Embed Size (px)
Citation preview
//
ftNftLVSIS OF ELECTROPHORESIS PERFORMANCE
Final ReportRAI-84-E-3
Period Covered* 11/27/83 - 4/6/84Contract: NAS8-35912
Prepared fort
George C. Marshall Space Flight Center
flSFC, AL 35813
Prepared by*
Glyn 0. Roberts
Date: 4/6/84
Roberts Associates/ Incorporated1726 Pine Ualley Drive
Vienna* UA
https://ntrs.nasa.gov/search.jsp?R=19840016648 2018-08-12T06:07:53+00:00Z
ABSTRACT
Our computer code SAMPLE models electrophoresis separation
in a wide range of conditions. Applications include steady three-
dimensional continuous-flow electrophoresis (CFE), time-dependent
gel and acetate film experiments in one or two dimensions and
isoelectric focusing in one dimension. Sample results for all
these are included.
The code evolves N two-dimensional radical concentration
distributions in time, or distance down a CFE chamber. For each
time or distance increment, there are six stages,^successively
obtaining the pH distribution, the corresponding degrees of ion-
ization for each radical, the conductivity, the electric field
and current distribution, and the flux components in each direc-
tion for each separate radical. The final stage is to update the
radical concentrations.
The model formulation for ion motion in an electric field
ignores activity effects, and is valid only for low concentrations;
for larger concentrations the conductivity is therefore also in-
valid.
We have duplicated one-dimensional results obtained with
a similar model by Bier et al.
We have successfully modeled eight experiments done with
hemoglobin in a barbitol buffer on acetate film.
(i)
TABLE OF CONTENTS
Abstract (i)
Table of Contents (ii)
Section 1. Introduction 1-1
2. Description 2-1
3. Results 3-1
Appendix. Graphs 1-47 A-l
Section 1
INTRODUCTION
This report is brief, and references the numbered view graphs
in the Appendix.
Graphs 1-6 are an introduction to the presentation and to
electrophoretic separation processes, and are not further described.
Section 2 and graphs 7-22 are a description of the model
formulation, assumptions, and applications, and of the numerical
methods, coding and use.
Section 3 and graphs 23 - 46 present results of analytic
validations, acetate film experiment simulations, continuous flow
electrophoresis simulations, and simulations of electrophoresis
cases modeled by Bier et al.
This is a continuing program. Some plans for future work
are presented in graph 47.
1-1
Section 2
DESCRIPTION
Graph 7 is a code summary, listing the six stages in updating
N two-dimensional radical concentration distributions* and describing
some available applications of the code in its present form.
Graphs 8 and 9 describe the ionization model and assumptions.
All salts are fully ionized. The ionization equilibrium of each rad-
ical (or ampholyte, including proteins and cells) is a function only
of pH. Four distinct ionization models are used. The coefficients
are determined to fit titration measurements of the mean ionization.
Graph 10 presents the mean ionization for three hemoglobin types;
the root mean square difference from measurements is of order 0.1,
which,is excellent for magnitudes of order 20.
Graph 11 describes the calculation of the pH distribution
from local charge neutrality. Newtons iteration gives very high
efficiency and accuracy. .
Graph 12 describes the model for ion motion and radical flux,_
and for the resulting changes in radical concentration. The radical
flux is obtained by summing over the degrees of ionization, and has
terms representing transport by the mean flow, transport proportional
to the local electric field, and a diffusive flux.
The electric field distribution is calculated from charge
continuity, the computed total conductivity distribution, and the
computed ion diffusion current potential, as described in Graph 13.
Graph 14 discusses an improved model for ion motion, including
activity effects of varying concentration. This model has not yet
been finalised or implemented, but is a requirement for quantitative
agreement with measurements, except at the lowest concentrations.
Applications to continuous flow electrophoresis (CFE), moving-
wall CFE, and to time-dependent one and two-dimensional cases are
described in Graphs 15 through 19, with the corresponding requirements
for initial conditions, computational domain and boundary conditions.
2-1
Output options are listed in Graph 21.
Graph 22 show the input data, and demonstrates the code's
flexibility. This data file can be modified in a few moments to
make appropriate changes in the problem, the method or the-output.
This data is for a 3-radical computation with hemoglobin in a
barbitol buffer.
2-2
Section 3
RESULTS
Analytic code validations are discussed in Graph 23.
In certain simple cases and for some components of the problem,
analytic solutions and simple measurements are available. The
success of the code in duplicating these encourages us in app-
lying it to much more complicated situations. Graphs 24 and
25 are contour plots showing how a weak circular hemoglobin
sample preserves its shape after 40 seconds of advection in
a uniform barbitol buffer with pH 8.5, with a field of 100
V/cm. This is only one of the wide range of validations we
have performed.
Graphs 26 through 36 describe our successful simulations
of eight heraoglobin-barbitol experiments with.:a circular sample
in acetate film, performed in May i1983. Graph 27 is a poor
reproduction of.some of the original films, with the hemoglo-
bin "fixed" by standard processing. Graphs 28 through 36
(including some plots without graph numbers) are concerned
with one case, with sample 8.8% hemoglobin in a 2.5X MACDAC! • - - • • • • " .
barbitol solution, and with a 0.75X buffer. The mean field
is 12 V/cm to the left, and the computation went to 360 sec-
onds. Graphs 31 to 3 show successive center-line concentrations
of hemoglobin, barbiturate sodium; the hemoglobin spreads to
the right with a low constant concentration of approximately
circular shape, while the sodium arid >batbittrEate build up >on
the left and are depleted on the right. Graph 34 shows the
successive conductivity profiles on the center-line. The
ratio of maximum to minimum increases from 2 to 50; the max-
imum moves to the left and a minimum forms to the right.
3-1
The following plots show the successive pH profiles, the elec-
tric field along the film (varying from 2.8 to 25 compared
with a mean of 12 volts/cm), the current lines, and the trans-
verse electric field, with peak 6 V/cm, which spreads the
hemoglobin sample.
Graphs 35" through 39 are contour plots of continuous
flow electrophoresis (CFE) tests, with a weak circular hemoglo-
bin sample so that the conductivity and field remained effec-
tively uniform. The tests include a moving wall case where the
sample remains circular and CFE with electroosmosis respectively
zero, negative and positive. The distortions due to nonuni-
forra flow down the chamber and to positive electroosmosis approx-i
imately cancel in Graph 39, as predicted by theory.
Finally our successful duplication of the seven one-
dimensional computation of Bier et al. is described and demon-
strated in Graphs 40 through 46. ,
3-2
APPENDIX
Computer-generated viewgraphs numbered from 1 through 47 are
presented on the following pages.
A-l
(AHaUJ
>j£
00o»
3-a«3QC
I"
aui
o
o
O
S u
1 §*• o
OLO
oUJ
UJ
en
UJX
£I
hi
(AZO
t-o
(A Q.Z Q.O «1-4(- OQ. V)Z
3LJ J
<r <rzujQ.<IX
Z <E Ooov>oi-i zo:I-OOI-
J<TI-UJEUJQUJ
oo3zu.o<xoLJ UI UI
t-U>(Ali!HH
LUOOO.O
00 Io
ZO
oo
(L>UJEorcn3<r 3(AO(AZOCC3Z
0<C
Ifi XZ^JI-o<ruil-i-<tt-t J3Z<X0»-10UI •-!
-COUJZZE003
(0 ^
(AUI
o,(AQ.1-0zo:UJt-
oo: ji-l UI UII-Q. (A<IX3UIO U J O U >
JEU.O<r j^HHtAQ.
U.33O OO
UII-l
oHUlZl-UJ<T
' I-3MZitLE Jt-«
UJQ:Zl-
ou.<r o(A
•-1I-IOH3 • •O
<A~Ji-i-a:
3<r <co»
o:•
(A a a.z uioo a. aHOUQ.a. i-i ao*-xCCHU.
Q-E (Af-o:zo3OOIH
t-<LUUIOZ
000:0Qiauja.a.t-o
a.•
tni-QLUJCOo
<r
00
<traOf
aui
uio
a
OZi-t_1
u UJOoo
s u
.oUJjUJ
rt
UJN
o >*•
S"
n
io
orj'Q
-O
UJU.O
LUC£OXa.o
oUJ
cU.
wcz:1-1-z.o
ftfr
0UJU.
l-OUJtoentooa:o
UJ
O»-ia:t-oUlJUJ
oooooooooooooooooooooo
aoa.
oUl
vft
Xoa.
o
oUl
UJ
oOf.
o
00o>
tt<ca«
o
E OE O•~| CA
' 8 3u £
Is£ Ul
I ouUJ
enr-UJ
paN
I
O
o-o
ce
UJ
Q.I- EM <C
<A
KO
Ul
Q:<EQ.
