Citation
Isolation and characterization of a variant human H2B histone gene expressing alternative mRNAs regulated differentially during the cell cycle and differentiation

Material Information

Title:
Isolation and characterization of a variant human H2B histone gene expressing alternative mRNAs regulated differentially during the cell cycle and differentiation
Creator:
Collart, David George, 1961-
Publication Date:
Language:
English
Physical Description:
xvii, 196 leaves : ill. ; 29 cm.

Subjects

Subjects / Keywords:
Bacteriophages ( jstor )
Cell cycle ( jstor )
Cells ( jstor )
Complementary DNA ( jstor )
DNA ( jstor )
Hela cells ( jstor )
Histones ( jstor )
Messenger RNA ( jstor )
Plasmids ( jstor )
RNA ( jstor )
Gene Expression Regulation ( mesh )
Histones -- genetics ( mesh )
RNA, Messenger -- genetics ( mesh )
Genre:
bibliography ( marcgt )
theses ( marcgt )
non-fiction ( marcgt )

Notes

Thesis:
Thesis (Ph. D.)--University of Florida, 1991.
Bibliography:
Includes bibliographical references (leaves 178-195).
General Note:
Typescript.
General Note:
Vita.
Statement of Responsibility:
by David George Collart.

Record Information

Source Institution:
University of Florida
Holding Location:
University of Florida
Rights Management:
Copyright [name of dissertation author]. Permission granted to the University of Florida to digitize, archive and distribute this item for non-profit research and educational purposes. Any reuse of this item in excess of fair use or other copyright exemptions requires permission of the copyright holder.
Resource Identifier:
25487850 ( OCLC )
ocm25487850
001645434 ( ALEPH )

Downloads

This item has the following downloads:


Full Text



















ISOLATION AND CHARACTERIZATION
OF A VARIANT HUMAN H2B HISTONE GENE
EXPRESSING ALTERNATIVE mRNAs REGULATED DIFFERENTIALLY
DURING THE CELL CYCLE AND DIFFERENTIATION







By

DAVID GEORGE COLLART


A DISSERTATION PRESENTED TO THE GRADUATE SCHOOL
OF THE UNIVERSITY OF FLORIDA IN PARTIAL FULFILLMENT
OF THE REQUIREMENTS FOR THE DEGREE OF
DOCTOR OF PHILOSOPHY

UNIVERSITY OF FLORIDA
1991




ISOLATION AND CHARACTERIZATION
OF A VARIANT HUMAN H2B HISTONE GENE
EXPRESSING ALTERNATIVE mRNAs REGULATED DIFFERENTIALLY
DURING THE CELL CYCLE AND DIFFERENTIATION
By
DAVID GEORGE COLLART
A DISSERTATION PRESENTED TO THE GRADUATE SCHOOL
OF THE UNIVERSITY OF FLORIDA IN PARTIAL FULFILLMENT
OF THE REQUIREMENTS FOR THE DEGREE OF
DOCTOR OF PHILOSOPHY
UNIVERSITY OF FLORIDA
1991


Dedicated to Mary
the mother of God


ACKNOWLEDGEMENTS
I would like to thank Drs. Gary and Janet Stein for
their support and guidance during my graduate studies.
Their dedication as scientists has provided us with a
laboratory which has always been well equipped and supplied,
allowing us to carry out the experiments necessary for the
completion of our individual projects. In addition, I would
like to thank Gary and Janet for all their support on a
personal level during all the trials which life has offered
throughout my journey from a single, and sometimes
foolhardy, new graduate student to a married, and hopefully
more mature, scientist. I would also like to thank Drs.
Harry Nick, Philip Laipis, Harry Ostrer, Carl Feldherr, Lee
Weber and Eileen Hickey for their helpful advice as members
of my graduate committee. I am also grateful for the
support and help of Dr. Richard Boyce. In addition, I thank
Anna Ramsey-Ewing, Dr. Paul Romain, Suzie Pilapil, Dr. Jane
Lian, Shirwin Pockwinse, Dr. Rita Bortell, Dr. Carlo Croce,
Dr. Kay Huebner and Dr. Linda A. Cannizzaro for their
collaborative efforts on various aspects of this research.
I would like to acknowledge Christine Dunshee for her
photography.
iii


I would like to thank all of the members of the Steins'
lab for their companionship over the years. I thank Linda
Green, Farhad Marashi and Mark Plumb for their friendship
and helpful discussions during the early years of my
graduate studies. I thank Andr van Wijnen for his
friendship and helpful suggestions on writing manuscripts
and Anna Ramsey-Ewing for her friendship and scientific
expertise in tissue culture and transcriptions. I would
also like to thank Tim Morris for his friendship and
discussions on everything in the heavens and on earth, and
Gerry Zambetti for his friendship and for being the
godfather of little Timothy Gerard. In addition, I thank
all my roommates during my pre-marriage days in particular
Charlie for his Scottish outlook on life and money, and Andy
for all of his friendship and support during my years as a
graduate student.
Finally, I would like to thank my family. My parents
and brothers and sisters have always supported me and have
taught me so much about caring. I thank my wife Kathy,
whose prayers and constant love have always been a source of
joy and strength for me. I thank our son Timothy, whose
smiles, giggles and squiggles have taught me what happiness
is and for his suggested addition to this dissertation, "aah
ahh ook." Lastly, I give thanks to God for his constant
love, forgiveness and blessings.
IV


TABLE OF CONTENTS
PAGE
ACKNOWLEDGEMENTS iii
LIST OF FIGURES ix
KEY TO ABBREVIATIONS xii
ABSTRACT xv
CHAPTERS
1) INTRODUCTION
General Background 1
Histone Proteins 2
Structure and Organization of Human Histone Genes... 4
Coupling of DNA Synthesis and Histone Gene
Regulation 4
Transcriptional and Post-Transcriptional Regulation
of Human Histone Genes 5
H2B Histone Gene Expression 7
Overview of Project 7
2) MATERIALS AND METHODS
Materials 9
Materials and Biochemical Reagents 9
Antibiotics, Growth Factors, Inhibitors and Media 10
Enzymes and Kits 11
Nucleic Acids and Nucleotides 12
Propagation and Maintenance of Bacterial Strains.... 13
Growth of Bacterial Strains 13
Storage of Bacterial Strains 13
Propagation and Maintenance of X Bacteriophage 14
Growth and Titration of X Bacteriophage 14
Preparation of plating bacteria 14
Obtaining a titer of X bacteriophage 15
Small scale liquid cultures of X bacteriophage 15
Large scale liquid cultures of X bacteriophage 16
Isolation and Purification of X Bacteriophage.... 17
Storage of X Bacteriophage Stocks 18
Recombinant Phage DNAs 19
Isolation and Purification of Bacteriophage DNA 19
Large Scale X Bacteriophage DNA Isolation 19
Small Scale X Bacteriophage DNA Isolation 20
v


Single-Stranded M13 Bacteriophage Template
Isolation 21
Recombinant Plasmid DNAs 2 3
Isolation and Purification of Plasmid DNA 23
Rapid, Small Scale Plasmid DNA Isolation 23
Large Scale Plasmid DNA Isolation 24
Isolation and Purification of Eukaryotic Genomic DNA 26
Isolation and Purification of Mammalian RNA 27
Selection of Poly A+ RNA 29
Spectrophotometric Quantitation of DNA and RNA 30
Preparation of Radiolabeled DNA 30
Random Oligonucleotide Primed Labeling 30
Labeling the 3' Termini of DNA 31
Labeling the 5' Termini of DNA 32
Analysis of Recombinant DNA Clones 34
Mapping by Single and Multiple Restriction
Endonuclease Digestions 34
Indirect End-Labeled Mapping 34
Southern Blot Analysis 36
Recovery of DNA Fractionated Electrophoretically.... 37
Analysis of Mammalian RNA 38
Agarose-Formaldehyde Denaturing Gel
Electrophoresis 38
Northern Analysis 38
SI Nuclease Protection Analysis 39
In Vitro Nuclear Run-on Transcription Analysis... 41
Library Screening 4 3
Lambda gtll cDNA Library Screening 43
Lambda EMBL4 Genomic DNA Library Screening 49
Cloning and Construct Preparation 51
Subcloning into Plasmid Vectors 51
Construct Preparation 51
Subcloning into Bacteriophage M13 Vectors 59
Transfection of DNA into Bacterial Cells 59
Preparation of Competent EL. coli cells for
Transfection 59
Transfection of Recombinant Plasmid DNA 60
Transfection of Recombinant Bacteriophage M13 DNA 60
DNA Sequencing 61
Sanger Dideoxy-Mediated Chain Termination Method. 61
Chromosomal in situ Hybridization 61
Mammalian Cell Culture 62
HeLa Cell Culture and Synchronization 62
HL60 Cell Culture and Differentiation 63
3T3L1 Cell Culture 63
Transfection and Transient Expression of
Recombinant DNA in 3T3L1 Cells 63
Selection of Stable Polyclonal 3T3L1 Cell Lines.. 64
Induction and Differentiation of 3T3L1 Cells 65
Cell Culture for Chromosome Localization 65
THP-1 Cell culture 66
vi


3) ISOLATION AND CHARACTERIZATION OF A cDNA FROM A HUMAN
HISTONE H2B GENE WHICH IS RECIPROCALLY EXPRESSED IN
RELATION TO REPLICATION-DEPENDENT H2B HISTONE GENES
DURING HL60 CELL DIFFERENTIATION
Introduction 67
Results 68
Isolation of an H2B cDNA Clone From a Igtll
Poly A+ cDNA Library 68
Restriction Endonuclease Mapping of Positive
Clones Isolated From the Agtll Poly A+ cDNA
Library Screening 71
Seguencing Strategy for the Positive Clones
Isolated From the Agtll Poly A+ cDNA Library
Screening 71
Sequencing of a Poly A+ H2B cDNA 78
Comparison of the A.HHC289 Predicted Amino Acid
Sequence With Other H2B Amino Acid Sequences. 83
Comparison of 31 Non-translated Sequences From
AHHC289 and Other Histone cDNAs and Genes.... 86
HHC289 mRNA is Expressed Throughout the HeLa
Cell Cycle and During Inhibition of DNA
Synthesis 86
Transcription of the HHC289 Histone Gene During
the HeLa Cell Cycle 93
HHC289 mRNA is Expressed in a Reciprocal
Relationship With Replication-Dependent H2B
mRNAs During the Onset of HL60 Cell
Differentiation 93
Discussion 100
Structural Analysis of the AHHC289 H2B cDNA 100
Cell Cycle Regulation of the HHC289 H2B Histone
Gene 103
Regulation of HHC289 mRNA During Differentiation
in HL60 cells 105
4) ALTERNATIVE 3' END PROCESSING IS INVOLVED IN
MODULATING EXPRESSION OF A HUMAN HISTONE H2B.1 VARIANT
GENE, LOCATED ON CHROMOSOME 1, IN RESPONSE TO CHANGES IN
THE PROLIFERATIVE STATE OF THE CELL
Introduction 109
Results 110
Isolation of an H2B Genomic Clone From a A.EMBL4
Human Adult Lymphocyte DNA Library 110
Restriction Endonuclease Mapping of Positive
Clones Isolated From the A.EMBL4 Genomic DNA
Library Screening Ill
Cloning and Nucleotide Sequence of the H2B-GL105
Gene 118
Genomic Organization of AHHG5E 12 6
Chromosomal Localization of the H2B-GL105 Gene.. 131
H2B-GL105 mRNA Levels in Proliferating THP-1
Cells 141
vii


H2B-GL105 mRNA Levels in Synchronized HeLa Cells 144
Mapping of the H2B-GL105 5' mRNA Start Site 145
Mapping of the H2B-GL105 mRNA 3' Termini 150
Discussion 155
Structure and Organization of the H2B-GL105 Gene 155
Transcriptional regulation of the H2B-GL105 Gene 156
Post-Transcriptional Regulation of the H2B-GL105
Gene 157
5) SUMMARY AND FUTURE CONSIDERATIONS 160
APPENDICES
A) REAGENTS FOR PROPAGATION AND MAINTENANCE OF BACTERIA
AND BACTERIOPHAGE 169
B) REAGENTS FOR NUCLEIC ACID ISOLATION AND PURIFICATION. 175
C) BACTERIAL STRAINS USED 177
REFERENCES 178
BIOGRAPHICAL SKETCH 196
viii


LIST OF FIGURES
Figure Page
2-1 Schematic diagram of histone gene probes used for
the Agtll poly A+ cDNA library screening 46
2-2 Outline of the screening scheme for the Xgtll
poly A+ cDNA library 48
2-3 Outline of the cloning scheme for the construction
of the H2B construct pGL105SV 54
2-4 Outline of the cloning scheme for the construction
of the H2B 3' deletion construct pGLHOSV 56
2-5 Outline of the cloning scheme for the construction
of the H2B hairpin motif minus construct pGL109SV.. 58
3-1 Plaque dot hybridization analysis of putative
positive clones from the Agtll poly A+ cDNA
library screening 70
3-2 Restriction endonuclease Southern mapping of
A.HHC185 73
3-3 Restriction endonuclease maps of positive clones
isolated from the Agtll poly A+ cDNA library
screening 75
3-4 Sequencing strategy on the positive clones isolated
from the Agtll poly A+ cDNA library screening 77
3-5 Sequencing strategy for the human H2B histone cDNA
AHHC289 80
3-6 (A) Nucleotide and deduced amino acid sequence of
the human H2B histone cDNA A.HHC289. (B) Restriction
map of the cDNA A.HHC289 82
3-7 (A) Comparison of the A.HHC289 histone H2B amino
acid sequence with the sequences of other H2Bs.
(B) DNA sequence comparison of the 3' untranslated
region of selected histone sequences 85
ix


3-8 A.HHC289 mRNA levels during the HeLa cell cycle 88
3-9 H2B mRNA levels during the HeLa cell cycle 90
3-10 Transcription of the A.HHC289 histone gene during
the HeLa cell cycle 95
3-11 A.HHC289 mRNA levels during proliferation and
differentiation of HL60 cells 97
3-12 H2B mRNA levels during proliferation and
differentiation of HL60 cells 99
3-13 Densitometric analysis of the 500 nt replication-
dependent H2B mRNA and the 2 3 00 nt A.HHC289 mRNA
levels during differentiation of HL60 cells 102
4-1 Third round screening of a AEMBL4 human adult
lymphocyte DNA library with a A.HHC289 3' non-
translated trailer probe 113
4-2 Indirect end-labeled mapping of the putative
genomic H2B clone A.HHG6F 115
4-3 Restriction map of putative genomic H2B clones 117
4-4 Southern blot analysis of the genomic H2B subclone
pGL201R 120
4-5 Sequencing strategy for the human histone gene
H2B-GL105 122
4-6 Nucleotide and deduced amino acid sequences of the
human H2B and H2A histone genes from the genomic
clone AHHG5E 124
4-7 Restriction data for the genomic clone A.HHG5E 128
4-8 Southern blot analysis of the genomic clone A.HHG5E
with probes for histones HI, H2A, H2B, H3 and H4... 130
4-9 Southern blot analysis of HeLa genomic DNA with a
H2B-GL105 probe 133
4-10 Southern blot analysis of DNAs from mouse-human
somatic cell hybrids probed with a 32P-labeled
DNA fragment from the human histone gene H2B-GL105. 135
4-11 Presence of the human histone gene H2B-GL105 in a
panel of 21 rodent-human hybrids 137
x


4-12 Regional localization of the human histone gene
H2B-GL105 on human chromosome 1 140
4-13 Northern blot analysis of H2B-GL105 mRNA levels in
proliferating THP-1 cells 143
4-14 Northern blot analysis of H2B-GL105 mRNA levels in
synchronized HeLa cells 147
4-15 SI nuclease mapping of the H2B-GL105 5' mRNA start
site 149
4-16 SI nuclease mapping of the H2A-GL101 5' mRNA start
site 152
4-17 SI nuclease protection analysis of the 3' termini
of the H2B-GL105 mRNA 154
5-1 SI analysis of the 3' termini of H2B mRNA expressed
in 3T3L1 cell lines 165
xi


KEY TO ABBREVIATIONS
APS:
Ammonium Persulfate
ATP:
Adenosine-5'-triphosphate
bp:
Base pair
BSA:
Bovine serum albumin
CIP:
Calf intestinal alkaline phosphatase
O
O

Degree centigrade
DMEM:
Dulbecco's minimal essential medium
dATP:
2'-Deoxyadenosine 5'-triphosphate
dCTP:
2'-Deoxycytidine 5'-triphosphate
dGTP:
2'-Deoxyguanosine 5'-triphosphate
DNase I:
Deoxyribonuclease I
DNA:
Deoxyribonucleic acid
DEPC:
Diethyl pyrocarbonate
dpm:
Disintegrations per minute
DTT:
Dithiothreitol
ddH20:
Double-distilled water
EDTA:
Ethylenediaminetetraacetic acid
g:
Gram
g:
Gravitational force
GAPDH:
Glyceraldehyde-3-phosphate-dehydrogenase
HBSS:
Hank's balanced salt solution
IAA:
Isoamyl alcohol
xii


IPTG:
Isopropyl-0-D-thiogalactopyranoside
1:
Liter
Mg:
Microgram
/m:
Micrometer
mg:
Milligram
ml:
Milliliter
mm:
Millimeter
mM:
Millimolar
M:
Molar
MOPS:
Morpholinepropane-sulfonic acid
ng:
Nanogram
nt:
Nucleotide
Q
o
Optical density
PBS:
Phosphate buffered saline
pfu:
Plaque forming unit
Poly (A):
Polyadenosine
PEG:
Polyethylene glycol
PVP:
Polyvinyl-pyrrolidone
PVS:
Polyvinylsulfonic acid
RF:
Replicative form
RNase A:
Ribonuclease A
RNA:
Ribonucleic acid
rpm:
Revolutions per minute
SDS:
Sodium dodecyl sulfate
TCA:
Trichloro-acetic acid
TEMED:
N,N,N',N'-tetramethylethylenediamine
xiii


tH2B:
TPA:
TTP:
U:
X-gal:
Testis specific H2B
12-0-tetradecanoyl phorbol-13-acetate
Thymidine 5'-triphosphate
Unit of enzyme
5-bromo-4-chloro-3-indolyl-/?-D-galactoside
xiv


Abstract of Dissertation Presented to the Graduate School
of the University of Florida in Partial Fulfillment of
the Requirements for the Degree of Doctor of Philosophy
ISOLATION AND CHARACTERIZATION
OF A VARIANT HUMAN H2B HISTONE GENE
EXPRESSING ALTERNATIVE mRNAs REGULATED DIFFERENTIALLY
DURING THE CELL CYCLE AND DIFFERENTIATION
By
David George Collart
May 1991
Chairman: Gary Stein
Major Department: Biochemistry and Molecular Biology
We have isolated and characterized a cDNA (A.HHC289)
corresponding to a variant human histone H2B gene. The H2B
protein coding region of A.HHC289 is flanked at the 3' end by
a 1798 nt non-translated trailer that contains a region of
hyphenated dyad symmetry and a poly(A) tail. Nuclear run-on
transcription analysis revealed a two fold increase in
transcription of the HHC289 gene during S phase. Northern
blot analysis indicated that the levels of the 2300 nt
HHC289 mRNA species were constant during the HeLa cell
cycle. Northern blot analysis also revealed that the levels
of the 2300 nt HHC289 H2B species increased 10-fold during
HL60 cell differentiation whereas the levels of replication-
xv


dependent H2B mRNAs decreased to less than 1% of those in
proliferating cells.
Using a probe from the AHHC289 cDNA that detects only a
single copy region of the human genome we isolated a variant
human H2B histone gene (GL105) which expresses alternative
mRNAs regulated differentially during the HeLa cell cycle.
This H2B gene encodes both a 500 nt replication-dependent
mRNA and the 2300 nt HHC289 constitutively expressed mRNA.
The 3' end of the cell cycle regulated mRNA terminates
immediately following the region of hyphenated dyad symmetry
typical of most histone mRNAs, whereas the constitutively
expressed HHC289 mRNA has a 1798 nt non-translated trailer
that contains the same region of hyphenated dyad symmetry
but is polyadenylated. The cap site for both H2B-GL105
mRNAs is the same and is located 42 nt upstream of the
protein coding region. The H2B-GL105 histone gene was
localized to chromosome region Iq21-lq23 by chromosomal in
situ hybridization of a H2B-GL105 specific probe and by
analysis of rodent-human somatic cell hybrids. The
H2B-GL105 gene is paired with a functional H2A histone gene
and this gene pair is separated by a bidirectionally
transcribed intergenic promoter region containing consensus
TATA and CCAAT boxes and an OTF-1 element. These results
demonstrate that replication-dependent and constitutively
expressed histone mRNAs can be encoded by the same gene and
indicate that alternative 3' end processing is a major level
xv i


of regulation by which cells can modulate the synthesis of
variant histone proteins during the cell cycle and at the
onset of differentiation.
xvii


CHAPTER 1
INTRODUCTION
General Background
All cells of higher animals and plants carry genetic
information in chromosomes in the form of long linear
molecules called DNA. Within these DNA molecules are
regions containing units of genetic information, called
genes, whose expression results in the production of
proteins or RNA molecules necessary for cellular function.
In the human being more than 1 meter of DNA is contained
within the cell nucleus. DNA must be packaged efficiently
within the cell in such a manner that it can be replicated
when the cell divides, and transcribed at most stages
throughout the cell cycle. This packaging process is
carried out primarily by a group of proteins, approximately
equal in mass to the DNA itself, called histone proteins.
When DNA is associated with histone proteins it is called
chromatin.
When a cell divides, the DNA within a cell is
replicated and the cell must therefore also double the
amount of its histone protein. Because the cell requires a
large quantity of histone protein to package its DNA, it
1


2
must carefully regulate the production of these proteins
according to the cellular needs. The cell must allow for
production of large amounts of histone protein during
periods of rapid growth and division but when DNA
replication ceases, such as outside of S phase or at the
onset of differentiation, the cell must downregulate histone
production to a low level but sufficient to provide for
replacement of histone proteins and for reorganization of
chromatin. Therefore, the study of histone gene expression
provides an excellent opportunity to understand the changes
in gene expression during the transition from a
proliferative to a nondividing state.
Histone Proteins
Histones are a complex family of highly conserved basic
proteins responsible for packaging chromosomal DNA into
nucleosomes (85,86). There are five major classes of
histone proteins (HI, H2A, H2B, H3, and H4) (85,86). Histone
proteins exhibit two levels of diversity: (i) evolutionary
diversity between species and (ii) subtype diversity in a
class (HI, H2A, H2B, H3, or H4) within a species (164). The
subtypes within a species are often referred to as variants
(187,188,195). Replication-dependent histone variants
represent the majority of the histone proteins synthesized
in proliferating cells, and it is well established that
their synthesis is functionally and temporally coupled to


