Sistema complemento e resposta de produção de anticorpos em ratos hipertireoideo...

(1)

!

!"#$ !

%

!

"! !

&

"

' $

(2)

!

!"#$ !

%

!

' $

())*

! "

+ ! #$ , #

(3)

' * (+* ' , *& -.* , &(/ 0 '-.* * '0 * ' ('0 &, ,

'1'02* * 3 '03 , 4,(* *5/,5 (*5'0 * ,0, 65( *

' ' (5 &, , &* , , 3 ( ' &, &, 3 , ( '&' ' *5 ,"

( 2' ' '0* 7 ( '

1

) " 899:"

;;; "

! <7

$ # =

"

* $ ' ) ' ( "

(4)

- .

Autora_{: Claudia da Silva Bitencourt}

Título_{: Sistema Complemento e Resposta de Produção de Anticorpos em Ratos} Hipertireoideos

1 ,B

% < =" & < ="CCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCC ( CCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCCC' CCCCCCCCCCCCCCCCCCCCCCCCCCCCCCC

' D CCCCCCECCCCCCECCCCCC

! "

(5)

(6)

! "

# $

! %

&

!

'

(

)

*

+ ,- ./

) 0

"

(7)

!

"

!

#

$

%

!

%

(8)

#

!

' (

)

*

)

'

)

(

"

!

(

+

)

+

(9)

, !

$ ) ! - .

) % , )++

% ! F ) "

F G # G " *

) ) "

, )

' G )

" ' F G % " '

" '

G B F % F "

) / , 0

' F G H G I G "

* G % F ) % ! " D

B ) "

!

' % " )

" D ) # ! " 5 F H

% "

1 2 +

' ) " 3

J " ' & ! F

(10)

/ )

/ ' K H & " ? %

% ! " * ) ) " ' #

% %# & " ) "

' ( 3 4 5

1 ) G % F # D H % # %

" ' J G G "

% ) ) H "

) 6

) D G ) % " ' % %

" * F G ) #

% "

) ' 4 4

4 ) ) G " ) %

H G

"

) , 7 8 ) , , 4 8 / 8

8 6 ) , 8 2

' % G " D

J " * " ) F "

) & ) ' ) ' ' ,& ' ,

9 ,"+ 2 ' ' 4 %

(11)

+ 4 5 4 , : 4 ;

6 < / 4 2

3 1 G ! "

G B L %

H " ' D G )

% " * ) "

) ' ) , ' + * 2 + '

2 ' 2 ; 7 ++

H 0 1 G ! " ' H

% F B J

F " 0 J

"

' ( ) , , ' ( , 9 5 2 ' (

, , ' ( , + ' ( 3

4 5

% " '

G G B ) " *

(12)

) " 9 / , & ! 9 <

' ' ( ; , 5 ; < 9 +

4 / ) , 7 , <

5 6 ; 6 =

' !% " 4 ) " '

) ) "

) " >! - ( + .

) / % 2 / 2

/ B! # N

) + + , F

) N

5 N

' ( " ); ?< ' ( ) 7 +

J J !% G G )

(13)

' ( " ) ! ! ' ( 9 $ % ' +

J "

' ( ! !

(14)

(15)

1( ,5 * " " ! !"#$ ! % ! " 899:" OP " <& = Q

899:"

J

) M < =

) ?

% " * # # ! #

G " G

) ? #

G ) G ) J

D ) % !

# ) ?

"

F ) R B

) ! H E

# ) ! N ! # % 1

# % % 1N

) H < 1 = ,0( ' <S FT U

TV= S G % V < =" * )

F J ! H E "

(16)

! % % ; A

A9 ;99µ B E899 ;@

9 ;Aµ E899 W ;8

" * ! # % % 1 % F J "

' #

> F 1 " , G !

) ? F %

) H

) " , %

GX !

# !

F " , H " '

J B )

% J ) "

& / % & $ ) ?

(17)

(18)

1( ,5 * " " T % % T

R ) )T )T " 899:" OP " ) <& = Q

"

) % ) T ) )

)T )T ) % ) T < =

) R U ) %% % ) ) % )T

) ) ) T "

) ) % R) )

) % % B R )

% ) % ) "

Y R ) )T ) R

B % )T )T ) T T %

E )R T % ) ) ) ) T T N )

% % 1 T % % ) < 1=N )

% ) < 1 = ) ) FT U

<,0( '= G % < = T "

)R T T R %% ) )T )T "

2 R ) ) )R T T R % T

% ;N AN A9 ;99 µ % )T B < @= E899 % R ) E T % ;@

T % % 9 ;A µ % )T < >= %

(19)

) " ( ) R % ) T

R ) > F R ) 1 "

) ) R ) ) % T % )R T

G % ) ) % )T ) T T

T T ) T ) T "

) ) % %

G ) % )

) T " '

G ) "

0 1 2 ! $ T )R T )T ) T

(20)

(21)

'5*/' H I

1 #

,@A*70 @

; O

>

> 1 >

> 1 > A

, B % %% C

, ' #

& S % B V

4 B

T

,2 8

; ;

8 8

0' @ % # W

(22)

,' F

) H

,& ' H #

, ' H #

%4* ) > +

% FH #

2 ) ? % !

2 ) ?

) % ) ? "

20' ! H )

( 5 Z % Z

( & &

( , ,

(

( 4 4

02 ) ? F

4' B G

4' 410

410

(23)

42 B )

5[ S U V

E E

1 % %

' A ( ( =

EF

'/, "

1 % %

J

& %

5

S ( ( V

2 , &

\ A9] ) H

>

(24)

1' B < C

1 B < C

,'

2 ) ? %

2 %

2

(25)

(26)

3 / .

343 5 )(

34( 5 ! & #$ )6

347 5 8" #$ ! 3(

349 5 " #: ' ;8 ! ! & #$ !

3(

34< 5 #$ ! !

!"#$ ! = " 3<

34* 5 ; ! > % ? !"@ ! " A" #: 3B

346 5 A" #: ! ; ! (3

34C 5 " ;8 > A" #$

! (7

( / D (C

7 / _E 7)

743 5 7)

74( / - ? 73

74(43 5 ! " $ ! & F G 9H

G 7H 73

(27)

747 / ! % ! 7(

74743 5 ! ! ! G H 77

!

"

77

74943 5 % ! 8" ! % ! " $

! @ ! ! % I 79

7494( 5 #$ ! & ! ! J ! & & !

5 K = ! = L 7<

74947 5 #$ ! & ! ! J ! & & !

5 K = ! I L 7*

74< 5 & #$ ! & ! ! J ! I M 76

74<43 5 #$ ! J " ! /% I !

76

74<4( 5 % ! 8" ! ! " $

! @ ! ! % I 7C

74<47 5 #$ ! & ! ! J ! & I M

5 K = ! = L 7B

# $ ! % &

(28)

74* 5 & #$ ! ! 9)

74*43 5 ! ! ! ! A G H 9)

74*4( 5 & #$ ! J& = A" ! A 93

746 5 #$ ! J& = ! % ? 93

74C 5 #$ ! N ! = " !" !

G H 97

74C43 5 ! " @ #$ 97

* + , % 99

74C47 5 ! = " ! ' # 99

74C49 5 ! 9<

- . / " 8 ! O @ H 9*

74B43 5 ! J8 & ! 9*

- + 0 96

- . 0 9C

74B49 5 ! #$ ! 8P 9B

743) 5 I ! " ! <)

(29)

1 2 $ 30

, 0

943 5 " 9Q <(

94( / Q <6

947 5 ! *)

1 + ! 0

943 5 " #$ ! % *(

94( 5 & ! ! J ! & I M *9

947 5 & ! ! J ! & & *<

94743 5 " 9Q *<

9474( 5 Q G 7H *C

94747 5 ! G 7H 63

949 5 & #$ ! J& = A" ! A 6(

94943 5 ! ! A ! A G H 6(

9494( 5 & #$ ! J& = A" ! 6<

1 1 0 4 56

, 7

943 5 & #$ ! N ! = " !" ! 66

(30)

9434( 5 F & #$ !

