2/T/nT=nnI=nnr&V 2 nrn'BrnCn&nrnn2nnI=& n\nn'("rn nrr rnr'rr rr+rn n&rrn&r rnnrn rnnrn')$, 9:%;&n r
PAGE 57

n nn n ) ) ) ) ) ) ) ) ) )) ) ) )) ) & && &0*# 0*#0*# 0*# -B:;C6r 2 / > 8 =/ ==> ='? = /'? / 3/'? 3= 3='? 3Er ; n nn n ) ) ) ) ) ) ) ) ) )) ) ) )) ) & && &0*# 0*#0*# 0*# -B:;C6r 2 n nn n ) ) ) ) ) ) ) ) ) )) ) ) )) ) & && &0*# 0*#0*# 0*# -B:;C6r 2 / > 8 =/ ==> ='? = /'? / 3/'? 3= 3='? 3Er ; #=6(r r
PAGE 58

)$1$' ;%!&nrnrCr '(nrrn 2n"rr"n&n&[n&VI=J[ "&IVJBN\NN]/Cr&I=J")&rnn&]/"rn \I-NN&2nn;+'n"+nrnrn rnn r+rn<"nn" <')$1$) !!"r!#=>n#=?nr
PAGE 59

n ) )) ) ) )) ) ) )) ) ) )) ) ) )) ) ) )) ) ) )) ) ) )) ) ) )) ) ) )) ) 1 2+%$3# 1 2+%$3#1 2+%$3# 1 2+%$3#1 243# 1 243# 1 243# 1 243# )567(6 )567(6 )567(6 )567(6 %# %# %# %# %# %# %# %# n%# n%# n%# n%# %# %# %# %# %# %# %# %# %# %# %# %# %# %# %# %# %# %# %# %# n n n n n n n n n n n n n n n n %$2+%$3 %$2+%$3%$2+%$3 %$2+%$3%$243 %$243 %$243 %$243 )567(6 )567(6 )567(6 )567(6 %# %# %# %# %# %# %# %# n%# n%# n%# n%# %# %# %# %# %# %# %# %# %# %# %# %# %# %# %# %# %# %# %# %# 1 8 ) )) ) ) )) ) ) )) ) ) )) ) ) )) ) ) )) ) ) )) ) ) )) ) ) )) ) ) )) ) 1 2+%$3# 1 2+%$3#1 2+%$3# 1 2+%$3#1 243# 1 243# 1 243# 1 243# )567(6 )567(6 )567(6 )567(6 %# %# %# %# %# %# %# %# n%# n%# n%# n%# %# %# %# %# %# %# %# %# %# %# %# %# %# %# %# %# %# %# %# %# n n n n n n n n n n n n n n n n %$2+%$3 %$2+%$3%$2+%$3 %$2+%$3%$243 %$243 %$243 %$243 )567(6 )567(6 )567(6 )567(6 %# %# %# %# %# %# %# %# n%# n%# n%# n%# %# %# %# %# %# %# %# %# %# %# %# %# %# %# %# %# %# %# %# %# 1 8 #=>Cr
PAGE 60

n'(9!"rn "2nr nBnnCnr nn')$5 )$5$# !!#=0nnn Br Crr3rrn rnn2r r2nn#=8'#rrrnr" r rBC&nnnrr n# =0' #=8n
PAGE 61

=/3=// =/=/> =/8 =// =/= =/=/6 =/> =/? =/8 =/0 =/. =/7 ,rrnn I=1 I=/1 I=//1 I= I=/ =/3=// =/=/> =/8 37/ 3./ 30/ 38/ 3?/ 3>/ 36/ 3-/ 3=/ / ,rr#nn;+ nn I=1 I=/1 I=//1 I= I=/ #=0rrr3r rnrnn 2r'(n&/'=L#&nI=/M'

PAGE 62

r / -/// >/// 8/// ./// =//// / =/// -/// 6/// >/// ?/// 8/// 0/// ./// 7/// =//// nBCn nB3nCn I=1 I='01 I6'=1 I?'81 I=/1 #=.r
PAGE 63

nn'nrr2n rnr rnrn&rn2n"r" rnn %nrnrn')$5$' !!nrr+rnr& n2 2n2rrnn r'n
PAGE 64

=/3=// =/=/> =/8 =// =/= =/=/6 =/> =/? =/8 =/0 =/. =/7 ,rrnn ,rrnn;+ =/3=// =/=/> =/8 37/ 3./ 30/ 38/ 3?/ 3>/ 36/ 3-/ 3=/ / nn nrn % rn =/3=// =/=/> =/8 =// =/= =/=/6 =/> =/? =/8 =/0 =/. =/7 ,rrnn ,rrnn;+ =/3=// =/=/> =/8 37/ 3./ 30/ 38/ 3?/ 3>/ 36/ 3-/ 3=/ / nn nrn % rn #=7rrr&I=30&I=8&I=38&I=-' #-/n
PAGE 65

/ > 8 =/ =<=/? / > 8 =/ =<=/? nBCnnB3nCn #-=(r0Cr9r BrCDrrBrC &2rn2 <'(rnrn2nr rr r"rnrr'(rrn nrn& rr"'(nr +rn2 rrrnrrrr rrrnn

PAGE 66

2r'0& r2nrrnnr r nr+rn')$< /!rrrnrrn 2rn rn'(rnr nr< rnnrn& rnr rnnn< "r'nr 2r&nrnrrr n rH'nrrn n rn"nn& nr nnPr2nn r'r nnrnn
PAGE 67