3
DNA replication (1,133,164). Replacement variants are
expressed in nonproliferating cells and are often expressed
in proliferating cells, although normally at a lower level
than their replication-dependent counterparts (188,195).
The third type of variant within a species is the tissue-
specific variant (17,40,83,84,172). Tissue-specific
variants are generally not synthesized in a replication-
dependent manner (40,83,84).
Histone H2B proteins have been studied in a variety of
speciesincluding chicken, mouse, rat and humanwith
respect to their subtype diversity (17,88 and reviewed in
164,195). Although the replication-dependent and tissue-
specific H2B proteins are easily detected in most species,
the replacement histone H2B proteins are not always observed
in humans, possibly due to the lack of sensitivity of the
assay (164). In mouse the H2B.2 variant is the replication
dependent subtype and is abundant in rapidly dividing cells
(195). The H2B.1 variant is the constitutively expressed
subtype and is present in small amounts in rapidly growing
tissues but increases sharply as proliferation slows,
especially after the postnatal growth phase (195). The H2B
subtype specific amino acids of the H2B.1 mouse protein
(Seru, Ala21, Asp25, Lys27, Gly60, Asp68, Gly75, Glu76) differ
from those of the H2B.2 protein only at Ser75 (195),
however, the functional significance of this change is
unclear. In general, replacement histone variants are less


4
hydrophobic than replication-dependent or tissue specific
variants.
Structure and Organization of Human Histone Genes
Human histone genes are a family of moderately
reiterated sequences (75,91,154,164) and are arranged in
clusters; they show no evidence of a simple tandem repeat
organization (75,154,164,194) as do the histone genes of sea
urchin species (65,164,186) and Drosophila (102,164). In
humans, these clusters are localized on chromosomes 1, 6,
and 12 (62,164,171) and are associated with a series of
repetitive DNA sequences (36). Cell cycle dependent histone
genes do not contain introns (91,164), and they encode mRNAs
which are not polyadenylated and have short 5' leader and 3'
trailer sequences (77,164). Although replication-dependent
histone mRNAs are structurally simple, they do have a
characteristic 3' stem-loop motif (164). In contrast,
constitutively expressed histone genes are structurally more
complex. They may contain introns (20,43,182,183,185), and
they encode mRNAs which are polyadenylated and frequently
contain long 5' leader or 3' trailer sequences (16,20,28,45,
46,71-73,78,96,100,110,116,143,144,175,182,183).
Coupling of DNA Synthesis and Histone Gene Regulation
Replication-dependent histone genes are coordinately
expressed during the cell cycle and their expression is


5
coupled with DNA synthesis (3,74,126,130,131). The
abundance of their mRNAs is regulated at both the
transcriptional and post-transcriptional levels
(3,74,130,131,176 and reviewed in 113,150,151,163,164). In
comparison, the mRNAs for replacement histones
(10,19,25,45,71,89,152,155) as well as tissue-specific
histone mRNAs (27,34,64,83,84,93) are not always regulated
in a replication-dependent manner.
Transcriptional and Post-Transcriptional
Regulation of Human Histone Genes
Transcriptional regulation of human histone genes
involves a 2 to 5 fold enhancement in the rate of
transcription within the first 2 hours of DNA synthesis
followed by a return to basal level by mid to late S phase
(4,74,113,131,151,163). However, steady state levels of
histone mRNA are elevated 15-to 20-fold during S phase of
the cell cycle and parallel the rates of DNA synthesis
(3,74,130,131,176). Although the rate of histone gene
transcription peaks within the first 2 hours of DNA
synthesis, histone mRNA levels do not reach a maximum until
4-5 hours into S phase. Taken together these observations
suggest post-transcriptional mechanisms play a role in the
regulation of histone mRNA levels.
Post-transcriptional regulation of histone genes
involves a rapid and selective destabilization of histone
mRNAs toward the end of S phase or in response to inhibition


6
of DNA synthesis (3,76,117,127,128,130,142). However, there
is little or no effect on histone gene transcription by
inhibition of DNA synthesis (4,130,155). This
destabilization of histone mRNA upon inhibition of DNA
synthesis is dependent upon protein synthesis (3,22,76). In
contrast, the mRNA levels of replication-independent histone
genes are not regulated in the same cell cycle dependent
manner as their replication-dependent counterparts and
remain relatively constant upon inhibition of DNA synthesis
(10,19,25,35,45,71,89,152).
The 3' end of cell cycle dependent mRNAs contains a
region of hyphenated dyad symmetry (77) which is essential,
although not sufficient, for coupling histone mRNA stability
with DNA replication (2,101,106,125,159). Graves et al.
(61) observed that moving the 3' stem-loop motif affects
coupling of histone mRNA stability with DNA replication.
Destabilization of histone mRNA initiates at the 3' terminus
(140,142), and is continued rapidly in a 3' to 51 direction
by a ribosome complex-associated endonuclease (141). In
addition, subcellular localization may play a role in
histone mRNA stability (192,193).
The 3' end of most eukaryotic mRNAs is formed by an
endonucleolytic cleavage reaction followed by
polyadenylation of the 3' terminus (12,53,122,129). This
cleavage reaction requires two sequences, a highly conserved
AAUAAA sequence and a downstream GU rich sequence, which


7
flank the cleavage site (12,53,122,129,179). Most histone
mRNAs are not polyadenylated and their mRNA 3' end formation
requires two elements: the stem-loop motif and a purine rich
sequence 3' to the stem-loop motif (9,12,159,178). An
endonucleolytic cleavage reaction occurs 31 to the stem-loop
motif (12,55,95,118,167) and is dependent upon the
interaction of the U7 snRNA with the purine rich sequence
(12,39,118-120,156,167).
H2B Histone Gene Expression
To date, H2B histone gene organization and expression
have been studied most thoroughly in the chicken; five
replication-dependent, one partially replication-dependent,
one uncharacterized and one testis-specific chicken H2B
genes have been described (27,59,60,83,84). Human H2B
histone gene expression has not been reported in such
detail. Although several human histone H2B replication-
dependent genes and one pseudogene have been described
(74,112,131,134,154,164,194), no human replacement or
tissue-specific H2B genes have been reported.
Overview of Project
The overall aim of this work was to isolate and
characterize a variant human histone gene and study its
regulation in response to changes in the proliferative state
of the cell. When we began these studies, information in


8
the literature on nonhuman variant histone genes indicated
the genes for human variant histone proteins would be
structurally more complex than their replication-dependent
counterparts (20,43,71,185). This indication was supported
by the observations of Borun et al. (16) that a fraction of
human histone mRNAs contained short tracts of poly(A). In
addition, because constitutively expressed histone proteins
are synthesized throughout the cell cycle (164,188,195),
their mRNAs must be stable in the absence of DNA synthesis.
Presumably, structural features of constitutively expressed
histone mRNAs could render them resistant to the mechanisms
which rapidly and selectively destabilize replication-
dependent histone mRNAs toward the end of S phase or in
response to inhibition of DNA synthesis (3,76,117,142).
Alternatively, the absence of structural elements endogenous
to replication-dependent histone mRNAs could have the same
result. Therefore, we have focused our attention on the
cloning of a variant human H2B histone gene, and
characterization of its structural features and their effect
on expression in response to changes in the growth state of
the cell.


CHAPTER 2
MATERIALS AND METHODS
Materials
Materials and Biochemical Reagents
The majority of chemicals, reagents and solvents were
purchased from either Fisher Scientific, Springfield, NJ or
Sigma Chemical Co., St. Louis, MO., unless otherwise
indicated. Formamide and X-gal were purchased from Bethesda
Research Laboratories (BRL), Gaithersburg, MD. Bio-Gel
A-1.5m, A-5m, and A-15m (100-200 mesh) agarose beads for gel
filtration, Dowex 50W-X8 cation exchange resin (200-400
mesh) hydrogen form, ultrapure electrophoresis grade agarose
and Zeta-probe nylon membranes were purchased from Bio Rad,
Richmond, CA. Acrylamide (crystallized), APS, ultrapure
bis-acrylamide (N,N'-methylene-bis-acrylamide), DTT
(Cleland's reagent), glycogen (molecular biology grade), and
IPTG were purchased from Boehringer Mannheim Biochemicals
(BMB), Indianapolis, IN. Cronex x-ray film was manufactured
by Dupont, Wilmington, DE. Dimethyl sulfate,
dichlorodimethyl silane, hydrazine, PEG 8000, PVS, XAR-5
x-ray film and X-Omat duplicating film were manufactured by
Eastman Kodak Co., Rochester, NY. SPECTRA/POR
9


10
molecularporous membrane tubing, cellusolve (ethylene glycol
monoethyl ether), dimethyl sulfoxide, ethidium bromide,
2-piperidine, TCA and toluene were purchased from Fisher
Scientific, Springfield, NJ. Cesium chloride (special
biochemical grade 99.9%) was purchased from Gallard-
Schlesinger Industries Inc., Carle Place, NY. OPTI-FLUOR
scintillation cocktail was purchased from Packard Instrument
Co., Inc., Downers Grove, IL. Ficoll 400 was purchased from
Pharmacia Inc., Piscataway, NJ. Type 55 and type 57 instant
sheet film were purchased from Polaroid, Cambridge, MA.
Concentrated liquid scintillator (Liquifluor) was purchased
from Research Products International, Elk Grove Village, IL.
Elutip-d mini-columns, 82 mm and 132 mm nitrocellulose
circles (pore: 0.45 /m) and nitrocellulose sheets were
purchased from Schleicher and Schuell, Inc., Keene, NH. BSA
(fraction V), DEPC, N,N-dimethyl formamide, PVP and TEMED
were purchased from Sigma Chemical Co..
Antibiotics. Growth Factors. Inhibitors and Media
Antibiotics and media were prepared as described in
appendix A. Fetal bovine, calf bovine and horse sera were
obtained from Gibco Laboratories, Grand Island, NY or Flow
Laboratories, McLean, VA. Maltose was purchased from Becton
Dickinson and Co., Cockeysville, MD. Bacto-agar, bacto-
tryptone, bacto-yeast extract, and vitamin-free casamino
acids were purchased from Difco Laboratories, Detroit, MI.


11
Dextrose was purchased from Fisher Scientific, Springfield,
NJ. Geneticin (G418 sulfate) was purchased from Gibco
Laboratories. Chloromycetin sodium succinate
(chloramphenicol sodium succinate) was purchased from Parke-
Davis, Morris Planes, NJ. TGF-/31 was purchased from R&D
Systems, Inc., Minneapolis, MN. Ampicillin (D[-]-a
aminobenzylpenicillin) sodium salt, dexamethasone,
hydroxyurea, insulin (from bovine pancreas), 3-isobutyl-
1-methyl-xanthine, NZ amine (casein enzymatic hydrolysate),
penicillin-G (Benzyl penicillin) potassium salt,
streptomycin sulfate (streptomycin sesquisulfate),
tetracycline, TPA and trypsin (from porcine pancreas)
prepared in IX HBSS without calcium or magnesium were
purchased from Sigma Chemical Co.
Enzymes and Kits
Restriction endonucleases were purchased primarily from
BRL, BMB, and New England Bio Labs (NEB), Beverly, MA.
Enzyme reactions were carried out using the buffer and
incubation conditions described by the manufacturers unless
otherwise indicated. E_*. coli DNA polymerase I large
fragment (Klenow enzyme) was purchased from BRL.
Proteinase K, CIP, nuclease SI and T4 DNA ligase were
purchased from BMB. Trypsin (0.25% in buffered saline) was
purchased from Hazleton Biologies, Inc., Lenexa, KS. The
EXOMETH DNA sequencing kit with reverse transcriptase and


12
the PRIME-IT random primer kit were purchased from
Stratagene, La Jolla, CA. DNase I (DN-EP), DNase I (DN-25)
and RNase A (Type III-A) were obtained from Sigma Chemical
Co.. The Sequenase 2.0 DNA sequencing kit and T4
polynucleotide kinase (cloned) were from United Stated
Biochemical Corp. (USB), Cleveland, OH. Lysozyme was
purchased from Worthington Biochemicals, Freehold, NJ.
Nucleic Acids and Nucleotides
[methyl-3H]Thymidine (20.0 Ci/mmole) [a-32P]dATP
(-3,000 Ci/mmole), [a-32P]dCTP (-3,000 Ci/mmole), (y-32P]ATP
(-3,000 Ci/mmole), [a-32P]UTP (-3,000 Ci/mmole) and
[a-35S]dATP (>1,000 Ci/mmole) radionucleotides were
purchased from Amersham Corp., Arlington Heights, IL, or NEN
Research Products, Wilmington, DE. Oligonucleotides were
synthesized in the recombinant DNA core facility of the
Department of Cell Biology at the University of
Massachusetts Medical Center using an Applied Biosystems
(AB) 380A DNA synthesizer purchased from AB, Foster City,
California. ATP (from equine muscle), dATP (sodium salt),
dCTP (sodium salt), dGTP (sodium salt), TTP (sodium salt),
oligo dT-cellulose and salmon testes DNA (type III) were
obtained from Sigma Chemical Co. RNA molecular weight
markers were purchased from BMB and from BRL. M13mpl8 RF
DNA and M13mpl9 RF DNA were obtained from BMB.


13
Propagation and Maintenance of Bacterial Strains
Growth of Bacterial Strains
Escherichia coli (E. coli) bacteria (see appendix C)
were grown at 37C in YTN medium with vigorous shaking or on
plates containing YTN bottom agar (see appendix A) unless
otherwise indicated. If necessary, ampicillin was added to
the media to a final concentration of 50 /g/ml or
tetracycline to 15 /xg/ml immediately prior to inoculation of
liquid cultures or pouring of YTN agar plates. Liquid
cultures of bacteria were usually grown such that the volume
of the airspace above the culture was at least 3 times that
of the medium, unless otherwise indicated.
Storage of Bacterial Strains
Bacterial colonies were stored for short periods of
time on the surface of bottom agar plates at 4C. These
plates were wrapped in parafilm to prevent moisture loss and
stored inverted.
For long-term preservation of bacterial strains
bacteria were stored in medium containing 50% glycerol at
-70C. To prepare bacterial stocks, 5 ml of YTN was
inoculated with a single colony of bacteria and grown
anaerobically for 18 hours. A 0.5 ml aliquot of the
anaerobic culture was then placed into a vial containing
0.5 ml of sterile glycerol, the contents were mixed and the
vial stored at -70C.


14
Propagation and Maintenance of X Bacteriophage
Growth and Titration of A. Bacteriophage
Lambda Charon 4A (13) and AEMBL4 (54) recombinant phage
were grown in the LE392 strain of EL. coli (15,44,121) and
Agtll recombinant phage in the Y1088 strain of EL. coli
(191). Lambda bacteriophage were grown either on NZCYM
plates as described by Maniatis et al. (108), in small scale
liquid cultures as described by Leder et al. (98) or in
large scale liquid cultures (13,109).
Preparation of plating bacteria (108). A single
bacterial colony (or 1 loop of bacteria from a glycerol
stock) was used to inoculate 50 ml of NZCYM medium,
supplemented with 500 /i 1 of 20% maltose (see appendix A) ,
and grown for 10-15 hours at 37C with vigorous shaking.
Maltose induces the maltose operon, containing the gene
coding for the A receptor which is essential for efficient
absorption of A bacteriophage to bacteria. The cells were
then centrifuged at 3000 rpm in an IEC rotor (2000 X g) for
5 minutes at 4C. The supernatant was discarded and the
cell pellet resuspended in 20 ml (0.4 X the volume of the
original culture) sterile 10 mM MgS04. These bacteria were
stored at 4C and used for up to 3 weeks; however, normally
the cells were used within 1-2 days of their preparation.


15
Obtaining a titer of 1 bacteriophage. Titration of
bacteriophage was carried out essentially as developed by
Felix d'Hrelle in 1920 (165) and described by Maniatis
et al. (108). Serial dilutions of bacteriophage were
prepared in SM and 100 /xl aliquots of each dilution to be
assayed were placed into 13-mm x 100-mm culture tubes. A
100 Ml aliquot of plating bacteria was placed into each
tube, and the tube was vortexed gently and incubated at 37C
for 15-20 minutes to allow the bacteriophage to absorb.
Upon completion of the incubation period, 4 ml aliquots of
NZCYM top agar (4 5C) were added to each tube, and the
contents were vortexed gently and then poured onto NZCYM
bottom agar plates (see appendix A for preparation of NZCYM
top agar and NZCYM bottom agar plates). Upon solidification
of the top agar the plates were inverted and incubated at
37C for 12-18 hours to allow plaque formation.
Small scale liquid cultures of X bacteriophage (98).
Approximately 1/3-1/2 of a resuspended bacteriophage plaque
or 3 X 106 bacteriophage from a stock was mixed with 100 nl
of a fresh bacterial overnight culture in a sterile, 50-ml
polypropylene tube and incubated for 20 minutes at 37C.
Following absorption of the bacteriophage, 4 ml of NZCYM
medium was added and the tube incubated at 37C with shaking
for 6-12 hours until lysis occurred. After lysis, 40 nl of
CHC13 was added and the incubation continued for 15 minutes.


16
Upon completion of the incubation period, the culture was
centrifuged at 5800 rpm in a Beckman JA20 rotor (4000 X g)
for 10 minutes at 4C. Next, the supernatant was recovered,
12 jul of CHClj added and the stock stored at 4C. The
titers were normally in the range of lO8-lO10/ml.
Large scale liquid cultures of X bacteriophage
(13.109). A single bacterial colony (or 1 loop of bacteria
from a glycerol stock) was used to inoculate 100 ml of NZCYM
medium, which was grown for 10-15 hours at 37C with
vigorous shaking. The OD600 of the culture was measured and
the cell concentration calculated, assuming that
1 OD600=8 X 108 cells/ml (108). An aliquot containing 101
cells was removed and centrifuged at 3000 rpm in an IEC
rotor (2000 X g) for 5 minutes at 22C. After
centrifugation the supernatant was discarded and the cells
were resuspended in 3 ml SM. Approximately 5 X 108
bacteriophage were added to the cell suspension and the tube
was incubated at 37C for 2 0 minutes with intermittent
shaking. Upon completion of the incubation period, the
contents of the tube were added to 500 ml of prewarmed NZCYM
and incubated at 37C with vigorous shaking for 7-12 hours
until lysis occurred. After lysis, 10 ml of CHC13 was added
and the incubation continued for 30 minutes. Upon
completion of the incubation, the A bacteriophage were
purified as described by Yamamoto et al. (189).


17
Isolation and Purification of A. Bacteriophage (189)
After treatment of the large scale preparation of
X bacteriophage with CHC13, the culture was chilled to room
temperature, pancreatic DNase I and RNase A were added to a
final concentration of 1 /xg/ml each and the culture was
incubated for 30 minutes at room temperature. Upon
completion of the incubation, solid NaCl was added to a
final concentration of 1 M (29.2 g), dissolved by swirling
and the culture incubated for 1 hour on wet ice. The
culture was then poured into a 500 ml centrifuge bottle, the
CHClj was carefully left behind, and the culture centrifuged
at 9500 rpm in a Beckman JA10 rotor (11,000 X g) for
10 minutes at 4C. The supernatant was poured into a clean
flask; solid PEG 8000 from Eastman Kodak (chemical source is
important) was added to a final concentration of 10% w/v
(50 g), dissolved by slow stirring, and incubated for at
least 1 hour on wet ice (often this precipitation step was
carried out at 4C overnight). After the incubation, the
precipitated bacteriophage particles were recovered by
centrifugation at 9500 rpm in a Beckman JA10 rotor
(11,000 X g) for 10 minutes at 4C. The supernatant was
poured off and the bottles were drained in a tilted position
for 5 minutes. The bacteriophage pellet was gently
resuspended in 8 ml of SM, using a Pasteur pipette and a
rubber bulb to dislodge the bacteriophage particles from the
wall of the centrifuge bottle. The bacteriophage suspension


18
was extracted with 8 ml CHC13/IAA (24:1, v/v) and
centrifuged at 2500 rpm in an IEC rotor (1400 X g) for
15 minutes at 4C. The aqueous phase was recovered and the
above extraction repeated until no PEG interface remained.
Upon completion of the CHC13 extractions 0.75 g of solid
CsCl was added for every ml of solution and mixed gently.
The bacteriophage suspension was then centrifuged at 38,000
rpm in a Beckman 50Ti or 70.1T rotor (135,000 X g) for
24 hours at 4C. The band of bacteriophage particles was
collected using an 18 gauge needle and stored at 4C.
Storage of A Bacteriophage Stocks
Bacteriophage were stored in one of three ways: (i)
Pasteur pipette plugs of bacteriophage plaques were placed
into 1 ml aliquots of SM (initial titers of approximately
106-107/ml) and stored at 4C for 2-4 years, (ii)
supernatants obtained from small scale liquid cultures of
bacteriophage (initial titers of approximately lO8-lO10/ml)
were stored at 4C for 3-5 years, and (iii) suspensions of
bacteriophage in CsCl, obtained from large scale liquid
cultures of bacteriophage purified over CsCl gradients
(initial titers of approximately I01-l011/ml) were stored
at 4C for over 5 years. Stocks of bacteriophage generally
experienced a drop in titer of one order of magnitude during
the first month of storage and an equivalent drop during
each year of further storage.