" %" 6C

94( 5 #$ ! J& = ! / C(

94(43 5 & #$ ! J& = ! 8P C(

94(4( 5 & #$ ! J& = ! 8 C(

< / C6

* / R BC

6 / P + 3))

(31)

(32)

3 5 !"#$

343 5

* # D ! #

G %

" * %

,) ) 4 ) < *0,N 4* '5 899>N * 0, ;OW:=

% H G

# ) H < ' *0 899@=" ' %

% # ^(^ 1 1X ) % %% % <1* &,

;O99N ' *0 899@N * 0, ;OW:=" &# # #

H% % G D ! G %

" 4 D ! D

% ) % ) H

" ' % G

S V < (00,4, " ;OA@=" #

% ) M H <* 0, ;OW:="

4 % S V G ) L

#

< K( ' 8998="

, S% H V

(33)

G % F < ; 8 >=" ,

% J # F B

X! D )

!%

% <1' (5 *5 " 899;N ' *0

(34)

' 3Q

J H

"

GT 5 J H

#$ I

Gµµµµ8M H

" ! & ! & #$

> ;PA <α ;;9Eβ WA= ;899 ;>99

&

1 O> 899

& 8@ 8

_ ;;9E;:AE899 < ? Q AA= 8A

I

;G @:9 ;A9

; PA A9

@ 89A <α OWEβ WAEγ >>= >99 :99

8 ;98 89

!

410__ 899E>99E@99 ; <9 9; 89=

4' ; ;99 ; A ;8

4' 8 W:

A ;O9 <α ;;AEβ WA= P9

: ;;9 @A

W ;99 O9

P ;A9 <α :@Eβ :@Eγ 88= AA

O W9 :9

410 Q N 4' Q 410N Q N

(35)

' ( Q !

J H

"

GT 5 J H

#$ I

Gµµµµ8M H

( PP >A

2 ;A9 >99 @A9

; (52 ;9A 8@9

@ AA9<WBW9 ;B@A= 8A9

! </ = P@ A99

< @9 @9= W9 <>A `>A= A9

B 5 _ <8BO9 8BA9= >A

' " '" #$ !"

; < &>A= ;O9 4 % %

% # 1

&' < &AA= W9 # X! %# < B

5[= #

# "

4 < &@:= @A W9 __ ( &' < B =

&AO ;P 89 ( &'

; (52 Q ( ;N Q N &' Q % N

4 Q ! % N & Q S % % VN <_= Q

(36)

' 7 ' $ % %

5 J 8" ! ! ' !

" J

8" !

A ! ! " #$ ' ;8

; > @

( %

"

4 > @ % % ( >

@

&' @ 8 > 1 '

2 E P PE O ( ! <4' =

&AO O ( ! <4' =

5 J 8" ! I

J " #$ ' ;8

( ; 1 G ; ; "

@ ' B @ 8 %

@ % (

2 ' B >1 %

> % ("

( ( @ > "

B ' ( % B > @ A "

! 0 A :W B %!

0 A :W B %!

"

, F > "

(37)

34( 5 ! & #$

) $ H

" & G

F F ! #

" ,

! ' ! I

# ! <[0,(5 ;OO9N ' 0 ;OOO=" ' G H

H F G !

" # Q

% > > < ;=" * G

% >

"

/ H

' H # F "

' )

%! F

" * ( ( 4

G # ( 4 M %! G %

(38)

# ( I % B >9 @9 <K'5,Y'a

" 899>=" * # B ;

;G ; ; %

# ! ``_{<[0,(5 ;OO9=" ;G} _M

% % G B J ! !

; " ; % H F ; G F

@ G M %! " ' 8 # #

; % %! > / H

< @ 8 = <K'5,Y'a " 899>N [0,(5 ;OO9N ' 0 ;OOON ,(&N * , ;OP;="

/

' # %

! ! I > < ,' *5 ;OP;N

2 4'5N 2*0, 899:="

% > $ % % " '

% % # % # >

# H % ><28*= #

" , B ;] >

) < 2 4'5N 2*0, 899:=" * ><28*= # >

S > U V M 1" 3 #

(39)

F 1 1 1 %

% % < >28*1 =" 5 ! 1 # B

> > % % B %! " * > #

! 1 ! & G

1 1 " ' % > < > 1 =

< ,' *5 ;OP;N ,(&N * , ;OP;N 2 4'5N 2*0, 899:="

' % # #

% > H " ,

S % V

< ,' *5 ;OP; ,(&N * , ;OP;N

2 4'5N 2*0, 899:="

/

' G # <410=

G #

4 7 <K'5,Y'a " 899>= ! <[0,(5 ;OO9N

2(,0 ;OOW=" ' 410 < = H

4' ; 4' 8 B # )

B ; / H " ' G B

%! 4' 8 @ 8 % >

(40)

> 4' ; >

< K( ' 8998="

/

> "

* % M / G %

< *0, N 4* '5 899>="

' > # F # >

G < ;=" * > #

> > G % B M %! " 5

> M > % A

> 1 > @ 8 > H " ' A

A A A " * A # % 4'

< B ' G 4 =" 5 % % ! A : W" ,

% B J ) % W G

" * P G F

F O" ' # O % 4' "

* 4' <0' 24'55N

' [(5 *5N ,(& ;OO;= S UT )V G M

(41)

8" 3 Q , G "

' K'5,Y'a " <899>="

+ +

4 J8

/ ! J 8 " AJ;8 !

, > 9> (

7> 3U> 3 > 3 V

9V (

7 &

@ > A

! ! J! ! A #$

! !

J! !

A #$ ! ,

<'> *> 6> C B

! , <'> *> 6>

C B F ! U" ' F ! U" O

' G H

" ! A 8 ;

" ! A 8;

8 #$ ! 8 #$ ! > > ! ;8 = " M & #$ "

! ;8 = " M & #$ "

@ #$ ! ;8 @ #$ !

;8 F_F#$ !#$ !"_"

+ +

4 J8

/ ! J 8 " AJ;8 !

, > 9> (

7> 3U> 3 > 3 V

9V (

7 &

@ > A

! ! J! ! A #$

! !

J! !

A #$ ! ,

<'> *> 6> C B

! , <'> *> 6>

C B F ! U" ' F ! U" O

' G H

" ! A 8 ;

" ! A 8;

8 #$ ! 8 #$ ! > > ! ;8 = " M & #$ "

! ;8 = " M & #$ "

@ #$ ! ;8 @ #$ !

(42)

347 5 8" #$ !

F H

%

G " * # < ' *00 899@N

*0,N 4* '5 899>N K'5,Y'a " 899>N ' 0 ;OOO=" * %

%! %

" ' # # !

# %! %

<K'5,Y'a " 899>N [b20 899:=" , ! M

) < ' 0 ;OOO=

< (=N #

< 2=N % <4 2=N F

< =N % <&' =

% 4' M %! < &AO= < 8

>= < ' *00 899@N *0,N 4* '5 899>N K'5,Y'a " 899>N [b20 899:N

' 0 ;OOO="

349 5 " #: ' ;8 ! ! & #$ !

' % G F

(43)

L " ' % N B N

N % N N )

"

* F $ > > G M

# % " * > M

> > > G

D F

% < '2 "N 0'41 ( 899;=" * @ @

B ; # # <K'5,Y'a " 899>N ' 0 ;OOON

'2 N 0'41 ( 899;="

4' $ # % ! % B G M

G G

# <4,/* ' 2 899>=" * 4' % #

G ! F J

% J # < *0,N 4* '5 899>=" %

% 4' !

# H G ? <[0,(5 ;OO9N ' 0 ;OOO=

# % % B # )

(44)

B $ B %

! % % "

' B % B

/ H "

* > B ; <

; = ) H " ' ) H

B # %! ) H

;

% % " ' # B

" ( % >

J # ! ! " <K'5,Y'a " 899>N

[0,(5 ;OO9N ' 0 ;OOON="

% $ % > @

A < > @ A = % B

) G % H ( ,

< ,' *5 ;OP;N K'5,Y'a " 899>=" ' % B

# % " > A

) " *

F )

(45)

! #

B G % " * % @

J G > A

B % < '5[N (, ;OO;=" > F

L % % %

< '5[N (, ;OO;=" A # G HB G %

% !