)$<$# n&rrr ==? =. == ==6 =-=8==/: ; < = > 2J3 ==? =. == ==6 =-=8==/: ; < = > 2J3 #--#nrnnrr (rrnn 2nn n'(rr rn nr2nG0?H'(rn%nr2n ^&_&n` rnr"rnrnrnr nrr nn'(`rnn rnn 2&2_rnr< 23)nn r'(^rnrrr2 nnnn rnrn
PAGE 68

nn"rnnrnrn r'n nnr%rrnn r"' ;n&n2r%rrrn<
PAGE 69

n #-6rnr"nrr nnrrrn r3rn'n3"r&nrnnr n rn')nr"r "& rnrrnnn rnr' 9n&nrrrnrn n&2 nrrnnrnr '(n n#->'nnrnnrnnn 2nB3 $nrnnCn 2"'(rr nnnrrrn n"rn "n2G8>H'rnrn rr &rrn2nnr nrG00H& G0.H&+G07H&nr rnrnG./H&G?8H' <3 %% n n r r n ?#

PAGE 70

0% 0% 0% -$% @# % '%( 0% $%#( ( 0% 0% 0% -$% @# % '%( 0% 0% $%#( ( #->rnrrnrn rr ')$= rnnrnrr nr3n rr"2'(rnr nnrnr n+2<'( rrn rrr3nn
PAGE 71

, -rr%nn r ,$# !n&rn&nr nr '(" rr2 rnr2nr rrn'( "rnr(,9!"rr rn& rn&n&+rnn"+ rnn',$' (rrnn %rr' V 6///n"nrnr2' (r% r
PAGE 72

r nr r r r nr r r r #-?.3r"'(r rnrn.3 r'nr "&nr rnr rrrr'( % nr 2nr<-Lrr'!rnr rnr" rnnn rr nr2r3r<'(" r2nn# -?',$) % ; n r n r #-82nrnn rn'

PAGE 73

n (2nrnrC rn350>7/5C6r. rP30>,=6.5Cn"3;0>,/> C C,' nrn&B?ECr rnr J:rr 9((n'(r r nrrn'n&"r rnra///Qn6 nnrn'(rrrn rn6r. rrn=======/'("n" nn"r ///////='(nrrnr nr &2rnnr2r%n r'(ra=Qn rrnr"22B:5C nnr 9((rr 2r%nr nnr'nr nn& B?ECrnr rn'(rn nr///r//='rnnn" n&rrnn r2//////=/'(22rnrn nn rn2r%nr'nr n#-n n+rrnr n r.r'2 2nn rrrn',n "nn',$)$# %nn ."rr>7"8 !rrnrr2rr"n nr-?;+r =;+nnrnn+B#-C rr 9(( 6/rnrJ9"nr'(r".3r nP# rnn+'n#r n& rrn&'' nnr= ;+r=/;+'r$rrrnr r2 rJ:'#rn.3rn&n2r$2 r r.'nrn& B?ECrr

PAGE 74

n nrJ:rr 9( (n'(r rnr.3r2 '( r2nrrnb=rnrr nrnrn rn'(rrrn!(:,# r"n n<',$, r%r2n"rrnrn' rrn rnrr2'!rnn nr.3 r"&n=/nn r&2n n2'("n?/Y nrr=-r' (rnrr2rrr r "rrnn"r"n rnn "+rnrr
PAGE 75

n rn2"rrrrn &n&+rn n"+rn'#-0rnr %r2r(, 9!' ((17-18%" 4% & % !&%##+, %& %% 0% *#6 ((17-18%" 4% & % !&%##+, %& %% 0% *#6 #-0:nrrrrnn nr' (rrr2rnn rr2& =Crn&-CnrrnBC& 6C"n Crn"rnr nnr&?C -n6"+rn'rr!(:,r BrCrn nr(,rBnrnC'(r nn rnnrrnnrnr '(nr"r r3nnrnr"n" nnn(,'( nrn"nr' rnr5Z. <&25nrnrn n.nr

PAGE 76

n r'rnr
PAGE 77

n r'(nrr nnr &r2"&2nr 2n'( rnnn"rnr nrnr r'("n rrr nnnrr2nn#-. '"nr2r r2nrrr rn&r #-.nrnn nnr' r r r ( @-$% r r r r r ( @-$% BC BC #-."nr+ rnBCn BCn'((,nrn+rr
PAGE 78

n (nrn+nrnn n r'rnr&r2"&r"r nnn"r '(rrnnn "2%r ,nrnnn,"nrn' C,nrnn ,nrnnr"nrnn3Bnrnn 3nCr' &r"nrnn3 "nrr r"nn'B>C' r nrnn3nnrnrr "r3" nrn (n&n"r&r+rnr n r (nrn,nrnnn && nr+&n"rn$rn' C," ,"r"nrnn"3nB3 nCrn3 n'(rnnn rn<& rn r

PAGE 79

nn "nn<&nr"r &2-7n'(ra# Q2 rnn#n'( raQ 2rnn, nrnn'r" r,c5:5,5n,c !E#(" rrr'(r2rr "rrr "rnn%3rnnrnrn +'

PAGE 80

n n nn n n nn n n nn n 7.%#@A 7.%#@A 7.%#@A 7.%#@A -+""%B$ -+""%B$ -+""%B$ -+""%B$A#+ +%# A#+ +%#A#+ +%# A#+ +%# (#% (#% (#% (#% 9 9 9 9 0# 0# 0# 0# ,rr#n nn # n nn n n nn n n nn n 7.%#@A 7.%#@A 7.%#@A 7.%#@A -+""%B$ -+""%B$ -+""%B$ -+""%B$A#+ +%# A#+ +%#A#+ +%# A#+ +%# (#% (#% (#% (#% 9 9 9 9 0# 0# 0# 0# ,rr#n nn # #-7&nr: ">-7' C"+rn "+rnrnrr rnrnrr n"r"n'E+r n"r n"'(9!nB9! C"rr rnn"+rnr2n n#-0'rn& r"nrr"'