U) U) 0.HH <rOU) JE o (A »JV) W>3 <LOUJM 3
O O«3Oo: tto-zHO XM
Ul 030
OZXQJMOHUJJ Cv^ •• ^1"
P"* f* 4&B
(/>MO,J
Z(AHK
ounnoZQ^QCUJOO Qi«i-t O LU
UJ0)
Ul
OH
OHOZO
UJUJ
U. J
<zz
o<x>^J COUJZMOZU.H4O
HI-HOOH
oercoo
JOQUJCK
I>ZMO
3<ICL
ZHOOZ3OUIO
oo:EZXaoHOOU. (A
zooi o:
OM(AZ UJ
LUXUI
<r3 QiCOO
U.Q
a:HC3ZZUJ t-t
ZUiZ<xxo
^^ - B^
MEUI(AXEUIOMO OH
O 3ZOt-ttQ.EZ UIX3<EU«O^J rDE3
OQ. Xtt<X ««OX
C3XZ HZ(AO_1o »-«_nn< T U J O E O3E^ Of
U.OO UJQQi
E»UI
M o
1 1 1 1OZ I 1 :.l :l
zoocnoZ I I I I
zo«
<r oo
<rzuiEQ£3tt
OOOH
UIQf^J<CQ.UI
(AZUI
-JHE
OQ(A
Q.UJZ
>U.<C
E I I
tni-Q£UJ03Oo:z,jC5
-
«
CO0)•»H
%o*"*J<caM
_ LJH U»<r
C5Z
o- •"•
i |s»- £ °a °•* (A-i j. UJ
tft *^o Ul g:
8 8 «<" g tn
3 o u1 tt H1 o
Ul
UJ
•10N
I :
RA
I/E
S7
3
a.^^ -zST.8" ' ' -
Lf>
1
OD
UC
TIO
N
oQi
H" Z!|»H
O
Q£y*
1||
U.
UJ
<rh-LJO
H
13
yh-
UJ
£Q_XUJ
<c ~UJ Jz <tM O>J frH
0.x •>1- H
3<n ^UJ (/)
- J UJ(DO. -J
UlOil *~^"McnUJod0
Q_OcroUJ
UJ
«-l<cr .0M</iZU.E:»-icA
|iW)
h-» E Q.
Ul V) <Ca: en
CL j <roa zao 1-1f— QC U.OMUIO
A J* . > ^^
o A ^
!.
r«-*
Q9
JLUl
•^«^
LU .1. .«- " . - '
»J « 3UJ Z( H O *U, • IH W)
cnuO Z»HM LUQ
O Q V)UJ |-J UIUIUl ZU.
OO
A oca:ooU.U,
OH- UlUl IH ,Jz a.oo s:IH UJ ^V) (f) Cftiii 3o -J
u <c<E<E 1-
EW sUJ=3HV) •
! > (f)^ cnuj
CDOL«_ ^s
•
*
; • • .,'j/'-
*" - *'..('•'
-' ' "•".. . .
'.- /-;/.-..w '
t-tn
COI-ctHIee»oOf
o
COO)
01<r
OZ
Q LU0 O °«- E °S °1 « ...
Lu ^
lioUl,JLU
LU\
IoMI-o
- O
o
oo
o»-lo:oUJ
1
Ho
zo»-«t-<to
a.a.<r
<to
LUXOoh-t03
Ul
OOI
ot-HI-
o<r
z<rzoOHH
LJZI--j I-H <r«QN<r rji-tLU_JZEOOazi-iLU »-iCL ZE ^<EM(/)U
us:•H Z t-
3OLJI-XXLUO.I-«E:
<rx»j oLU *H4
<r LUH-IH-
ZCKC»-«E
QTLUuorDa: uZ)X<Cf- PO
<rLU LULUl-l-X> Wl-JO<C(-X ' •<£CLWE LU Q<C XUI
CO CO
Q t-i
cooLu<TLuOH-
COLULUQ:
"
LUt-
(-OCOQiMQ.Q
LULUO
ZLU
LUt-H «.Ht-H-IZr>i-<MOV)CLIDU.ELu=}»-( IHOtt
£Qoct-iLU JOMarj<r o_1LU
r>MQ<r LULUXX(rt
t-n(ft OL U O O.JZli.
OQ>LUZJ
»-t:3 ooQi
LU3H-(ft
LU
V).":>Lu ^-J QC/Jo ozQC Qit-Hh-i-LUo ot-LULUO
OXZOL<I:
.O<E
LU
t- LU LUQ.i-1
LU J ONoJCZUJ
<E LU J> O
w *o *<C u
t
DCUI
Z
*Jo
000>
>0£<t
Q£M
UJ
is*• oM uu) s!£ ix »
- £Q£ £
UJ1-1UJ
(*)(AUJX
~a
o
x°i<ft
oUI=>oUI
UI
<E
COzoHIH=3CO
COCK
ocoz«ci-iO
ecu.t-ozui<roZZ0=3OO
Q
<EUI00
0<r<EH(KCO
»HJO
ZZOi-i
zoUI
U
a:=3.
CO0£
O
E<r
uiH<ri- •UICOout<r ja.ui<ri-x<ru
=30
•> «EZ=3 HIHI COOOO JCOO
OUIECOUI
X>EO
CO «.JUI
»HUI OnO E HiO
OH=9I-Z
H UIX
«f-ctco
<rzO»H
Z<ECLoao.H4 «I-UI=3l->
HIMQiM
•* CO ZUIHIO
Z«-i
t-oo»H COOZUIouiOt-OOQKtCO
<CUIH-i|-Q.Z •UI N O f-CO UI O CO
i-i<c a i- o co ui
<C CO HI 0£ (— UIHO oa:a:co=>ZOZ =30«-i»H
E UIHU I U I E H Q Z Z Xax r>zoo=3Z ZH HHU.
O =3 JUIQiUI
o u. <c z i-i IH z: H<E UIOU.OUJUI Z E O O O
O O X Z H
Of-ZZt t<EOOU. =3 t-* »-i H UI O
COOOO JXCO »-i Z Z UI Z <C H
OHQ.Q.UI t-iE<r co co co cooH H-I UI UI UI UI<C H
Q£Q£H=3 OX X O O JUIMM aooaiu.x,-ico o t-a.
UIUIUIU.UI EU I X X X XU1H4
<L => <CZHHHJHQ«ui ui ui ui o ui a.aooocoo=3
xo«e«o©
zo«-(
0.oz<r
uizooUICO
z<EO
COUICO<ro
<rzoI-ICOzUI
oUI
o
UIM
0=3•* •* xo
COCO O.OHH OU.zz aUIUI HO
CO* MM UIQCz »<KQ: JHo co uiui «.uio1-iZQ.Q.CO UIH O X X M > , J=3 M UI UI CO QC UI_1F- UI<IO
O»-«»HX=3UICOU.U.O.O J
***lt C^ CO CCOO UIUI UIX Z
O ,-> O<E <X <C Z MM J HHHHHCO
I <r<KCt-HEEOO3 »-i
1-1 <j <r <i <r <r iU I = > Z Z Z Z Z U IU . O M O O O O Z
ococococoJJEZZZZH<x<r uiuiuiuizZZ J E E E E U IOO<Ei-ii-i>-i>-iOCHi-iZOOOOZCO COO I I I I UI
• •ZZMOUIUIUIQ.UIUIUICOI3ZZZUIO E E Z H O O O O=)tHHUI I
O IZUiM UIUI
I HIZZZZEUIUI UI UI HI
I O O O O H
coxZHo ,H>
<c<rOUJHlHJCO
Q.O<r
X 3 UI UI UI UI QiHHQ.Q.Q.Q.O
UIUI UIUI>>oooo>00 I I I I O<r <i ui ui ui ui <rU I U I E E E E U I(- H HI 1-4 H4 M HC O C O H H H H C O
ooooeoe
COHLUtt0K;
z•*J
O
uu"
00
*H
*^u
3
ttLJ i
i
O
a£ t*^
1 g LU
" i §x Oo "j CO ^ju. HI .
ftRS
HA
LL
SP
AC
E
TR
OP
HO
RE
S
THE
S
AM
P1
LU»JLU
COr-coUJ
- M
I.Ko
00
1
H>
OL
2
i°llJ
i°
OzI-H • \±
o oz
5
09
daUi
J
•J*LU CO • • • COQ. Z •LU OO » t-t •
Q HZ ~ LU <CO LU N ft: -HI J M H
<r <L o LU +N J M O + (UHI »-t Z <t XZ « >• 00 => J 0
Z3 LjJ ** *u. co u. r>o ,j «co
LU LU XOLU on i-t +
,; LU- o COH <t xSit ft: Z E O LU CI*JLr • ! O O t— I— ttJg ; UJHi . Z >O QH- X 0 0 JLUE <C 0. HI LU OX -
<zoa i i- E xi-Z t- <S> <t 3 0.O L U Z < r 4 N C O E Z»-l I>LU H i C O <THi <E «i_ <ro • z <r<r ^z o QZS 0 X Hi LU ZOrjj • ZO oc d HI — •5 Q O K « 1 -Z LU HiZ 3 • *<CO N 1- O O COLU CON 1 «-iM HI 3Hi Lu LULU LUHH <L V
z -J coo: cozo oz t- cro <roHI COLU Z «LU COM
O LU Q — •> COO CO Q VLuJ 3 Di LU Z O a O | (U• - 1 O O t t T <ELU L U | V
;.< 3 LU ^ Q. CO 3 LU d *^: LU 3x coo LU v
o f- on. %aLU d LU <C HIE COOQi X OHI QLU *•<r Zf- LU <r HIQ
HI Of Z O •• .. • • .r „ CO Z <t O « <t X
>••!- COO Z O: _i i-i LU o o v «: •<t c: > a: at- a: LUCO 0>J LU f- LU
, HIZ X CO 3
• • Hz «.