19
Recombinant Phage DNAs
The A Charon 4A recombinant bacteriophage used in this
study were previously isolated (24,154,196) from a A Charon
4A genomic DNA library. Also used in this study were Agtll
recombinant bacteriophage isolated (see chapter 3) from a
Agtll human liver cDNA library and AEMBL4 recombinant
bacteriophage which were isolated (see chapter 4) from a
AEMBL4 adult human lymphocyte genomic DNA library.
Isolation and Purification of Bacteriophage DNA
Large Scale A Bacteriophage DNA Isolation (109)
DNA was isolated from purified A bacteriophage
as described by Maniatis et al. (109). The purified
bacteriophage preparation was dialyzed twice at room
temperature for 1 hour against a 1000 X volume of 10 mM
NaCl, 50 mM TrisCl (pH 8.0), 10 mM MgCl2, to remove cesium
chloride. The bacteriophage suspension was transferred to a
glass, 50 ml extraction tube and EDTA added to a final
concentration of 20 mM. Proteinase K was added to a final
concentration of 50 /xg/ml, SDS to a final concentration of
0.5% (w/v) and the solution was mixed gently and incubated
at 65C for 1 hour. The solution was extracted with an
equal volume of equilibrated phenol (see appendix B) and
centrifuged at 2500 rpm in an IEC rotor (1400 X g) for
10 minutes at room temperature. Next, the solution was
extracted with an equal volume of a 1:1 mixture (v/v) of


20
equilibrated phenol and CHC13/IAA (24:1, v/v) and
centrifuged at 2500 rpm in an IEC rotor (1400 X g) for
10 minutes at room temperature. The aqueous phase was
recovered and extracted once with an equal volume of
CHC13/IAA (24:1,v/v). Upon completion of the extractions
the aqueous phase was transferred to a dialysis sac and
dialyzed sequentially for 6 hours, at 4C, against 3 1000 X
volumes of 10 mM TrisCl, 1 mM EDTA (pH 8.0). The purified
DNA was quantitated and stored at either 4C or -20C.
Small Scale k Bacteriophage DNA Isolation (149)
Small scale k bacteriophage DNA preparations were
carried out as described by Schweizer (149) A high titer
lysate of k bacteriophage, 0.5 ml, was transferred to a
1.5 ml microcentrifuge tube containing 10 /I 10% SDS (w/v) .
Subsequently, 1 ¡il DEPC was added and the tube mixed by
inversion. Next, 100 jul of 1 M Tris Cl (pH 8.0) and 25 /I
0.5 M EDTA were added, the tube mixed again by inversion and
incubated for 5 minutes at 70C. After the incubation,
80 /I 3 M sodium acetate (pH 5.2) was added and the
incubation continued for 30 minutes on wet ice. The
solution was then centrifuged at 12,000 X g for 15 minutes
at 4C. Following centrifugation, the supernatant was
decanted to a fresh microcentrifuge tube. The tube was
filled with 95% ethanol (-20C) mixed by inversion and
centrifuged at 12,000 X g for 5 minutes at room temperature.


21
The supernatant was discarded and the pellet washed with
1.0 ml 70% ethanol at room temperature. The pellet was
dried briefly under vacuum and resuspended in 200 /l TES
buffer (20 mM TrisCl (pH 7.5), 10 mM NaCl, 0.1 mM EDTA).
The DNA was applied to a 2 ml column of Bio-Gel A-0.5m
agarose beads and eluted with TES buffer. Fractions were
collected and analyzed on a 1% agarose plate containing
5 nq/ml ethidium bromide. Fractions containing
A. bacteriophage DNA were pooled and precipitated with 0.3 M
NaCl and 2.5 volumes of 95% ethanol (-20C) mixed by
inversion and centrifuged at 12,000 X g for 5 minutes at
room temperature. The supernatant was discarded and the
pellet washed with 1.0 ml 70% ethanol at room temperature.
The pellet was dried briefly under vacuum, resuspended in
50 /I 10 mM Tris Cl (pH 8.0), 1 mM EDTA containing DNase-
free RNase (50 /ig/ml) and incubated for 30 minutes at 37C.
Following incubation the DNA was stored at -20C.
Single-Stranded M13 Bacteriophage Template Isolation
Single-stranded M13 Bacteriophage templates for
sequencing cloned inserts were prepared as described by
Sanger et al. (146) and Messing (115). An exponentially
growing culture of host cells, XLl-Blue (21) (see
appendix C), was diluted 1:100 in 2 ml of YTN medium,
infected with a purified M13 plaque and incubated for 6-8
hours at 37C with vigorous shaking. Following the


22
incubation, 1.5 ml of the culture was transferred to a
microcentrifuge tube and spun in an Eppendorf centrifuge at
12,000 X g for 10 minutes at 4C. The supernatant (1.2 ml)
was transferred to a microcentrifuge tube containing 300 ill
of 20% PEG, 2.5 M NaCl and incubated at room temperature for
15 minutes. The remaining supernatant was placed into a
clean microcentrifuge tube and stored at -20C as a phage
stock. After the incubation, the phage suspension was
centrifuged at 12,000 X g for 5 minutes at room temperature
and the supernatant was carefully removed and discarded.
The phage pellet was resuspended in 150 /I TES buffer (20 mM
TrisCl (pH 7.5), 10 mM NaCl, 0.1 mM EDTA) by vortexing for
2 seconds. Equilibrated phenol (50 ¡xl) was added to the
virus suspension, vortexed for two seconds, incubated at
room temperature for 5 minutes, vortexed for an additional
2 seconds and centrifuged at 12,000 X g for 4 minutes at
room temperature. Single-stranded template DNA was
precipitated from 130 ¡xl of aqueous phase with 5 fil 3 M
sodium acetate, 1 /I glycogen (20 mg/ml) and 4 05 /I 95%
ethanol on crushed dry ice for 15 minutes and pelleted by
centrifugation at 12,000 X g for 30 minutes at 4C. The
supernatant was decanted and the pellet washed with 100 /I
70% ethanol at room temperature. The DNA pellet was dried
under vacuum, resuspended in 8 0 ¡xl TES buffer and stored at
-20C.


23
Recombinant Plasmid DNAs
DNA fragments containing HI, H2A, H2B, H3 and H4 genes
from recombinant A. Charon 4A phage (24,154,196) were
subcloned into pBR322 or pUC8 ((131,154,196) and unpublished
data). The plasmid RGAPDH-13 (52) was generously provided
by Dr. Ph. Jeanteur and the plasmid G/32m (38) by Dr. Kenneth
Soprano after preparation from a 550 bp Pst I fragment of
human /32-microglobulin kindly provided by Dr. K. Itakura,
Harvard University.
Isolation and Purification of Plasmid DNA
Bacteria containing plasmids were grown in YTN medium
containing 50 /zg/ml ampicillin and amplification was induced
in the presence of 200 /g/ml chloramphenicol. Plasmid DNA
was isolated and purified using the following methods.
Rapid. Small Scale Plasmid DNA Isolation (80)
Small scale plasmid isolation was carried out using the
rapid boiling method of Holmes and Quigley (80). An
overnight culture (1.5 ml) of plasmid-containing bacteria
was transferred to a microcentrifuge tube and centrifuged in
an Eppendorf centrifuge at 12,000 X g for 2 minutes at room
temperature. The supernatant was decanted and the pellet
vortexed for 2 seconds and resuspended in 150 n1 STET buffer
(8% sucrose, 5% triton X-100, 50 mM EDTA, 50 mM TrisCl
(pH 8.0)). After resuspension, 38 /x 1 of freshly prepared


24
lysozyme (10 mg/ml) was added to the cell suspension, the
mixture boiled for 60 seconds and centrifuged at 12,000 X g
for 10 minutes at room temperature. The pellet was removed
from the microcentrifuge tube using a toothpick and the DNA
precipitated from the supernatant with 188 /I isopropanol at
room temperature for 10 minutes. Following precipitation,
the DNA was pelleted by centrifugation at 12,000 X g for
5 minutes at room temperature, the supernatant decanted and
the pellet washed with 100 ¡j.1 70% ethanol at room
temperature. The DNA pellet was dried under vacuum,
resuspended in 40 /il 10 mM Tris Cl, 1 mM EDTA (pH 8.0)
containing DNase-free RNase (50 /ng/ml) and incubated for
30 minutes at 37C. Following incubation the DNA was stored
at -20C.
Large Scale Plasmid DNA Isolation
Large scale isolation of plasmid DNA was carried out
using the alkaline lysis method described in Birnboim and
Doly (11) and modified by D. Ish-Horowicz (87) A 500 ml
culture of plasmid-containing bacteria was centrifuged at
5000 rpm in a Beckman JA10 rotor (4400 X g) for 15 minutes
at 4C. The supernatant was decanted and the cell pellet
resuspended in 10 ml alkaline lysis solution #1 (see
appendix B) containing 50 mg lysozyme and incubated for
5 minutes at room temperature. After the incubation, 20 ml
of alkaline lysis solution *2 (see appendix B) was added,


25
the solution was mixed gently by inverting until no clumps
remained and further incubated for 15 minutes at 4C
followed by the addition of 15 ml ice cold 5M potassium
acetate (pH 4.8) and incubation at 4C for an additional
15 minutes. Upon completion of the incubation the solution
was transferred to a 50 ml polycarbonate centrifuge tube and
centrifuged at 16,000 rpm in a Beckman JA20 rotor
(31,000 X g) for 20 minutes at 4C. The supernatant was
transferred to a 150 ml corex centrifuge bottle containing
28 ml isopropanol, incubated at room temperature for
15 minutes and the DNA collected by centrifugation at
6000 rpm in a Beckman JA7.5 rotor (6700 X g) for 30 minutes
at 20C. After the centrifugation, the supernatant was
decanted and the pellet washed with 10 ml 70% ethanol at
room temperature. The DNA pellet was dried under vacuum and
resuspended in 8 ml 10 mM TrisCl, 1 mM EDTA (pH 8.0).
The DNA collected after isopropanol precipitation was
purified by CsCl buoyant density gradient centrifugation as
described by Clewell and Helinski (33). Solid CsCl, 8 g,
was added to the resuspended DNA (8 ml) and the solution was
mixed gently until all of the salt was dissolved. A
10 mg/ml stock of ethidium bromide, 0.9 ml for every ml of
DNA solution, was added and the final solution transferred
to Beckman quick seal ultracentrifuge tubes. The plasmid
DNA was banded by centrifugation at 53,000 rpm in a Beckman
70.1T ultracentrifuge rotor (270,000 X g) for 24 hours at


26
20C. The band of supercoiled plasmid DNA was collected
using an 18 gauge needle and the DNA passed over a cation
exchange column of Dowex 50W-X8 followed by chromatography
through a 30 X 1.5 cm BioGel A-15m column, developed with
10 mM TrisCl, 1 mM EDTA (pH 8.0). Fractions containing
plasmid DNA (Vo) were pooled and ethanol precipitated. The
purified DNA was quantitated and stored at either 4C or
-20C. Alternatively, after collection of the supercoiled
plasmid DNA band, the DNA solution was transferred to a
dialysis sac and dialyzed sequentially for 6 hours, at 4C,
against 3 1000 X volumes of 10 mM TrisCl, 1 mM EDTA
(pH 8.0). The purified DNA was then quantitated and stored
at either 4C or -20C.
Isolation and Purification of Eukaryotic Genomic DNA
Eukaryotic genomic DNA was isolated and purified
essentially as described by Gross-Bellard et al. (66) and
modified by Blin and Stafford (14). Approximately 2 X 107
eukaryotic cells were collected by centrifugation at
2500 rpm in an IEC rotor (1400 X g) for 5 minutes at 4C.
The supernatant was decanted and the cells gently
resuspended in cold PBS and recollected by centrifugation at
2500 rpm in an IEC rotor (1400 X g) for 5 minutes at 4C.
After centrifugation, the supernatant was decanted and the
pellet was vortexed gently, resuspended in 10 ml DNA lysis
solution (0.1 X SSC (20 X SSC: 3 M NaCl, 0.3 M sodium


27
citrate; pH 7.4), 0.5% SDS (w/v) 200 jug/ml proteinase K)
and incubated for 24 hours at room temperature with very
gently shaking. Upon completion of the incubation the
solution was extracted with an equal volume of equilibrated
phenol and centrifuged at 2500 rpm in an IEC rotor (1400 X
g) for 10 minutes at room temperature. Next, the solution
was extracted with an equal volume of CHC13/IAA (24:1, v/v)
and an equal volume of equilibrated phenol and centrifuged
at 2500 rpm in an IEC rotor (1400 X g) for 10 minutes at
room temperature. This extraction was repeated until a
clear interphase was visible. The final extraction was with
an equal volume of CHC13/IAA (24:1, v/v) followed by
centrifugation at 2500 rpm in an IEC rotor (1400 X g) for
10 minutes at room temperature. Upon completion of the
extractions the aqueous phase was split into two fractions
of equal volume and transferred to 30 ml corex centrifuge
tubes. The DNA was precipitated with 0.5 ml 3 M sodium
acetate and 15 ml 95% ethanol overnight at 4C. The
precipitated genomic DNA was hooked out with a bent glass
rod, resuspended in 10 mM TrisCl, 1 mM EDTA (pH 8.0) and
stored at 4C. In addition, cellular DNAs used for
chromosome localization were isolated as described (81,82).
Isolation and Purification of Mammalian RNA (131)
Isolation of total cellular mammalian RNA was carried
out as previously described (131). Cells growing in


28
suspension culture were harvested by centrifugation at 2500
rpm in an IEC rotor (1400 X g) for 10 minutes at 4C and
washed twice with cold 1 X spinner salt solution (GIBCO).
The cells were lysed in a solution (1.3 X 07 cells/ml lysis
solution) containing 1.6 mM TrisCl (pH 7.4), 0.8 mM EDTA,
5.0 /xg/ml PVS, 2.0% SDS (w/v) and 500 /g/ml proteinase K for
15-30 minutes at room temperature. Following the
incubation, NaCl was added to 0.25 M and the aqueous phase
was extracted once with an equal volume of a 1:1 mixture
(v/v) of equilibrated phenol and CHC13/IAA (24:1, v/v) and
twice with an equal volume of CHC13/IAA (24:1, v/v). Upon
completion of the extractions the total nucleic acid was
precipitated using 24 fil 5M sodium acetate (pH 5.5) and
three volumes of 95% ethanol overnight at -20C. The total
nucleic acid was collected by centrifugation at 6000 rpm in
a Beckman JA7.5 rotor (6700 X g) for 30 minutes at 20C.
After the centrifugation, the supernatant was decanted and
the pellet washed with 10 ml 70% ethanol at room
temperature. The pellet was dried under vacuum, resuspended
in DNase I digestion buffer (20 mM TrisCl (pH 8.0), 10 mM
CaCl2) and incubated at 37C for 20-40 minutes with
0.1 mg/ml DNase I (electrophoretically pure) which had been
pretreated with proteinase K for 2 hours (174). Following
the incubation, NaCl was added to 0.25 M, SDS to 2.0% (w/v)
and extractions were carried out as described above. Upon
completion of the extractions the RNA was precipitated using


29
a final concentration of 25 mM sodium acetate and three
volumes of 95% ethanol overnight at -20C. The total RNA
was collected by centrifugation at 6000 rpm in a Beckman
JA7.5 rotor (6700 X g) for 30 minutes at 20C. Following
the centrifugation, the supernatant was decanted and the
pellet washed with 10 ml 70% ethanol at room temperature.
The pellet was dried under vacuum, resuspended in ddH20,
quantitated and stored at -20C.
Selection of Poly A* RNA
Poly A+ RNA was selected using a single pass over oligo
dT-cellulose as described by Maniatis et al. (108). A
single oligo dT-cellulose selection greatly enriches for
poly A+ RNA but does not totally remove all of the poly A'
RNA. Total cellular HeLa RNA in loading buffer (10 mM
Tris-HCl (pH 7.6), 500 mM LiCl, 1.0 mM EDTA, 0.1% SDS (w/v))
was heated to 65C for 5 minutes and applied to prewashed
oligo dT-cellulose columns. The columns had been prewashed
with i) three column volumes ddH20, ii) three column volumes
0.1 N NaOH, 5 mM EDTA and iii) three column volumes loading
buffer. After addition of the sample to the column the
flow-through was collected, heated to 65C for 5 minutes,
cooled and reapplied to the column. The column was then
washed with 5 to 10 column volumes of loading buffer,
followed by 4 column volumes of loading buffer containing
100 mM NaCl. The poly A+ RNA was eluted with 2 to 3 column


30
volumes of elution buffer (10 mM TrisCl (pH 7.5), 1.0 mM
EDTA, 0.05% SDS (w/v)) and precipitated with 300 mM sodium
acetate (pH 5.2) and 2.2 volumes 95% ethanol overnight at
-20C. RNA was collected by centrifugation at 10,000 rpm in
a Beckman JA20 rotor (12,000 X g) for 30 minutes at 20C.
Following the centrifugation, the supernatant was decanted
and the pellet washed with 10 ml 70% ethanol at room
temperature. The pellet was dried under vacuum, resuspended
in ddH20, quantitated and stored at -20C.
Spectrophotometric Quantitation of DNA and RNA
Nucleic acids, both DNA and RNA, were routinely
quantitated by OD at 260 nm. An OD at 260 nm (OD260) of 1
corresponds to approximately 50 ng/ml for double-stranded
DNA, 40 /xg/ml for single-stranded DNA and RNA, and 20 ¡jlg/ml
for oligonucleotides (108). The purity of the nucleic acid
was often assessed by calculating the 260 nm:280 nm optical
density ratio (OD260:OD2g0) Pure preparations of DNA and RNA
have an OD260:OD280 of 1.8 and 2.0, respectively.
Preparation of Radiolabeled DNA
Random Oligonucleotide Primed Labeling
A random oligonucleotide priming technique (47,48,169),
simplified by Roberts and Wilson (139), employing T7 DNA
polymerase (57,168), was carried out using a random primer
kit. The volume of a sample, containing 25 ng of DNA


31
template (linearized plasmid DNA or an isolated DNA
fragment) to be labeled, was adjusted to 23 ill with ddH20.
The solution was then heated in a boiling H20 bath for
5 minutes and centrifuged briefly to collect the
condensation. After heat denaturation, 10 /I of 5 X primer
buffer (supplied by the manufacturer) 5 /il labeled
nucleotide (~50 /xci) ([a-32P]dCTP (~3,000 Ci/mmole) ) and
1 ill diluted T7 DNA polymerase (2 units//il in enzyme
dilution buffer supplied by the manufacturer) were added to
the template DNA and the mixture was incubated at 37C for
2-10 minutes. Upon completion of the incubation period, the
reaction was quenched by the addition of 2 ill of stop
mixture. The volume of the reaction mixture was adjusted to
1.2 ml with Elutip-d low salt buffer (200 mM NaCl, 20 mM
TrisCl (pH 7.4), 1 mM EDTA) and the labeled DNA purified
using a Schleicher & Schuell Elutip-d column as described by
the manufacturer. The labeled DNA was stored at 4C until
use.
Labeling the 31 Termini of DNA (124)
DNA hybridization probes labeled at the 31 termini were
prepared as described by O'Farrell (124). The volume of the
solution containing the DNA fragment to be labeled was
adjusted to 12 /il with ddH20. BSA, 2.5 ill of a 1 mg/ml
stock, was added to the DNA solution along with 2.5 ill of
Boehringer Mannheim restriction endonuclease 10 X digestion


32
buffer L and 2.5 m! T4 DNA polymerase (1 unit/Ml) and the
solution incubated for 2-5 minutes at 37C. Immediately
following the incubation, 1 Ml of each of the following was
added: 2 mM dGTP, 2 mM dATP, 2 mM TTP and 2 1 (-2 0 ¡jlci) of
[a-32P]dCTP (-3,000 Ci/mmole), and incubation was continued
for 1 minute at 37C. Upon completion of the incubation,
0.5 Ml of 2 mM dCTP was added and the incubation continued
for 10 minutes at 37C. The reaction was stopped by heating
the solution for 5 minutes at 70C followed by the addition
of 1 Ml of 250 mM EDTA. The volume of the quenched reaction
mixture was brought to 1.2 ml with Elutip-d low salt buffer
and the labeled DNA purified using a Schleicher & Schuell
Elutip-d column as described by the manufacturer. The
labeled DNA was stored at 4C until use.
Labeling the 5' Termini of DNA
DNAs to be labeled at the 51 termini and used as
hybridization probes were treated with CIP as described by
Chaconas and van de Sande (26) and modified by Maniatis
et al. (108), followed by 5' end labeling with T4
polynucleotide kinase and [y-32P]ATP (-3,000 Ci/mmole) as
described by Maxam and Gilbert (114) and modified by
Maniatis et al. (108). The volume of the solution
containing the DNA fragment to be labeled was adjusted to
44 Ml with ddH20. CIP 10 X buffer (500 mM TrisCl (pH 9.0),
10 mM MgCl2, 1 mM ZnCl2, 10 mM spermidine), 5 Ml/ and 1 nl


33
of CIP (0.2 units/Ml for DNA fragments with protruding
5' termini or 1 unit/Ml for DNA fragments with blunt or
recessed 5' termini) were added to the DNA solution and
incubated for 15 minutes at 37C followed by incubation for
15 minutes at 56C. Upon completion of the 56C incubation
a second aliquot of CIP was added to the reaction mixture
and the successive incubations were repeated. The reaction
was quenched by the addition of 4 0 Ml ddH20, 10 /I 10 X STE
buffer (100 mM Tris*Cl (pH 8.0), 1 M NaCl, 10 mM EDTA) and
5 /x 1 10% SDS (w/v) and the CIP was heat inactivated for
30 minutes at 68C. Upon completion of the incubation the
reaction mixture was extracted twice with an equal volume of
equilibrated phenol and twice with an equal volume of
CHClj/IAA (24:1, v/v) Following the addition of 6.3 /il 5 M
NaCl, 1 Ml glycogen (20 mg/ml) and 330 Ml 95% ethanol, the
DNA was precipitated on crushed dry ice for 15 minutes and
pelleted by centrifugation at 12,000 X g for 30 minutes at
4C. After centrifugation, the supernatant was decanted and
the pellet washed with 100 ¡il 70% ethanol at room
temperature. The pellet of DNA was dried under vacuum,
resuspended in 44 Ml 10 mM Tris*Cl, 1 mM EDTA (pH 8.0) and
stored at -20C until the 5' end labeling reaction was
carried out.
Spermidine, 1 Ml of a 100 mM stock, and 5 Ml 10 X
kinase buffer (660 mM Tris*Cl (pH 9.5), 100 mM MgCl2, 100 mM
2-mercapto-ethanol) were added to the solution of CIP


34
treated DNA and incubated for 5 minutes at 70C. After the
incubation, the solution was snap frozen in crushed dry ice.
The frozen solution was quickly thawed, added to 10 /I
(-100 Mci) of [y32P] ATP (~3,000 Ci/mmole) and incubated for
30 minutes at 37C. The reaction was stopped by the
addition of 1 /xl 100 mM EDTA and 1.2 ml Elutip-d low salt
buffer, and the labeled DNA was purified using a Schleicher
& Schuell Elutip-d column as described by the manufacturer.
The labeled DNA was stored at 4C until use.
Analysis of Recombinant DNA Clones
Mapping by Single and Multiple Restriction Endonuclease
Digestions
Single and double digests were carried out on plasmid
and A. bacteriophage DNAs using restriction endonucleases and
analyzed electrophoretically on horizontal agarose gels
(108) .
Indirect End-Labeled Mapping
Restriction mapping of recombinant A. phage by indirect
end-labelling was carried out as described by Rackwitz
et al. (137) using hybridization conditions defined by
Little and Cross (103). Oligonucleotides complementary to
the cos sequence of A. phage were 51 end labeled using
T4 polynucleotide kinase. Oligonucleotides (33 ng) in 1 /I
were combined with 1 /il 10 X kinase buffer, 2.5 ¡jlI (~25 nci)
of [y-32P]ATP (-3,000 Ci/mmole), 0.4 ¡jlI 100 mM spermidine,