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n # 9 ; 1 # 9 ; 1 ,#6/(,nr nn "+rn'("rrnnrn rr2 (rrnrn rn2rnr CnrnrnB,"C
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nr rrnCrn&Cr&Cr
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1 !rr*(,'@*( 1$# !n&nnr:">-7 rn rnnr2 B.)=Cn' .)=2nrnr3 nn'( rrnnn rnrrnn n3"rnnrr' nr "n2"&nn rrn n"'(nnn"rn -7 n.)=nrr nnrrnr rnn" rrnn'1$' rAr 1$'$# =3#3%n&.)=22 nn>-7> nn+rrnn '(.)= n.32"'2 2-?/L2r%n rr2&n rrnn

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rn'(2r%nrr r nn-?/LB-72r" r%;" rr'(2r2nn=77n =/?B==C B&',r&:Cn2=/d BC& -,3n&=//nJnn&n=//L Jrnn ?d:-&n60r3r'2 n"-36&nnrnr r2&n2 3./drnnn=Znr rn'#r3r nrnnrr2.e= />J-&rnn 2rnnr2; n%Qn rrnB;P&n'&;nrn&EC2r nr' n2%nnrr r--r'

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1$'$) n,'@,rrr(n3rnrnn>-7> nn+n n2Frrrnr2n n#6-'( n>-7>nn+r>rn r2 rn'nnrn3r3r rn2 rrnrnnrnn< 'n2 rr>/;+r=//;+&2n" r-?E'(n "2rnnrnr nrr' #6-nnn+ n'1$) (nrr3rn -n#r-L %rn2""r?&2 nrn')n nnr&n r2 %nrnrrn'(r "rn 2rrnnr n'( r2n&n2n r"r
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rnnrrn&"n rrr rn2&%nrr nr-7
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n#66&rnr r&2r nrnrnnnn n" nB9;+C'rrrnn&r n&nn&rr n&rrn'1$, "rr 1$,$# rr Br7B 8#rr3r&nnrrrr nrnr n&rrr:">-7 &;' <&r2nrnrr nrr rnn';r2"& nrnrr2 r r&nr< rrnn '(rnrrn 2rn-?/ L2r%nr'nrnr r"rn n"r2nn#66'

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=/=/6 =/> =/? =/8 =/0 =/=/6 =/> =/? =/8 ,rrnn rr#nn;+ =/=/6 =/> =/? =/8 =/0 37/ 3./ 30/ 38/ 3?/ 3>/ 36/ 3-/ 3=/ nn = 6 ? 8 0 #6>rnnrr0" r.)=2 'r">2nrnn rrn' rnnrnrr rnr" rnr<6<=/?;+'(nrrnn r"nnrrr"r "B#6=C' #6?r
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n r&#&n"nr rrrn2n n'r& n"rnn r"rr;rnnn.)= "n (-' /'? = ='? -'? 6 6'? > <=/36 /'? = ='? -'? 6 6'? > <=/36 nBnCnnnBCnn =//;+(r7;+ = 6 ? 8 0 #6?r
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rrnr'(r rnnrn 2nrnnr'(n nn2rnn2nr "rrr r' r rn n r rn n #68"nnrn ;r.)= "'(
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/ = 6 > ? <=/36 / /'? = ='? -'? 6 6'? > >'? ? <=/36 nBCnBnC # #60n"r;n n.)="?' nr
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(r2rrrn "rr n Br-7rnnrr 2r"r rnrnn2r%n rBrn-7n BCn.)= "'!n%;3rr& nr2 nr"rnnr:E>-7'( r rrnrnrn2n n'rr"& nnr&2
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"r"r"'(& n&r 3rnnn& rn' =/=/6 =/> =/? =/8 =/0 =/=/6 =/> =/? =/8 ,rrnn rr#nn;+ =/=/6 =/> =/? =/8 =/0 3./ 30/ 38/ 3?/ 3>/ 36/ 3-/ 3=/ nn = 6 ? 8 0 #67rnnrr0" r.)=2 'r">2nnn r' #r#67r"n rn& nn2nrrr nrnr rn&nn2n;n :E'( rrn2 nrnr; '#nnn
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#>/r<"rr:E>-7 n0"r .)='#>/r<"rr:E> -7' rnnnnrr< nnnn' (nrnnnn "n'(r2 nrnBCrr &3n rnBCrn nn rnBCrrrrn rr' / /'? = ='? -'? 6 <=/> / /'? = ='? -'? 6 <=/> "BnC3n "BC3n =//;+(r7;+ = 6 ? 8 0

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/ /'? = ='? -'? <=/> / /'? = ='? -'? <=/> "BC "B3nC # #>=r<"rr nn "nr#>-' $ $ r rn r $ $ r rn r #>-#nr+ :E>-7n'

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(-7"nrr2n< rn n n n n 2nr&nn r'(rnnr"nr< n"rn #>='(r2n3r@ n%A"n rnnr'# >-n rr:">-7rr' <& rnnrn3 nr r"nr n%"2n'#rrrn &#>/r2 r<"rr:">-7rn.)= "'(n rBnrr2nCnrnrn rnrn& 26rnnn n"n' (n"rn n=L#J-' nrrr rr2 nr&
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n&rn 2"n nnr2r n' (6 r:E>-7n n0"r.)= %%0##!?'% n nr 4# 4# 4&% @0# &A# @0 &A @0 &A #>6 "rnr:E>-7n nrnr "'(6+-'#>6rr r"n '(rnrnrrn rn-rn nnr&rr r rnnrn '(rn$ "rrrnrn nr" r ! ! ! 4#D0)r)n)r)r))))n)r)) n))n) 4#D0))))))nr)nn))n) ))r)rr) 4&%D0)))))r)))n)r) )n))) @02#3D0r)))r)n)nn)nn)))n )n))n)) :2#3D0)r))r)r)))r))) r))r))r @02 3D0rr))))))rr)r)) )))r)) :2 3D0)n)))n)n))n))) r))r)) @023D0r ) ) )n ) ) r) ) )n ) ) n) ) ) ) :23D0)r)nr)n))nn)rr))n)) ))r)r) -D0))r)nr))))))n)) n)n)) +9 !0*@0 !0*@0!0*@0!0*@0!0*@0n!0*@0!0*@0 )n))r)))n)