• - • LUH-
S5 ->OCH- X3CO -9COZ 0
ai 01 <ro \JO X LUO v*
<* CXJ E S0 0 L U
EZ -0OO X
• •» QJ •>Lu * 0
_1 (U
*1 •»-! ZO I/J0 X LU y
, -rn :>co •IO f* ' ^
O ELU
« ** LjJ Q£QiLU<xx «.
^ Q XQHI r>ZO 0
* •• c; <c \^ i1 OV XX <£
X *-t COLU -^1 1-3 X
0 Z -»<CI- 0HZ . 7
ru COLU ]/J.1 ZJ K^
(U X O<E IX (U OZ>
I 0 E0 LUZ
XO- HE
• • LUC: z« • 0£ LU
LUQ: x• • x o H r
3 Lu
J 00 (-J ^ • • • •<r ft: «-i ni
•*•to
<A O>1- «40£ »UJ ^« *-•oQ£ >ocz <r> 3.-) DC
.UJc £U-i i£ O
OZ
OJ _1
£ « ui0 o 2«- E °5 o_> yj uj
CA ""o UJ °-<E QC E
ft 1 S»J M
5~ — 1 •!<I ^3 Sr
5 cc fcEi o
UJ
O)
1n i& oy*i •-•^J a__V *"^
-M °-Z >v ^^
1 iw« .UJ
1
•»
y
1n»UJ
<r tn t»-l HI *
id >CO |* ^
1 (OV) V) J
® (— UJ UJid z => o
Ul HI 0HI » C3 E
. .••* o o<cQZ<C f
X
H nil- «) Q£u.<r M 3
N U.O X O•H <&• UJ I- U.
O V) A «~ OZ<r uj E x *H oo - 1-1UJ E
*M
C•H
. coc:
1
^M1,ugjG
ZOh-»t-<c^_^"
cH1
f~* HI 4XUJ X HOC
•H A M UJ Ja E *^ C*) l~ OL • ^^ O O) ^4 ^ ^ O
!-:> -f O C Oi 0 N I-T-I Q£HKC -o uju. rf U. ' UJ HI • O3x ^ Q: i xz z® u.
^ o w x , > K- o M^A ^^ ^v £• 2*1
1— UJ ^ Q£ Hi '>3C \j.J Oh- -H_j ^« /N <Q ^3
^5 ^^ ^& ^Jx i cnr> +Q.UJ EEX 0.0: xI ii '
OU. •+• UJ 1Z O "^ N O -<r o HIHI
e- ^ ZH
-1 M 09 *
! • - . - I-H uiz ^<r x•HO O C O X "O
ja o u ot- z xE^ OE E O E
+ V) TH Z U J t A H l T3^3 <EQ XW 1-
X (A^ U U <t XI UJ » Of UI.J Ni »r> ft & "•» TT i—i 4-Ml v * «J «» ^1 ~
S «^ ^ - ( A O U - Z- UJ I C O O OJ
a * u tr HI E• H O E O Q £
«Z X O> Id UJE Z U.O U. •cnui x H; I .Snz W zz ^<C UJ * •'X 4C«Q_ O
UJ E UJ
<CZ Z3UJHI HI3 UJUJ
. 00 0 QC 0V.H o^ H tnoc EN U. UJ (-LU UJ
E (At- Z UJ(A 3Ht H»U GCUJ U. OQC O
| « . ( A U J O L < £ U I
uj u <ro® DC UJ QC 1- (A UJ
Z h-Oi Jd "0 O<E <E DC*o oo
(AHi CKU.Z M3 QZ <LHI i-o <r 3
Qf- X UJ a Z O V) CD U OHI Q£ON XUJ
<C -r^E V)X OCI-
<c HI uj x E • - x o. Q. tn xi- zZ rJH
^ O Q-<THi EUJUJ 033 .O
• • •."n •«•
i c- <n f-o <E; /;,Hi<A ZUJ HI O UJ
3 <C HI J 3 QC E
UJ• • x
1™ •
-•• ,.
to
UJCftOK
Z^»,j<9
jjJ~
00O)
*4
^»o:3o:A
-uSu.ib4
C9
|v IH£ ,J5r UJ . .••• itf
* 2 1r O"* (A— j .. ujj
3s 1
g UJ &<E QJ 585 ® $-J Q_
^ QC »^
I OUJ
UJ
(*)^~WJUJX
— IH
o f-«!>•HI
iKO
*^*"*
zov~a.a
2°ZUJgo£
u
iO o
ee>
S 5o iUJ Mo t-r «UJ (Zf. H
Z .• UJto o
•• . i- z • •Z • 0 Q.
^^1-* j*J^cQ^1-
UJ^^
bJv3
<E3C0g*O1.\ .
••M
O1-4t-ZJ«
-•' • HHQ£
U)h"Q
XP
UJ ^ O UZ *i 1-O 1 J tOo. «> aC <H 0 • X HO 0 0 O T3 tOO * ,-J M X Of.
a Z E MtO • UJ UJ O T3 U.1- 1- X 1-M <C 1- O tO UJ
X 3 Z UJ Xx^ \ u. o o: HH 3 a: o E: M;t-i S^ 0 3" 1-3: U. Z tO O <C
O M UJO 1 1- »-i Qi I-UJ Z 1- E 0-Q£ <C O 0£ X UJ<£ X H- Z UJ O Oa. to 3 t- MXE Z U. X UJO + 0 > 3o o <r a i-
LU *• ZUJ ~ Z tO D Z UJ-j x o <r UJ o 1-1« ^ HH MOl-l -H t- UJ J- Mo E « «. o <r u.M -i M x Z Q£ U.^» O M M UJ 3o . . z to i- toUJ zlAM O tO MZ; ^H M UJ •» tO
Z UJ tO >to;; uj M 3 >. «:
" «;••"• x E o z 3i- a: i-i o J
> UJ Z H <ti 1- tO t- O 3
M H-l UJ UJ tOtO Q tO Z •-•
u; 3 z to zO' . ^ O Z UJ O»-• o to »-iUJ. 1- »-« 3 t-O UJ <E 1- <IG£ a 3 3 LU Ol<c uj o -i a LUX X UJ O 0 t-0 3 tO 0 1-1
tnUJ HI UJ UJ UJX X X X Zf- (- t~ HO
ent-IVhfc
UJCQOfWK
z>l-l0
LU
i
^00O)vl
^^>*H^J^"Of<C
O£«
.. UJujiTf- UB
S
o^^4a «1,1 *
£ UJ1 « 0* i ox *" Oo»H (A~> 2 uj^ 5 *J8 S £<t IX E0- S d«» 2 5
MAR
SHAL
L
ELE
CTR
OP
f
THE
:
COr-enUJN
> M
o«
rM
*.
(UM^^^
iX.oW '
i-ftuu
A/
-oico
Iso
p>n
£
t sJ- Tt-l • mO ff0 CO I*" ^^ ^UJ^> « Oi
2O1-41-«o:2^LUO
OO
<ALUM0LUa.wIU=T=XI—
Q
«
U.
Q°3^<L
2TOM
O£
yOhS
-- - ^^
X P <Et- zM yr . .xt <k3 H ' Z
CO O O0 Z Z HI*-« O t-i H13 0 E o <E*J E OU. Q ID i-i
UJ > CO UUJ E UJ --H D.X 13 M > -st) 0.i- co o at r> <t *
COX « ^ > Q - H U J * ^O ^ o UJ O XUJ O3 O Z i-U. ->0 Z \ M r0 0
a o o < t i x EX l-l -- . 1- i H K E1- 1- O «. « »-«O N
« E O ^ 3U. O)> N E O•- I t - t . ^ - H C O -«(0l- « . t JH « . Z Q i U . l » - i 3 U J Q . \z * zoo co rsj -H H-»LJ> «UJ On- i l -Z r • E . Q S O K U . 0»Q I D M O O - H C O U J I ^ X M . X - -Z C. I-HLU »H | <E ^UJ^ I - 3^UJ Ldl <IfH^ |- 4- H-t a 0 Oa. NZ a: • ^jz «oco •* - --uj H«3Q a <co v >tK3 3 — oO Z i - l i - -i O»-t P! t-UJU. EZ OBUJ Z • O hHK i-»NU. 1- EIH MUJi-i LJ Q<E >^ O M ZS
Q.U. O <TN -H OHO <E «co u. z a:»-» u i-iz t-joUJ O>O O Z UJUJZ CO O^ *-»-! O U.O OCOt-i Z'-'O UJl-lQi O»H. • UJUJ OE UJi- lh-UJ OQ£t- OQ
Qi«-«O X XU. T1 *-«CLCO «Z O t t U J t - 3 O U . I 3 Z COQiO UJ O i-I U _J f- Hi <E <X*-• OEUJ - U.CO LL<CUJ OU.