35
3.8 /I ddH20 and 1.5 ill of 10 unit//xl T4 polynucleotide
kinase and the reaction mixture was incubated for 30 minutes
at 37C. The reaction was stopped by adding 0.4 ill 250 mM
EDTA and 100 ill 10 mM Tris Cl, 1 mM EDTA (pH 8.0) and
heating for 10 minutes at 70C. The reaction mixture was
extracted with an equal volume of a 1:1 mixture (v/v) of
equilibrated phenol and CHC13/IAA (24:1, v/v). Following
the addition of 6.7 jul 5 M NaCl, 1 ill glycogen (20 mg/ml)
and 350 ill 95% ethanol, the oligonucleotides were
precipitated on crushed dry ice for 15 minutes and pelleted
by centrifugation at 12,000 X g for 30 minutes at 4C.
After centrifugation, the supernatant was decanted and the
pellet washed with 100 ill 70% ethanol at room temperature.
The pellet was dried under vacuum, resuspended in 100 ¡il 10
mM TrisCl, 1 mM EDTA (pH 8.0) and stored at 4C until the
hybridization was carried out.
To 10 nl of partially restricted A phage DNA
(0.5-1.0 /iq) was added 1 /xl of a solution containing: 2% SDS
(w/v) 50 mM EDTA, 1.0 iiq/iil proteinase k and 0.026 ng of
5' end labeled oligonucleotides, and the mixture was
incubated for 10 minutes at 75C. Immediately following the
incubation, the mixture was transferred to 45C and
hybridized for 60 minutes. Following the hybridization, the
samples were placed on wet ice and 1 /xl TPE tracking dye
(36 mM TrisCl (pH 7.7), 30 mM NaH2P04 (monobasic), 1 mM
EDTA, 0.05% bromophenol blue and 0.05% xylene cyanol,


36
10% glycerol) was added. The samples were then loaded onto
a 0.5% agarose TPE (36 mM TrisCl (pH 7.7), 30 mM NaH2P04
(monobasic), 1 mM EDTA) gel and electrophoresed for 24-28
hours at 40 watts with buffer recirculation. Following
electrophoresis, the gel was dried under vacuum for 1 hour
at 80C. Autoradiography was performed using pre-flashed
Kodak XAR-5 X-ray film and a Cronex Lightning Plus screen at
-70"C. I would like to acknowledge Dr. Paul L. Romain and
Suzie Pilapil for their work on the mapping analysis of the
A.HHG5E genomic clone.
Southern Blot Analysis
Restriction endonuclease digested DNAs were separated
by electrophoresis through agarose gels and transferred to
Zeta-Probe blotting membrane as described by Reed and Mann
(138). The agarose gel was soaked in 25 mM HC1 for
10 minutes at room temperature, rinsed briefly in ddH20 and
the DNA was transferred with 0.4 N NaOH overnight at room
temperature. Following transfer, the membrane was baked in
a vacuum oven at 80"C for 1 hour, prewashed in 0.5% SDS
(w/v), 0.1 X SSC for one hour at 65"C and then prehybridized
and hybridized using a modification of the method described
by Wahl et al. (180). Prehybridization was carried out for
two hours in 50 mM sodium phosphate (pH 6.5), 0.45% SDS
(w/v) 47% formamide, 9% dextran sulfate and 500 /xg/ml
double stranded E^. coli DNA. After prehybridization,


37
32P-labeled probe was added to the mix to a concentration of
1 X 106 dpm/ml and hybridized for 6-18 hours at 48C. The
hybridized membrane was then washed three times as follows:
1) in 100 ml 100 mM potassium phosphate (pH 7.4) for
30 minutes at room temperature; 2) in 100 ml 100 mM
potassium phosphate (pH 7.4) for 30 minutes at 60C; 3) in
100 ml 1 X SSC and 0.2% SDS (w/v) for 30 minutes at 60C.
Autoradiography was performed using pre-flashed Kodak XAR-5
X-ray film and a Cronex Lightning Plus screen at -70C.
32P-labeled probes used for Southern analysis of the genomic
H2B clone A.HHG5E were as follows: HI, 1445 nt Pst I/Eco RI
fragment from pFNC16 (131); H2A, 980 nt Sst I fragment from
pFF435 (112) ; H2B, 340 nt Eco RI/Xho I fragment from A.HHC289
(Figure 3-6); H3, 2100 nt Eco RI fragment from pST519 (111);
H4, 1740 nt Hind III/Eco RI fragment from pF0108A (131).
Recovery of DNA Fractionated Electrophoreticallv
An agarose slice containing the DNA fragment of
interest was cut into very fine pieces, using a clean
scalpel, and the resulting agarose puree was then placed
into the pocket formed by folding a large piece of parafilm
into guarters. The puree was frozen on crushed dry ice
followed by thawing while applying pressure with the thumb
and forefinger. The resulting liquid which was forced out
of the puree was drawn off with a pipette and placed into a
clean tube. This process, including the freezing step, was


38
repeated until no additional liquid was squeezed out of the
puree. The DNA fragment was purified using a Schleicher &
Schuell Elutip-d column as described by the manufacturer and
was stored at -20C.
Analysis of Mammalian RNA
Agarose-Formaldehyde Denaturing Gel Electrophoresis
RNA was size fractionated in a 1.5% agarose, 1 X MOPS
(20 mM MOPS (pH 7.0), 5 mM sodium acetate, 1 mM EDTA), 6%
formaldehyde gel using a 1 X MOPS, 3.7% (v/v) formaldehyde
running buffer, a modification of the method used by Lehrach
et al. (99) .
Northern Analysis
Northern blot analysis was carried out as follows: the
RNA was size fractionated as described above and transferred
as described by Thomas (170) to Zeta-Probe blotting membrane
using 20 X SSC as the transfer buffer. Following transfer,
the membrane was baked in a vacuum oven at 80C for 1 hour,
prewashed in 0.5% SDS (w/v), 0.1 X SSC for one hour at 65C
and then prehybridized and hybridized using a modification
of the method described by Wahl et al. (180).
Prehybridization was carried out for two hours in 50 mM
sodium phosphate (pH 6.5), 0.45% SDS (w/v), 47% formamide,
9% dextran sulfate and 500 /g/ml double stranded E_¡_ coli
DNA. After prehybridization, 32P-labeled probe was added to


39
the mix to a concentration of 1 X 106 dpm/ml and hybridized
for 6-18 hours (the temperature of the hybridization was
dependent upon probe size and percent GC content and ranged
from 38C-48C). The hybridized Zeta-Probe membrane was
then washed five times as follows: 1) 10 minutes at room
temperature with 5 X SSC, 1 X Denhardt's; 2) 30 minutes at
60C with 5 X SSC, 1 X Denhardt's; 3) 30 minutes at 60"C
with 2 X SSC, 0.1% SDS (w/v); 4) 30 minutes at 60C with
1 X SSC, 0.1% SDS (w/v); 5) 30 minutes at 60C with
0.1 X SSC, 0.1% SDS (w/v). Autoradiography was performed
using pre-flashed Kodak XAR-5 X-ray film and a Cronex
Lightning Plus screen at -70C. The developed X-ray films
were analyzed by laser densitometry using a LKB 2400 GelScan
XL densitometer. 0-particle analysis of hybridized
membranes was carried out using a 0etagen (Waltham, MA)
Betascope 603 blot analyzer.
SI Nuclease Protection Analysis
SI analysis was carried out according to Berk and Sharp
(6) as modified by Haegeman et al. (67). DNA fragments used
as probes (preparation described above) were treated with
CIP as described by Chaconas and van de Sande (26) and
modified by Maniatis et al. (108) followed by 5' end
labeling with T4 polynucleotide kinase and [y-32P]ATP
(-3,000 Ci/mmole) as described by Maxam and Gilbert (114)
and modified by Maniatis et al. (108). Alternatively, DNA


40
fragments were 3' end labeled with T4 DNA polymerase and
[a-32P]dCTP (-3,000 Ci/mmole) as described by O'Farrell
(124). The RNA to be analyzed was co-precipitated with
labeled DNA probe using 0.3 M NaCl, 1.0 /il glycogen
(20 mg/ml) and 2.5 volumes of 95% ethanol for 15 minutes on
crushed dry ice and pelleted by centrifugation at 12,000 X g
for 30 minutes at 4C. After centrifugation, the
supernatant was decanted and the pellet washed with 100 /i1
70% ethanol at room temperature. The pellet was dried under
vacuum, resuspended in 5 |il 5 X SI hybridization buffer
(5 X: 2 M NaCl, 200 mM Pipes (pH 6.4), 5 mM EDTA) and 20 /il
deionized, recrystallized formamide. Hybridization
mixtures, 25 /il, were heated at 90C for 10 minutes and then
annealed at 55C for at least three hrs. Annealing
reactions were guenched with eight volumes, 200 /il, SI
nuclease buffer (25 mM NaCl, 30 mM sodium acetate (pH 4.6),
1.0 mM ZnS04) followed by incubation at 37C for 30 minutes
with 900 units of SI nuclease in the presence of 20 /ig
salmon sperm DNA. After SI digestion the samples were
extracted with an equal volume of a 1:1 mixture of
equilibrated phenol and CHCl3/IAA (24:1, v/v) and ethanol
precipitated using 0.3 M NaCl, 1.0 /il glycogen (20 mg/ml)
and 2.5 volumes of 95% ethanol overnight at -20C. The
protected fragment was pelleted by centrifugation at
12,000 X g for 30 minutes at 4C. Following centrifugation,
the supernatant was decanted and the pellets were washed


41
with 100 ill 7 0% ethanol at room temperature. The pellets
were dried under vacuum, resuspended in 6 nl loading buffer
(95% formamide, 20 mM EDTA, 0.05% bromophenol blue and 0.05%
xylene cyanol) and the samples were heat denatured and
loaded onto a 6% polyacrylamide sequencing gel containing
7.8 M urea (108). After electrophoresis the gel was soaked
in 10% acetic acid, 12% methanol for 15-30 minutes to remove
urea, dried under vacuum at 80C, and placed against
preflashed Kodak XAR-5 X-ray film and a Cronex Lightning
Plus screen at -70C.
In Vitro Nuclear Run-on Transcription Analysis
The in vitro nuclear run-on transcription assays were
performed using a modification (4) of the method used by
Flint et al. (51). Cells were harvested by centrifugation
and the cell pellets washed twice in cold isotonic buffer
(125 mM potassium phosphate, 30 mM Tris-HCl (pH 7.9), 5 mM
MgCl2, 10 mM /3-mercaptoethanol) Cells were disrupted by
homogenization (6-12 strokes) with a Dounce homogenizer,
Wheaton type A pestle. After >90% of the cells had been
lysed, nuclei were pelleted by centrifugation at 2000 rpm
for 10 minutes in an IEC centrifuge at 4C and resuspended
in nuclei storage buffer (50 mM Tris-HCl (pH 8.3), 5 mM
MgCl2, 0.1 mM EDTA) containing 40% glycerol. Nuclei were
aliquoted and used fresh in the in vitro transcription
reactions. Reactions typically contained 107 nuclei,


42
100 nCi a-32P-UTP (3000 Ci/mmole), 1 mM ATP, 0.25 mM GTP and
0.25 mM CTP in a final volume of 130 ¡jl 1 and were incubated
for 30 minutes with intermittent shaking at 30C.
Radiolabeled RNAs were isolated by treatment of nuclei with
DNase I (100 /xg/ml) in the presence of 0.6 M NaCl, 50 mM
Tris-HCl (pH 7.5), 20 mM MgCl2 for 15 minutes at room
temperature. The mixture was then incubated with
proteinase K (200 /g/ml) for 30-60 minutes at 37 C in the
presence of 150 mM NaCl, 12.5 mM EDTA, 100 mM Tris-HCl
(pH 7.5) and 20 mM MgCl2. Sodium acetate (pH 5.5) was added
to 200 mM and nucleic acids were extracted several times by
the hot phenol method (32,148). To the aqueous solution of
32P-labeled RNAs, 150 nq of yeast RNA and 2.5 volumes of 95%
ethanol were added. Precipitation was overnight at -20C.
Radiolabeled transcripts were resuspended in 1 ml of 10 mM
Tris-HCl (pH 8.0), 1 mM EDTA and an aliquot of each sample
was precipitated with 150 /xg yeast RNA and cold 10% TCA.
TCA-precipitable counts were determined by liquid
scintillation spectrometry.
Following preparation of Southern blots (157) of
electrophoretically-separated restriction endonuclease-
digested plasmid DNAs or slot blots (prepared using a slot
blot apparatus with conditions as described by Schleicher
and Schuell Inc.) of linearized plasmid DNAs, DNA excess
hybridizations were carried out. Southern or slot blots on
nitrocellulose were prehybridized in 1 M NaCl, 20 mM


43
Tris-HCl (pH 7.4), 2 mM EDTA, 0.1% SDS (w/v), 5 X
Denhardt's, 25 jug/ml denatured E_ coli DNA and 12.5 mM
sodium pyrophosphate at 65C for at least 6 hours.
Hybridizations were conducted at 65C for 72 hours in
1 M NaCl, 20 mM Tris-HCl (pH 7.4), 2 mM EDTA, 0.1% SDS
(w/v), 2.5 X Denhardt's, 25 /g/ml E_¡_ coli DNA with
32P-labeled transcripts at 5 X 105-1 X 106 TCA-precipitable
dpm/ml of hybridization solution. Blots were washed at 65C
for 15 minutes in fresh prehybridization solution, 1 hour in
2 X SSC, 0.1% SDS (w/v), overnight in 2 X SSC, 0.1% SDS
(w/v) and 1 hour in 0.2 X SSC, 0.1% SDS (w/v).
Autoradiography was performed on air dried filters with pre
flashed XAR-5 or Cronex film and Cronex Lightning Plus
screens at -70C. The developed X-ray films were analyzed
by laser densitometry using a LKB 2400 GelScan XL
densitometer. I would like to acknowledge Anna Ramsey-Ewing
for her work on the transcriptional analysis of the
H2B-GL105 gene.
Library Screening
Lambda qtll cDNA Library Screening
A Agtll human liver cDNA library, made from total
cellular poly A+ RNA and generously provided by Dr. S. L. C.
Woo (Baylor College of Medicine, Department of Cell
Biology), was screened with the 32P-labeled human histone
probes indicated in Figure 2-1. The library screening was


44
carried out as indicated in Figure 2-2 according to the
procedure described by Benton and Davis (5) with the
following modifications in the hybridization conditions.
The filters were prehybridized at 60C for two hours in a
solution containing 5 X Denhardt's (100 X: 2% ficoll, 2%
polyvinyl-pyrrolidone), 5 X SSPE (20 X: 3.6 M sodium
chloride, 200 mM sodium phosphate, 20 mM EDTA; pH 7.4), 0.2%
SDS (w/v), 0.1% w/v BSA and 500 nq/ml double stranded
E. coli DNA. Hybridization was carried out at 60C for 24
hours in a solution containing 0.5 X Denhardt's, 5 X SSPE,
0.2% SDS (w/v), 0.01% w/v BSA, 500 ng/ml EL coli DNA and
2 X 106 dpm/ml heat-denatured probe DNA fragment.
Hybridized filters were then washed at 24C for 30 minutes
in 100 mM potassium phosphate (pH 7.4) followed by two
30 minute washes at 60C in 2 X SSPE, 0.3% SDS (w/v).
Autoradiography was performed using Kodak XAR-5 X-ray film
and a Cronex Lightning Plus screen at -70C.
Plaque dot analysis was carried out according to the
procedure described by Powell et al (132) A 100 /il
aliquot of plating bacteria was placed into a tube and 4 ml
of NZCYM top agar was added to the tube, vortexed gently and
poured onto a NZCYM bottom agar plate. Upon solidification


Figure 2-1. Schematic diagram of histone gene probes used for the Acrtll poly A+ CDNA
library screening
A Agtll human liver cDNA library was screened with the 32P-labeled human histone
probes A, B, C, D, and E indicated by the dotted lines, set apart by arrows, beneath
each restriction map. The human histone probes are as follows: A, 2100 nt Eco RI H3
fragment; B, 1740 nt Hind III/Eco RI H4 fragment; C, 550 nt Sst I H2B fragment; D,
980 nt Sst I H2A fragment; E, 1445 nt Pst I/Eco RI HI fragment. A.HHG41, A.HHG55 and
A.HHG415 are X Charon 4A recombinant bacteriophage which were previously isolated
(24,154) from a X Charon 4A genomic DNA library.


H3
H4
^ t XHHG 41
Eco Rl
:L
*
Hind III
XHHG 55
XHHG 415
a\


Figure 2-2. Outline of the screening scheme for the Acrtll poly A+ cDNA library
A Agtll human liver cDNA library was screened as described by Benton and Davis
(5) The A.gtll cDNA library was grown on 100 mm diameter petri dishes. Following
overnight incubation, duplicate nitrocellulose filters were lifted from each plate
and prepared as described (5). One set of filters was hybridized to the H3 and H4
probe at 60C and the other set to the H2A, H2B and HI probes at 58C as described by
Lawn et al. (97). Hybridized filters were then washed and autoradiography performed.
The positive plaques (or plaque areas) were picked and screened a second time.
Single well isolated positive plaques were picked and screened a third time. Upon
completion of the third round of screening 151 H3/H4 positives and 158 H2A/H2B/H1
positive plaques had been obtained. Positive plaques were separated according to
individual histone class, using the human histone probes described in Figure 2-1, by
plaque dot analysis.


xgt-11 Poly A+
cDNA Library,
(Human)
. Plate Positives and
^ Repeat Two Times.
i
Pick
Positives
1
Check For
Positives
Upon Completion of
Third Round of
Screening:
H3. HA = 151 Positives
ILL, H2A, H2B = 158 Positives
Separate Plaques,
03


49
of the top agar, 0.5 /I aliquots of A bacteriophage
dilutions were spotted onto the plate and allowed to dry.
The plate was inverted and incubated at 37C for 5-12 hours
to allow plaque formation. Up to 3 nitrocellulose filter
lifts were prepared from the plate and hybridized to various
probes as described in the previous section.
Lambda EMBL4 Genomic DNA Library Screening
An adult human lymphocyte genomic library, containing
inserts of between 15 and 20 Kb, was prepared by inserting
Sau3A I partially digested adult human lymphocyte genomic
DNA into the Bam HI site of A.EMBL4 and was generously
provided by Dr. Paul Dobner (University of Massachusetts
Medical Center Department of Molecular Genetics and
Microbiology). A complete equivalent of this AEMBL4 human
genomic DNA library (8 X 105 phage) was grown on 14 150 mm
diameter petri dishes and screened with a 32P-labeled H2B 3'
non-translated probe (900 nt Sac I fragment isolated from
AHHC289) according to the procedure described by Benton and
Davis (5). LE392 (121) plating bacteria, 0.3 ml, were mixed
with 50,000 pfu of recombinant phage and incubated for
20 minutes at 37C. Next, 6.5 ml melted top agarose (46C)
was added and the mixture poured onto a 150 mm NZCYM bottom
agar plate. The top agarose was allowed to solidify at room
temperature and then incubated 10-12 hours (Or until the
plaques reach a diameter of approximately 1.5 mm) at 37C.


50
Following overnight incubation the plates were chilled for
60 minutes at 4C. For filter lifting, plates were removed
from the cold box and nitrocellulose filters carefully laid
on top of the agarose for 2-3 minutes. The orientation of
the filters was marked in three asymmetric positions using a
nitrocellulose marking pen. The filters were then removed
and soaked in denaturing solution (1.5 M NaCl, 0.5 N NaOH)
for 30-60 seconds and transferred into neutralizing solution
(1.5 M NaCl, 0.5 M TrisCl (pH 8.0)) for 5 minutes.
Subseguently the filters were rinsed in 2 X SSPE, blotted
dry on Whatman 3 MM paper and baked for 2 hours at 80C in a
vacuum oven. Hybridization of these filters was carried out
essentially as described by Lawn et al. (97) with the
following modifications in the hybridization conditions.
The filters were prehybridized at 60C for two hours in a
solution containing 5 X Denhardt's, 5 X SSPE, 0.2% SDS
(w/v) 0.1% w/v BSA and 500 /g/ml double stranded E_¡_ coli
DNA. Hybridization was carried out at 60C for 24 hours in
a solution containing 0.5 X Denhardt's, 5 X SSPE, 0.2% SDS
(w/v), 0.01% BSA, 500 /g/ml EL. coli DNA and 2 X 106 dpm/ml
heat-denatured probe DNA fragment. Hybridized filters were
then washed (10 ml wash solution/filter) at 24C for 0.5
hour in 100 mM potassium phosphate (pH 7.4) followed by two
0.5 hour washes at 60C in 2 X SSPE, 0.3% SDS (w/v).
Autoradiography was performed using Kodak XAR-5 X-ray film
and a Cronex Lightning Plus screen at -70C. Following


51
hybridization, the positive plaques (or plaque areas) were
picked and placed into 1 ml of SM containing a drop of
chloroform. These positive plaques were screened a second
time by hybridization. Single, well isolated, positive
plaques were picked and screened a third time.
Cloning and Construct Preparation
Subcloning into Plasmid Vectors
DNA fragments were isolated from agarose gels, and
ligations into the appropriate vectors were carried out
using T4 DNA ligase as described by the manufacturer. The
ligation products were used to transfect competent E_*. coli
DH5a (8) (see appendix B) bacteria. The 6100 nt Pst I
fragment, 4100 nt Eco RI fragment and 6100 nt Eco RI
fragment from A.HHG5E (Figure 4-7) were isolated and cloned
into pUC19 and the resulting subclones termed pGL105, pGLIOl
and pGL102, respectively.
Construct Preparation
Construct I (pGL105SV) is the control construct
containing the entire A.HHG5E H2B gene, promoter and 2000 nt
of 3' untranslated region (Figure 2-3). The 6200 nt Pst I
fragment from A.HHG5E was isolated and ligated, using T4 DNA
ligase, into the Pst I site of pUC19 to obtain the
intermediate (pGL105). Construct I was obtained by
isolating the Eco RI/Eco RI SV40 enhancer element from


52
pSVE108A (153), blunt-ending its termini with the Klenow
fragment of DNA polymerase I and ligating it into the Bam HI
site of pGL105 which had previously been blunt-ended with
the Klenow fragment.
Construct II (pGLHOSV) is similar to construct I but
has the 3' Eco RI/Pst I fragment, containing the poly(A)
addition seguences, deleted (Figure 2-4). The 4100 nt Eco
RI fragment from A.HHG5E was isolated, blunt-ended using the
Klenow fragment and ligated using T4 DNA ligase into the
Hie II site of pUC8SV40E, which contains the SV40 enhancer
element from pSVE108A (153) resulting in pGLHOSV.
Construct III (pGL109SV) is similar to construct I but
has the 170 nt Xho I fragment, containing the histone 3'
hairpin motif, deleted (Figure 2-5). Preparation of
construct III was carried out as follows. The intermediate
pGL106 was prepared by isolating the 4200 nt Hind III
fragment from pGL105 and ligating it, using T4 DNA ligase,
into the Hind III site of pUC19. Next, the intermediate
pGL107 was prepared by digesting pGL106 with Xho I to remove
the 170 nt Xho I fragment containing the hairpin motif
region and recircularizing the remaining fragment using T4
DNA ligase. The final construct was obtained by isolating
the 4200 nt Hind III fragment from pGL107 and ligating it
into the 4300 nt Hind III fragment from pGL105SV to give
pGL109SV.