% < ,' *5 ;OP;N '5[N (, ;OO;= #

F # " A ' < A

% H = % G B < ,' *5 ;OP;=" #

< *0,N

4* '5 899>= % < ( 2,0 *5N

' '0(N 4,/* ' 2 899;=" A

<[b20 899:= %!

B H < ,a " 899>="

34< 5 #$ ! ! !"#$ !

= "

' M % 1 # %

(46)

, Y'5 N 1, ;OP@N [b20 899:N 4' ,00* " 899AN ( [, 899AN

'2 N 0'41 ( 899;N * ( 899A=" ,

% G > M %

1 <5 ,5+Y,( " ;OW;= G > !

! < , a ;OW8=" ) <;OWA= % G

H H !

% " G

% S D V > <&,4 ,a " ;OO:="

& <899@=

% % 1$ # 1 N

% 1 N N

# 1 % 1

"

* % 1

%! ( 4 ( & G B %

! # 1 < # 1=

<K'5,Y'a " 899>N ' 0 ;OOO=" 3 1 ) !

#

! # ! #

%! # 1 ! # B

) <42 = (( <K'5,Y'a " 899>N ' 0

(47)

= # H # B < ! !% =

G ) B ! $42 (( %! #

! H G

# 1 % B <K'5,Y'a " 899>=" ,

% > ! < ' *0 899@N ( [,

899AN '2 N 0'41 ( 899;N [b20 899:=" % B

% # %! &8;$ &;O$ &P;

# 1 1 G ! " *

! > > &8; < 8

= G

G % F <[b20 899:N

4* '5 899AN ( [, 899A=" KH ! F

# 1 % !

!% " ! G F

( 4 F % # <K'5,Y'a "

899>="

* % # 1 #

! % % G ;A 8;

F " c G % % B

) H " ' J %

B # % ! B % 1

(48)

! ! F B

% 1 < ( [, 899A=" F )

! # 1 % %

% % " *

# )

# G F

% < ' *0 8999N ' *0 899@N K'5,Y'a " 899>N

( [, 899A="

* F % 1

% 1 ; G G F

! # &8; &>A < 8 ;=

B # < ' *0 899@ [b20 899:="

* % G F

# B < ' *0 899@=" & % ;G @

0

G I # 1

" & 8; &>A % #

(49)

34* 5 ; ! > % ? !"@ ! " A" #:

' ) ? $ < >= B < @=

" * > @ L

) <1('5 * ;OO;N 0' ,5N (5 1' ;OO>="

! ! H " * @ > W9] WA P9]

1 <S )T V= " ' H

1' <S )T V= G G %

<9 >] > 9 98 9 9>] @= # F # B

% <1' ' " 899>N 0' ,5N (5 1' ;OO>N 0( 0, 899:="

5 # ) ?

% ! ) ? " ,

J &5' B

% J F 5' " * !

) ? G

! F " * %

$ %

# B N J

! # %! N

! # " * > # ) ?

F ;9 ;A F G @

(50)

* @ # ) ? B H % G F

> Ad " , #

%# G % @9] @ > " ' Ad

# F @ > < > = G " ,

# ) ? " &

H I O>] # @ > G

> H # @ %# <'203/(

;OW:N 0' ,5N (5 1' ;OO>N 0( 0, 899:="

* ) ? % F G

% " *

<e/f> = G J

! <(5 , ( 899A=" , J

< '5(5 , " ;OP:="

' ! ) ? H

" ' ) % # !

2 <S )T 2 V=" * 2 !

# I G

! ) ? " & 2 #

(51)

346 5 A" #: ! ; !

' % J I ) ) % " *

) # %

% ;9] <0( 0, 899:= #

% * F 4 L L

L <'5&, *5 " 899A="

& ) % J :9 P9] M & D

# ) " *

8A >9 ;9"999 ) <0' ,5N (5 1'

;OO>N 0( 0, 899:=" c % ) I M G

2 I " *

% ) < 2 =

) ? " ' H

@ A] < *00(5 N * 2 8998="

2H G B ) ?

< @=" 3 ! # ) ? B

X! B G

) ? B % ! <0( 0, 899:="

* ) # % D

(52)

KH ) # % ) ?

# "

' B < @ # = # %H )

) % L B

" @ # > #

0 B ) ? " '

0 < > # = # G B

F H )

< ,,5 '5N &*5 ;OOA="

' 9 % L )

# "

2 < B = 2 % H

&

1 B

' B

2 % H

' H Q 2

2

% H )

) ? <% =

I H

( < ) ? =

2 Q 2 ? N ) Q % ) ? "

(53)

34C 5 " ;8 > A" #$ !

* "

* ) ? F

< '/(5*N &' &,55, ;OOA=" * J

) G

<1, ,&*/ [aN * [(5 ;OWA=" , ! %! % F

) ? G #

% " * ) ? ) % F

! ) I J " '

! % 1 %

Z <( 5 Z=" * ) ? < 2= # B

) ? F <02= % ! < 2=

< 4( 2N 10'0* [ ;OPP="

* ) ? % < 2 2=

! I < > @= #

" 0 % ) ?

<'203/( ;OW:N [,' N 'a , ;OP9N Y'00N Y*2( N 2' (,

;OP;=" * 2 2 ! <[ , "

;OPO= G 2 # J % % 1

<10'0* [ " ;OP@N [ , " ;OP:=" ' # [ <;OPO=

(54)

G %

H % "

' G H

!% G % #

" 5 B B 20'

0' @ G

B # # I #

& @9 ! <Y,, 4'5 899>=" G

G % %

I ! I <

B 2 =" 5 ) 2

G H 42 (( (

% # <Y,, 4'5 899;=" 5 # %

% G

I <5'['( " ;OO8=" , &

@ % G # 5[

! ! @ < ' ( " ;OPW=" ' # )

& GX !

F ! )

<' Y' " ;OOAN 2' 4'5 " ;OPA="

5 % G A9] )

(55)

G " ,

<*[* (,4, " 8998=" * )

% $ %

N L # 5[N 20' < (= 8

% &@` !

</*0 g 899;="

#

# " G 4'

# & 2 ) " ' B

< &@: &AA &AO=

F % B # < '5&*5 "

;OO@=" * UT <;OP:= % G ! O &

G ! " O

% 4' #

G % "

4 B F

" ) % :

) 4' M

<(5* , " ;OO>=" * ) )

% % # " 5 @

(56)

[ U <;O@9= %

) G

) F )

F " *

% G %

% J F " , F

G M F )

H "

, ) F <& ' , 8998= % G

< = G G !

) ? G )

J !

H " '

G % # %

1 % G # # F

B ! ) ? J <& ' ,

" 8999=" ' ) H

) H <& ' ,

899>= G J G

(57)

(58)

(4 D

B D $

• ( % )

) ? < > @=

! H E N

• ,% H H ) ?

J !

H ) ? " , #

% !

# ) ? N

• ( ! J J

) N

• ! J

(59)

(60)

7 5 E

743 5

* % % 1 # %

# $

• R ) D <;P9± ;9 =

B < @= < >= E

F ) H N

• ) 5 R + % D <±>[ =

) H / '

N

• 4 ) H

H E !

F % ) ?

N

• D F

L # < ="

# % % 1 # <S % % V=

(61)

• R ) D <;P9± ;9 =

F ) H "

% # #

# Q , ' Q

L 9A";";;:9"A>"O" ' G D % %

F "

74( 5 - ?

74(43 5 ! " $ ! & F G 9H Q G 7H

B < @h % T ) = %

H G %H

J 9 ;AN 9 >9N ; 9N A 9N A9 ;99 µ E 0" '

% F h H

J 9 ;AN ; 9N ;9 A9 µ E 0" ' % B ; 0

(62)

74(4( 5 ! " $ ! ! G 7H

* %H % <0 % = %

G B < 4 =

% B $

Q $

B < 4 =""""""""""""""""""""""""""""""""""""9 A]

5 """""""""""""""""""""""""""""""""""""""""""""""""""""""""""""""""""9 ;P]

5 F """""""""""""""""""""""""""""""""""""""""""""""""""""""""""""""""""9 98]

7 G """"""""""""""""""""""""""""""""""""""""""""""";99 0

' %H % !