Z UJX LULULJ O Q:UJ UJ UJ UJUJo xxx *H oa x x xx<C H-l-l- 1- I-O 1- 1- t-t-ui <r ouj
co co co ^ ua co co cocot- l-ll-ll-l Qi ^ HH l-l »-l l-l^ UJ UJ f
X CO UJX1- C 3 Ul Z X J- «t« -«
~ 0 K Si « QiU.UJ O ttE UJ UJU13 a: E ' - a^ " ujCO UJ O UJ O O£CO X Qi X UJ<C 3 U. 3 X
enHOfUJ
o
z_1
oM
It
I
^00O)v4«
•T-l
3»Q!
3a
- ijjpa.'.2
oa HI
t u , ,£ Q ui0 o §S E °\&
~j i i LJ"" 3 Jw ..I Q.u IU ~«. OS *-&- S <T•* g 5-j n§ o ui
Of, *~CK 1— Hi o
UJ
UJ
enIVCftUlS
~ HI5 G.^S
iog
CO
ioHI|-a.Of,
^0
fco^5
ro
f-«
CO• O Ul i
> Z X wH M H • tHI iii H CO 3"CO X.J UJ <C <E UJ HZ HOC X Z CO HIUl 3 H Of CO 3 .-1o u.o uj ui ui <r <r
o a: H HI of. o aH Ul UJ J 0 <E Ul HZ HX X <I Ul 00 3UJ ZH H Q. J UJQ £ U I H i Z C O ^ * Z
(Al?VW
» __r~^3CQ>Hcc«r\.t
• <. ^^"™^
2UlQ^aoQ55;^ r
^2»jUlH4u.
.
QB
OUl
1
Ul
Of. HICO UJ « HI >3 0 HI Q H H UJ^^ ^r o£ c^ ^^ 2^ • * ^^ ^3
0:0 o z <r uj o az oz u. « H ~ ^> o <c<E « CO 0 J XUJ UJ CO CO Z Q . X X U J OE XUJ Z Ul O X X ^
HO 0 0 HI H X X EUJ Z HI z Z O O Q OX COUJ H UJ Of Ul 3 3 HI Of.H3O <C Of UJ Of. 333 U.
za 3 uj => a ^ ^ JQC HiUJ G» U. O * 3 U. «.O E^ Ul U. • O .J
HI I-IQ o a. ' + i uj cO COO Ul OO Z > Z X 3
UJ O HI LUh- E HI X X UlHI CHQf. i-l HUJ E -f CO 3 3 EU. ZUI 1 HIE 3 , 3 X X 0 »J
<CN Z Z C O U l l U . Of -to o • HIUJ v, u. u. <rHI «,CO COZ U.^ > D HI + +c* i-i coo Hin Q z uiH OCX HI HI OH O Of.O 3 OH Z^ O • Z -H -H HI CEUl Of* Q.O H4Q£ LJ O 3 3 H•-1 O Z COUJ Z HI -H -H 3 COUJ OH 03 3H HI -i E E « UJ
CEO HU. HI « O H H HI HIZ UJUI 0 H Z 0 0 Of H< T N O « Q H U J ^ f i O « H HiUJ Z <TZ I>HI O Z • JE C O H O U J U J » J Q > ^ - ^ O O H I
<czn ^>Q£ o<r co H H oo: Muj XUJH a: CO" HI HI u. a: ui oX QJO CO3 I> HN EH OQCZ ZO COH Of KH • • Z
.-133 O HIHIO H i U J C O U JOf. uiou. HIU OH o o a. a: HI xUJ HI |-X ZHI O 3 Of. Hx U.UIH an oj ui o 3>H XZ 3 HIQ. ^ Z OH U.HI o H ui o a: HE o HI »-iUl HI QJ UO <EHi H O OCO
ac •(* 3 z zz o. . UJ H J 3 O J 0 Z UJ UJ UJ QT> • C O O « O 3 < I U J O Q r X CO O UJ
, : . O U J H I f-Hi H I Q : H HINQ . J H U J H O : H 3 U J
; E U J Z X U J Z U J O O U J U J C 9 C OHI UH COUJ XUJ Of. Of.Qf. HI
LU H UJ H H Of, Ul Ul Ul <£U I X O U . X O H I H I X X X X3 HQ.O HO. UJQ H 13 HO Q.
•
eftQJ
UI(AOOf.
Z
jj
O„f/£*•
00O)
«
^4
>•
<rttn
t? u.<i
C9z
"x ^^
g UJ u
o S Qf- E O
1 W °~i .. UI(A ""
H UJ °-2 * c
T y)
i ft-| o uj
« ouiUI
nr-UIN
f 3
TOC.
^»^
zoM
Q.M
l&
1°
5*I
• H 1z o io oi-i a:(ft «.^^ i.i ***
O»H1-om~
r^
oucQJ
1—uQirisOLOu.•—iucE«wu^UC^cU)z:i•
^^• *Uw
o__z
1UI
60UIc.
UJ 1— »
UJ Q£t- <L o . aw ID 1- 0(ft O U.O (ft , Qa. . »-i QQ. UJ UJ Q • =D UJOX f— 3 Q i-J IL
I- 1-1 O J U. OUJ (ft ^J U -1 •XX O O •-» UI UI (ftHI- Q. U. X ^ Z
*-» a. Q t- u oX 3 • 0 U UI Q »-iH 0 X X H(-1 (ft U I Q £ t - O «. O <T3 ui x <t ^ z a •
1-1 *• HX> O 3 »-<(-(ftUJ Qi (ft O flft (^ Ull UJ Z HC3 <E HZ X C ttUIZoc:> o »-• LJ Q ^ - ^ > oui<C UJ X UI M Z EX(ft Lu 0 H 0 > | UJOUJo ID u . z < C * H V > < a o az Q :> o a. o o ui (n« <r oo ( f t i - i o (ft x <cLU a: 3 E U I M .j z H uE H O Q UI LU O E
ui (ft a x ^ . t-i u.OH « U - « - » Z H « ( f t
Z HO 1-1 J ZU J « . O OUJ U-« M ^3H>-»
UJ Z Q H UI Q UJ H C K O C 9x o » oe uj (Ai-i x o o o: <ca.t-«z <r x u. H ui H <r u.( f t H O H . H O M M UJ Qo <r »-i ui u. ui LU o a: uiE ID H Q: HUJ O 'O X » H « HH G» <C « (ft Z»^UJ Z H H 3 (ft^C LxJ Of CO ^D ' G) 1C ^C ^^ G^ tjJ
H n Q < C > J H X (f t»J(f tHZ (ft Z » H O O O *H«<r ^ LU z 3 LU (ft z LU ui
Z O O - J t t U I U J ( f t < C X M( f t <E Z • -" U. XOH «H« E O H Z H Z 3 O | U J JX N O O UJO Q UJ ^HHCKiJ
H E X » H . * < U J U. <E LU »-«ZJUI H H : • - '> 02 U. EQ3O O X Z O ;; J UJ UJ HI QiM « H O » LU Q (ft QC. X O >
n H a Z •-• UI UJ O UIX E U. ( f t » - « O X (ft X Q £ U . XOOO UI Q J O O - I - UI 3 0 _ O f r -<x a x HH uj x a.UJ U. O- U. (ft t- «
<nt-ttUJ
o
zJ
_
£r
00a«H
*'<'
3»Of<r3QJM
w LlJ£u.aa
O
i5 j£ U . .ui a y0 o Si- C 05 • °•"* fcil l
"• S JLlJ yj Q.
<c ty E« § gH Q. . .•= !sr LUi 2 x«» *c §_K |— r^
I oUJ
UJ
to1tnUJX
~ HI§<E^S*-«
1Of5 K9
in
ioi-Q.M
..Q!
^0^UJ1°xo
si
t- 5* *
•w 5p *P- Z 3M OO MXO <A
(AH4V)LUQ£
CLOo:§••oLU
LU«KO*••LL,
SO
M
OO
o1-y»
ok^^^
^H»<EOH-
Q.a.