Figure 2-3. Outline of the cloning scheme for the construction of the H2B construct
PGL105SV
Construct I (pGL105SV) is the control construct containing the entire A.HHG5E H2B
gene, promoter and 2000 nt of 3' untranslated region (Figure 2-3). The 6200 nt Pst I
fragment from A.HHG5E was isolated and ligated, using T4 DNA ligase, into the Pst I
site of pUC19 to obtain the intermediate (pGL105). Construct I was obtained by
isolating the Eco RI/Eco RI SV40 enhancer element from pSVE108A (153), blunt-ending
its termini with the Klenow fragment of DNA polymerase I and ligating it into the Bam
HI site of pGL105 which had previously been blunt-ended with the Klenow fragment.
Restriction endonuclease sites are as follows: A, Asp 718; B, Bam HI; E, Eco RI; H,
Hind III; P, Pst I; S, Sac I; S', Sph I; X, Xho I; a letter containing a semicircle
hat indicates a restriction endonuclease site which has been destroyed due to a
blunt-ending reaction. AHHG5E is a A.EMBL4 recombinant bacteriophage which was
isolated from a A.EMBL4 genomic DNA library (see chapter 4) .


2Kb
E 410Ont E 61 OOnt
I PS X XH XX I b PSP A
LI I I I imcd I|L IU 1
51OOnt
B
I
2Kb
E PS X XH
| 11 i M
o
XX
E
s*
PH
44_
pGL 105
E 3150nt E
I A
HHG5E
EPH
pUC 19
2Kb
2 E
E^ 'SPS' X XH XX i
PH
E E
SV 40 ENHANCER
pGL 105SV
U1
4*


Figure 2-4. Outline of the cloning scheme for the construction of the H2B 31
deletion construct pGLHOSV
Construct II (pGLHOSV) is similar to construct I but has the 3' Eco RI/Pst I
fragment, containing the poly(A) addition sequences, deleted (Figure 2-4). The
4100 nt Eco RI fragment from HHG5E was isolated, blunt-ended using the Klenow
fragment and ligated using T4 DNA ligase into the Hie II site of pUC8SV40E, which
contains the SV40 enhancer element from pSVE108A (153), resulting in pGLHOSV.
Restriction endonuclease sites are as follows: A, Asp 718; B, Bam HI; E, Eco RI; H,
Hind III; P, Pst I; S, Sac I; S', Sph I; X, Xho I; a letter containing a semicircle
hat indicates a restriction endonuclease site which has been destroyed due to a
blunt-ending reaction. AHHG5E is a A.EMBL4 recombinant bacteriophage which was
isolated from a A.EMBL4 genomic DNA library (see chapter 4) .


2Kb
E
41 OOnt E
s
61OOnt
E
51 OOnt
E
31 50nt
E
|
PS X XH^XX
PSP A
iii i
1
B
i
1
A
i
1


Figure 2-5. Outline of the cloning scheme for the
construction of the H2B hairpin motif minus construct
PGL109SV
Construct III (pGL109SV) is similar to construct I but
has the 170 nt Xho I fragment, containing the histone 3'
hairpin motif, deleted (Figure 2-5). Preparation of
construct III was carried out as follows. The intermediate
pGL106 was prepared by isolating the 4200 nt Hind III
fragment from pGL105 and ligating it, using T4 DNA ligase,
into the Hind III site of pUC19. Next, the intermediate
pGL107 was prepared by digesting pGL106 with Xho I to remove
the 170 nt Xho I fragment containing the hairpin motif
region and recircularizing the remaining fragment using T4
DNA ligase. The final construct was obtained by isolating
the 4200 nt Hind III fragment from pGL107 and ligating it
into the 4300 nt Hind III fragment from pGL105SV to give
pGL109SV. Restriction endonuclease sites are as follows: B,
Bam HI; E, Eco RI; H, Hind III; P, Pst I; S, Sac I; S',
Sph I; X, Xho I; a letter containing a semicircle hat
indicates a restriction endonuclease site which has been
destroyed due to a blunt-ending reaction. Plasmid GL105SV
is described in Figure 2-3.


58
c
2Kb
Eft ftPS' X XH ^ J
LMJJI 1. i ill 1 lL
pGL 1Q5SV
PH
=w_
3
c
C
4
2Kb
EPH
PH
=U
Gycjo)i
2Kb
Eft ftPS- X XH
Maiy 1 11
3
2Kb
EPH
PH
rU_
PGL1Q6
3
c
l
$ 2Kb
EPH X
' I
PH
pG L107
3
c
I
2Kb
EftJPS X
J>
H X
2Kb
PH
dJ
2Kb
Eft ftPS X XH X
U I I 1
PH
pGL1Q9SV
3


59
Subcloning into Bacteriophage M13 Vectors
DNA fragments to be sequenced were isolated from
agarose gels and ligated into M13mpl8 and/or mpl9
replicative form (123,190) using T4 DNA ligase as described
by the manufacturer. The ligation products were used to
transfect competent XLl-Blue (21) (see appendix C) bacteria.
Transfection of DNA into Bacterial Cells
Preparation of Competent E. coli Cells for Transfection
Competent E_i. coli cells were prepared using a
modification of the method described by Mandel and Higa
(107). An overnight culture of E^_ coli was diluted 1:100
into 100 ml YTN medium and grown at 37C with vigorous
shaking until the OD at 590 nm reached approximately 0.37.
The cell culture was chilled on wet ice for 5 minutes, split
into two 50 ml tubes and then centrifuged at 2500 rpm in an
IEC rotor (1400 X g) for 10 minutes at 4C. Following
centrifugation the supernatant was discarded and the cell
pellets vortexed gently, resuspended in 10 ml cold sterile
transfection buffer (60 mM CaCl2, 10 mM Pipes (pH 7.0), 15%
glycerol) and centrifuged at 2500 rpm in an IEC rotor
(1400 X g) for 10 minutes at 4C. Next the supernatant was
discarded and the cell pellets were gently resuspended in 10
ml cold sterile transfection buffer and incubated for
30 minutes on wet ice. Upon completion of the incubation
the cells were centrifuged at 2500 rpm in an IEC rotor


60
(1400 X g) for 10 minutes at 4C, the supernatant was
discarded, and the cell pellets were gently resuspended in
2.5 ml cold, sterile transfection buffer. The competent
cells were stored at 4C and used within 24 hours of
preparation.
Transfection of Recombinant Plasmid DNA
Transfection of recombinant plasmid DNA was carried out
using a modification of the method described by Mandel and
Higa (107). A fraction of a ligation mixture (1/10-1/2) and
100 ill competent Eh. coli DH5a (8) cells were combined in a
1.5 ml microcentrifuge tube, incubated for 10 minutes on wet
ice and then incubated for 5 minutes at 37C. Following the
incubation the cells were diluted with 1.0 ml prewarmed YTN
medium and incubated with shaking for 1 hour at 37C. An
aliquot of the transfection mixture was plated onto the
appropriate bottom agar plate and incubated inverted for
12-18 hours at 37C.
Transfection of Recombinant Bacteriophage M13 DNA
Transfection of recombinant bacteriophage M13 DNA was
carried out as described by Messing (115). A fraction of a
ligation mixture (1/10-1/2) and 200 fil competent Eh coli
XLl-Blue (21) cells were combined in a 14 mm X 100 mm glass
culture tube, incubated for 40 minutes on wet ice and then
heat-shocked for 2 minutes at 42C. Immediately following


61
the incubation, 4 ml of YTN top agar (45C) was added to the
tube, vortexed gently and poured onto a NZCYM bottom agar
plate. Upon solidification of the top agar the plate was
incubated inverted for 12-18 hours at 37C to allow for
plaque formation.
DNA Sequencing
Sanger Dideoxv-Mediated Chain Termination Method
Dideoxy sequencing (146) was carried out using a
Sequenase 2.0 DNA sequencing kit according to conditions
described by the manufacturer. After the sequencing
reactions were completed, loading buffer (95% formamide,
20 mM EDTA, 0.05% bromophenol blue and 0.05% xylene cyanol)
was added and the samples were heat denatured at 72C for
2 minutes and loaded onto a 6% polyacrylamide sequencing gel
containing 7.8 M urea (108). Following electrophoresis the
gel was soaked in 10% acetic acid, 12% methanol for 15-30
minutes to remove urea, dried under vacuum at 80"C using
3 sheets of Whatman 3MM paper as backing, and placed against
preflashed Kodak XAR-5 or Cronex X-ray film at room
temperature.
Chromosomal in situ Hybridization
Chromosomal in situ hybridization was performed as
described (23). Probe DNA (871 nt Eco RI/Sac I fragment
isolated from A.HHC289 (35)) was labeled with 3H to a


62
specific activity of 2-4 X 107 dpm//ng. Slides containing
metaphase chromosomes from normal male (46 XY) peripheral
blood lymphocytes were aged at 4C for 7-10 days and
pretreated with ribonuclease A (Sigma) for 1 hour at 37C.
The chromosomal DNA was denatured at 70C for 2 minutes in a
70% formamide: 2 X SSC mixture (pH 7.0). The probe DNA was
denatured in a hybridization mixture containing 50%
formamide, 2 X SSC and 10% dextran sulfate (pH 7.0).
Hybridization was carried out at 37C overnight. After
rinsing at 39C in 3 changes of 50% formamide: 2 X SSC and 5
changes of 2 X SSC, slides were dehydrated, air dried,
subjected to autoradiography and banded with Wright1s-Giemsa
stain solution mixed with 1-3 parts of a pH 9.2 borate
buffer (23).
Mammalian Cell Culture
HeLa Cell Culture and Synchronization
HeLa cells were grown in suspension culture (at
3-6 X 105 cells/ml) in Joklik-modified Eagle's minimal
essential medium supplemented with 7% calf serum and were
synchronized by two successive treatments with 2 mM
thymidine (162). The two, 14 hour, 2 mM thymidine
treatments were spaced 9 hours apart. Rates of DNA
synthesis were monitored by measuring the incorporation of
[3H]-thymidine into TCA-precipitable material in a 30 minute
pulse (162). DNA synthesis was inhibited by treating the


63
cell culture with 1 mM hydroxyurea for 1 hour, beginning
four hours after release from the second thymidine block.
Cells (hydroxyurea treated or untreated) were harvested at
various times after release from the second thymidine block.
HL60 Cell Culture and Differentiation
Human promyelocytic leukemia cells (HL-60 cells) were
cultured in liquid suspension in RPMI 1640 medium
supplemented with 10% fetal calf serum. HL60 cells were
plated at a cell density of 2 X 105/ml, grown for 24 hours
to a cell density of 3 X 105/ml and harvested by
centrifugation. For differentiation of HL60 cells,
cultures at a density of 3 X 105/ml were treated with TPA at
a final concentration of 16 nM, and harvested after periods
of 1, 2, 3, 4, and 5 days.
3T3L1 Cell Culture
Mouse 3T3L1 cells were maintained as subconfluent
cultures in DMEM containing 5% calf serum in a moist,
5% C02, 37C incubator.
Transfection and Transient Expression of Recombinant DNA in
3T3L1 Cells
Transfection of 3T3L1 cells was carried out as
described by Gorman et al. (56). Four hours prior to
transfection, 3T3L1 cells in monolayer at 20-40% confluency
were refed with fresh medium. The calcium phosphate/DNA


64
complex was prepared as described by Graham and Van der Eb
(58) using 10 /zg of plasmid DNA and 10 /zg of salmon sperm
DNA as carrier. Four hours after the addition of the
calcium phosphate/DNA complex the medium was removed and the
cells subjected to a one minute shock with 15-20% glycerol
(v/v) in medium containing 10% fetal calf serum. Incubation
was continued in fresh medium for 40-48 hours. Following
incubation, the cells were harvested and total cellular RNA
was isolated and analyzed by SI nuclease protection
analysis.
Selection of Stable Polyclonal 3T3L1 Cell Lines
3T3L1 cell monolayers were transfected as above using
20:1 molar quantities of construct and pSV2neo (158) without
using carrier DNA (20 /zg total DNA quantity) Selection of
cells was carried out as described by Southern and Berg
(158) using Geneticin (G418 sulfate). The medium plus drug
was changed every three to four days until resistant
colonies grew to approximately 1 cm in diameter. Polyclonal
cell lines were established by combining resistant colonies
as follows: the medium from each plate (containing from 2-20
resistant colonies) was removed, and the plates were treated
with 2 ml trypsin (0.25% in buffered saline) for 2-5
minutes. The cells were rinsed from the surface of the
plate and a 0.2 ml aliquot of the heterogeneous cell
suspension was transferred to a plate containing fresh


65
medium and was maintained as a subconfluent culture in DMEM
containing 5% calf serum in a moist, 5% C02, 37C incubator.
Ten polyclonal cell lines were established from the
transfection of each of the constructs described above and
are numbered PI through P10. Cell lines 105-P7, 110-P10,
and 109-P7 were chosen for further analysis (see chapter 5)
based upon their ability to differentiate to adipocytes.
Induction and Differentiation of 3T3L1 Cells
Induction of 3T3L1 cells to adipocytes was carried out
as described by Bernlohr et al. (7). 3T3L1 cells were grown
to confluence and 3 days post confluence were given fresh
DMEM containing 10% fetal calf serum, 0.22 /nM insulin,
0.6 /nM dexamethasone and 0.5 mM 3-isobutyl-l-methyl-
xanthine. Forty-eight hours later the medium was replaced
with fresh DMEM supplemented with 10% fetal calf serum.
Adipocyte conversion was detected within 8 days of
induction.
Cell Culture for Chromosome Localization
Isolation, propagation and characterization of parental
human and murine cells as well as the human-mouse somatic
cell hybrids was carried out as previously described
(81,82,136) in Dr. Carlo Croce's laboratory at Temple
University School of Medicine, Fels Research Institute,
Philadelphia, PA. The human chromosomes (either partial or


66
complete chromosomes) retained in the rodent-human hybrids
used in determining the chromosomal location of the human
H2B histone gene are schematically illustrated in Figure
4-10B. I would like to acknowledge Shirwin Pockwinse, Dr.
Kay Huebner, Dr. Linda A. Cannizzaro and Dr. Carlo Croce for
their work on the chromosomal localization of the H2B-GL105
gene.
THP-1 Cell culture
Acute monocytic leukemia cells, THP-1 (173), were
maintained in suspension cultures in RPMI medium 1640
supplemented with 5% fetal calf serum and kanamycin (60
Mg/ml).


CHAPTER 3
ISOLATION AND CHARACTERIZATION OF A cDNA FROM A HUMAN
HISTONE H2B GENE WHICH IS RECIPROCALLY EXPRESSED IN RELATION
TO REPLICATION-DEPENDENT H2B HISTONE GENES
DURING HL60 CELL DIFFERENTIATION
Introduction
Replication-dependent histone genes do not contain
introns (164) and their mRNAs are structurally simple; they
are not polyadenylated and have short 5' leader and 3'
trailer sequences (77). However, replication-dependent
histone mRNAs do have a characteristic 31 stem-loop motif
(164). Replication-dependent histone genes are coordinately
expressed during the cell cycle and their expression is
coupled with DNA synthesis (3,74,126,130,131,176). The
abundance of the replication-dependent human histone mRNAs
is regulated at both the transcriptional and post-
transcriptional levels (reviewed in 150,164).
In contrast, replacement histone genes may contain
introns (20,182,183) and their mRNAs, which are
polyadenylated and frequently contain long 5' leaders and 3'
trailers, are structurally more complex than their
replication-dependent counterparts (20,28,45,71,72,182,183).
Although the processes governing replacement histone mRNA
levels are not well understood, it is clear that their mRNAs
67


68
(10,19,25,45,71,89,152,155) as well as tissue-specific
histone mRNAs (27,34,64,83,84,93) are not always regulated
in a replication-dependent manner.
In these studies we cloned and characterized a cDNA
from a variant human histone H2B gene which has a complex
pattern of regulation with respect to the HeLa cell cycle
and HL60 cell differentiation. Our results reveal a
reciprocal relationship during the onset of HL60
differentiation between the expression of the HHC289 H2B
gene and the replication-dependent H2B genes.
Results
Isolation of an H2B cDNA Clone From a Iqtll Poly A+ cDNA
Library
A Agtll human liver cDNA library was screened with the
32P-labeled human histone probes indicated in Figure 2-1.
The library screening was carried out as indicated in Figure
2-2. Upon completion of the third round of screening 151
H3/H4 positives and 158 H2A/H2B/H1 positive plaques had been
obtained. Using the human histone probes described in
Figure 2-1, 51 H3/H4 positives and 51 H2A/H2B/H1 positive
plaques were separated according to individual histone class
by plaque dot analysis (Figure 3-1). Two HI clones, 9 H2A
clones, 22 H2B clones, 26 H3 clones and 5 H4 clones were
identified. No clone gave a positive signal with more than
one histone probe (Figure 3-1).


Figure 3-1. Plaque dot hybridization analysis of putative
positive clones from the Agtll poly A+ cDNA library
screening
Upon completion of the third round of Agtll library
screening (see Figure 2-2) 51 H3/H4 positives and 51
H2A/H2B/H1 positive plagues were separated according to
individual histone class by plaque dot analysis. Spot assay
analysis was carried out using 0.5 nl aliquots of
A bacteriophage dilutions ~103 pfu. Upon plaque formation
2 filter lifts were prepared from the H3/H4 plate and 3 from
the H2A/H2B/H1 plate and each hybridized to an individual
histone 32P-labeled probe (see Figure 2-1) as indicated
above each panel. The filters were then washed and
autoradiography performed using Kodak XAR-5 film.


70


71
Restriction Endonuclease Mapping of Positive Clones Isolated
From the Igtll Poly A+ cDNA Library Screening
Twelve clones suspected of containing human histone
genes were picked for further analysis (two from each
histone class as well as two randomly picked additional
clones) and mapped using a combination of restriction
endonuclease double digestions and southern blot analysis
(using the probes in Figure 2-1) as illustrated in Figure
3-2 for clone A.HHC185. A restriction map of AHHC185 is
illustrated at the top of Figure 3-2. Restriction
endonuclease maps of the 12 positive clones isolated from
the Agtll poly A+ cDNA library and chosen for further
analysis are illustrated in Figure 3-3. Insert sizes for
the positive recombinant clones ranged from 490 nt for
AHHC176 to 2160 nt for AHHC289.
Sequencing Strategy for the Positive Clones Isolated From
the Aqtll Poly A+ cDNA Library Screening
To evaluate whether any of the positive recombinant
clones contained histone coding sequence, the largest clone
in each of the histone classes was chosen for sequence
analysis. The arrows in Figure 3-4 represent regions of the
various Xgtll clones which were sequenced by Sanger dideoxy
sequencing. Three of the clones (A.HHC289, 1HHC227 and
1HHC4) were found to contain histone protein coding sequence


Figure 3-2. Restriction endonuclease Southern mapping of
A.HHC185
Each lane of a 0.8% agarose gel was loaded with 3 /g of
AHHC185 DNA restricted with: E, Eco RI; X, Xho I; S, Sac I;
H, Hind III; EH, Eco RI/Hind III; XH, Xho I/Hind III; SH,
Sac I/Hind III; EB, Eco RI/Bam HI. Lane U was loaded with
3 nq uncut A.HHC185 DNA and lane M with 3'-labeled A. DNA
digested with Eco RI and Hind III and with 3'-labeled pBR322
DNA digested with Hinf I. Following separation of the DNA
fragments electrophoretically, the DNA was transferred to
Zeta-probe nylon membrane and hybridized to a 32P-labeled
H2B probe (see Figure 2-1). A photograph of the above gel
stained with ethidium bromide is shown to the right of the
figure for reference. A restriction map of A.HHC185 is
illustrated at the top of the figure. Agtll arms are
represented by closed boxes at the termini of the
restriction map. Restriction enzyme abbreviations are as
follows: E=Eco RI; X=Xho I; S=Sst I.


73
1200 bpl
U E X S H EH M XH SH EB
AhHC 185 MAPPING


Figure 3-3. Restriction endonuclease maps of positive
clones isolated from the katll poly A+ cDNA library
screening
Restriction endonuclease maps of 12 positive clones
isolated from the Agtll poly A+ cDNA library screening are
illustrated. Restriction endonuclease sites are as depicted
in the top right corner of the figure. Agtll arms are
represented by closed boxes at the termini of the each
restriction map. A denotes a fragment giving a positive
hybridization signal with a histone coding probe.


75
"X DNA FLANKING ECO Rl INSERTS
Xhhc 2271
H1 I
I 300 bpr I ECO Rl T
HIND III f
XHO I t
SST I ¡t
BAM HI
KPN I f
239 H-
H2AV
1 I


Figure 3-4. Sequencing strategy on the positive clones isolated from the Aqtll
poly A+ cDNA library screening
Arrows represent regions of the Igtll clones which were seguenced by Sanger
dideoxy sequencing. A "c", below a map, indicates a restriction fragment which
hybridized to a histone protein coding probe (see Figure 2-1) and a "C" to the left
of a map marks the clones identified, by sequence analysis (146), to contain histone
protein coding sequences. An "A" to the left of a map marks the clones identified,
by sequence analysis, to contain poly(A) stretches at the end of the insert.
Restriction endonuclease sites are as depicted at the top of the figure. Agtll arms
are represented by closed boxes at the termini of the each restriction map.