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n nrr#>6'(r &"n-7nB# >6C2nr 2;nB#6.C'(r &"r0" '( "n"r7-C rnn r n'#>>rr"6r.)= 22 :">-7n2'n &n rnrrnr;n:"'( rn rnnnnr ';nr rr"nrr':n rr &n&nBCnr2 rBnC'r %nrrn"" &r rr '(nrrn rrnn rrrB"rCrrn'(nr rr r =/? =/8 =/0 =/=/6 =/> =/? =/8 ,rrnn,rrnn;+ 65r 6 =/=/6 =/> =/? =/8 =/0 =/=/6 =/> =/? =/8 ,rrnn;+ 65r 6 =/=/6 =/> =/? =/8 =/0 37/ 3./ 30/ 38/ 3?/ 3>/ 36/ 3-/ 3=/ nn 65r 6 #>>rr"62n2r #>>rrnrrr:" >-7n;' nrnr& rn &r2nrn&''r nn rrnnrn'( n nr'( rr"r rnnrnn"n 'nrn& "nrnn '(rn"rrnr rnnrr"nr< nrn#>?'

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(rnn23n r< "nB#>=C'("n nr2"r 2nrr# >-' /'? = ='? -'? 6 <=/36 /'? = ='? -'? 6 <=/36 =-'.6 => =? 6'=? =8 -'88 =0nBnCnnnBCnn =? 6'=? =8 -'88 =0 =//;+(r=/;+ 65r 6 #>?r-7r"nn2 +nr "nn'(n3 nn n=?6'8=M3-n.'0??L#J-&"'"n r"rr"rnnn; n:">-7r"n nnrrr nrnr nr"nrr< =//1;+'

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r nn" "rnr r2rnr"r nnnrnn'(r2 r"r rrrrnn 2F nr'

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r 5 !( rr5$# !n&rrrr rnn rnrrnr"nrrr r<' ('-.JCr rrr&"rr rr rnnn9n&;n! Ennr B;!ECn
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r rnnnrnr nrrn r'rnn&n rnnn rrr':+rnr rrn rrnr rn"n $rnnG.>3.8H "n&rr nrn r+rnnrn'n r+rnrr nrrr"r r3'( rrnr nrnrn rnrr'(3 rn"r "'nn rrrnr&n r+rnnnnr'n rrr& nrnrrn nr rnrnn& r2n' r&nnnr nnrnrn3 n'5$' rArC rrB1C (nrnr "/'.?d rrB1Cnrrn2 rn"r=>'-. J'1&"&nr rn"n' C;!E ;nnr&;!E&Brn r'&nr&C 260rn?d:-nnrr2B9P rnr'C22n2r r2n=/nJ nrnnr2r&=LJ rrrn&?/ LJn&?/nJrn&=-LJ r"nn<& n-dr"nBrnnn rnnrnC';!E 22n20/drnn&n 2

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r n">.r';!E n< 2n//fCrnr -3n2'5<&rrr2 rrn rnn=.=6r"rr' "rn& %n2nrr rr
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r %nrr<='0L'(22 r"nnn rnnnr&2nrrr rr2 rrn>nr BrC'( "r%nrn2r< >L'r"n J&-r=.=62 nrnr2r nrrrrr'(r r"rr "8&C'( rnn2r"rJnr '( "r2nn#>8'(r"r "rr +nn#>8BC'(r ?//LBC& -?/LBC&=//LBCn?/LBC'!rnr rnr" rn&?nnrnnn r rr"B#>8&Cr +rnr1n;!E BC BC BC BC BC BC #>8BCnrr-r rnrr2n rnr'BC rr r>r3 rr"rn2'BC rr rr" 2rnnnnrr "'5$)$' !rrnn>-7>nn+nr> rnr2 rn&nn2 (rr rn2rnn'' nnrn3r3r rn2rrnrnnrn n<'

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r n2rr>/;+r=//;+& 2n"r -?E'(nr2nn#6-' #&n2r2% n%n rnrB#>8&C'rrn r%n n&F2nrnr rrr n'1n;!En2r rn "'5$, "rr 5$,$# >0#>0rrr1rr r+&3n#>8& BC'(nr+rnn r2n BT=/%;+Cr22"rrnn '(r 22nrr n"rn 2nrnnn' nBg=/ %;+C&rnnrnr"& nrr rrnB#>8&C'(rrn rn rrBrCr nrr rnrrnnr rn' rnrrnnrr n&r rnrrnn%nr 2nrn n'(rnrnr"n rr& 2rnnnrr +
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r r<.M&rrrr2"B r<=//1MCr rB?//LC'(r" nrr "' =/=/6 =/> =/? =/8 =/0 =/. =/=/6 =/> =/? =/8 =/0 ,rrnn,rrnn;+ ?// -?/ =// ?/ =/=/6 =/> =/? =/8 =/0 =/. 37/ 3./ 30/ 38/ 3?/ 3>/ 36/ 3-/ 3=/ / nn,rr#nn;+ ?// -?/ =// ?/ #>0rr1nr"r" nr' (r.'nnn&nnr r'( r3r2nr r nnrrnn'(3r 3r rnnrn'2rrnr n""rr rnnrn'#r" 2r 3rrr3 rrn