J % > 9 ;AN ; 9N ;9N 8AN A9 i E 0" ' % B

; 0 "

747 5 ! % !

* % B

<@ E = H j j

<≅8@k =" * ! % > W ;@ 8; 8P

(63)

* H G 4

B B " '

%H G % F H

# I !

F ) H % B " ' # !

% D D P ;9 ) " &

H % ) B

% M H @9 "

' ! % % 8A99 ;9 " * % G

QW9k H "

74743 5 ! K L ! ! G H

% < l89=" *

) % )

S V < 5 = G % G

(64)

749 5 & #$ ! & ! ! J ! & &

' ! % #

) ! # $ # < *02(00N ( N K*25 *5 ;OWP=

H <4'a, ;OW;="

74943 5 % ! 8" ! % ! " $ ! @ !

! % I

* ) % ) ' # <2* 4'5N

4'a, ;OWW= ;$; ;A >Wk

,'E,& ' < Q 8P 4N H # Q

;9 4N 2 W @N 9 ;] < E = =" ' ! % %

;9 8999 % G )

% ) H ,' 4 8` _< _Q

8P 4N % # Q 9 9984N 2 W @N 9 ;] < E = = " ' %

F L

' # " ' ) H % % @k

# ;A F "

5 ! F

(65)

8999 " ' ) H % F

,'E, 'E4 8`_< _{Q 8P 4N H} _# _{Q ;9 4N}

% # Q 9 9984N 2 W @N 9 ;] < E = =" '

) H % F I W99

9 W9 9 P9 G ! ; A F "

7494( Q #$ ! & ! ! J ! & & !

5 K = ! = L

* QW9k % >Wk !

,'E, 'E4 8` _J _;$> _{;$@ <899i =}

F % ? < & W9 1 U '= >Wk

8d>9dd"

F ) H ) >Wk <@99i =

# ! ;9 < *02(00N ( N

K*25 *5 ;OWP="

' H% # \ #

A9] ) H <

8=" ' ! # \ G

G \ H ! G \

(66)

8" ( Q H% # ) H )

! / ' " '

A9] F H \" ' Q !

;$> 1 Q ! ;$@"

74947 Q #$ ! & ! ! J ! & & !

5 K = ! I L

* QW9k % >Wk !

,'E, 'E4 8` _H _J _{<>99i =" ,} ₎

) F ) H

% A99i0 >Wk >9 " * %

) A99i0 1 <

% % = " % ;9 8999

Tempo (min) A b s o rv â n c ia (7 0 0 n m ) 1.0 1.0 0.0 0.0

0.5 (B) 0.5

t½ (A) t½ Tempo (min) A b s o rv â n c ia (7 0 0 n m ) 1.0 1.0 0.0 0.0

0.5 (B) 0.5

t½ (A)

(67)

I % % ? @;8 " '

I % M I

) H H <;99] ="

74< 5 & #$ ! & ! ! J ! & I M

74<43 5 #$ ! J " ! /% I !

* # F ) ) H

% ' % <;OW:="

) % G

J ;$A9 # ;$P99 < E = % F F

) H ,' < F

) H =" * ,' % ) !

H H < =

) " ' ! ) D

G H F % G ) H

(68)

74<4( 5 % ! 8" ! ! " $ ! @ !

! % I

* % ) ' #

;$; % @k # ;A F "

! H E F

!G % ' # % %

;9 8999 " ' % G )

% ) H F

& <S % B V Q H L Q 9 9;AA 4N

Q 9 99@A4N Q 9 W8A 4N # Q 9 99@;A 4N

H Q 9 99;8A 4N F Q 9 99A 4= <2' ( *5N 0' 24'55

;OP:=" ' ) H % F 9 W9 9 P9

% ? I W99 G ! @9 F "

,' %

) H ! % ! #

F ) H " ' % @k ;A

F 9 W9 9 P9 % ?

(69)

74<47 5 #$ ! & ! ! J ! & I M !

5 K = ! = L

' / H E0 % F

,R " <;O:;= <;OO9="

* % ! & % 899i0

! J ;$89 ;$@9 F

% ? < & W9 1 U '= >Wk 8d>9dd"

( ,' <@99i0=

>Wk # <;9 ="

' \

F / ' "

74<49 5 #$ ! & ! ! J ! & I M 5 K = !

I L

* QW9k % >Wk ! & H

J <>99i0=" , ) ) ,'

<899i0= % A99 i0 >Wk >9 " *

(70)

A99 i0 1 " % ;9 8999 I

% % ? @;8 "

' I %

I ) H H <;99] ="

74* 5 & #$ ! !

74*43 5 ! ! ! ! A G H

#

< = <2' ( *5N 0' 24'55 ;OP:=" ' !G 9 A 0 5

, %h ; A 0 % G )

B A:k " ' >9 :9 i0 G %

% ) 8@9 i0 ,'E, 'E4 8`

) " ' F ) H

) <899 i0= >9 >Wk " ' A99 i0 1

% ;9 8999 " ' I

% % @;8 " *

% F !

A:k H

" ' % 1 %

! % 1

(71)

74*4( 5 & #$ ! J& = A" ! A

* ! # % % 1 % ) !

H % 1

! 1" <>99 i0

! ;$A= ,'E, 'E4 8` _% _{:9 i0} ₁ _{899 i0}

) H ) F "

746 5 #$ ! J& = ! % ?

! # > @ 2 #

G ) ( h ;999 <& =

F U !% ) ? " %

) S V" ' S

V % ) G "

* ! > # % F U > G

! ! H " 8A i0

% >Wk >9 )

# > % %

< = D >" '

(72)

G < B = % %

% ? ;9 >Wk " '

% % % < 4 =

G < = % >

< E 0= F "

* @ 2 % % "

* ! @ # % F U @ G

! B ! H " ;A i0 %

>Wk >9 @

% % < = D @" * %

@ <i E 0= F "

* 2 # % # U 2

F WA i0 >Wk :9

2 % % < = D

2" * % 2 <i( E 0=

F "

(73)

74C43 5 ! " @ #$

% < lP=

4 N B 4

H ;8 " 5 % F 9 : 0

) H 5 # 9 O] <;$A= %

B I " ' ! A :

F % % )

"

F !G B A

) ' <;$;= @k %

% 5 # 9 O] < E = ;9 8999 " *

% ) H % F

J " * ) H % 5 #

9 O] < E = ;$A < $5 $ = %

(74)

74C4( 5 ! ' ! & ! % I !

D %

! " ' @9 H %

% 8999 ;9

) S V"

) H # % J 8999 1

# % ;$;9 < $ $ =" '

% ) ;9 % @k A

;A99 < ,&' " ;OO;=" * % !G

W9k "

74C47 5 ! = " ! ' #

* %

) H > # 2 U

< 2 W"@=" ' ; 0 # 2 U % %

h ;A 0" ' ;9 G

% % % h A9 0

@9 0 52@ 9 PA] < E N 2 W 8= ) H "

( >Wk ;9 ! % %

(75)

2 U % J F " *

% > 0 # 2 U " &

I 5 L 0

F F G F # H

< , 25,a ;OPW=" ' % ! 8 AB;9A _{8 AB;9}:

# E 0"

74C49 5 !

F F $

− 0I <8 ABW A = 9 A]

H % @P ) N

− ) @99i B %

< (1 * h= 9 A] < E = # 2 U N

− 1 ) H #

5 # 9 O] < E = %

# 2 U A] < E = ) H N

− # J 8 AB;9A _{8 AB;9}:

# E 0"

(76)

5 B % >Wk

A9i0 1 ;99i0 " 2 F L

% I %! " ' %

I % % ! %

B " ' # 2 U

! " * % % I L

% *8 <A]= >Wk ;A9 " ' !