J Z XM LJ UJ
^ ECU MBX CL Q^ M |~H J O-P
J LlJ*bj UJ
m . i *w C"iU toM ^^ CM
x 3 ze-ro o» <n u
F> M 30^ v> v> oz
• 3 «O UJ ZUJZ 3 LUE E Qo N z<n o ' u>MM uj <r o o M U ) c u
• H - N J I U J ^ U J X•G£O «> •> Q. O Oft Q£ ** '
a: uj LU a> i a: -» <uLUCQO: o c n o t - z - H UJ >CO E M U J M O O U . I X XE <r a * ZEU •"•rs! -* *~ w
^c ^c b^ ^J i"™ V^ ^ -^xoo ai oujuj c ocnO J X MX ID 1 MM |
UIUJ ^ Xt-t- O QUIXM OJ t- Z UJ « <XU ^-txi-u- > 3 E«: aj ~1- X O Q £ O H » Z - H 00 P)
CO UJ ro J iii Q£ y) O • O U. otnx a U.MU.Z HI « OMZ0(- | E 0 (- 3) QC •O Of *•< Z Q O ^ "^ X ^^>oz ^ <TXUJ oc. y 3^<C<£M 3 UJOaUJ 1- J 3
n E 13 X Z U, UJV) (/> V) LU CO fr- iii E»-<i-ii-i -^ UJX^ O - Qi HIXXX XI-UJU> Z Ot-<I<E<C H EM O {-
• tO O O3(AMU) UJUJ
X 3)N i*. M tH 3 •-» I>Z
> H O LJUJQUJUJUJ I3(U 3»U M XXZ
^ X X X Q ,HH<Ei-t-t- <C
UJ Qi UJujujuj z a: a:^ ^ ^ UJ UJ UJ UJ<r <t <r x x x xh-l-H- 1- 3 H 3
s
tOt*Of.UlpaoQ£
>.»JC9
~Sc&
00a> .
*»H
'a3aM
— Ulp U.g
Ok_J
fy »^^
UJ ,J
5 o ^0 o 9^ E g« ^^
§ ^ uiLL. ^^ i
? 1 3i o si f fI o
Ul
Ul
rv_
S>UX
,H
U)v4
oHI1-OLM
~o
*•
•r* CO
.A
cy> *UlV)<coui
* XO (-Qi •ui a. uiN Ul 1-^^ i rUl <E Crt »-im ui t-
E Ul Z> OJ W E Ul<X X Ul UJ i-t QiE ^ X U) 1- U)
»•« H- « U.1- O O U.H-I | 0 1- *-*
t- ~i Q
Z Q > o z *-< <nO Ul X Ul O Z '-h-l U)(U Ul f-i a O|— O 3) Q. O) HH<C OL n tn z uat-^ E ^ Uj 3 »-«
I"j C^ .J M Zt~ j <x a ui og- J 1 3 | W> O «•. - •Q- M o z)^ t- J3UI 3 -J
Ql -» t- 0 »-•Ul v> o a i-X »-• I*- Z ui •-•|Z -t- 0 <C N Z
° 3 O | W>Z3 «H 1-4 LJ *~ • O
M" 1- Z ZUl Ul <C 0 3- <EU. • Q£ O0 1- .J >
Q ui r u. aj ui 3 x ui <rj ui i- a o o<r Q. z 1-1 z3 «A tA Ul 3 3
1 H4 Q, -1 0O i-l Ul U. COZ J O»-• <r JD i ui ui:> 3 D ui x xO E H 1-E 1 1 1 1-1• t- 1 1
•
(/)HttUJ
O
Z
»Jo
u?
c
00at
3
>
•<co:
£iPo
OZ
£ *"*i S ^° o 91 = 8j JJJ UJ
8 S ^£ 2 1«> o s
^p
^ °- ui2 ^9 ^p
a ^ ~E P.
UJ•JUJ
n(Au
~ Ht
2 f>>
1
s;
^"*^
O
HQ.M
M ~»
igj
o
««•
O! ill a><E X 0 SU. H Z I
O U. CDUJ •U) O i-i OUI
v> H ujoa.* z z a: E
O UJ UJUIC*) M Q XXW
t , HZ HHuj <r uj uix u) o a. v) *xH 3* MUI i- i Ld H
M »_1 O 10 Oo: Q n a. z u <r (/)o « P U Q . M O:U.Mu. o: <i oo:
en xuio:«. X C5 J M Q.HUJ
Q H Z NJ OZU.uj »-( i-« xx uj on-tu.t-t 3 U) H H 3
riNl l-l «. O H UIZXf O ^ Q U. U) J<IH~ ui ® ui • ui iJ u.° a. .* x j > a. ui1— » o o: <x o: <xHI UI — < O LJ UJ < C U JQ o: E z H oxxZ « a> uj tn ZHHO H C5 3MO O <X V) i-l O 3 •
Z M > « M« o Q _i z i (/>n
^ M • uj > H M oo: o
£- au H 3 M u ^u.H« HI> Z Z OX 03 *Z ZM UJ M 1-1 H EMl-l UJH O H U. ®
OO Z U. O OOUZUJ * 0 3 ZZ Z3O Q- Q£ O Wt M O ^ HOW) « W> OiM
UJ .J M UJ 3H (AUIZjo: 3 Q a 03 MtKM^ o <x r on to ^MUJ o: z U.M uj Ej^ i fc«4 f^ t is ^v rv 1 1 1 ^"^!•• »-J r ^ UJ it IX, UJ ^MQ. O M Z OH OttQZE H 0 MU) XUI 1»-• <x v> <x M o: M QL x N
tn M o: H HO OHU. H M 0 X UJOO, . UIZ O UJ_! O«X
Z ^1 UJ Z _1<E <EUHo>« a. o o uji-i HJH EZ 0 OH • 00.0)uo: <c o .(AM tncujwuj o>o uj MZ M<r<n
u. in »-• o)3ou. LU uj uj o: of.33 XX X O OH« H H H U. U.
<Ot-OTUl«Oor^^.
^jo
Ul£*
.09O)
^^»
^4
Or
f^CO ,
lu J~«S
_{* Jj
.g g Ul
H 1 §z *" Oo „ v
3 2 ui"• S ->o UJ £
2 g i« § gJ O- i.ii ° *5 0£ |E
« 6c ui•J
.., .
roCOUlN.
- HI
|*
i5S.
CO*^
ioHO.
o:^0
fcu
«5
• ptn •UJ ^.. 009^ 1
* s* u.
• o 4-UJ Or— «J»— « UJZ ^h^u. a HZ M ZM 111 <L
XI-> HO)U Z •
• UJ H 0 Ul»-i ^> « o a.M i- a o xo • o uj ui v>M t- UJ X U)u. z u. M o ujMM u. u. a. •->f. 0 Ul E 0.a o. ui M E
Z <t w) co <rM • X " X <A
p M Ul Z C M O "
| z c 3 ° 3 5
° M or E z uj :>. <c o o M ^ j x
%J EM Q «OO 0.<C o or i E E :> <cZ o uj N o uj orO It- Q Z • . 0 <• - •t-i 3) Z UJ <T Ul 3f-. M X «-I O X UJ
2 x uj z > :>E f- Z «. 0 UJ U.S3 O O M X O Hg. ^ z 1- t- XE: (A ui M a: z uio ui v) a) t - o r o zO <A 3 . 3 3 0 M
<t 0 CL t- UJO Q O E * '3 X
Z U, O <A J 1-j <r o uj o<C O 3 (A Zz * M or *J uj M • • • • ' . ,o J or a « or^ j • •__ ^^ *^. y^M i-I t— O ^ OCA ^ O «-l Ulz E uj u. *J <r i-Ul (A J O <C E <IE uj M M orM > O <A »- t- fr-o or <A uj M a. <A| UJ M M Z O Z3'
uj ^ or M _j :z or <L <A j 'O > O O Q Ul M .
u. z uj :>UJ Ul 3 (A M V)X ^ 1- O O O Mi- <c a. co o.
E UJ E U> (Aor o ui M M M0 UJ X X X XU. 3 Ul f- I- *-
aUJ«o
C3
00
<r=3
QO
« <ftIHO>
o Ui<x OJ& oj Q.
ouJu
§o
UJ\
P.*<xN .
IoMI-CL1-1QL
"O
c•H
OO
<E
O
o1-4
Oo
.
<rE:o
<xzhH I—I
K <t»-iEZOI-H Q
<rz
ce
Q
OU
OOOooooooooooooooooooo
V)
atoa.