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rn nrr'#r< rrrP rrrnn nnrn .r
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r #>7"nrr r1'(n rnrnrF'("nr2nn#> 7r+ 31'(< n(>'( n2nrn+rnr?//L r' (rrnn2nr nrnrn rr&'(r2rr rnrn Q"'(nn32 nr '(nrnrn3nrn rn'rnn n3nn3rr&rnBn3Cn rBn3rC n'r&rr n" rr&nrn r2 rBn3rC n':nrn&2n r&n rn"nnnrrnnr rr'( rr2"rn3rnrn3nr nrn' (nn"n'B?'=C&
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r nnnnn rnn rnn'(>"nr nr1 r' Jr ?//L-?/L=//L?/L rBMC 0'=7-0'0.-0'8-BMC='666-'=860'6/6='68> VB'nC-'/-3.>'8=370'083=/?'683=/ n/'7>/'7?/'.7/'.8 VrB'nC>'?/37>'>63=/>'-/3==>'0=3== nr/'?0/'00/'.0/'.? (+rnrnnnr nnrrn n2nr"rr& rnrn"' nrrnn'n nrr3r &nn rrrnn' rnn&nrn 2rr+rn nB"C2nr9;+n'#r nr"r nrrnrn nn rnrn2 r'nn nr&r2n&rrn nr
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rr (r<-/r8/L#J-rBrCnr "-rrrnnr'( r -/'8L#J-B?//LrC&=.'.L#J-B-?/L rC&=7'.L#J-B=//LrC&n?>'8L#J-B?/L rC'("rr nr?/L rrrnnn '5$,$' >C9(nrrr+ nF nr1,n nn#?/'( nrn=6;+#JB"rC&]" "&&nnn BrnnC ''&nrr2 rnr=J rrnr'

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n n ,rrnn ,rrnn;+ n r n nn #?/rr68/r rr"r nrr'nrnF21&nr Fr2n 1'(nrr.C'rnn&rnrnr n nrF'(n< nrn"r& rrr'

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n n nBnCnnnBCnn =//;+(r=6;+ -?/ ?// =// ?/ #?=r.C'( ?+
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(?"nr nrF r' Jr ?//L-?/L=//L?/L BMC8'--68'786='?6>-'=6> VB'nC='/?306'=630?'8.3.0'7.3. n0'>63=0'>=3=8'.73=8'0/3= VrB'nC>'0.3=-='=838-'7?3=6-'/73=6 nr7'//3=='7>3=='//='// 5$,$) >BA?(;!Ernr r"n r'(rrnnn rnr'( rrrB3 rnCrr 2nrnnr'(nnnn rr rrrnnrnrr r1'(< n"rr2rrr nn rrnn&rrnnnrn rrnn' #rrr;!EB#?-C& nBi=/%;+C rnrr" nrnr2r rnr"n1,'(n rnr3 rn3rrr2 rrnn' n
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n n ,rrnn-+""%B$6 n r n nn #?-rr;!Enr"r "nr' (nr"r& nrnn r'(nn2rnrr &rnn nnrnnrn rnn nnr Q'nn &n rrr" '"n nBr
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rnnn&nnr nr nBg=/;+C2n nn' nn2rnnnn&2 n"r r"rrBrn
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#?>"nrr nrnr ;!Ernrr"'rrnn& nn& Or rnnn B C& Orrn & O &n n'#?6nr;!E rnrn rr"nr'(r nnnn -?/n?//Lr2n nr2rr +'nr2rrnr 3nn nnrn2nr n', r+rr2nn2 3n r'( n n"rrnnn '( n2r""rrrn n&<&r nr'rn nnrn3 n&rnn r+rnr rnrrnnn"n rr'

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n (8"nr nr;!E' Jr ?//L-?/L=//L?/L rBMC?'7=-8'0.-8'.7-?'./BMC6'?6-='>-66'8=6>'/06 VB'nC-'=73.?'>7378'?-3=/>'/>3=/ n/'78/'7?/'7>/'7= VrB'nC-'->3==?'=.3==-'>?3==6'.-3== nr/'7?/'7//'76/'7= BMC8'0-66'-->='67?-'.6? VB'nC>'?63.='7>376'/=3=/6'/63=/ n/'87/'.=/'.6/'07

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5$,$, >B n n ,rrnn,rrnn;+ n n nn #??rrn%n rn"r nr'#??rnrr< n%n 2r"r"nr'( nrr%n %r"r&n &1&F n;!E'(nrrn2n rnn r%r'(n nn"rn rr
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nr+nn rnrnnr %nnn"nrr'!n rrnr rrnr&rnn r%n rn&n2 nrnrr rrn'5$1 (r+&nrnr1n Fr2n r+rnnrrnn'( n3r2 nn2r+&n r2=//Lr &nnnrn3nnr n'(;!E r
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r < r!<$# !n&n rrn& n&rnnnnr:">-7:" nn' rnrr" n6':">-7 rnrn nnnnn &rrrnrrnnnnn nn'( nrB;Crnr r=8/rr "rnr':">-7nrnr r==8r'( r$"r2r%nn Crn3"r "rr:">-7&Crn3 n3 nrn& n&r&rnn nnr:">-7rnn Crr2 nrrnrnr< n rrr:">-7' <$' ; nrB;C2rn rnrn n2'n2 rrn. r"r'rr8rn 2rr" rr=8/r'(rnnnr 2 n60Ynr2nn'( 2rnr r22n r'

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:">-7"r2rnrnrn n2 =/8J-rnnrnr:">-7'(nr r2nnrr rr-rrr2 r+'+rnr 2nrrr2& 2rnr"r2nrnB
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n r r n r r #?8rr;rnr r=8/r' (2rn%n2n rrnnP n+rrrnn'

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/ /'= /'/'6 /'> /'? -8/ -0/ -./ -7/ 6// 6=/ 6-/ 66/ 6>/ 6?/ 68/ (n;rnnn ;= ;;6 ;> ;? ;8 ;0 ;. #?0nnr B;Cnrn rr.r'nr<-/Mn r"r n"rr"rr=8/r'#r#?0&2rrn nrr nrnr&r"n nnn "2nnrr<-/M' (r<8d "rnr""rn nrr' ("rnnrrnnr rn"rnn