) H

! ;$A 2 U " ( I L

% *8<A]= >Wk @A "

* ) H % % B

"

74B 5 G " 8 ! O @ H

74B43 5 ! J8 & !

* ! ) H % %

<;OO> ;OOA= % J "

% ) ' ;$;

P99 0" ' % % ;A ;999 "

& % ) H %

(77)

% F J " &

) H % < 2 9 9A4

,& ' 9 ; 4 2W":=" * % % 8A"999 >9

% # " * % % F

@k " ' % 8A"999 >9

% F %

< & = 9 ;] < E = 9 98] < E = F " * ! F

% F & 9 ;] < E = 1

G 89k "

' ! !

% # 1 % <;OW:= 9 W

E 0"

74B4( 5 & !

,0( ' ( 821h O: % !

<;8A i0E = ) H P i E 0

(78)

74B47 5 & #$ ! 8

H

! % > F ;8Ai0E 1 2 W @ 9 9A]

< E = R 89 < 1 =" ' 1 A] < E =

8 ) " ' %

1 <;8Ai0E = > F " ' %

1 ;$;: % ;$89@P" '

% ; ) "

J <

( 4 Q i ) % Q ) 1 )h= ! 1 ;$;999 ;8Ai0

" ' ; ) % >

F 1 < m ) T

) ) =" ( ;A

% % @9A % ? ,(' R Q

& " , H % % S V

F N S V

(79)

74B49 5 ! #$ ! 8P

! % > F

;8Ai0 1 2 W @ 9 9A] < E = R 89 < 1 ="

' 1 A] < E =

8 ) " ' % 1 <;8Ai0 =

> F " ' % %

1 ;$:@" ' % M

; ) " J

< ( Q Z ) % Q ) 1 )h=

! 1 ;$;A99 ;8Ai0 " ' ; ) M

% > F 1 <

m ) T ) ) =" ( ;A

! % % @9A

% ? ,(' R Q & "

H % % S V

(80)

743) 5 I ! " !

! % F

"

* '5*/' & 55, % F J L

% % G # "

* 4 Y) T % F "

# G

! < # = #

% ! "

% # % R )

>"9 Y R ) % R & % '

(81)

(82)

94

1 2 1 9 30 , 0

) ? %

% J $

− @h B < @= % N

− h < >= % N

− < >= Q % !

%H F % "

943 5 " 9Q

F % !

% ) ! 2

< >= @ < @= > < A="

* % B ! ;@ " *

A9 i B %

(83)

A i ! ) ? G F !

"

* ) ? % A9 i

B < @h= ;@ " * ! # @ %

% % %

% A i " * ! > <) ? % =

% A i # G

< @= A9 i B

(84)

0.0 0.1 0.2 0.3 _A

Controle 5µg

T S H ( µµµµ IU /m L ) 0.0 0.1 0.2 0.3 _B

*

T S H ( µµµµ IU /m L )

Controle 50µg

8" 7Q ,% B < @= ! # 2"

5! # 2 H < = Ai <'=

A9i <1= @ ;@ " & ! 4

Y) T $ ' l 9 W8@8 l;9" 1 l 9 9;9W l;9N <_=

(85)

0 5 10 15 _A

Controle 5µg

T 4 ( µµµµ g /d L ) 0 5 10 15 _B *

Controle 50µg

T 4 ( µµµµ g /d L )

8" 9 Q ,% B < @= ! # @"

5! # @ H < = Ai <'=

A9i <1= @ ;@ " & ! 4

Y) T $ ' l 9 ;8W@ l;9" 1 l 9 99P; l;9N <_=

(86)

0 50 100 150 200 A

Controle 5µg

T 3 ( n g /d L ) 0 50 100 150 200 _B

Controle 50µg

T 3 ( n g /d L )

8" < Q ,% B < @= ! # >"

5! # > H < = Ai <'=

A9i <1= @ ;@ " & ! 4

Y) T $ ' l 9 W8@8 l;9" 1 l 9 9W8W l;9"

' F G

B ! 2 @ >

(87)

94( / Q

* Q < >= J ! @

< := > < W= " * 2 < P= %

; ;9 A9i "

/ % G ! # @ % !

; ;9 A9i Q" & ;9 A9 i

! @ ! H F

) ? "

-2.5 0.0 2.5 5.0 7.5

Controle 1µg 10µg 50µg

T

4 (

µµµµ

g

/d

L

)

8" * Q ,% < >= ! # @"

5! # @ H < = ; ;9 A9i

(88)

* ! > M G

> ! W " ,

HB F G

H "

0 500 1000 1500

*

T

3 (

n

g

/d

L

)

8" 6 Q ,% < >= ! # >"

5! # > H < = ; ;9 A9i

> W " & ! '5*/' & 55,

(89)

* h ! 2" * J

G %

% I ! J "

0.000 0.025 0.050 0.075

T

S

H

(

µµµµ

IU

/m

L

)

8" C Q ,% < >= ! # 2"

5! # 2 H < = ; ;9

A9i > W " & ! '5*/'

& 55, J L l;: )

(90)

947 5 !

* G > < O=

! ) ? % ! # 2 <%

;9=" * ! @ % F

H # ! F < G ;i E 0="

0 250 500 750 1000

*

Controle 25µg

T

3 (

n

g

/d

L

)

8" BQ ,% < >= ! # >"

5! # > 4 < =

8Ai > ;8 " & !

(91)

0.00 0.05 0.10 0.15

*

T

S

H

(

µµµµ

IU

/m

L

)

Controle 25µg

8" 3) Q ,% < >= ! #

2" 5! # 2 4 < =

8Ai > ;8 " &

(92)

1 + ! 6 0

& )

F H ) ? H !

% ! J " '

% % ! #

) ? E 2 G G !

% "

I

943 5 " #$ ! %

F ! H

) < ) H = F

F F ) H "

' ) % # ) ! H %

" 5 <% ;;=

) M % !

F ,'" * G ) # F

% ! ;$P9 < =" ' %

(93)

) ! F ! ;$A9 )

"

8" 33 Q ) ) ! H

H E " )

B C ) H F

% ! ) H <) =

% $ ;$89 ;$@9 ;$P9 ;$;:9 ;$>89 ;$:@9" , F "

0 80 160 240 320 400 480 560 640 0

25 50 75 100

recíproco da diluição da hemolisina

%

d

e

h

e

m

ó

li

s

(94)

94( 5 & #$ ! A ! ! % ? !

' & ! ! J ! & I M

' ! H E %

B " % %

;8 % B "

% >"

8" 3( Q ,% B < @= !

H E " * \ < =

H < = ;i Ai @ " '

) F # B

" ! $ W ;@ 8; 8P < % ="

& ! '5*/' & 55, J

L ) J % "

Controle 1µg 5µg

0 1 2 3 4 5 7 dias t 1 /2 ( m in u to s )

Controle 1µg 5µg

0 1 2 3 4 14 dias

Controle 1µg 5µg

0 1 2 3 4 21dias t 1 /2 ( m in u to s )

Controle 1µg 5µg

(95)

947 5 & #$ ! A ! ! % ? !

' & ! ! J ! & &

94743 5 " 9Q

* ! ;@ % )

9 ;A i B < @h=" ' % H

! B # F ) G

) " ' % ;> "

8" 37Q ,% B < @= !

" # < lP =

\ < = H < =N 9 ;AN 9 >9N ; 9 Ai

@ " ! $ ;@ " & !

'5*/' & 55, J L <_= % "

Controle 0,15µg 0,30µg 1µg 5µg 0.0

2.5 5.0 7.5 10.0

*

t

1 /2

(

m

in

u

to

s

(96)

& % ;> %

! ; 9 A 9i B " '

% H "

' % ;@ !