ECO Rl I
> DNA FLANKING ECO Rl INSERTS
HIND III t XHO I ¥
I 300 bpr I
SST I
KPN I ?
289 H2B CA


J SL
227 H1 CA
239 H2A
J!L
h
4 H3
41 H4


78
and 3 clones (A.HHC289, A.HHC227 and A.HHC41) to contain a
stretch of poly(A) at the terminus of their inserts (Figure
3-4) .
Sequencing of a Poly A+ H2B cDNA
Of the 22 cDNA clones containing H2B histone sequences
obtained from the initial screening of the lambda library
four were mapped using a combination of restriction
endonuclease double digestions and southern blot analysis
and the clone with the largest insert (Figure 3-3, AHHC289 -
2119 nt) that also contained a poly(A) tract (Figure 3-4)
was then selected for detailed sequence analysis. Regions
of the human H2B histone cDNA AHHC289 which were sequenced
are indicated by arrows in Figure 3-5 and the nucleotide
sequence is presented in Figure 3-6(A). The AHHC289 cDNA
encodes H2B amino acids 25-125 and the translation stop
codon. Also contained in the cDNA sequence is the poly(A)
addition sequence, AATAAA, located 1776 nt downstream from
the translation stop codon, and a poly(A) stretch located
16 nt downstream of the poly(A) addition sequence. The
protein coding region is GC rich (63%) whereas the 3' un
translated trailer is relatively GC poor (45%). Shown in
Figure 3-6(B) are pertinent restriction sites and the
AHHC289 fragments used as probes for northern blot analysis.


Figure 3-5. Sequencing strategy for the human H2B histone cDNA A.HHC289
Arrows indicate regions of the human H2B histone cDNA 1HHC289 which were
sequenced by Sanger dideoxy sequence analysis (146). The H2B protein coding region
is illustrated by a hatched box. Restriction enzyme abbreviations are as follows: E,
Eco RI; X, Xho I; S, Sst I. Agtll arms are not represented in this illustration.


200 nt
80


Figure 3-6. (A) Nucleotide and deduced amino acid sequence
of the human H2B histone cDNA A.HHC289. (B) Restriction
map of the cDNA A.HHC289
(A) The 1HHC289 cDNA encodes H2B amino acids 25-125 and
the translation stop codon. Also contained in the AHHC289
cDNA sequence is the poly A+ addition sequence, AATAAA.
located 1776 nt downstream from the protein stop codon and a
poly A+ stretch located 16 nt downstream from the last
nucleotide of the poly A+ addition sequence. Solid double
underlined ( ) sequences represent potential splice sites.
(B) The H2B protein coding region is illustrated by a closed
box and the Agtll arms are represented by the open boxes.
Solid bars below the restriction map illustrates AHHC289
probes used for northern analysis. Restriction enzyme
abbreviations are as follows: E=Eco RI; X=Xho I; S=Sst I.


82
GAC
GGC
AAG
AAG
CGC
AAG
CGC
AGC
CGC
AAA
GAG
AGC
TAC
TCC
ATC
45
?5Asp
Gly
Lys
Lys
Arg
Lys
Arg
Ser
Arg
Lys
Glu
Ser
Tyr
Ser
He
TAC
GTG
TAC
AAG
GTG
CTG
AAG
CAG
GTC
CAC
CCC
GAC
ACC
GGC
ATC
90
4 Val
Tyr
Lys
Val
Leu
Lys
Gin
Val
His
Pro
Asp
Thr
Gly
He
TCG
TCC
AAG
GCC
ATG
GGC
ATC
ATG
AAC
TCC
TTC
GTC
AAC
GAC
ATC
135
55Ser
Ser
Lys
A1 a
Met
Gly
lie
Met
Asn
Ser
Phe
Val
Asn
Asp
He
TTC
GAG
CGC
ATC
GCG
GGA
GAG
GCT
TCC
CGC
CTG
GCG
CAC
TAC
AAC
180
7CPhe
Glu
Arg
lie
A1 a
Gly
Glu
A1 a
Ser
Arg
Leu
A1 a
His
Tyr
Asn
AAG
CGC
TCC
ACC
ATC
ACA
TCC
CGC
GAG
ATC
CAG
ACG
GCC
GTG
CGC
225
esLys
Arg
Ser
Thr
He
Thr
Ser
Arg
Glu
He
Gin
Thr
A1 a
Val
Arg
CTG
CTG
CTG
CCC
GGC
GAG
CTG
GCC
AAG
CAC
GCC
GTG
TCC
GAG
GGC
270
ioc^eu
Leu
Leu
Pro
Gly
Glu
Leu
Ala
Lys
His
Ala
Val
Ser
Glu
Gly
ACC
AAG
GCG
GTC
ACC
AAG
TAC
ACC
AGC
TCC
AAG
TGA
GTC
CCT
GCC
315
usThr
Lys
A1 a
Val
Thr
Lys
Tyr
Thr
Ser
Ser
Lys
GGACCTGGCG CTCGCTCGCT CGAGTCGCCG GCTGCTTGAC TCCAAAGGCT CTTTTCAGAG 375
CCACCCACCT AATCACTAGA AAAGAGCTTG TTCACTTATT CCCTTAGTTT CTTTTCATAA 435
AGTAAGTTAT CTTAGTGTGA AGGTCATGGG AAATGGCATA CGTAGCTTTT TAACTATTTG 495
GAACTtGAGG TCCCCAGTGC GTCATTGGAT TTGCTTTTGA ATCTAGAGCG TGTCTTTACT 555
CATTGTGCTG CTTAGCCTTC CCAGGAGTCG GTTCTCAATT AGGCTGTTGG GAATCCGCCT 615
CTTTACCCGC CCCCACTCCC GCCCCACACG CGCCCTGGTG GCTCCTTGGG TCTGTTTCAT 675
TCTAAAACGA AGTGGCTGAG TTCGGCTGTC ATTTAAGAGA ACTCCAGGAC ACAATTCAGC 735
CCGGGTTCCG CAAACACTGC GTGACAGCTC TGTATGACTG ACGCTTGGCA GCAGCTTTTG 795
TGTCCGGTCA CCAGTTCTGC CGTGCGATGG GGCCTCCTGT GGATACCAGC CGTTCTGTGT 855
ATTTTGGACG AAGGCGGCAG CCGGGTCCCA GCCTTGTCCT GATTGGGCGA CAAGAATATT 915
CAAAATTCTG CGCCTTTTTC TAATTTGTAG ATTTCAGTTT CCGTCGTTCA CTTTGAGACT 975
TTGAAATTCC TATTTCTCAT TTTGTTGATA ATTTCTGCAT TTAATGGTCT GTGCTTTAAA 1035
TGGTAACGCT ACGGCCCCAG GTCACTGCGA GGCACTTACC ATGTAGATAC GGGCTCAAAA 1095
GTCACCTCTC AGAGACCTAC GTCATCCACT CAGGAATTCG CGCCTCTCAT ACTTGCCTGT 1155
CTCATTTTAT CTTCCTTCTA GCAGCTGTCT GAAATTGGTT CGTCTGTTTT CTTGTTTATG 1215
GTATTCTCAA GCCCTTGACA GACCGGCTAG TGTGGTTTTC CCGTGCATCT TCAGCCTGGC 1275
ACATTATGGA CACTTAAATA CTACGTATTG ATCTAATATT GTTGGGTTAA TTTTTCCATC 1335
CCACCCTTTT CTTAATCGCT TCCGTGGATG GATGAAGGGT GCTGTTCATT TCCATTAGAT 1395
GTATGTGAAG GCACAGTGAA AATGGAAATG TTCTTGGAGC TACTTCCTCA AAATGTATCC 1455
TTAGTCACCT CAGTGCAACA GCTGGGAGGG GGCCGTGTTA AGATTTTTTT TGCTACAAAG 1515
AGGAGGTGGC AATGGTAGAT CCACCCTTAT GCTTCTCAGT TTAGCATAAC CTCTTATGGA 1575
TTTTCATCAA ATTCAGCGTG TTGGTCACTG GAAAGAGCCT TTTCCTTCTC CTTTTCTTAC 1635
TCTCCCCTCA TGGTGTTCCC CTCTTAAAGG AGAGGAGCTT TTAATTTACA CTTACCACCT 1695
CATTTGCTTT TCTGGAGGCC ATGCAATATA GGCGGGACTA CAGAGTTAAT CTCCTTTTTA 1755
CAAATGAGGC CAAGAGAAGC CTCATTGGTT CACAGTCATG CAGCTCATAC TGTCCACCCT 1815
TGTATTCTCA GATGCAGGAC AATTGCATTT TAGTTTTATT TTGTGGAGGT GCAGAATATT 1875
TACTCTTTCT GTCCAACCCT TGATTCTGCC GAGGAAGACA CTGATGGTTT GATGAGTGAT 1935
TCAGCTGTTT TTGGCTAAGG GCTTTTGGAG CTGATGGCAG GGGTTTGATG AATCCAAATG 1995
AGCTCTAGAC ATTATCACAG ACTGAATAGA TCTTAACTGT CTCCTACATG TGTGTTTTCA 2055
AATGTGTATA GATGCTATTG TTATTAATAA AGTTACCAAT TAATTTAAAA AAAAAAAAAA 2115
AAAA 2119
B
PROTEIN
CODING
E
S E
24.1
Kb
HHC 289
340 nt 5 PROBE
870 nt 3 PROBE


Full Text

PAGE 1

,62/$7,21 $1' &+$5$&7(5,=$7,21 2) $ 9$5,$17 +80$1 +% +,6721( *(1( (;35(66,1* $/7(51$7,9( P51$V 5(*8/$7(' ',))(5(17,$//< '85,1* 7+( &(// &<&/( $1' ',))(5(17,$7,21 %\ '$9,' *(25*( &2//$57 $ ',66(57$7,21 35(6(17(' 72 7+( *5$'8$7( 6&+22/ 2) 7+( 81,9(56,7< 2) )/25,'$ ,1 3$57,$/ )8/),//0(17 2) 7+( 5(48,5(0(176 )25 7+( '(*5(( 2) '2&725 2) 3+,/2623+< 81,9(56,7< 2) )/25,'$

PAGE 2

'HGLFDWHG WR 0DU\ WKH PRWKHU RI *RG

PAGE 3

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

PAGE 4

, ZRXOG OLNH WR WKDQN DOO RI WKH PHPEHUV RI WKH 6WHLQVn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

PAGE 5

7$%/( 2) &217(176 3$*( $&.12:/('*(0(176 LLL /,67 2) ),*85(6 L[ .(< 72 $%%5(9,$7,216 [LL $%675$&7 [Y &+$37(56 f ,1752'8&7,21 *HQHUDO %DFNJURXQG +LVWRQH 3URWHLQV 6WUXFWXUH DQG 2UJDQL]DWLRQ RI +XPDQ +LVWRQH *HQHV &RXSOLQJ RI '1$ 6\QWKHVLV DQG +LVWRQH *HQH 5HJXODWLRQ 7UDQVFULSWLRQDO DQG 3RVW7UDQVFULSWLRQDO 5HJXODWLRQ RI +XPDQ +LVWRQH *HQHV +% +LVWRQH *HQH ([SUHVVLRQ 2YHUYLHZ RI 3URMHFW f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

PAGE 6

6LQJOH6WUDQGHG 0 %DFWHULRSKDJH 7HPSODWH ,VRODWLRQ 5HFRPELQDQW 3ODVPLG '1$V ,VRODWLRQ DQG 3XULILFDWLRQ RI 3ODVPLG '1$ 5DSLG 6PDOO 6FDOH 3ODVPLG '1$ ,VRODWLRQ /DUJH 6FDOH 3ODVPLG '1$ ,VRODWLRQ ,VRODWLRQ DQG 3XULILFDWLRQ RI (XNDU\RWLF *HQRPLF '1$ ,VRODWLRQ DQG 3XULILFDWLRQ RI 0DPPDOLDQ 51$ 6HOHFWLRQ RI 3RO\ $ 51$ 6SHFWURSKRWRPHWULF 4XDQWLWDWLRQ RI '1$ DQG 51$ 3UHSDUDWLRQ RI 5DGLRODEHOHG '1$ 5DQGRP 2OLJRQXFOHRWLGH 3ULPHG /DEHOLQJ /DEHOLQJ WKH n 7HUPLQL RI '1$ /DEHOLQJ WKH n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

PAGE 7

f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f $/7(51$7,9( n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

PAGE 8

+%*/ P51$ /HYHOV LQ 6\QFKURQL]HG +H/D &HOOV 0DSSLQJ RI WKH +%*/ n P51$ 6WDUW 6LWH 0DSSLQJ RI WKH +%*/ P51$ n 7HUPLQL 'LVFXVVLRQ 6WUXFWXUH DQG 2UJDQL]DWLRQ RI WKH +%*/ *HQH 7UDQVFULSWLRQDO UHJXODWLRQ RI WKH +%*/ *HQH 3RVW7UDQVFULSWLRQDO 5HJXODWLRQ RI WKH +%*/ *HQH f 6800$5< $1' )8785( &216,'(5$7,216 $33(1',&(6 $f 5($*(176 )25 3523$*$7,21 $1' 0$,17(1$1&( 2) %$&7(5,$ $1' %$&7(5,23+$*( %f 5($*(176 )25 18&/(,& $&,' ,62/$7,21 $1' 385,),&$7,21 &f %$&7(5,$/ 675$,16 86(' 5()(5(1&(6 %,2*5$3+,&$/ 6.(7&+ YLLL

PAGE 9

/,67 2) ),*85(6 )LJXUH 3DJH 6FKHPDWLF GLDJUDP RI KLVWRQH JHQH SUREHV XVHG IRU WKH $JWOO SRO\ $ F'1$ OLEUDU\ VFUHHQLQJ 2XWOLQH RI WKH VFUHHQLQJ VFKHPH IRU WKH ;JWOO SRO\ $ F'1$ OLEUDU\ 2XWOLQH RI WKH FORQLQJ VFKHPH IRU WKH FRQVWUXFWLRQ RI WKH +% FRQVWUXFW S*/69 2XWOLQH RI WKH FORQLQJ VFKHPH IRU WKH FRQVWUXFWLRQ RI WKH +% n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f 1XFOHRWLGH DQG GHGXFHG DPLQR DFLG VHTXHQFH RI WKH KXPDQ +% KLVWRQH F'1$ $++& %f 5HVWULFWLRQ PDS RI WKH F'1$ $++& $f &RPSDULVRQ RI WKH $++& KLVWRQH +% DPLQR DFLG VHTXHQFH ZLWK WKH VHTXHQFHV RI RWKHU +%V %f '1$ VHTXHQFH FRPSDULVRQ RI WKH n XQWUDQVODWHG UHJLRQ RI VHOHFWHG KLVWRQH VHTXHQFHV L[

PAGE 10

$++& P51$ OHYHOV GXULQJ WKH +H/D FHOO F\FOH +% P51$ OHYHOV GXULQJ WKH +H/D FHOO F\FOH 7UDQVFULSWLRQ RI WKH $++& KLVWRQH JHQH GXULQJ WKH +H/D FHOO F\FOH $++& P51$ OHYHOV GXULQJ SUROLIHUDWLRQ DQG GLIIHUHQWLDWLRQ RI +/ FHOOV +% P51$ OHYHOV GXULQJ SUROLIHUDWLRQ DQG GLIIHUHQWLDWLRQ RI +/ FHOOV 'HQVLWRPHWULF DQDO\VLV RI WKH QW UHSOLFDWLRQ GHSHQGHQW +% P51$ DQG WKH QW $++& P51$ OHYHOV GXULQJ GLIIHUHQWLDWLRQ RI +/ FHOOV 7KLUG URXQG VFUHHQLQJ RI D $(0%/ KXPDQ DGXOW O\PSKRF\WH '1$ OLEUDU\ ZLWK D $++& n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

PAGE 11

5HJLRQDO ORFDOL]DWLRQ RI WKH KXPDQ KLVWRQH JHQH +%*/ RQ KXPDQ FKURPRVRPH 1RUWKHUQ EORW DQDO\VLV RI +%*/ P51$ OHYHOV LQ SUROLIHUDWLQJ 7+3 FHOOV 1RUWKHUQ EORW DQDO\VLV RI +%*/ P51$ OHYHOV LQ V\QFKURQL]HG +H/D FHOOV 6, QXFOHDVH PDSSLQJ RI WKH +%*/ n P51$ VWDUW VLWH 6, QXFOHDVH PDSSLQJ RI WKH +$*/ n P51$ VWDUW VLWH 6, QXFOHDVH SURWHFWLRQ DQDO\VLV RI WKH n WHUPLQL RI WKH +%*/ P51$ 6, DQDO\VLV RI WKH n WHUPLQL RI +% P51$ H[SUHVVHG LQ 7/ FHOO OLQHV [L

PAGE 12

.(< 72 $%%5(9,$7,216 $36 $PPRQLXP 3HUVXOIDWH $73 $GHQRVLQHnWULSKRVSKDWH ES %DVH SDLU %6$ %RYLQH VHUXP DOEXPLQ &,3 &DOI LQWHVWLQDO DONDOLQH SKRVSKDWDVH 2 2 f f 'HJUHH FHQWLJUDGH '0(0 'XOEHFFRnV PLQLPDO HVVHQWLDO PHGLXP G$73 n'HR[\DGHQRVLQH nWULSKRVSKDWH G&73 n'HR[\F\WLGLQH nWULSKRVSKDWH G*73 n'HR[\JXDQRVLQH nWULSKRVSKDWH '1DVH 'HR[\ULERQXFOHDVH '1$ 'HR[\ULERQXFOHLF DFLG '(3& 'LHWK\O S\URFDUERQDWH GSP 'LVLQWHJUDWLRQV SHU PLQXWH '77 'LWKLRWKUHLWRO GG+ 'RXEOHGLVWLOOHG ZDWHU ('7$ (WK\OHQHGLDPLQHWHWUDDFHWLF DFLG J *UDP J *UDYLWDWLRQDO IRUFH *$3'+ *O\FHUDOGHK\GHSKRVSKDWHGHK\GURJHQDVH +%66 +DQNnV EDODQFHG VDOW VROXWLRQ ,$$ ,VRDP\O DOFRKRO [LL

PAGE 13

,37* ,VRSURS\O'WKLRJDODFWRS\UDQRVLGH /LWHU 0J 0LFURJUDP P 0LFURPHWHU PJ 0LOOLJUDP PO 0LOOLOLWHU PP 0LOOLPHWHU P0 0LOOLPRODU 0 0RODU 0236 0RUSKROLQHSURSDQHVXOIRQLF DFLG QJ 1DQRJUDP QW 1XFOHRWLGH 4 R 2SWLFDO GHQVLW\ 3%6 3KRVSKDWH EXIIHUHG VDOLQH SIX 3ODTXH IRUPLQJ XQLW 3RO\ $f 3RO\DGHQRVLQH 3(* 3RO\HWK\OHQH JO\FRO 393 3RO\YLQ\OS\UUROLGRQH 396 3RO\YLQ\OVXOIRQLF DFLG 5) 5HSOLFDWLYH IRUP 51DVH $ 5LERQXFOHDVH $ 51$ 5LERQXFOHLF DFLG USP 5HYROXWLRQV SHU PLQXWH 6'6 6RGLXP GRGHF\O VXOIDWH 7&$ 7ULFKORURDFHWLF DFLG 7(0(' 111n1nWHWUDPHWK\OHWK\OHQHGLDPLQH [LLL

PAGE 14

W+% 73$ 773 8 ;JDO 7HVWLV VSHFLILF +% WHWUDGHFDQR\O SKRUERODFHWDWH 7K\PLGLQH nWULSKRVSKDWH 8QLW RI HQ]\PH EURPRFKORURLQGRO\O"'JDODFWRVLGH [LY

PAGE 15

$EVWUDFW RI 'LVVHUWDWLRQ 3UHVHQWHG WR WKH *UDGXDWH 6FKRRO RI WKH 8QLYHUVLW\ RI )ORULGD LQ 3DUWLDO )XOILOOPHQW RI WKH 5HTXLUHPHQWV IRU WKH 'HJUHH RI 'RFWRU RI 3KLORVRSK\ ,62/$7,21 $1' &+$5$&7(5,=$7,21 2) $ 9$5,$17 +80$1 +% +,6721( *(1( (;35(66,1* $/7(51$7,9( P51$V 5(*8/$7(' ',))(5(17,$//< '85,1* 7+( &(// &<&/( $1' ',))(5(17,$7,21 %\ 'DYLG *HRUJH &ROODUW 0D\ &KDLUPDQ *DU\ 6WHLQ 0DMRU 'HSDUWPHQW %LRFKHPLVWU\ DQG 0ROHFXODU %LRORJ\ :H KDYH LVRODWHG DQG FKDUDFWHUL]HG D F'1$ $++&f FRUUHVSRQGLQJ WR D YDULDQW KXPDQ KLVWRQH +% JHQH 7KH +% SURWHLQ FRGLQJ UHJLRQ RI $++& LV IODQNHG DW WKH n HQG E\ D QW QRQWUDQVODWHG WUDLOHU WKDW FRQWDLQV D UHJLRQ RI K\SKHQDWHG G\DG V\PPHWU\ DQG D SRO\$f WDLO 1XFOHDU UXQRQ WUDQVFULSWLRQ DQDO\VLV UHYHDOHG D WZR IROG LQFUHDVH LQ WUDQVFULSWLRQ RI WKH ++& JHQH GXULQJ 6 SKDVH 1RUWKHUQ EORW DQDO\VLV LQGLFDWHG WKDW WKH OHYHOV RI WKH QW ++& P51$ VSHFLHV ZHUH FRQVWDQW GXULQJ WKH +H/D FHOO F\FOH 1RUWKHUQ EORW DQDO\VLV DOVR UHYHDOHG WKDW WKH OHYHOV RI WKH QW ++& +% VSHFLHV LQFUHDVHG IROG GXULQJ +/ FHOO GLIIHUHQWLDWLRQ ZKHUHDV WKH OHYHOV RI UHSOLFDWLRQ [Y

PAGE 16

GHSHQGHQW +% P51$V GHFUHDVHG WR OHVV WKDQ b RI WKRVH LQ SUROLIHUDWLQJ FHOOV 8VLQJ D SUREH IURP WKH $++& F'1$ WKDW GHWHFWV RQO\ D VLQJOH FRS\ UHJLRQ RI WKH KXPDQ JHQRPH ZH LVRODWHG D YDULDQW KXPDQ +% KLVWRQH JHQH */f ZKLFK H[SUHVVHV DOWHUQDWLYH P51$V UHJXODWHG GLIIHUHQWLDOO\ GXULQJ WKH +H/D FHOO F\FOH 7KLV +% JHQH HQFRGHV ERWK D QW UHSOLFDWLRQGHSHQGHQW P51$ DQG WKH QW ++& FRQVWLWXWLYHO\ H[SUHVVHG P51$ 7KH n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n HQG SURFHVVLQJ LV D PDMRU OHYHO [Y L