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/ /'= /'/'6 /'> /'? 6 > ? 8 0 <=/37 (n;r 3,nn'n ;= ;;6 ;> ;? ;8 ;0 ;. #?.nrn r B;C nrnrr.r'nr< -n#nr" rn"rr"rr=8/r 'r< nrnrnr"'(nrn rnrnrB# ?.C
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/ /'= /'/'6 /'> /'? /'08 /'00 /'0. /'07 /'. /'.= /'./'.6 (n;rnr 3, ;= ;;6 ;> ;? ;8 ;0 ;. #?7 r2rrn r B;C nrnrr.r'nr< /'/-nr" rn"rr"rr=8/r 'r< nrnrnr"'( r2rB 3nCr2nnrn rn#?7' ( 3nr2nrr 3n B#?.C'( "r 3nnnr/'.r rn
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(rnrnrnr2nr nr" rnrn nrnrnnr&"nr r" n'rnnn nn &rBrC3Br Cn rnn+r3 rnrn'<$)$# r-n-& *(,'@*(r:">-7nrn2nnr 2 r.3r"'n2 rr" rrrn==8rrrnr" r'( -7 nr nrnnnnr nrn'/'-?d (n3(B(nC2r2 8.rr nrrn'(2r"r rnn'( n"2rB/n0=r Cnn rn"r"nr r3 rnB#?8C'nn"nn r2n/n0=& &&r"&rrnnn '("n nr"' n"rrnr rnn"rn rnr r<>/1;+n'(n % r<>.r&2%nr2nr rnnr:">-7 '(rn"r rr n>n?&22rn rrn rnn"rrr'

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n #8/rrnnr nrnn' ("nnr"rnn nr n"rnr 'n/1;+n' (%rnrn2% nr2nrrnn r:E>-7'rnrn% n&r nrnnrn'(n2 r32 r<8.rrnrrn'(r n" nn
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n r r n r r #8= nrnnrnr n&n rrrnrnrr==8r'

PAGE 131

1 8 @ 1 8 @ #8-"rnr#8=&rn rn"nn rrn'36rnrrn& 3-/rnrrnP n"nrnnr&3>7r nrrnP"n rnnr&3rnnrnP 1 8 1 8 #86rrrrrnr' 3rnn:">-7 &3nrn'

PAGE 132

r (nnr2r:">-7 nn '#8/r r2rn nrr='( r"rnr r2nn&2n n" rrnrrn%rnr '(nr n3Bn "& nr+rnCrB"nr nC'"n nrrn:">-7n rn nn3' rn"n rnr"nn rnrnr 2n2rnn nn' n"(,rr+ nr 3n3nrn'(r2 n3 nrn&2n 3 r')nG.0Hrr2 nn r3nrn'(%n rr 2n'(rrr r2 nrnQ'rnn&r"nr n nnr2nnBT=1;+ C&2 nn$nrnn n'(rnrnr n&2 nnr%n rr n'23rrnnn rnrnr nn$nrn n'n2rr &nn2 nn' (rr3 rnr rnrnrnn &n2 rnrnr nn '(rnn n

PAGE 133

n'( r+rnr r"rr2nn#-.BC'<$, %>/r/! r"rnrrn"r '#8> r#87r "nr #-.BC&r+nr r"r' (rrn#8>r# 87n rnnn"r' ( 2nrrnrnB( I0=rCn (I/'(,r2r< rn r2nn#8='2rrn 2r +nrnr rnn nB#-.C'nr& nrnr '(rn#-.BC rn% rr"nrn#-.BC rn rnnn'('(n nr2n n2rrnr'( r" rrnrnrrn"r '(nn r2nnrnnrr2 rnn nn'#rr"n& nn rnrnrr
PAGE 134

rnnrnrrn nn"B>/3?/ rC':2nr2B-/38/rC&2 nnrnr r7 BrnnCn (I0=B(nrnC' / =/ -/ 6/ >/ ?/ 8/ 0/ >// >-/ >>/ >8/ >./ ?// ?-/ ?>/ ?8/ ?./ 8// (n;rnnn ;= ;;6 ;> ;? ;8 ;0 ;. #8>nnnrnr r.r' (n2(I8.r'2rrnnnrn nn#?0B3 Cn#8>B3;C' (nrn rBr<=?/MCnr 3

PAGE 135

rrrr<-/Mr3; '(nr nnnrnrrr n3; nnnr& nr rnn"rnnn r'<$,$' / =/ -/ 6/ >/ ?/ 8/ 0/ /'? /'8 /'0 /'. /'7 = ='= ='='6 ='> ='? <=/3. (n;r 3,nn'n ;= ;;6 ;> ;? ;8 ;0 ;. #8?nrrn nrnr r.r'rnn n 3nr rrrnnnr 'nr rn!3; rnrn rrrn3 '(

PAGE 136

rrBrnrr %C rn "rr'(rn rrr nrnnn':n rnB rC3nrrn n' nrnr#?.2 3n r3;& 3nr3 r "'rr "r& rrnnn'rnrnn2nrrr nrr nrnrB#88C'rnn nnrn+
PAGE 137

/ =/ -/ 6/ >/ ?/ 8/ 0/ /'7 /'7= /'7/'76 /'7> /'7? /'78 /'70 /'7. (n;rnr 3, ;= ;;6 ;> ;? ;8 ;0 ;. #88 r2rBnCrr nrnr'<$,$) 7n /D8"rr#80rrnB $nrnCnrnr '(nn n'(3 $nrnnnnrnr nrnn& n'#80n nnrn ?/rrnrrn'(rnn2 %nr2nrn r:">-72nn nnrrn'( nnnrnn n' nrn'!rnnrn&$n rnn"n'