M F

# ! H

"

! #

B % ! " *

A9 ;99i ! ;@

" ' ! 8; 8P

(97)

7 14 21 28 60 70 80 90 100 110

5

0 µ

g

100 µ

g

Controle

*

Período (dias)

a ti v id a d e l ít ic a ( % )

8" 39 Q ,% B < @= %

! "

F \ " * %

H < = A9 ;99 i @ ! W ;@ 8; 8P "

& ! '5*/' & 55, J

L " ' ! # %

(98)

9474( 5 Q G 7H

' h %

F H ! ) "

' ! > W ;@ <% ;A=" * ! ;@ %

M J !

B % J ! "

* > 9 ;Aµ h J

! # W %

% " '

! W % )

% %H "

' % ;: ; A9µ "

/ % G H

! % A9µ "

& A9µ G \ ;9

< F B =" , )

! ' &

G \ H

(99)

0 1 2 3 4 5

Controle 0,15µg

A t 1 /2 ( m in u to s ) 0 1 2 3 4 5 *

Controle 0,15µg B t 1 /2 ( m in u to s ) 0 1 2 3 4 5

Controle 0,15µg C t 1 /2 ( m in u to s )

8" 3< Q ,% < >= !

" * \ < =

H < = 9 ;Ai > > <'=N W

<1= ;@ < =" ' ) F #

B " & ! 4

(100)

1µg 10µg 50µg 0.0

2.5 5.0 7.5

t

1

/2

(

m

in

u

to

s

)

Controle

8" 3* Q ,% < >= !

" * \ < =

H < = ; ;9 A9 i > W " '

) F # B

" & ! '5*/' & 55,

J L ) % % 4

(101)

94747 5 ! G 7H

' h

! # ! A9 i F

H <8Ai =

< ;W=" % ! ;8 "

* % \ G

% ! >" 0.0 2.5 5.0 7.5 10.0 * t 1 /2 ( m in u to s )

Controle 25µg

8" 36 5 ,% < >= !

" # < l P =

\ < = 4 < =

8Ai > ;8 " &

(102)

949 5 & #$ ! J& = A" ! A

!

) ? < > @= H !

! # 1 % J "

94943 5 ! ! A ! G H

1 G A:k %

) ! H < ;P=" * G

>9 ) B :A]

>9] :9 G " * G !

] ) # ! M G #

B G " #

% 1 % ;P9

" ' G %

% 1" !G G A:k ;P9 %

% 1 ) % " F J

% 1$ 9 ;>> µ Eµ0 B G N 9 8;9 µ Eµ0

M G N 9 >99 µ Eµ0 G

(103)

8" 3C Q # !

G S V A:k "

# ! "

0 30 60 90 120 150 180 210 240 0

25 50 75

Tempo (segundos)

%

d

e

h

e

m

ó

lis

(104)

8" 3BQ ' H !

G A:k " * $ 1 ! G A:k

% J % 1 ) % <

% =" 1 Q ! G A:k

1" ; Q ;99] ) < H =" 8 Q !

G A:k "

& % ;O !

G A:k # % 1 ) %

G 9 8;9 µ Eµ0 ! ) M

G " ' % G G A:k ;P9

% 1" & S V

% G ;P9 1

F " 0 25 50 75 100 Fator B RB Controle 1

µg/µL µg/µL µg/µL

0,133 0,21 0,30

(105)

9494( 5 & #$ ! J& = A" ! A

* ! 1 % !

% % %

>"

* !

! W < 89=" ,

% % 1 J "

* ! ;8 G

<% 8;= # ! #

% 1" , % %

! G % F

(106)

0 10 20 30 40 *

% d e l is e

8" ()Q ,% < >= ! # %

1" # )

1 H < =N ; 9 ;9 A9 i

> " ! $ W " & !

'5*/' & 55, J L <_= % "

0 25 50 75

*

Controle 25µg

%

l

is

e

8" (3 5 ,% < >= ! #

% 1" #

) 1 4

< = 8Ai > ;8 " &

(107)

1 0 , , 7

943 5 & #$ ! N ! = " !" ! G H

94343 5 ! @ #$ ! J ! ! " @ #$

! F G F

% F 1 A]

% % % <% 88=" &

F L #

1 G ! " * !

% ) H G G M

(108)

4 5 6 7 0 50 100 150 200

Período de imunização (dias)

P F C ( 1 0

6 c

é lu la s /m L )

8" (( 5L # < = F "

' # % @ W F

1 A]" ' # %

! F " < l> E ! ="

9434( 5 F & #$ ! "

%"

D %

) ? " 5 <% 8>

G ;= ) ? % W

% F G %

) " 5 <% 8> G 8=

% > ;8 F

(109)

8" (7 / & B

) " * G L

F "

* # B

G ; ) ? W

% % > <

=" ' % %

G 8 >"

' % 8@ I

# F > < =" *

1 7 8 12

PFC Imunização

1 7 8 12

PFC Imunização

Tratamento

Dia 1 7 8 12

PFC Imunização

1 7 8 12

PFC Imunização

Tratamento Tratamento Dia

Dia 1 7 8 12

PFC Imunização

1 7 8 12

PFC Imunização

Tratamento

Dia 1 7 8 12

PFC Imunização

1 7 8 12

PFC Imunização Tratamento Tratamento Dia Dia

Esquema 1:

Esquema 2:

1 8 12

PFC Imunização

Tratamento

Dia ₁ 8 12

(110)

G

L # 1 "

' L # %

> < A=" / % G W; PW]

% "

' : L

# % "

8" (9 5 % H% I <8 ABW A =

#

" , % 8 A";9A _# _{E 0}

8Ai ;8 F

1 " ' " 1 Q "

(111)

' < H # L "

U" !"#$ ! N

!

N !

F & ! G >'H

N " GWH

>8 8> W; PW]

< = 8Ai ;8 F 1

"

< = L G %

' )* 55L # "

=! ! N ! G X ! & HY

; 8O 99 o; @; 9

8 8W 99o> A> 9

> >; A9oO ;O WP 99o@ 8@

@ 9 9

Q > F

<_= # # I "

,B F 8 A";9A _# _{E 0"}

, % Ak F "

(112)

94( 5 #$ ! J& = ! /

/ F ) ) ?

! # ( ( 4 F % F

1 % ,0( '"

94(43 5 & #$ ! J& = ! 8P

* > F

B % G M

( ,0( '" 5 % % ! # (

1 B F F N

"

94(4( 5 & #$ ! J& = ! 8

! # ( 4 ,0( ' % %

J G F

H " ' % 8A S V

F 1 F " ' J %

H % F " F S V

(113)

* > ! #

1 <% 8: 8W=" 3 % 8A 8: G

( 4 1 % !

!

F "

10 100 1000 10000

0.00 0.25 0.50 0.75

recíproco da diluiçao do soro

D

.O

.

(4

0

5

n

m

)

8" (< ( 4 1 ,0( '" ! % ) < )

I S V " !

<

),

I S V

(114)

10

100 1000

10000

0.00

0.25

0.50

0.75

Controle 25µg

recíproco da diluição do soro

D

.O

.

4

0

5

n

m

8" (* 5,% < >= ! #

( 4 1 " # I

% J 4

< = 8Ai > <

)

;8 lP"

* % M ! # ( 4 1 %

I

% 8W" / % G !

(115)

0.0 0.1 0.2 0.3

*

Controle 25µg

E

L

IS

A

(

u

n

id

a

d

e

s

)

8" (6 5,% < >= ! #

( 4 1 "

I ,0( ' L

J % " 4 < =

8Ai > ;8 " & 4

(116)

(117)

< 5

* D ! J

! % J %

) *

) % F @h < @=

h < >= < >=" ' !

H E N ! # %

1N L # ! ( 4

F "

* B )

F @ > %

I <3 ,( *+ N 2'*N ( 4'(0 1,( ( 899@N a'4'&'

" 899@N 4' '5 " 899;=" , G

G J %H %

G

H D H " J D

# G D E

% G "

G G %

H %

(118)

F ) F

H ! % ) "

5 )

< @h h = I F %

%H H ! " ) F A9µ

@h < ,,5 '5N &*5 ;OOA=" '

9";Aµ E 899 M F )

W9U <A9µ E =" ' > % ) "

G ) % G )H )

I F G ) ? A ;A F

G < a *5N 2'00 ;OO:=" 5 )

I F ) ? G 2

< 2 ) ? ;8 )

G G 2 ) = !