o
o
ou
U)1-QCUln0
Z^»,_JC9
—Hi
000)
^^H.a3OfM
- UlK U.S
C3tj
5 ^5 g u: i §i OM tft
J M tft
o U g
2 r> C
3 £ ui 2 i^^ u* ^^a u. r~i 0
Ul
Ul
O)
V)Ul\
- t-4
o ^^Sr
4D(U
1-•a
- w• Ul£a
§
••
mZ i1-4 Ul
i I0
2 Q
<TLUEXOt-Q
U.utoXHl-
OO1-ZV)t-l l-l
•HOt-H
zt-tft 00z: , £E
2 « . - ozP O EO
»-* Q ZO <C OZZ tt >-(<£o ui0 > ZE
V. Ul U1ZfC I> EUlSF ui > x•T Oi ^^ ^3o u. <r uiZ o o x~3 Z 3UJO x n ^tt =3 O Oi-i
U. O </>tftZ N O<C
O M Q-CLQ£ tfl Q£. E ^^Ul IDU1 MN OX V) Ul
•» ' U. M (/) Z H-«(ft UJ <COI-Ul O ' O >-•
a Qi <L (£ <C «-i^ 1— uj p; QgQ OX XC-U1Z UI3 I-ZO=) _1_1 OU1O UU. OOtt 0 -JZCt
cn o j o uj3) M Z <C O N
-
o e • . . : - . - •
1-Q£UJaoa^^.jijj
-r
00
T-l
^
*H
>tt
3aeM
-Ul
ao
Oz
a: Mii *™
1 g|IT • ^™" )O
^RS
HA
Lt
SPSC
E
FLI
TR
OP
HO
RE
SIS
TH
E
SA
MP
LE
E OUl
. Ul
co
Ul
»*«s
w
Ai-i
gata:
i
$
00
u1*
UJ^J
tt^rv
^
IH
o ~•-I EOO OXu. x y)Ul HE^i _)<£H O ^>
21 a B^jO 0 ^ <E ^
1- Ei-lH UJo ui ~ oujz v>O J E XiHUJ Z X
C O ^ > O V ) U . H O ^ Ei_ « E XH 0 »H EUIOC ^ J OUJCnJOQ. UJZ H OO^S* Q i-l XOO.OM Q.O « XZg- CU CI 0<EE^QiH >•-• W) 0<tZt O) ^ XHXt^-HZ HH i-i XXO3 o a <r <n £:_)•«- ouj <rzz>EO»-tOl M Z Ul "^ O Q Ul tt i-l Ct O O H -^ H
H « UJ -1~ ^C/iJJCK «|-H-II-II-I ZO(A Of. OQ X >• Ul LJ UJ ZD OZH 0£>*OZO Q « «^HUIZM O i-iUI «UJ«HM3z f- «* i-i ex-H o >H u. uj oocno: JMHU.<t CL ^ H <r--i-i<r ooco aozaE»-ia>M H X 0 :> §-flOHi-«UJ>H OOO HHHO ~^ IH <*•» i_I | O Q£ Z Q£ 1^ VJ X i— i (A Ul O Z *Ho a: z (/> a. J ZJIDUIHVJZD oo£ o^uioXT »»> il. S ** »«s S 5 rls S II i ii /i ^ IT ^ JjO O Z H Z> O Ul X O HH 3 -1 tt i-i OUIU1XOOUIi-i i-i i-i ^ o M zcioaouiHa O:EEEOOOI>O : X Q : U J E X U I oui o_ a. H ~ a. o i i i i i i i u . i i i i i i iE <c <rZD o: • • • a • •z o oA A A
"
(f)
CQO
OJ
E
TCOO)
CO
Q:<r
zLJ
C5Z
UJ
o
(ft
(ftUJ
i
ro(ftUJ
<ta:
tro
<xQ.
2
£ Z
OUJJUJ
ruru
Iu>t-raUJ
GOOi
QUI
fcr •
w> U)®eu nrufi*
s . rv'z-.-.-. §2* s.l-g
® © W 3- ® * J O l£ O (w l£ l£ l£ (£s i£ tD U l£ l£ (£— K — U«Z --.i--— „.«,.,.,..».,
UJ J Z — Ct Bl P. Pl P. P. P, P, Pi Pi P, P> D> »
iS <£ Pi • • Or LJ 15• • • Z D & — UJ
e « ID o o: ct o C-Zt-uMftinif tuM/i in in in in in in inin uj K &— o — x. •v> h- h- z • o o • '— MX o r uo c. w ot. ca
u c-j —— — —— — —_ — —^- ,—r Q c P. p. pi PI P, p. p. pi p. p. PI p> PiUJ ® — c? Pt Pi 0; Pi P. pi PipiPiPiCTiPiPiD T —
u ® O U . ' * ~ £ P)O)3O><£> • O PlOtt-inin o ts^* • • — c tc. inao.r: or> — --. — --— _^ ,_^^_ —<r z - v ^ i i i i i i i i i i i i i
• I 19 'I- C5 «Z zco M w uj ru ru ru ru ru ru rururururururu£ 9 ® r u *^oo o ^< & * - « 9 o n X H i - a
3 IAZ • • * >-<UJUJ Z~ u.en« c o> ru ru ru ru ru ru rururururururu© WG *M
g soco z ect... UJ « O i
O *^ & V t^ C O tj(& t~ . UJ Pl ' — ** *^ ** *^ — ** *^ * *^ •** ^* ** *^O"« Ct Pi Ui —S Z »d p> u i B U Jess o i-i sen » t* s ->u9«s o u. . . . M in M — f f k r uJnjrunJ — ~i — ru ru ru ro ru ru ru
a . . . . a vi o in ^o et"*j»»— o ce CE K zUJ — Z 9U>T UJ f- US O— m —— — — ~— ^i —— —— ——I 5 sect- K Z i- z— — i I I I I I I I I I I I I
* * £ • • « UJ O^-i* UJ *«
in c s? "Q: •« u/« ®oc» c ZN> a w«»- in-11-0-0.0. o Z ees c. >-<uu a. LU inina:c OLUCCC a o ... xxx . ca zuuJuj cu)««c ui M n —io fi — — — i- — to ru ru ru ru ru ru rururururururu_> c Jin win N i i r> o u. u. u 3 Q . I I / H - C — « - . — — — ,*.*,.........g WUXEO. uj i E. LJ rws i i t i I i i i i i i i iK uj 3 tnouzCL Z esso c o ocot-
O C ' S O M M L J U l•»>t-S® « »» t-eao — i—t - ruiD— K «« —— aoetUJr-«l/>r)9 ff UJ Q.
z ca s s Q ' in t£
•eo
a
Z
SS5S!
V)
I SC *+ m 9 t t
in zr«- in• ui» cX
ru
toc
UJ
• £ - • •»- Qor x u czuee&eos esossse— PI o> en a PI 01 c
P, (71 P> Ol Pl O> CO LJ t- Pi O) Pl P) Pl PI Pi 0> Pi O PI O>
CZOO)O)PiPiPlUJ CCLLl —tn OCKUJ — K Z t
5 - B «•« u.—
o x uj «n i- t-D U J C M Cj c jct-itta u £wJ CO CD Q O N *^ 23UJ N N N N N O O C»-(j LJ 5 —
in OCEoc
c.uj-
a. aUJUJUJ
--£
MOK a£3-— moo —- c u —SrZ — ~. 5N>QOUJU)S —c^n CL u uj LJ o u c E u inin
(A
i » iUJ
O
>
(9tug
z
_00O)
*^"
^_a:
Of«
UJ i
«a
OZ
ot. *~*UJ ft.J
i_ ?T
W Q UJ
° g 0%
0 ...3 J5 uu r? O_o UJ =
ft TJJ ^^ft O 5IVJ
_• m.3 9- ...? ° ¥5 or *Q£ £•» ^^
i 0UJ.-1UJ
Dfs-
1,1V
— *HZ ^pa|5j»2° -.
(*)(U
1inH*J3
~.UJzQfHa;fo
(0r>r-«
iii <>r. • s: o»
o S 1 4UJ O ^ +N »H "> H Z_j <r oz H cn<X Z 3
Ul U.cn o u.LU Z Mc n o Q<t oo a
3 • UJ U.Of OZ 00O JO ZU. M 3 Z
an o>- O <L • UJ w •Q£ u. oc *J H enO H • . 0 - < C Zuj >z en E 0:0
cn x JLU o <r <xw•»£ HZO o e n o . H0 OZ •! LU<CJL o o enn « z e n o i :P z e n o LULU < E H51 <c H enE »-« azSb zz z <r en ouiO H LUi-t o oj a en u.o,j ui LU of H ui i— • eno ^ z oOI E t t ^ ^ J H ZZ OOO-> LU 3LU i-l Q.Z O 3 •-»
a: enz 3 ELU M Q as H3O Z 3 <CIH 0 MH H LUZJ <E 3O
*~* t-^ ^c E ^! Q_ <v Q_ ^ n 3 O5 j 3 H L U e n o o r H E E U - O W H^J t* t^ 1^ li ^^ 1 1 1 tft £^ g* ^; /N
Ou 3 x t ^ > - z ^ o o j o » ^ e nZ Q. H =3H UJ > • Z OCOO HLU^ E >-4 M a Q£H> O <XZU- U. >OL
O 3 W3 Of OZO O .-J»-n-i O »J>O LU »-t 3 LU LU Z 3 I> Z CH
> LUH O XKUJ i-J OO3 C3 Z> _ i a : o HQJH <r CKE z: <EZH H O3 Q 3(n 0 •-« - M Mi-i O <CO Z XO»H »-» O «. LU O X «r > < r z <c H en Q Q J < T H OIH X >0 MLuZ <T ZJQ. LU t-UH UJ HO O 300 (£ »-<LUE K 3Oo i-i >j o I-KC a. o3 en ^_i u HH u. Luu.cn en HEO LU i-iUJ t-i ZZJ O O ZLUZ Ul HO U. LU LU <Z ZO^l H LUX0 0 : 0 0 E E Z Z < E Q : < C O EO O 3E O LU LU O: O X LU LU UJ LULU • , 5 •
<T O t-t LU LU LU i-i O N 3 u£ LULUO ZLU Q£ QTQiH H t£ QC. (£Z X OX H OOZ 3 0 1 1 0 OX<C O. OH O <C<Ei-i »J Z O «H
LU 0x » » _ > • • • : > • » •CU LU UJ
• O 0
tn
o:LJCO0cc
. ^.