PAGE 17

RI UHJXODWLRQ E\ ZKLFK FHOOV FDQ PRGXODWH WKH V\QWKHVLV RI YDULDQW KLVWRQH SURWHLQV GXULQJ WKH FHOO F\FOH DQG DW WKH RQVHW RI GLIIHUHQWLDWLRQ [YLL

PAGE 18

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

PAGE 19

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f 7KHUH DUH ILYH PDMRU FODVVHV RI KLVWRQH SURWHLQV +, +$ +% + DQG +f f +LVWRQH SURWHLQV H[KLELW WZR OHYHOV RI GLYHUVLW\ Lf HYROXWLRQDU\ GLYHUVLW\ EHWZHHQ VSHFLHV DQG LLf VXEW\SH GLYHUVLW\ LQ D FODVV +, +$ +% + RU +f ZLWKLQ D VSHFLHV f 7KH VXEW\SHV ZLWKLQ D VSHFLHV DUH RIWHQ UHIHUUHG WR DV YDULDQWV f 5HSOLFDWLRQGHSHQGHQW KLVWRQH YDULDQWV UHSUHVHQW WKH PDMRULW\ RI WKH KLVWRQH SURWHLQV V\QWKHVL]HG LQ SUROLIHUDWLQJ FHOOV DQG LW LV ZHOO HVWDEOLVKHG WKDW WKHLU V\QWKHVLV LV IXQFWLRQDOO\ DQG WHPSRUDOO\ FRXSOHG WR

PAGE 20

'1$ UHSOLFDWLRQ f 5HSODFHPHQW YDULDQWV DUH H[SUHVVHG LQ QRQSUROLIHUDWLQJ FHOOV DQG DUH RIWHQ H[SUHVVHG LQ SUROLIHUDWLQJ FHOOV DOWKRXJK QRUPDOO\ DW D ORZHU OHYHO WKDQ WKHLU UHSOLFDWLRQGHSHQGHQW FRXQWHUSDUWV f 7KH WKLUG W\SH RI YDULDQW ZLWKLQ D VSHFLHV LV WKH WLVVXH VSHFLILF YDULDQW f 7LVVXHVSHFLILF YDULDQWV DUH JHQHUDOO\ QRW V\QWKHVL]HG LQ D UHSOLFDWLRQ GHSHQGHQW PDQQHU f +LVWRQH +% SURWHLQV KDYH EHHQ VWXGLHG LQ D YDULHW\ RI VSHFLHVf§LQFOXGLQJ FKLFNHQ PRXVH UDW DQG KXPDQf§ZLWK UHVSHFW WR WKHLU VXEW\SH GLYHUVLW\ DQG UHYLHZHG LQ f $OWKRXJK WKH UHSOLFDWLRQGHSHQGHQW DQG WLVVXH VSHFLILF +% SURWHLQV DUH HDVLO\ GHWHFWHG LQ PRVW VSHFLHV WKH UHSODFHPHQW KLVWRQH +% SURWHLQV DUH QRW DOZD\V REVHUYHG LQ KXPDQV SRVVLEO\ GXH WR WKH ODFN RI VHQVLWLYLW\ RI WKH DVVD\ f ,Q PRXVH WKH +% YDULDQW LV WKH UHSOLFDWLRQ GHSHQGHQW VXEW\SH DQG LV DEXQGDQW LQ UDSLGO\ GLYLGLQJ FHOOV f 7KH +% YDULDQW LV WKH FRQVWLWXWLYHO\ H[SUHVVHG VXEW\SH DQG LV SUHVHQW LQ VPDOO DPRXQWV LQ UDSLGO\ JURZLQJ WLVVXHV EXW LQFUHDVHV VKDUSO\ DV SUROLIHUDWLRQ VORZV HVSHFLDOO\ DIWHU WKH SRVWQDWDO JURZWK SKDVH f 7KH +% VXEW\SH VSHFLILF DPLQR DFLGV RI WKH +% PRXVH SURWHLQ 6HUX $OD $VS /\V *O\ $VS *O\ *OXf GLIIHU IURP WKRVH RI WKH +% SURWHLQ RQO\ DW 6HU f KRZHYHU WKH IXQFWLRQDO VLJQLILFDQFH RI WKLV FKDQJH LV XQFOHDU ,Q JHQHUDO UHSODFHPHQW KLVWRQH YDULDQWV DUH OHVV

PAGE 21

K\GURSKRELF WKDQ UHSOLFDWLRQGHSHQGHQW RU WLVVXH VSHFLILF YDULDQWV 6WUXFWXUH DQG 2UJDQL]DWLRQ RI +XPDQ +LVWRQH *HQHV +XPDQ KLVWRQH JHQHV DUH D IDPLO\ RI PRGHUDWHO\ UHLWHUDWHG VHTXHQFHV f DQG DUH DUUDQJHG LQ FOXVWHUV WKH\ VKRZ QR HYLGHQFH RI D VLPSOH WDQGHP UHSHDW RUJDQL]DWLRQ f DV GR WKH KLVWRQH JHQHV RI VHD XUFKLQ VSHFLHV f DQG 'URVRSKLOD f ,Q KXPDQV WKHVH FOXVWHUV DUH ORFDOL]HG RQ FKURPRVRPHV DQG f DQG DUH DVVRFLDWHG ZLWK D VHULHV RI UHSHWLWLYH '1$ VHTXHQFHV f &HOO F\FOH GHSHQGHQW KLVWRQH JHQHV GR QRW FRQWDLQ LQWURQV f DQG WKH\ HQFRGH P51$V ZKLFK DUH QRW SRO\DGHQ\ODWHG DQG KDYH VKRUW n OHDGHU DQG n WUDLOHU VHTXHQFHV f $OWKRXJK UHSOLFDWLRQGHSHQGHQW KLVWRQH P51$V DUH VWUXFWXUDOO\ VLPSOH WKH\ GR KDYH D FKDUDFWHULVWLF n VWHPORRS PRWLI f ,Q FRQWUDVW FRQVWLWXWLYHO\ H[SUHVVHG KLVWRQH JHQHV DUH VWUXFWXUDOO\ PRUH FRPSOH[ 7KH\ PD\ FRQWDLQ LQWURQV f DQG WKH\ HQFRGH P51$V ZKLFK DUH SRO\DGHQ\ODWHG DQG IUHTXHQWO\ FRQWDLQ ORQJ n OHDGHU RU n WUDLOHU VHTXHQFHV f &RXSOLQJ RI '1$ 6\QWKHVLV DQG +LVWRQH *HQH 5HJXODWLRQ 5HSOLFDWLRQGHSHQGHQW KLVWRQH JHQHV DUH FRRUGLQDWHO\ H[SUHVVHG GXULQJ WKH FHOO F\FOH DQG WKHLU H[SUHVVLRQ LV

PAGE 22

FRXSOHG ZLWK '1$ V\QWKHVLV f 7KH DEXQGDQFH RI WKHLU P51$V LV UHJXODWHG DW ERWK WKH WUDQVFULSWLRQDO DQG SRVWWUDQVFULSWLRQDO OHYHOV DQG UHYLHZHG LQ f ,Q FRPSDULVRQ WKH P51$V IRU UHSODFHPHQW KLVWRQHV f DV ZHOO DV WLVVXHVSHFLILF KLVWRQH P51$V f DUH QRW DOZD\V UHJXODWHG LQ D UHSOLFDWLRQGHSHQGHQW PDQQHU 7UDQVFULSWLRQDO DQG 3RVW7UDQVFULSWLRQDO 5HJXODWLRQ RI +XPDQ +LVWRQH *HQHV 7UDQVFULSWLRQDO UHJXODWLRQ RI KXPDQ KLVWRQH JHQHV LQYROYHV D WR IROG HQKDQFHPHQW LQ WKH UDWH RI WUDQVFULSWLRQ ZLWKLQ WKH ILUVW KRXUV RI '1$ V\QWKHVLV IROORZHG E\ D UHWXUQ WR EDVDO OHYHO E\ PLG WR ODWH 6 SKDVH f +RZHYHU VWHDG\ VWDWH OHYHOV RI KLVWRQH P51$ DUH HOHYDWHG WR IROG GXULQJ 6 SKDVH RI WKH FHOO F\FOH DQG SDUDOOHO WKH UDWHV RI '1$ V\QWKHVLV f $OWKRXJK WKH UDWH RI KLVWRQH JHQH WUDQVFULSWLRQ SHDNV ZLWKLQ WKH ILUVW KRXUV RI '1$ V\QWKHVLV KLVWRQH P51$ OHYHOV GR QRW UHDFK D PD[LPXP XQWLO KRXUV LQWR 6 SKDVH 7DNHQ WRJHWKHU WKHVH REVHUYDWLRQV VXJJHVW SRVWWUDQVFULSWLRQDO PHFKDQLVPV SOD\ D UROH LQ WKH UHJXODWLRQ RI KLVWRQH P51$ OHYHOV 3RVWWUDQVFULSWLRQDO UHJXODWLRQ RI KLVWRQH JHQHV LQYROYHV D UDSLG DQG VHOHFWLYH GHVWDELOL]DWLRQ RI KLVWRQH P51$V WRZDUG WKH HQG RI 6 SKDVH RU LQ UHVSRQVH WR LQKLELWLRQ

PAGE 23

RI '1$ V\QWKHVLV f +RZHYHU WKHUH LV OLWWOH RU QR HIIHFW RQ KLVWRQH JHQH WUDQVFULSWLRQ E\ LQKLELWLRQ RI '1$ V\QWKHVLV f 7KLV GHVWDELOL]DWLRQ RI KLVWRQH P51$ XSRQ LQKLELWLRQ RI '1$ V\QWKHVLV LV GHSHQGHQW XSRQ SURWHLQ V\QWKHVLV f ,Q FRQWUDVW WKH P51$ OHYHOV RI UHSOLFDWLRQLQGHSHQGHQW KLVWRQH JHQHV DUH QRW UHJXODWHG LQ WKH VDPH FHOO F\FOH GHSHQGHQW PDQQHU DV WKHLU UHSOLFDWLRQGHSHQGHQW FRXQWHUSDUWV DQG UHPDLQ UHODWLYHO\ FRQVWDQW XSRQ LQKLELWLRQ RI '1$ V\QWKHVLV f 7KH n HQG RI FHOO F\FOH GHSHQGHQW P51$V FRQWDLQV D UHJLRQ RI K\SKHQDWHG G\DG V\PPHWU\ f ZKLFK LV HVVHQWLDO DOWKRXJK QRW VXIILFLHQW IRU FRXSOLQJ KLVWRQH P51$ VWDELOLW\ ZLWK '1$ UHSOLFDWLRQ f *UDYHV HW DO f REVHUYHG WKDW PRYLQJ WKH n VWHPORRS PRWLI DIIHFWV FRXSOLQJ RI KLVWRQH P51$ VWDELOLW\ ZLWK '1$ UHSOLFDWLRQ 'HVWDELOL]DWLRQ RI KLVWRQH P51$ LQLWLDWHV DW WKH n WHUPLQXV f DQG LV FRQWLQXHG UDSLGO\ LQ D n WR GLUHFWLRQ E\ D ULERVRPH FRPSOH[DVVRFLDWHG HQGRQXFOHDVH f ,Q DGGLWLRQ VXEFHOOXODU ORFDOL]DWLRQ PD\ SOD\ D UROH LQ KLVWRQH P51$ VWDELOLW\ f 7KH n HQG RI PRVW HXNDU\RWLF P51$V LV IRUPHG E\ DQ HQGRQXFOHRO\WLF FOHDYDJH UHDFWLRQ IROORZHG E\ SRO\DGHQ\ODWLRQ RI WKH n WHUPLQXV f 7KLV FOHDYDJH UHDFWLRQ UHTXLUHV WZR VHTXHQFHV D KLJKO\ FRQVHUYHG $$8$$$ VHTXHQFH DQG D GRZQVWUHDP *8 ULFK VHTXHQFH ZKLFK

PAGE 24

IODQN WKH FOHDYDJH VLWH f 0RVW KLVWRQH P51$V DUH QRW SRO\DGHQ\ODWHG DQG WKHLU P51$ n HQG IRUPDWLRQ UHTXLUHV WZR HOHPHQWV WKH VWHPORRS PRWLI DQG D SXULQH ULFK VHTXHQFH n WR WKH VWHPORRS PRWLI f $Q HQGRQXFOHRO\WLF FOHDYDJH UHDFWLRQ RFFXUV WR WKH VWHPORRS PRWLI f DQG LV GHSHQGHQW XSRQ WKH LQWHUDFWLRQ RI WKH 8 VQ51$ ZLWK WKH SXULQH ULFK VHTXHQFH f +% +LVWRQH *HQH ([SUHVVLRQ 7R GDWH +% KLVWRQH JHQH RUJDQL]DWLRQ DQG H[SUHVVLRQ KDYH EHHQ VWXGLHG PRVW WKRURXJKO\ LQ WKH FKLFNHQ ILYH UHSOLFDWLRQGHSHQGHQW RQH SDUWLDOO\ UHSOLFDWLRQGHSHQGHQW RQH XQFKDUDFWHUL]HG DQG RQH WHVWLVVSHFLILF FKLFNHQ +% JHQHV KDYH EHHQ GHVFULEHG f +XPDQ +% KLVWRQH JHQH H[SUHVVLRQ KDV QRW EHHQ UHSRUWHG LQ VXFK GHWDLO $OWKRXJK VHYHUDO KXPDQ KLVWRQH +% UHSOLFDWLRQ GHSHQGHQW JHQHV DQG RQH SVHXGRJHQH KDYH EHHQ GHVFULEHG f QR KXPDQ UHSODFHPHQW RU WLVVXHVSHFLILF +% JHQHV KDYH EHHQ UHSRUWHG 2YHUYLHZ RI 3URMHFW 7KH RYHUDOO DLP RI WKLV ZRUN ZDV WR LVRODWH DQG FKDUDFWHUL]H D YDULDQW KXPDQ KLVWRQH JHQH DQG VWXG\ LWV UHJXODWLRQ LQ UHVSRQVH WR FKDQJHV LQ WKH SUROLIHUDWLYH VWDWH RI WKH FHOO :KHQ ZH EHJDQ WKHVH VWXGLHV LQIRUPDWLRQ LQ

PAGE 25

WKH OLWHUDWXUH RQ QRQKXPDQ YDULDQW KLVWRQH JHQHV LQGLFDWHG WKH JHQHV IRU KXPDQ YDULDQW KLVWRQH SURWHLQV ZRXOG EH VWUXFWXUDOO\ PRUH FRPSOH[ WKDQ WKHLU UHSOLFDWLRQGHSHQGHQW FRXQWHUSDUWV f 7KLV LQGLFDWLRQ ZDV VXSSRUWHG E\ WKH REVHUYDWLRQV RI %RUXQ HW DO f WKDW D IUDFWLRQ RI KXPDQ KLVWRQH P51$V FRQWDLQHG VKRUW WUDFWV RI SRO\$f ,Q DGGLWLRQ EHFDXVH FRQVWLWXWLYHO\ H[SUHVVHG KLVWRQH SURWHLQV DUH V\QWKHVL]HG WKURXJKRXW WKH FHOO F\FOH f WKHLU P51$V PXVW EH VWDEOH LQ WKH DEVHQFH RI '1$ V\QWKHVLV 3UHVXPDEO\ VWUXFWXUDO IHDWXUHV RI FRQVWLWXWLYHO\ H[SUHVVHG KLVWRQH P51$V FRXOG UHQGHU WKHP UHVLVWDQW WR WKH PHFKDQLVPV ZKLFK UDSLGO\ DQG VHOHFWLYHO\ GHVWDELOL]H UHSOLFDWLRQ GHSHQGHQW KLVWRQH P51$V WRZDUG WKH HQG RI 6 SKDVH RU LQ UHVSRQVH WR LQKLELWLRQ RI '1$ V\QWKHVLV f $OWHUQDWLYHO\ WKH DEVHQFH RI VWUXFWXUDO HOHPHQWV HQGRJHQRXV WR UHSOLFDWLRQGHSHQGHQW KLVWRQH P51$V FRXOG KDYH WKH VDPH UHVXOW 7KHUHIRUH ZH KDYH IRFXVHG RXU DWWHQWLRQ RQ WKH FORQLQJ RI D YDULDQW KXPDQ +% KLVWRQH JHQH DQG FKDUDFWHUL]DWLRQ RI LWV VWUXFWXUDO IHDWXUHV DQG WKHLU HIIHFW RQ H[SUHVVLRQ LQ UHVSRQVH WR FKDQJHV LQ WKH JURZWK VWDWH RI WKH FHOO

PAGE 26

&+$37(5 0$7(5,$/6 $1' 0(7+2'6 0DWHULDOV 0DWHULDOV DQG %LRFKHPLFDO 5HDJHQWV 7KH PDMRULW\ RI FKHPLFDOV UHDJHQWV DQG VROYHQWV ZHUH SXUFKDVHG IURP HLWKHU )LVKHU 6FLHQWLILF 6SULQJILHOG 1RU 6LJPD &KHPLFDO &R 6W /RXLV 02 XQOHVV RWKHUZLVH LQGLFDWHG )RUPDPLGH DQG ;JDO ZHUH SXUFKDVHG IURP %HWKHVGD 5HVHDUFK /DERUDWRULHV %5/f *DLWKHUVEXUJ 0' %LR*HO $P $P DQG $P PHVKf DJDURVH EHDGV IRU JHO ILOWUDWLRQ 'RZH[ :; FDWLRQ H[FKDQJH UHVLQ PHVKf K\GURJHQ IRUP XOWUDSXUH HOHFWURSKRUHVLV JUDGH DJDURVH DQG =HWDSUREH Q\ORQ PHPEUDQHV ZHUH SXUFKDVHG IURP %LR 5DG 5LFKPRQG &$ $FU\ODPLGH FU\VWDOOL]HGf $36 XOWUDSXUH ELVDFU\ODPLGH 11nPHWK\OHQHELVDFU\ODPLGHf '77 &OHODQGnV UHDJHQWf JO\FRJHQ PROHFXODU ELRORJ\ JUDGHf DQG ,37* ZHUH SXUFKDVHG IURP %RHKULQJHU 0DQQKHLP %LRFKHPLFDOV %0%f ,QGLDQDSROLV ,1 &URQH[ [UD\ ILOP ZDV PDQXIDFWXUHG E\ 'XSRQW :LOPLQJWRQ '( 'LPHWK\O VXOIDWH GLFKORURGLPHWK\O VLODQH K\GUD]LQH 3(* 396 ;$5 [UD\ ILOP DQG ;2PDW GXSOLFDWLQJ ILOP ZHUH PDQXIDFWXUHG E\ (DVWPDQ .RGDN &R 5RFKHVWHU 1< 63(&75$325

PAGE 27

PROHFXODUSRURXV PHPEUDQH WXELQJ FHOOXVROYH HWK\OHQH JO\FRO PRQRHWK\O HWKHUf GLPHWK\O VXOIR[LGH HWKLGLXP EURPLGH SLSHULGLQH 7&$ DQG WROXHQH ZHUH SXUFKDVHG IURP )LVKHU 6FLHQWLILF 6SULQJILHOG 1&HVLXP FKORULGH VSHFLDO ELRFKHPLFDO JUDGH bf ZDV SXUFKDVHG IURP *DOODUG 6FKOHVLQJHU ,QGXVWULHV ,QF &DUOH 3ODFH 1< 237,)/825 VFLQWLOODWLRQ FRFNWDLO ZDV SXUFKDVHG IURP 3DFNDUG ,QVWUXPHQW &R ,QF 'RZQHUV *URYH ,/ )LFROO ZDV SXUFKDVHG IURP 3KDUPDFLD ,QF 3LVFDWDZD\ 17\SH DQG W\SH LQVWDQW VKHHW ILOP ZHUH SXUFKDVHG IURP 3RODURLG &DPEULGJH 0$ &RQFHQWUDWHG OLTXLG VFLQWLOODWRU /LTXLIOXRUf ZDV SXUFKDVHG IURP 5HVHDUFK 3URGXFWV ,QWHUQDWLRQDO (ON *URYH 9LOODJH ,/ (OXWLSG PLQLFROXPQV PP DQG PP QLWURFHOOXORVH FLUFOHV SRUH Pf DQG QLWURFHOOXORVH VKHHWV ZHUH SXUFKDVHG IURP 6FKOHLFKHU DQG 6FKXHOO ,QF .HHQH 1+ %6$ IUDFWLRQ 9f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

PAGE 28

'H[WURVH ZDV SXUFKDVHG IURP )LVKHU 6FLHQWLILF 6SULQJILHOG 1*HQHWLFLQ VXOIDWHf ZDV SXUFKDVHG IURP *LEFR /DERUDWRULHV &KORURP\FHWLQ VRGLXP VXFFLQDWH FKORUDPSKHQLFRO VRGLXP VXFFLQDWHf ZDV SXUFKDVHG IURP 3DUNH 'DYLV 0RUULV 3ODQHV 17*) ZDV SXUFKDVHG IURP 5t' 6\VWHPV ,QF 0LQQHDSROLV 01 $PSLFLOOLQ '>@D DPLQREHQ]\OSHQLFLOOLQf VRGLXP VDOW GH[DPHWKDVRQH K\GUR[\XUHD LQVXOLQ IURP ERYLQH SDQFUHDVf LVREXW\O PHWK\O[DQWKLQH 1= DPLQH FDVHLQ HQ]\PDWLF K\GURO\VDWHf SHQLFLOOLQ* %HQ]\O SHQLFLOOLQf SRWDVVLXP VDOW VWUHSWRP\FLQ VXOIDWH VWUHSWRP\FLQ VHVTXLVXOIDWHf WHWUDF\FOLQH 73$ DQG WU\SVLQ IURP SRUFLQH SDQFUHDVf SUHSDUHG LQ ,; +%66 ZLWKRXW FDOFLXP RU PDJQHVLXP ZHUH SXUFKDVHG IURP 6LJPD &KHPLFDO &R (Q]\PHV DQG .LWV 5HVWULFWLRQ HQGRQXFOHDVHV ZHUH SXUFKDVHG SULPDULO\ IURP %5/ %0% DQG 1HZ (QJODQG %LR /DEV 1(%f %HYHUO\ 0$ (Q]\PH UHDFWLRQV ZHUH FDUULHG RXW XVLQJ WKH EXIIHU DQG LQFXEDWLRQ FRQGLWLRQV GHVFULEHG E\ WKH PDQXIDFWXUHUV XQOHVV RWKHUZLVH LQGLFDWHG (Br FROL '1$ SRO\PHUDVH ODUJH IUDJPHQW .OHQRZ HQ]\PHf ZDV SXUFKDVHG IURP %5/ 3URWHLQDVH &,3 QXFOHDVH 6, DQG 7 '1$ OLJDVH ZHUH SXUFKDVHG IURP %0% 7U\SVLQ b LQ EXIIHUHG VDOLQHf ZDV SXUFKDVHG IURP +D]OHWRQ %LRORJLHV ,QF /HQH[D .6 7KH (;20(7+ '1$ VHTXHQFLQJ NLW ZLWK UHYHUVH WUDQVFULSWDVH DQG