) ? < a *5N 2'00 ;OO:N ,,5 '5N &*5

;OOA=" ' % F G G DH

J ! ) G

) ) "

* @h % F

) ) " *

! # 2 ! @

(119)

! # @

2 ) % <0' ,5N (5 1' ;OO>N a *5N

2'00 ;OO:=" , ) 2 H ! M F #

H <0( 0, 899:=" ' ! G

@ # ! > G

"

) ? " B % J @ G F H >

# " , J )H B @ # ! >

F F > G

% <0' ,5N (5 1' ;OO>=" ,

@ > @h"

& B ! 2 @

! > ) ?

! "

G ! ) ?

% < >= % F h " * h

% F %

" * %

% G M ) @h

# "

F % <0 % h="

(120)

L H " ' %H L G

J G G " ' %

4 " ' 4 #

F H L %

L ! " F # F G

F % G %

% # <'5 ,0N * */( 2N '00,5

K " 899;=" ' > G

) !

" * 4

F "

* > !

B ! ! 2 @ !

>" * B ! @ M @ >

! ! H <0' ,5N (5 1' ;OO>="

' ! J %

) ? % % #

! H E " 5

J ! H E %

) ? " 4 J

(121)

O I <*0,, [a " ;OP:N 5'['(

" ;OO8N '5&*5 ;OO@=" # G O #

# H " " <899A= %

& F

) F )

% G B ) ? ! 410

4' 8 " 5 )

% J ! H E

B F " %

) & <8998= % G ) F

)H ! H

! "

' ! % !

) "

* G @h

; i ! # >" &

<9 ;A 9 >9 i = J G G

H <; A i = ! ! !

;@ " 5 W ;@ 8; 8P A9

;99 i B % !

;@ " ' % B %

(122)

! ! " (

# % ) F <& ' , 8998="

G % @

B J # " & % F G J

@h G

! M J ! @"

* #

! ! " * G

F > <9 ;Ai = !

W G % %

! <;@ =" & >

! W ! " * %

! G G F @"

% > ) ? %

F H % @" ' J

H ! "

, ! )

J ! # ) ? )

J % ! " % G

%

(123)

% G J ! G

F ! "

& M J ! %

1 % " , ) % G %

1 # F "

H ! F %

% 1" %

S V G

1 < ' 0 ;OOO=" * 1 % G

A:k 1" '

% % 1 ) %

1 ) ! " H

! D % % % 1 %

1 <2' ( *5N 0' 24'55 ;OP:= ' H

! ! #

% % 1 " , ! % !

) ? " ,

! " , # !

% # F G G

! % 1 ) H ! " ,

J

(124)

% DH H

# 1" ( !

) J

" * >

1 " / % G

! ! # % 1 F L

% "

, F % % G

> ! &5' G

G H <[,' N

'a0* ;OP;=" , ) F

) <& ' , " 899>= % G )

F 1

) ? " , %

) F " , F H

B % G >

% % 1 !%

<[,' N 'a , ;OP9=" [ " <;OPO= F 2

% I " ' R

(125)

% 1 G F 2"

2 <;OPA= % G D ) %

F @ D

) ? " *

G

% " * G

J )

"

/ p <899;= % G ) ) #

% ) G G )H

! ) ? " ,

G % %

" ' % G F @

) F " ,

> @

! F "

& M

H F # ,0( ' ( 4 1

" * ,0( ' %

" > ;8

( 4 1 G %

(126)

G

! % 1 ) # " ' #

% G )

! # 1 % F" ,

% !

% ! # %

(127)

(128)

* 5 R

• * > @

) G !

% !

H E " ' ! #

) ? ! "

• * ) ! #

% % 1"

• ' F 1

1 G %

) ? ) "

• * G J ! # ) ?

%

# G ) ? %

(129)

(130)

C4

'203/( K" ) )T $ " ! 8 >

) "89: " :@ W; ;OW:"

'04,(&' K" *"N ( , Kq5(* ,"2" 3 T F

% B ) % % % +"

4 4 - % 84 4 "' " *B% " >;O O8 ;OW:"

'5&, *5 4"N '[[* 2, 1"N , 0( ("N '00,5 2","N 1,5*( 1"

) R ) %

% T" " A % Z ! - % 8 @ "P> "W "

A;P 8A 899A"

'5 ,0 2" "N * */( 2 5" "N '00,5 K " 0" /" = ,

[" \ ! ' #$ ! I " : " ,

" 899;"

' Y' 4"'"N 0 [, a"&"N '0' ' ["N '1* 2' 2,4 ,"4"N ,0 ['55( 2a

4"2"N 1 4'5 ["&" ) % % )T )T B )T

) F )T 42 (( B " " 81

! " % 81 5 a U " W: "8 "89O 8;> ;OOA"

1' ' "1"N /,0' * 0" " "N , '52' " "N '4 * '"4"N 4* ' "5"N &(' "4" "N *0([' */ ("N (1,( * " "K"N (4,*5( 0"'"N 5,/, "'" "

4 ) ? " U" & ' !

! 8 ' 1 "@P "; "8A >O 899>"

1' (5 *5 "N +2'5 4"N ( 2, 4"N ' *00 4" " ) %

% T" " 8 & 2

) " ;P9 "A ;A 899;"

1, ,&*/ [a 2"N * [(5 ," ) ) )

" F 81 ! ! 5 R a U" " ;A9 " @::

(131)

1,/, 0,a " ) % > ) % T ) T " "

0 " >9@ "@ " >OP O ;OP>"

1('5 * '" " ' I " ( $ '( , 4"4"N 8 [

":O: W9A ;OO;"

10'0* [ K","N K*25 *5 2"4" 4( 2 ,"4" * , 1"'" , ) % )

T T )T " % ! ' %1 %

" 5 a U " ;8A "; " >9 @ ;OP@"

1*2'5' [' 2 '5 *"N +( * ,5 0"N &*5(5 5"N [ ' "N ( 2,0 *5 +"

T " " " 81 *B% "@; "

AP> OW 899@"

1* &, K" 0 )# T G B )# #

T T G " 4 " 4 " " " W " 8AW O: ;O99"

1 '& * & 4"4" ' ) % ) G % %

G % F ) % T " 4 % 4

5 a U "W8 "W "8@P A@ ;OW:"

' *0 4" " ) T 1 " "

" 81 *B% " @; " ;@; @: 899@"

' *0 4" " ) T % T"

" " 81" 5 R a U " A ";9 " OP; :" 899@"

' *0 4" " ) % 1 " !&

" 81 5 R a U "W@ ":; PO 8999"

*0, &" "N 4* '5 "1" 1 T T $ ) R % % "

(132)

*00(5 K"N * 2 " ' T )T " " D " A " ! 2 "8O "8 " @;W @8@ 8998"

* , *"N [' 5( (" ) % %

T" ! 81 % "8 " A>> @8 ;O@9"

&,4 ,a "Y"N '00( *5 4","N '[[' 'K "N **&5*Y " "N ,' *5

&" " > % D $ G

T" Y ) " 8W; " >@P A9 ;OO:"

& ' , " " & ! !

" , & 8 #$ ! ! #$ J& = !