COO)
"s
COft
.ceJ-P*
o
z Ul t-y Q -1
.0 3
Ul
S uj o£ o P, I <L
d o. oS O w5 tt J
1 o §Ul
Ul
enenui
ex.N
(U
IO)
O)Ul
I
I
oU.
t^inoarui i i i
UJU1UJUI
oujioincxs"-•(-r-r-aat- runoxsaz
zoLJ J I t •uoro »-OE ZOUlEEUJ
tn
LJCOO
CO
00
I >I CC! <ri ID
0 i
!i•- <E
c• Q
UJoo
« cn
UJXt-t<r<z.
|o^l"a
«I u; a.
cn
13cnua:
S o t—-> ^ <zj Q- QC O M5 o: Jg t- <rI o ^
UJJUJ
U)(U
Icnt-
cn
! a:i c
ar«•«»_i
OE I I IU. NLJLJUJ
cues©
K-flO IroUJ_) • I I Iooro Koc rOUJCCU
<T W—"-i_)<IXZ:OE:
V)HctLJca0 !Q? i
ir-. jo_
n1 c
j *~ i
C5Z
z LIUJ Q0 0*-• £x • . *•
-
-C ^IT
o; U.c- Q;fe c5 a•J •*« c5 QK- t-£ C
U>••
u
f.r-'inLJxh-t
_ Ct
5 <C
N mco£t
• : 0
T
00CDT-«
\COvH
5- '
ct3Dz
- <xH *^C
LJ^ .
OO
III_( jj
1 £:
1 ss LJJ X
^ H>J.1J
IOru1
v>H
**iin
". LJ0 Q/^
ccc0
(f)f—z:LJE:t— <1 1 1Q_XLJ
>*jl—iu_Hjt-<Ec~*LJO<rLJUion™^
*—LJ0
HzLJo:LJu.u.1— IQ
XMinozt— Iin3
«. inn o:00 LJCJ) U.»-( LL.
3«. CQ
>•tt »Js: o. H
1-1 CQo:
Q <ELJ .'£)s:Qi HO ZLL LJct ctLJ LJCL U.
U.LJ t-i(V Q
LJ3 o:
3in oH Lt-inLJ QH Z
<rzi— i mCQ LJ0 J_J CLo z:o <cH: inLJX
Hz •LJ inH XZ CLo <r0 Ct
oU. 00 H
O> XH CLzHH X
<E HH "-<Ct 3LJ0 QZ LJ3 Ct
<r0 CLH E:
oLJ • OID inQ H LJ
in CQin LJLJ H ZJ <XCL H Os: o^ z inin t-H z
H 0
X M HH 0 3
CQLL H H-«0 X Ct
O Hrv i— < cr<~n iii t— i0 QU. U-
O Z> »-,_j _j OQz <n o
O 0LJ H 0H E:<r <r LJJ X~~l QE: LJ LJ^ z> xin J H
o<=> r> u.j r o3 f-io ino in H
»-i OLJ X J3 H Q.
QOOC5
>JCO<czoin<xu££
LJCt<cinzo</-.f-^ct<xCLE:coLJXH
CtLJU. CtU. LJ13 U-CQ U.
3X CQHt-i Q3 Z
C5Z LJ
X CLin i-i E:LJ E: <rin in"1 Q
<r LJ uo in x
3 H0 <£Z 0 Z1— ( »H3 in s:o z »-j inJ O HI {-
-J 1-1 U. ZOH LJU_ <X LJ Z
Ct X OLJ H H CLx z E:H LJ O 0
OHOO ZH O LJ Ct. O .J LJ\n CL xLJ X El Ho o <x o
• z HI in•,LJ X U.ct "o oLJ Ct ZU_ O K-I inU. U. > H
O Ct CL LJo CL u.
LJ O <I LL.X CL LJH U.
in o LJLJ H-l J
H Q CQru o HICQ LJ x in*—< Q H inCt 0 LJ OH E: E: CLH^ I i i
LJ3
,
LJQo
LJX •H ,
z .• —
inLJMozLJi-<0i— iLuLJQ
CtLJXHO
LJJca»— ieninoQ.
1
ai
u
f-
£0
0)
COo i -*
>a:
o
£?is
•• ent-z:LJE:
C3 HIr a:*-» LJJ Q.LJ XO LJO
I ~ - tfl »-i
a.-'W)
LU LJo: t-0 <E1 t-0. LJO Oc£ <rt-O LJui en-j oL'J J
LJO
mr^enLJs
Of
isgi§c
Ien
en-. LJ
fc =?
k.-0-.i -?^x"t- -J.f':,-.-^ •'.. -
f *> 1
C
& -iiri
o
CSLUC=)
Occ
-1o
00
«,00
o:<r
-«UJ 1-j
co
inHZUIE:
K 1-4 U)UJ J Q.
£ u xUJ **S 111
I Jo »-»« in wr^ w u.
u UJo: t-oi0_o
UIo
a: <r(-o uU V)
UJ
_JUIo
cnu
00ru
Iin
inui
I CJ2I o
trc
ICO
exM
zc
g ca• i i lL.CDULJU
S cceruoe
c ctr(- — '»fui-ui irt>UJ C • • • •
ru K
_;>-< — a L.Jroc»Z>->
O
LJOOEli
CO
00
i aI =
r- <r
V)I-z:LJ
z at-l LU-1 Q.LU XQ LUO
* E 5O i-J
S « ttV)LU LU£E t-O <CXa_Oa:
uO<r
O UJLU V)U OU ->
LUO
V)LU
Qi
is§ao
0)cuItf)f-JZDcnLU
IIco
-O
xIU>1
5^axDir>i-r-K: •<B»s^nni
S061• i i i
L.COUJUJU1
r« no -i
LUC i • I I
s'« j«-i — crujjxrocOuJC>-<£<-iEOCE — I-
O
CDt-a:UJcaoor
! u! C
o"
COC7)
00
o:
o
; C: a
UJ
r Ki-( UJ_J CLU XQ Ulo
O —•« V) *•«t-i U.
UJ UJCC Ho <rx i-Q. LUO Oa <ct-o uuj tnj o
«jUJ
n.r-O)uxi-»<ro:
o:
UJo
n
en
UJ
: cci o
i »-• z
CO
a
cxu>
Kaeo • i ili.eOLJLJ
v£>ru
§«tOCOu>«
UJ« • I • •
£W tOIA Zopecu
in
LUCOoa:
C5
COO)
00
Of.<r
t-zLU£
O «-lr av-t LU*J 0.LU XO LUOE C
V) MH4 U.inLU LUa i-0 <r1 i-Q. LUo oae. <cO LULU V)J OLU -1
LUO
r-(ALU
<rcs:
+• a
isaocro
niin
Z)in
- LU
I!<x
V)HccUl«Oo:
0
uca
COO)
CO
ce<r13Z
a:UJ.
ui<•>
».
Ulo_
-J.
f-ZUl
z o:I-H UlJ CLUl Xa ui
en »-»t-H U.V)ui uia: t-o <tX I-Q. UlO Oex. <ch-O UlUl V)-1 OUl -J
Ulo
m
uiNt-«<ta\
OJtn
Ul
cUa:o 5
I2
« Norx
ru«• i
"Ul<9
mcu «~(ur- •-aroo<9<s<9o • i i iuu. oouui —\«®
OCV-—U4HUlzoUI<C t •oZUI
n
OUJ «>-•«
h-o
to
UCOOCK
z:
COO)
00
<rID2
- <rcQ
Uu
5
: Ul
inft
I-•z.UJ
W UJ-J Q.U XQ UOE: ^
»-i U.v>UU UJo: t-O <EI l-Q_ UO OQ£. <Lt-O UJUJ 0)J OUJ i-l
Z)
UJO
UJN
t—i<ro:\<x
CO
Ieni-_>13cnUJ
U
(»
£
iOB
-0
o:uCO
oa:
C3
I Z
TCOO)
00
cr<r
ec
cXinr-
u
O »-iz a:HI U
j a.u xQ UOE: E
M U.V)111 UJa t-o <rx H-D.Oa
O<z
O UJLJ V)J OUJ J
UJo
en
LU
<ro:
i3il
cn\
V)
V)
•runess i ». i i <u «
OCOS)V U)OlOoui»oet-X f- tfltf
00_
5jxrcLJ <£ ** ++
-0
.-r
X*4
•*
(V9• +
..UJe
ho
X
**. V
tt
(U
a
5UICO<-»- -in -at01- I I-JUJbJUf*c • • * •
""LI «-•uvi z:5S§iiE:t-3CEUJt>-J**»*<r yjwjxroe
• cXUlf-
t . I I IUCObJUi'UI\
U)UI I
£ < t I I I !