PAGE 138

/ =/ -/ 6/ >/ ?/ 8/ 0/ / /'/'> /'8 /'. = ='='> ='8 ='. <=/> (n;r(n3nn ;= ;;6 ;> ;? ;8 ;0 ;. #80(3$nrnnnr nrr. r'<$,$, %/n(nnnn n2nr nrnr#8.'nr Br< ?rCr2rnnn "&nnnrn rnnrnnr rnnr r'(n"&"n rnnnnrr 'rn?r& rnnn"& nnnrnnr" r'( 3 nr2nnnnn nrn'

PAGE 139

n / =/ -/ 6/ >/ ?/ 8/ 0/ / /'? = <=/3=/ (n;r 3nn'n ;= ;;6 ;> ;? ;8 ;0 ;. #8. nnrnr r.r' nnnnr"n 3 n"r r=&.n0&2rnr2 nrrr r'(rrnr rrrrn BC'(nrnr r =&.n0$nrrrn nr'n rr&" rrnnr rB(I>7rC'(n rr"nnrn3nrn rnr -nrn'

PAGE 140

/ =/ -/ 6/ >/ ?/ 8/ 0/ /'6? /'> /'>? /'? /'?? /'8 /'8? /'0 /'0? /'. (n;rnr 3 ;= ;;6 ;> ;? ;8 ;0 ;. #87 r2rBnCn rnrr. r'<$,$1 %%/("rrnrr2rB 3nCr r2nn #87'nrrnr"r 3nB#8.C n'rnn2r"rnn# 8.r 3n& 3nrr=&0n.%r2 'n2 3 n 3nr"rnn2 r rn>7r&2%nr2nr rnnr

PAGE 141

<$1 rBCrn rn& rn&n&"+rnn rnn nrrnrnn3"r"rr:" >-7r"nn '(nn nr2n nrnnnn'( nnr2 rrn"r nrn3nr'( rrn rr nr"n"r"rrnrn 'nrnr n"+rnr"r&" -n6 "+rnrrn"nr2 %'( rnnrr" rrn'

PAGE 142

r = rr3(nnrrr n nrrnrrn&n&rn nnnr :">-7n'cn" rr3 n3nrn2"rn "rnnr "nnn< rn rn'(nnr rnrn 2n2rrnnnn' ("rrnrr3 n3 nrnnn& $nrnnn n"nn+n rnr'( 3$nrnnnnrn rn nrnn&n' n2 r"rrnn nB Cnr2r' ( nrnn nrn3nrrn nrBT6/rC &22" rnrrn"' rn%nr2n rrnnBr<>.rCr:" >-7& n"rn r&nnrnn' n"rnr&n
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B>C'r
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"r=')rr&5'&&(n(r rr'-///& nP52Vr%)''n,' -'1n&''&'&rnE n rn '&-//>'=-8 B=C'=.83=7=' 6'Zr&'&'&nrrr< n3 nnnrnnrnnrn nrn r'n&-//-'0>B--C'?0>.3?6' >',r&''&''1&n'9"&;n n:nrrnnrrr '<& =77>'-=6B-C'67=30' ?'&V''&'&,3&3rn rnr3 "rnrnrrnnr '*nr r&-//>'-7-B=3-C'=7?3-/?' 8'Vn&;'&'&rn rrnrnn n3(;nrrnn;(3-7 n rr'n,&-//0'>/B=C'.?37 >' 0';rn&'*'&'&jj#rrrrr r3 rrnrrnnnrrn &r& nrr&RR' r(n'77'03=/' .'n&9''n'')&9r2r 5rn nr;nn' n&=708' 68B=/C'60>-360>0' 7'V&5'&'&3rnrnrrrr nn"n rnnrnr r9rn3 rr'n&-//8'0.B=C'6?3>6 =/' &'#'&'&"rnr rr& )r3&,3#(nrrrr" rn93r 9 '*rrn&-//0'

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=='n2&*''&'&nrn nr3 rnnnrnn nrr' nrrnnnnrn&= 77.'68B?C' 86/3860' =-'&1'&'&rrr rr' rrnrn&=778'>/B-C '=>=3=>?' =6'!n2rr&*'''&nrrnrrn rnn rr'=7.0n3E' =>'n'2%'r';rr3)%& nr'-//0 G-/-//0HP"rJJn'2%'rJ2%J;rr =?'#2&')'n)'rr&n k;52Vr%' =8'r3E&*''&(nrnr r' En rr:nn&-//?'>-B>C'>/?3>8' =0'r&1''n;'*'&n r9n7380/' =.'r&1''&nrnrnr '(*rn r9n rr&=7-.'=-B=C'-7368' =7'2n&;' '&r( 'rnn n"rnnr&=786'=B6C'=7.3-/.' -/'rn&')'&'&5rnnn r +rnr n r'rnnn&(nr nrn&=7./' =883=0=' -='&'('n*'*rn&(n rr""2' r&=77?'=8BC'=3=6' --';rn&'9'&'&:"rnr" n"rnn"' (nrr&=7/0'=B?C'==83=-.' -6')%r2%&*''&'"&5'#'& rr'=708r2 !n"

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-?')rn&*'V'&',n&n''n& rnn rn5rnn""rnrn9 r2r n' rnr5rn rn& =77>'7=B.C'6-/=36-/>' -8'&'*'n,''#%n&r r#nn nrn'-//=&)krn&52Vr%' -0'n1+&')'& rnrrn rnrn" nrn"nrr rr nrnr&53nr&nn+ rnr' rn&-//6'=?B==C'7=637>0' -.'9n&*'&V'!'c'r&nc'[n& rnr'*rn rrnnrnnn&-//>'70B>C'=73--8' -7'(%&;'&'&"rnrn rrr +rnrrrn 3 'nnnnnnrrr &-//>' -//>'rnn rn'-8nnnn rnrnn r&-//>'=' 6/'#2&*''n;'';&rr nr'n&=7.0' ?7B=?C'76637>>' 6='*rnrn&''&'&nn rn nnnrr'nn nnn nrrr&-//6' rnr-? nn nnrnrnnr&-//6'>' 6-'(&9'n'(%&r" rrrnr rrn'rrn &-//>'=0 nnrnrnnrn'BC&-//>'?3-.' 66',n&V''&'&rrrr rnrnr 'nrnr &-//6'=/. B=C'=-3=7' 6>'&('&'&63rr rnnnn nn'rnrkrr n&=777'=>B6C' ->03-?8' 6?'r&V';'&'&3rr +rnrn rr'-//?6J(rr nnrn rnrrnnnrrB' 5r'/?Z760C& -//?'8/386'