% ? ! ! !"#$ ! 4 8998"

& "

" "

& ' , " "N '++*0(5( '"," " N '5&* 2( '"("'" ,%% % ) %

R ) % T ) ) T 4 4

" % 4>' "> " ;@;O ;@8W 899>"

& ' , " "N '5 * "0"N '++*0(5( '"," " "N '5&* 2( '"("'" ) %%

% ) )T ) ) )R T %

" 4 D4 " % " *B% "88 "8A >> 8999"

, Y'5 " "N 1, '"&" ) % )

% " " 81 *B% " A; " 89W " 89W 8;O ;OP@"

, , '" "N ' 5 ++, "Y"N *40(5 *5 "N ' ' 0"&"N '50,a ["["

2 O$ T T B T" "

" 81 *B% " ">> " WEP " W8A >> ;OO:"

,Y'0& "'"N Y(00('5 K"2"N 1*Y&,5 &"2" ) R "

' % T % ) '

(133)

,' *5 &" " % % " ( $ ' ( '" " < =

( T T " ; "8 ;OP;"

, ('5( /" "N 1' 1* ' K"," &, ' /'02* (" " ) T

)R T % % T$ ) "

! 4 !4 "WO " >88 W ;OO9"

( 2,0 *5 +"N ' '0( "N 4,/* ' 2 &" "

" " 81 *B% " >P " 8 > "89W ;O 899;"

'5[ 4"4N (, 0" " ) % % ) T "

" 81 ! 1 ' " ;8 "O ;OO;"

K( ' " , % ) )R T

T" " & 2 " 81 0 "8 " >@: A> 8998"

'5(5 , Y"N ( ( 2 [" "N ,( , Y"N &, 2 ,"N Y'0&2' 0 Y"["

,%% % )T ) " D " A

! 81 ! ' % " :> "8 " @9W ;; ;OP:"

,,5 '5 " "N &*5 1"K" " ( $ [' + 5

1" " 8 'I J :" " [ "@>O @O ;OOA"

a *5 '" "N 2'00 K"," F ' T A ! %1 814 Y"1

T O" " ;OO: " O@A A:"

2' ( *5 "'"N 0' 24'55 "K" ) T" ( $ - !' T A

F " 81" " >O" Y &"4" , " 1 UR % *B% "; " >O"; @O ;OP:"

2' 4'5 "N Y,, 4'5 '" "N 55 "N (5 , 4"K"N Ya5 * &

2*4' &"N 2'00 "N 4 , * '"4" ) %% % )T )T

B )T ) " ! 81 % " ;;:

(134)

2* 4'5 0" "N 4'a, 4"4" ( ) T % B " ( $

% ! " 81 ! " % 1> Y "'" ) 4"Y"

< = ' ";@9 ;OWW"

(5* , ["N 5(, ,5 5"N 1(, , [, 5" "N 4(0 *5 "N '01(5( 1" %

B )T "

" 81 ! " % 81 5 a U " :: "; "; ;9 ;OO>"

(5 , ( " )T ) $ T T % T T" ! 81

% " ;@: "W " 8P:8 > 899A"

K'5,Y'a " '"N '/, "N Y'0 * 4"N 20*4 2([ 4" " ' 8 ,

" N! ! # " A" " " P" " P$>; P$>@ P$@@ P$@A O$;P 899>"

K*25 *5 ,"*"N ['4(0' ( " "N '0* , * '","N *0& "Y"N 2 * *

" " ,B T )T )T R ) % )

)T ) T B " " D " A ! 81> * "

;A> " ;WW PA 899A"

[,' &"N 'a , &"K" ' T T )T ) "

& ! & " 81, % "@ ">8> >>9 ;OP9"

[,' &"N 'a0* [" ) %% % T ) )T )

% T ) T " 4 F 4 " 4 *B% " @W " 8;W 889 ;OP8"

[0,(5 K" " ( , " 81" 1 UR % 1

" ;PP 88: ;OO9"

[b20 K" ) % " " 8

% 5 a U " >@ "8 ;AW ;W: 899:"

[ , ","N 10'0* [ K"," G % )T )

% T " % ] " A " 81 1 " ;>W ";

(135)

[ , ","N 4( 2 0" "N 2' 1* &"/"N 10'0* [ K"," )T $

% ) " % ] " A " 81

1 " ;@8 "> " W@@ W ;OPO"

0' 24'55 "K"N ' [(5 *5 K"N ,(& [" & $ ) ) % T " " 81 ! 1 ' ";8 "O " >;: 8; ;OO;"

0' ,5 " "N (5 1' "2" ) )T $ ( $ Y " K"&"

Z F ' T A ! 81" Y &"Y" , N P" " Y"1" T ":P8 P;A ;OO>"

0( 0, K"Y" )T " ($ 2T )T " " 8 1>

! > % 81> ! 81> ! ! ! 81 " 0 ";9; "> 8W: P> 899:"

4' '5 " "N (a'&' (5( &"N &2'a'[ 4' " "N ' 5'[' '5 K"N

' 0&2' 4"4" & %% %% % )T )T T

T " ] " A ! 81 ) "8; "@ " 89: ;A

899;"

4' ,00* &"N '5& *5( "N , (&( '"N 0'41 ( K"&" 5

R U T " " 81 * " ;;A " 88A

>A 899A"

4'a, 4"4" % B " ( $ ) " F

" % 14 ' ) ) ;OW;" ) @ ";>> ;@9" ;OW;"

4,/* ' 2 &" " ( $ 2, 4'55 4"N ['0&,5 " K"

! " " 1$ % " " Y T / 2

Y ) " AW WP 899>"

4* '5 1" "N 4' 21'5[ ["K"N 0*5 2( 4" "N 2' ( "0"N '00(4* ,

'"4" $ T "

(136)

4* 0 [a 2"K" ' TF & R ) ) % R ( " & ' RRR" ) " ;OOO"

5'['( 2"N 2( '['Y' "N 2'a'['Y' 5"N '4'5* "N * ' +" , )

B % T )T ) % d

" ! T 1 *U T "@: "A " >8> >9 ;OO8"

5 ,5+Y,( /"N 1('5 * "N & [* ","N ,&,5 '" % > 1

T ) T $ ) " ! 5 R a U"

;OW;"

*[* (,4, *" ,"N ' [, '" 1"N 4 00* 2 1"N 4* '5 1" "N

( 2' & "K"N 0'+' K"2" )T "

% ^" 1 ] " A ! *B% "OA ";W> ;WO 8998"

*0,, [a &"'"N ' '5' 2'(a'/*5 "N 0 & ' , 4"N 4* '5 1" "N

'4 1,00 '" ["N 4 , * '" 4" O

d " ! 81 *B% "8A ": " :8> >8 ;OP:"

* 0, '" " ( " ( $ * 0, '" "N Y,( 0" $ )

% " 5 R a U 2 ;OW: " ; A"

' '4( 2'(0 4"N (, ,+ "N ,410(5 "N K*25 *5 " N 2*01* *Y

,"K" N , a 4"1" % F % (

" ! & ] " A " 81 *B% " "@ " >@> ;OWA"

' ( 4"N ,(5 ,(0,(5 K"N +'['5K' 4" N 4 [,5+(, K"4 N ,, K" N

0, 2, 4"'" % ) U T T

B )T ) " % D " A & 8 5 R 2 "WO "

@9@ P ;OPW"

' 4,5 (, 2"["N 1',04'5 "N 5(, Y0'5& 4" "1" N &* 5a " N

(137)

% ) U T % T ) "

1 " 81 ! " % 81 ' " O9 "O; ;99 8998"

' 0 Y"," " ( $ " ! " 81" 0 Q

) " @" " " 8O ;OOO " O:W OOA"

, a 4"1" % % ) " " 2

' 81>0 " 8>W " ;AW O ;OW8"

(00,4, 0"N 10 4 0"N 0, *Y ("2"N * *"'"N *&& ,"Y"N Y' &0'Y '" "

) T T$ & % R

) " Y ) " ;89

" 8WO PA ;OA@"

*02(00 "1"N ( ["4"N K*25 *5 K "1" %

)R T ) " ,B

) T " % D " A " 81 1 ";8; "; ">:> W9

;OWP"

3 ,( *+ 4" "N 2'* a"N ( 4'(0 1,( ( " ,%% % )T )

> 5' B ) U "

%1 ! 5 a U ";@ "> ";WW PA" 899@"

,(& ["1"4"N * , " " ) T T % "

" & 2 A ' % 1 ' " A9 " @>> :@ ;OP;"

( [, " " % T ) T T > ) 1

B" " " 81 % " ;W " 8>W

8@> 899A"

(( '"0"N 2'5 ,5 "["N 2(,0 "N '/2*0 "2"N K,&&, "N * 4 ,5 0" "N K* ,5 ,5 K"*"N Y,,[, K"N 4*00, 5" )T )