-ltd (-VI Z
OEC —I-
I
_>eaUJ •U. S.X
O
ttt. . . . ru(- <c (vi in ru —
as 'UI<C • I • •
tnf—
UlCO0tt_?
uc
*
00o>
**>'jE
^2a-Ult i 1h* *••
o
uz
DC HHjj ,j
i §s^» ^^. ' MM ^^r— W^ ^*X Ou3 2 uiu. l-l 2
S " E
^ ° S1 | u,vt Q£ p<r JvE O
Ul1 . •Ul
tntnUlN
Z ^£2 «*H
|Of .5
li"5UlD•
f f\V)
J
^" 11
§a
iX .u
•• <r^^ CDz 7i-i gg J0
E a:Ul Ul 3X X 0
u. oO Ul H
otnuiuit- z uiZOQ.Z • M 3oz <iujuj tn o«-«<rxoo
^^S
^ft
UJQ£
CLOQ£h-OUJ
LL
O
LL.
CD^2G
2J™,
^3. iCOo
f-xtntnz » tna i-o:x uit-UIZOO O V)ZO.UI O »-iui<r oaui ui xoxtnuia. i-ztnuio<r uio azx x *oooo tn t- tn
z a: E^ 0(-O 1- Ulc:xi-tnz « ^JUJI- • CO3«-io ii tn ui o
3Z z i- aQ i-t tn tn o t-4 a.UIZOi— ii— i tH Z •^o<rtntn i- >-io>-•-I »-i Ul O O 3 U. QL H-OH-^JEE J ZUIM
zui -oo tn M»-i^tn i i >tn«
OUIOO O *Jt-«<Xuxroao: M ui t-I> Ql-i- 1- 2>Ztn
XX UIUI -1 h-3»-HOOf-U^J <X OJ Q
UI»-i<£UIUI Z UIOOO>-<L « U.OUIX<r cxzuiui u. OH-Qh-0=>=> X UIUIZU1
(/) »_| H-t •-! f- X Ul EO I-H-I- »-« UI»-»-<H I f\ Sf - • ^"^ /^x ^^ * _ *1 O ^ I— • 3 Ct O- r-
EOV) <tZUI"-<a j uio <n 30.0ui^uiza. ui tnoxHiE<ra: ui Q.QUIJasoxx a LJ a.O EI-H- O 1-3 *- E •u.<5 »-i»-i « tnoo»-«QCZ I 33 JZUl»-i U UIU. U.Q.r>LUUIUI J E UIO
O U, U. U. <E »-H Z I>tnEooo i-<t<cuiH tn uix tntnO««« HH UIE 3Ul X X Ul|_ t_ L_ ~3C K I ••*
ent-o:UJcaoo:
o! C
00C7>
».CO
a:<xZJr
- c
or
£ UJuj O0 Ot E
UIoco_V)
enUJo;oIo.oCKt-oUJJUJ
UJcc
ujU.o
nr -tnUJ
a:s<rDi
U)enIU)i-»j25. V>.UJ
zo
V)t-KUaOo:
O
ii
00
00
D;<tZJ2:<r-5
o•z.
u u>Q HO JE ZJ
U)cn uj1-1 Q:
i uu<ia.in
or
H(nu
u
o:oia.oo:B °UJeJu
nr^inLJ
<co:s
r-nI
en
tn
SI CI O
o
KU.«x»OU t i Ib. \UJUIU
— »»
«-n-r-eu»
1ES 'ld_) I • I •
28 -on z°£§5£K CCUt
!g15
t-o:LU
C3
00O)
00
<rOZ
- <E
a »-!u -Jz Ul tOS 2 *-.
toto uiw o:toUl t-cc toO UlX I-Q.O Ul£K U.t- OOUl"JUl
rortoui\
or\
00roIto
toUl
K<I
5VMi->KUIex roc
O UJC MZ »^E 3 C C •-• I-
CD
UJCOoa:
o
I j
COO)
03
a:<r
£S«a
O
ee »-•£ -1z uj <ns § 5
o-«Q.-V)
tftUJa:oxa.oa:t-oUJ
V)UJeg
(ftUJ
uu.o
nr-inUJ\•M<r
u?£ <tK a:
O)eni
V)
- v>
«5W00
g tj.e>~u l l
U. NUIUUIi- cons®OUJIOxOS
roUl J t I • I
O UJ C C UJZ33C
K CCUJ
(AI-
UJee>oo:
o
00
o:«Of.»
Z
6 g ": i§o
<ft ...
•MOLU
UJ
CO
LU H•J
-tf>
O
O.
m
nooO)
o •00
i-*<r
UJ JJID
ui
•O.OE
O00
Of.
xz
• o
K<£O
u
tt
z>oC9Z
o
UJ
<cX
a:UJ
<ra:
•z -0>
I-UJ
a:oz<rx
i-zUJ»Ho»-tu.u.
az
V)
WQCO OO3•2<XQ t-it-4O
V)OCL COQ JKI-IUJUJ z<rv>=i cos ujaOUJ E X
Zfi O OE
VJXOt-Z UJ
t-tJ-00 I-O<I*-
ZUJ I
Oil-003<ECU
V) O.
UJ %.a:o)
a: oujUJQiO
MOUJUJ »-i
•JOEujcn
u<rZ I— UJ ^ OHUJ N Q H HH V)f-ZU.»-»»-i COU>OOZZ3O»-iO
in i- V) t-i u. i- »-ZZUJ H4>UJ
i-t uj v> V)OQCUJUJUJf-*-iUJi-it-4
JZf-Q£0 QICCU.
OU.Q.UJJO EUJo z E o o o uj :D a:EOO UJ O Qi Z UJooujz <r 3or x a: a:UJ UJ UJ UJ UJ UJ UJ UJX X X X X X X Xfr-t-l- h-t-l-l-f-
wzo
I-Q.
O
Zo
UJ
o
oUJ
<r uj~Q.a
UJ
UJ
«>iZ)
tnxa enowQ. UJ U. 0) O
ZOOCK >UJ
=> I-Z) X »H-C <E
U)UJOQO(- O H4J I XOO -J
HUJOOUJQfCL
ou»-;f-ujoaQ.ZZ'OOUJuiootntn JUJa: N E: M «-i uj ^<r> i I I I I iUJX UJ
tA
auioa
O
v00
00
a<rr<r
5 wS *~w go g
~ ' ZI
is 2 ou v> t!
8 IO LJ«• ^>
o§u
cn
u
a:0)H
«Kao
(jetu/eiouiui) •3U03 »UIPHCIH
(ft
UJcaOK
O
CCN
TCR
c.VI
j
<E
00O)
CO
a<t
&i2
orMJUao
tnu)ui
uOf.
zo
*o: ^
QJ
U\
cur-
V)t-
S
oto
ot
uuV)
-jjf
• • *• •Ul• T
{5! «(0
s£-rr»Cinp-o
§ • • nj««e<0* • ixo «*u
•^ <3 • • •
S r w •«•M 3^oceSwwi c rsoa w wcc ooi o:»« « w •» ••^ujxrEi-*T
_
aUl«Ocez>jo
00
09
a<EDZ<I
Ul
0
5 2 2i in "iS
toUl
isMa ->H «,
ri
nrV)ui
K
«i
n
Wit-
u
1
OS• +
— UJon10
ozuo</><r ni
C 0>U3O
10
-w • • *uj nOZ
O C E O C iOO
• a — —• m •-• «
CUOCCt-MS
UJttOtt
000)
09
a
- <x
Qr " 1
w J ^= u SO Q SO isfc E a
| S O
S 2
u
o:
«
nwu\M<rcc
5i1-1 J?T
§
(A
UJa:
at9
to
CD
i
«CE
O
:iL1A
zosn Mcz • •« . i^-a. o»n ««uj
O 0 9> X -U>KU I • •
Hu>Co M D
OU(£ l-l >-• ill M l-«
o *^ *^ »••*UOCCI-ND
-0
KorUlcao
r
^-£I
trurUJo
o_ju.UiO
Q.
_1<T
ina«:
:•
nCDU)\M
^: a:fic^
«g
"T00
2s
CO**
V.<rz
- <rco
0zwgj
1 I.I
'HO
RE
SIS
MO
D
SA
MP
LE
CO
DE
i»iCD •»Of .
0UlJUl
Ul
0Ula.tf\
Ula:oE:
C/) x21 {-^1 i— t
COHcoUlH
o1— 1t—<rE:Ult—CO
r-
ICOr5
"CL§co
^«Cftv >
1
•
/_v
COUlHH
^
OP
TIO
NS
(H
O
(—230_fr-IDO
Ul25oo:a.E:
i
zo1— tf—oE;
ZO1—4
U.oz0t— 1
<rjIDE:.a:ouUl25OfX
Q.E:
i
t—jtnUla:
<xzUlE:Qi
flAL
OU
S
EX
PE
oz
Ul
<_3h-t
f-
V)Ul
z
1
- tot-zUl
orUlQ.XUl
I
T—O
sUlHZ
Q
ZUlE:E:o0Ul
1
.
CD0
E:<xZ ::
O
Ulf-
t—ia:OLoa:Q.CL<rQ , i','.".
1
Q 'C '
1
^
'"
• .- - -
5-oi
1CD**
-.