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68'9r&;','&'&rrn rr nrnr' rnrnr -//>B' 5r'/>;608/6C&-//>'=/==3=/=6Er-' 60';n&Z'&'nrnr rrnrn n"r'-//6' 6.'&'&r3r33n rn&rrnn r'"""&-//>'?8B==C'=?8 ?3=?.8' 67'9"&'n''1&rrrnr n rnr5rnr n&=77=' ..B=0C'0.78307//' >/'1&''&'&rnrrnr rrr2 'nnnnnnr r+n&& =77>'=6B6C'>/-3>/.' >='2+&''&'&9n0--3>0-7' >-'+n39r&'&(,"nnn' 5&=700'0>B0C' -.>>3-.>0' >6'1+&1''&'&(' :nr&-//8' >>',&5'&'(rr"%&n'#r&rrr n rrnrn"rr '" rn&-//6'6=B?38C'>-63..' >?'2'n''<8')n&5'n''n&rnrrr% n <<&&nn nrn'r *rn&=77>'88B8C'-=.=3-=.7' >0''n'r'(<<'-//> G=-*n-//0HP r JJ'n'rJ$%''nJrr JJ' >.'&1'&'&#rrn(n n*nrn 2n<B=C'>.03?-?'

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>7''n'r'*nrn2n'-//0 G=-*n-//0HP r JJ'n'rJ$%''nJrr J*J*nrn' ?/'n+&*''&'&nrnrnr rnn nrnrrnnr'*rr n&-//?' =/'07?3./?' ?='r%r&''&3rnrn rr'=777& nr!n"!n33rn' ?-'1&''n'9"'2rrn rrn3 nrn'=77/' & &! & 2&5*&!' ?6'Zr&'n*';'(',rn&:n3,nrnrnr 9r2n rr'nn&:'9'&'9n&n;' '2n& nnrn nrrrr'n rr nrrn&-//-'-8>6D?-' ??',&;'3'&'&nnnnn rrrnr 2(;+n3rnrr'r nrn rrn'n &r rr' ?8'r%r"%&'n'')rn&rnrnr n rnrrrn n rr' ''r&-//='>8'7.6D=// -' ?0';n&Z'&'&nrnr rrnrn n"r'nr&-//6' rn r&-//6'=' ?.'&1'&rnnrr rr7-D7' ?7',r&''&'&3rn rrn 'rnr*rn&=77.'-0B =C'73=0' 8/'&';';'&'&(J5.?J%n nnn = r2"r%n&n n#1&n rrn'*rnrn&-/ /6'==8B=>C' -.>?3-.??'

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n 8=';&(''&rrrrnrn 3 nrnnr"'n" rn (nrr&-//6'=B6C'>073>..' 8-'9r&5'&'&"rnrrrrnr "nr n3nnnB Crn r'*rnrnnn nrr& -//?'?B-C'6?0368.' 86'Zr&'&'&nn3nr 3 nnnrnn nr n'n&-//-'0>B8C '=6663=667' 8>',r&''&''1&n'9"&n nr1 3n nn' r*rn&=77?'87B8C'-.//3-./0' 8?')n&*'&'&n;'*'9&n nr nnrrnr rnr' *rnrrnrr&=77 8'6-B6C'=?=3 =0/' 88',r&''&''1&n'9"&3 rnnr rnnn nr rn'7.C&-//6'6/>36/7Er'=' 87'5n&''&nnnr n rrn+rn&n&rrn&n rn'-//>& nrn!n"!n33 nn "n' 0/'2&'&nrnrnrrn r"' r"&-//8'87' 0='[&*'&'&nn3rnr nrn% rr<"nrnnr '*nr r&-//8'

PAGE 150

0-'rn&9'*'&'& rr r2 52rr,3#n'*rn rrr nn&-//>'7B8C'>80' 06'*rn&'1'&'rn&*''&,n03-/7' 08'r&;''&nrnn 'rr& =77/'66'-?36?' 00'*rn&*'&Er"nnr nrr 'krrnnnkrn &=778'6>B?C' 6>836?/' 0.'&'&'&nn'r&&!& 2&5*&!' ./'9&*'&'&rrr nnr rrrnnrnr r'r *rn&=778'0=B=C'>7?3?/8' .='rrr&('&'&nr r3nr r'&=77/'--B-C'.837-' .-'r&'&'& nrnr rn C'=/=?3=/--' .6'"&''&rrnn' r'(n''r',rn' 'r'&=7.0'6=8'=8=3=80' .>'r&'n;'(%&nrr rn nrrnn' rr n&-//?' -8B-C'=73-8'

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.?'#n%&)'&'&n+rn nrnr rrrrn'r nnn& (nrnrn&-//?'?-B0C'=-7?3=6/-' .8',nr& '&'&n 'n& "rnr3 nrnrr' rr n&-//>'-?B6C'8>?38?.' .0')n&*'&''1&n'9"& 3 nnnBCnrnn""nr rnr %nrnr' <&-///'-?7 '=?.3=88'

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n"rrn nnn nrnnnnr!n "r&nn =770nrnnnnnn nr!n"r nnnn-//='!rnrrnr &" rnnnnrrnn#r #rnn nn"B-///D-//-C'; nB*n-.& -//0Crrrnn5n rnrr !n"rr#r';nn n rr&&nrn'