PAGE 29

WKH 35,0(,7 UDQGRP SULPHU NLW ZHUH SXUFKDVHG IURP 6WUDWDJHQH /D -ROOD &$ '1DVH '1(3f '1DVH '1f DQG 51DVH $ 7\SH ,,,$f ZHUH REWDLQHG IURP 6LJPD &KHPLFDO &R 7KH 6HTXHQDVH '1$ VHTXHQFLQJ NLW DQG 7 SRO\QXFOHRWLGH NLQDVH FORQHGf ZHUH IURP 8QLWHG 6WDWHG %LRFKHPLFDO &RUS 86%f &OHYHODQG 2+ /\VR]\PH ZDV SXUFKDVHG IURP :RUWKLQJWRQ %LRFKHPLFDOV )UHHKROG 11XFOHLF $FLGV DQG 1XFOHRWLGHV >PHWK\O+@7K\PLGLQH &LPPROHf >D3@G$73 &LPPROHf >D3@G&73 &LPPROHf \3@$73 &LPPROHf >D3@873 &LPPROHf DQG >D6@G$73 &LPPROHf UDGLRQXFOHRWLGHV ZHUH SXUFKDVHG IURP $PHUVKDP &RUS $UOLQJWRQ +HLJKWV ,/ RU 1(1 5HVHDUFK 3URGXFWV :LOPLQJWRQ '( 2OLJRQXFOHRWLGHV ZHUH V\QWKHVL]HG LQ WKH UHFRPELQDQW '1$ FRUH IDFLOLW\ RI WKH 'HSDUWPHQW RI &HOO %LRORJ\ DW WKH 8QLYHUVLW\ RI 0DVVDFKXVHWWV 0HGLFDO &HQWHU XVLQJ DQ $SSOLHG %LRV\VWHPV $%f $ '1$ V\QWKHVL]HU SXUFKDVHG IURP $% )RVWHU &LW\ &DOLIRUQLD $73 IURP HTXLQH PXVFOHf G$73 VRGLXP VDOWf G&73 VRGLXP VDOWf G*73 VRGLXP VDOWf 773 VRGLXP VDOWf ROLJR G7FHOOXORVH DQG VDOPRQ WHVWHV '1$ W\SH ,,,f ZHUH REWDLQHG IURP 6LJPD &KHPLFDO &R 51$ PROHFXODU ZHLJKW PDUNHUV ZHUH SXUFKDVHG IURP %0% DQG IURP %5/ 0PSO 5) '1$ DQG 0PSO 5) '1$ ZHUH REWDLQHG IURP %0%

PAGE 30

3URSDJDWLRQ DQG 0DLQWHQDQFH RI %DFWHULDO 6WUDLQV *URZWK RI %DFWHULDO 6WUDLQV (VFKHULFKLD FROL ( FROLf EDFWHULD VHH DSSHQGL[ &f ZHUH JURZQ DW r& LQ <71 PHGLXP ZLWK YLJRURXV VKDNLQJ RU RQ SODWHV FRQWDLQLQJ <71 ERWWRP DJDU VHH DSSHQGL[ $f XQOHVV RWKHUZLVH LQGLFDWHG ,I QHFHVVDU\ DPSLFLOOLQ ZDV DGGHG WR WKH PHGLD WR D ILQDO FRQFHQWUDWLRQ RI JPO RU WHWUDF\FOLQH WR [JPO LPPHGLDWHO\ SULRU WR LQRFXODWLRQ RI OLTXLG FXOWXUHV RU SRXULQJ RI <71 DJDU SODWHV /LTXLG FXOWXUHV RI EDFWHULD ZHUH XVXDOO\ JURZQ VXFK WKDW WKH YROXPH RI WKH DLUVSDFH DERYH WKH FXOWXUH ZDV DW OHDVW WLPHV WKDW RI WKH PHGLXP XQOHVV RWKHUZLVH LQGLFDWHG 6WRUDJH RI %DFWHULDO 6WUDLQV %DFWHULDO FRORQLHV ZHUH VWRUHG IRU VKRUW SHULRGV RI WLPH RQ WKH VXUIDFH RI ERWWRP DJDU SODWHV DW r& 7KHVH SODWHV ZHUH ZUDSSHG LQ SDUDILOP WR SUHYHQW PRLVWXUH ORVV DQG VWRUHG LQYHUWHG )RU ORQJWHUP SUHVHUYDWLRQ RI EDFWHULDO VWUDLQV EDFWHULD ZHUH VWRUHG LQ PHGLXP FRQWDLQLQJ b JO\FHURO DW r& 7R SUHSDUH EDFWHULDO VWRFNV PO RI <71 ZDV LQRFXODWHG ZLWK D VLQJOH FRORQ\ RI EDFWHULD DQG JURZQ DQDHURELFDOO\ IRU KRXUV $ PO DOLTXRW RI WKH DQDHURELF FXOWXUH ZDV WKHQ SODFHG LQWR D YLDO FRQWDLQLQJ PO RI VWHULOH JO\FHURO WKH FRQWHQWV ZHUH PL[HG DQG WKH YLDO VWRUHG DW r&

PAGE 31

3URSDJDWLRQ DQG 0DLQWHQDQFH RI ; %DFWHULRSKDJH *URZWK DQG 7LWUDWLRQ RI $ %DFWHULRSKDJH /DPEGD &KDURQ $ f DQG $(0%/ f UHFRPELQDQW SKDJH ZHUH JURZQ LQ WKH /( VWUDLQ RI (/ FROL f DQG $JWOO UHFRPELQDQW SKDJH LQ WKH < VWUDLQ RI (/ FROL f /DPEGD EDFWHULRSKDJH ZHUH JURZQ HLWKHU RQ 1=&<0 SODWHV DV GHVFULEHG E\ 0DQLDWLV HW DO f LQ VPDOO VFDOH OLTXLG FXOWXUHV DV GHVFULEHG E\ /HGHU HW DO f RU LQ ODUJH VFDOH OLTXLG FXOWXUHV f 3UHSDUDWLRQ RI SODWLQJ EDFWHULD f $ VLQJOH EDFWHULDO FRORQ\ RU ORRS RI EDFWHULD IURP D JO\FHURO VWRFNf ZDV XVHG WR LQRFXODWH PO RI 1=&<0 PHGLXP VXSSOHPHQWHG ZLWK L RI b PDOWRVH VHH DSSHQGL[ $f DQG JURZQ IRU KRXUV DW r& ZLWK YLJRURXV VKDNLQJ 0DOWRVH LQGXFHV WKH PDOWRVH RSHURQ FRQWDLQLQJ WKH JHQH FRGLQJ IRU WKH $ UHFHSWRU ZKLFK LV HVVHQWLDO IRU HIILFLHQW DEVRUSWLRQ RI $ EDFWHULRSKDJH WR EDFWHULD 7KH FHOOV ZHUH WKHQ FHQWULIXJHG DW USP LQ DQ ,(& URWRU ; Jf IRU PLQXWHV DW r& 7KH VXSHUQDWDQW ZDV GLVFDUGHG DQG WKH FHOO SHOOHW UHVXVSHQGHG LQ PO ; WKH YROXPH RI WKH RULJLQDO FXOWXUHf VWHULOH P0 0J6 7KHVH EDFWHULD ZHUH VWRUHG DW r& DQG XVHG IRU XS WR ZHHNV KRZHYHU QRUPDOO\ WKH FHOOV ZHUH XVHG ZLWKLQ GD\V RI WKHLU SUHSDUDWLRQ

PAGE 32

2EWDLQLQJ D WLWHU RI EDFWHULRSKDJH 7LWUDWLRQ RI EDFWHULRSKDJH ZDV FDUULHG RXW HVVHQWLDOO\ DV GHYHORSHG E\ )HOL[ Gn+UHOOH LQ f DQG GHVFULEHG E\ 0DQLDWLV HW DO f 6HULDO GLOXWLRQV RI EDFWHULRSKDJH ZHUH SUHSDUHG LQ 60 DQG [O DOLTXRWV RI HDFK GLOXWLRQ WR EH DVVD\HG ZHUH SODFHG LQWR PP [ PP FXOWXUH WXEHV $ 0O DOLTXRW RI SODWLQJ EDFWHULD ZDV SODFHG LQWR HDFK WXEH DQG WKH WXEH ZDV YRUWH[HG JHQWO\ DQG LQFXEDWHG DW r& IRU PLQXWHV WR DOORZ WKH EDFWHULRSKDJH WR DEVRUE 8SRQ FRPSOHWLRQ RI WKH LQFXEDWLRQ SHULRG PO DOLTXRWV RI 1=&<0 WRS DJDU r&f ZHUH DGGHG WR HDFK WXEH DQG WKH FRQWHQWV ZHUH YRUWH[HG JHQWO\ DQG WKHQ SRXUHG RQWR 1=&<0 ERWWRP DJDU SODWHV VHH DSSHQGL[ $ IRU SUHSDUDWLRQ RI 1=&<0 WRS DJDU DQG 1=&<0 ERWWRP DJDU SODWHVf 8SRQ VROLGLILFDWLRQ RI WKH WRS DJDU WKH SODWHV ZHUH LQYHUWHG DQG LQFXEDWHG DW r& IRU KRXUV WR DOORZ SODTXH IRUPDWLRQ 6PDOO VFDOH OLTXLG FXOWXUHV RI ; EDFWHULRSKDJH f $SSUR[LPDWHO\ RI D UHVXVSHQGHG EDFWHULRSKDJH SODTXH RU ; EDFWHULRSKDJH IURP D VWRFN ZDV PL[HG ZLWK QO RI D IUHVK EDFWHULDO RYHUQLJKW FXOWXUH LQ D VWHULOH PO SRO\SURS\OHQH WXEH DQG LQFXEDWHG IRU PLQXWHV DW r& )ROORZLQJ DEVRUSWLRQ RI WKH EDFWHULRSKDJH PO RI 1=&<0 PHGLXP ZDV DGGHG DQG WKH WXEH LQFXEDWHG DW r& ZLWK VKDNLQJ IRU KRXUV XQWLO O\VLV RFFXUUHG $IWHU O\VLV QO RI &+& ZDV DGGHG DQG WKH LQFXEDWLRQ FRQWLQXHG IRU PLQXWHV

PAGE 33

8SRQ FRPSOHWLRQ RI WKH LQFXEDWLRQ SHULRG WKH FXOWXUH ZDV FHQWULIXJHG DW USP LQ D %HFNPDQ -$ URWRU ; Jf IRU PLQXWHV DW r& 1H[W WKH VXSHUQDWDQW ZDV UHFRYHUHG MXO RI &+&OM DGGHG DQG WKH VWRFN VWRUHG DW r& 7KH WLWHUV ZHUH QRUPDOO\ LQ WKH UDQJH RI O2O2PO /DUJH VFDOH OLTXLG FXOWXUHV RI ; EDFWHULRSKDJH f $ VLQJOH EDFWHULDO FRORQ\ RU ORRS RI EDFWHULD IURP D JO\FHURO VWRFNf ZDV XVHG WR LQRFXODWH PO RI 1=&<0 PHGLXP ZKLFK ZDV JURZQ IRU KRXUV DW r& ZLWK YLJRURXV VKDNLQJ 7KH 2' RI WKH FXOWXUH ZDV PHDVXUHG DQG WKH FHOO FRQFHQWUDWLRQ FDOFXODWHG DVVXPLQJ WKDW 2' ; FHOOVPO f $Q DOLTXRW FRQWDLQLQJ r FHOOV ZDV UHPRYHG DQG FHQWULIXJHG DW USP LQ DQ ,(& URWRU ; Jf IRU PLQXWHV DW r& $IWHU FHQWULIXJDWLRQ WKH VXSHUQDWDQW ZDV GLVFDUGHG DQG WKH FHOOV ZHUH UHVXVSHQGHG LQ PO 60 $SSUR[LPDWHO\ ; EDFWHULRSKDJH ZHUH DGGHG WR WKH FHOO VXVSHQVLRQ DQG WKH WXEH ZDV LQFXEDWHG DW r& IRU PLQXWHV ZLWK LQWHUPLWWHQW VKDNLQJ 8SRQ FRPSOHWLRQ RI WKH LQFXEDWLRQ SHULRG WKH FRQWHQWV RI WKH WXEH ZHUH DGGHG WR PO RI SUHZDUPHG 1=&<0 DQG LQFXEDWHG DW r& ZLWK YLJRURXV VKDNLQJ IRU KRXUV XQWLO O\VLV RFFXUUHG $IWHU O\VLV PO RI &+& ZDV DGGHG DQG WKH LQFXEDWLRQ FRQWLQXHG IRU PLQXWHV 8SRQ FRPSOHWLRQ RI WKH LQFXEDWLRQ WKH $ EDFWHULRSKDJH ZHUH SXULILHG DV GHVFULEHG E\
PAGE 34

,VRODWLRQ DQG 3XULILFDWLRQ RI $ %DFWHULRSKDJH f $IWHU WUHDWPHQW RI WKH ODUJH VFDOH SUHSDUDWLRQ RI ; EDFWHULRSKDJH ZLWK &+& WKH FXOWXUH ZDV FKLOOHG WR URRP WHPSHUDWXUH SDQFUHDWLF '1DVH DQG 51DVH $ ZHUH DGGHG WR D ILQDO FRQFHQWUDWLRQ RI [JPO HDFK DQG WKH FXOWXUH ZDV LQFXEDWHG IRU PLQXWHV DW URRP WHPSHUDWXUH 8SRQ FRPSOHWLRQ RI WKH LQFXEDWLRQ VROLG 1D&O ZDV DGGHG WR D ILQDO FRQFHQWUDWLRQ RI 0 Jf GLVVROYHG E\ VZLUOLQJ DQG WKH FXOWXUH LQFXEDWHG IRU KRXU RQ ZHW LFH 7KH FXOWXUH ZDV WKHQ SRXUHG LQWR D PO FHQWULIXJH ERWWOH WKH &+&OM ZDV FDUHIXOO\ OHIW EHKLQG DQG WKH FXOWXUH FHQWULIXJHG DW USP LQ D %HFNPDQ -$ URWRU ; Jf IRU PLQXWHV DW r& 7KH VXSHUQDWDQW ZDV SRXUHG LQWR D FOHDQ IODVN VROLG 3(* IURP (DVWPDQ .RGDN FKHPLFDO VRXUFH LV LPSRUWDQWf ZDV DGGHG WR D ILQDO FRQFHQWUDWLRQ RI b ZY Jf GLVVROYHG E\ VORZ VWLUULQJ DQG LQFXEDWHG IRU DW OHDVW KRXU RQ ZHW LFH RIWHQ WKLV SUHFLSLWDWLRQ VWHS ZDV FDUULHG RXW DW r& RYHUQLJKWf $IWHU WKH LQFXEDWLRQ WKH SUHFLSLWDWHG EDFWHULRSKDJH SDUWLFOHV ZHUH UHFRYHUHG E\ FHQWULIXJDWLRQ DW USP LQ D %HFNPDQ -$ URWRU ; Jf IRU PLQXWHV DW r& 7KH VXSHUQDWDQW ZDV SRXUHG RII DQG WKH ERWWOHV ZHUH GUDLQHG LQ D WLOWHG SRVLWLRQ IRU PLQXWHV 7KH EDFWHULRSKDJH SHOOHW ZDV JHQWO\ UHVXVSHQGHG LQ PO RI 60 XVLQJ D 3DVWHXU SLSHWWH DQG D UXEEHU EXOE WR GLVORGJH WKH EDFWHULRSKDJH SDUWLFOHV IURP WKH ZDOO RI WKH FHQWULIXJH ERWWOH 7KH EDFWHULRSKDJH VXVSHQVLRQ

PAGE 35

ZDV H[WUDFWHG ZLWK PO &+&,$$ YYf DQG FHQWULIXJHG DW USP LQ DQ ,(& URWRU ; Jf IRU PLQXWHV DW r& 7KH DTXHRXV SKDVH ZDV UHFRYHUHG DQG WKH DERYH H[WUDFWLRQ UHSHDWHG XQWLO QR 3(* LQWHUIDFH UHPDLQHG 8SRQ FRPSOHWLRQ RI WKH &+& H[WUDFWLRQV J RI VROLG &V&O ZDV DGGHG IRU HYHU\ PO RI VROXWLRQ DQG PL[HG JHQWO\ 7KH EDFWHULRSKDJH VXVSHQVLRQ ZDV WKHQ FHQWULIXJHG DW USP LQ D %HFNPDQ 7L RU 7 URWRU ; Jf IRU KRXUV DW r& 7KH EDQG RI EDFWHULRSKDJH SDUWLFOHV ZDV FROOHFWHG XVLQJ DQ JDXJH QHHGOH DQG VWRUHG DW r& 6WRUDJH RI $ %DFWHULRSKDJH 6WRFNV %DFWHULRSKDJH ZHUH VWRUHG LQ RQH RI WKUHH ZD\V Lf 3DVWHXU SLSHWWH SOXJV RI EDFWHULRSKDJH SODTXHV ZHUH SODFHG LQWR PO DOLTXRWV RI 60 LQLWLDO WLWHUV RI DSSUR[LPDWHO\ POf DQG VWRUHG DW r& IRU \HDUV LLf VXSHUQDWDQWV REWDLQHG IURP VPDOO VFDOH OLTXLG FXOWXUHV RI EDFWHULRSKDJH LQLWLDO WLWHUV RI DSSUR[LPDWHO\ O2O2POf ZHUH VWRUHG DW r& IRU \HDUV DQG LLLf VXVSHQVLRQV RI EDFWHULRSKDJH LQ &V&O REWDLQHG IURP ODUJH VFDOH OLTXLG FXOWXUHV RI EDFWHULRSKDJH SXULILHG RYHU &V&O JUDGLHQWV LQLWLDO WLWHUV RI DSSUR[LPDWHO\ ,rOPOf ZHUH VWRUHG DW r& IRU RYHU \HDUV 6WRFNV RI EDFWHULRSKDJH JHQHUDOO\ H[SHULHQFHG D GURS LQ WLWHU RI RQH RUGHU RI PDJQLWXGH GXULQJ WKH ILUVW PRQWK RI VWRUDJH DQG DQ HTXLYDOHQW GURS GXULQJ HDFK \HDU RI IXUWKHU VWRUDJH

PAGE 36

5HFRPELQDQW 3KDJH '1$V 7KH $ &KDURQ $ UHFRPELQDQW EDFWHULRSKDJH XVHG LQ WKLV VWXG\ ZHUH SUHYLRXVO\ LVRODWHG f IURP D $ &KDURQ $ JHQRPLF '1$ OLEUDU\ $OVR XVHG LQ WKLV VWXG\ ZHUH $JWOO UHFRPELQDQW EDFWHULRSKDJH LVRODWHG VHH FKDSWHU f IURP D $JWOO KXPDQ OLYHU F'1$ OLEUDU\ DQG $(0%/ UHFRPELQDQW EDFWHULRSKDJH ZKLFK ZHUH LVRODWHG VHH FKDSWHU f IURP D $(0%/ DGXOW KXPDQ O\PSKRF\WH JHQRPLF '1$ OLEUDU\ ,VRODWLRQ DQG 3XULILFDWLRQ RI %DFWHULRSKDJH '1$ /DUJH 6FDOH $ %DFWHULRSKDJH '1$ ,VRODWLRQ f '1$ ZDV LVRODWHG IURP SXULILHG $ EDFWHULRSKDJH DV GHVFULEHG E\ 0DQLDWLV HW DO f 7KH SXULILHG EDFWHULRSKDJH SUHSDUDWLRQ ZDV GLDO\]HG WZLFH DW URRP WHPSHUDWXUH IRU KRXU DJDLQVW D ; YROXPH RI P0 1D&O P0 7ULV}&O S+ f P0 0J&O WR UHPRYH FHVLXP FKORULGH 7KH EDFWHULRSKDJH VXVSHQVLRQ ZDV WUDQVIHUUHG WR D JODVV PO H[WUDFWLRQ WXEH DQG ('7$ DGGHG WR D ILQDO FRQFHQWUDWLRQ RI P0 3URWHLQDVH ZDV DGGHG WR D ILQDO FRQFHQWUDWLRQ RI [JPO 6'6 WR D ILQDO FRQFHQWUDWLRQ RI b ZYf DQG WKH VROXWLRQ ZDV PL[HG JHQWO\ DQG LQFXEDWHG DW r& IRU KRXU 7KH VROXWLRQ ZDV H[WUDFWHG ZLWK DQ HTXDO YROXPH RI HTXLOLEUDWHG SKHQRO VHH DSSHQGL[ %f DQG FHQWULIXJHG DW USP LQ DQ ,(& URWRU ; Jf IRU PLQXWHV DW URRP WHPSHUDWXUH 1H[W WKH VROXWLRQ ZDV H[WUDFWHG ZLWK DQ HTXDO YROXPH RI D PL[WXUH YYf RI

PAGE 37

HTXLOLEUDWHG SKHQRO DQG &+&,$$ YYf DQG FHQWULIXJHG DW USP LQ DQ ,(& URWRU ; Jf IRU PLQXWHV DW URRP WHPSHUDWXUH 7KH DTXHRXV SKDVH ZDV UHFRYHUHG DQG H[WUDFWHG RQFH ZLWK DQ HTXDO YROXPH RI &+&,$$ YYf 8SRQ FRPSOHWLRQ RI WKH H[WUDFWLRQV WKH DTXHRXV SKDVH ZDV WUDQVIHUUHG WR D GLDO\VLV VDF DQG GLDO\]HG VHTXHQWLDOO\ IRU KRXUV DW r& DJDLQVW ; YROXPHV RI P0 7ULV}&O P0 ('7$ S+ f 7KH SXULILHG '1$ ZDV TXDQWLWDWHG DQG VWRUHG DW HLWKHU r& RU r& 6PDOO 6FDOH N %DFWHULRSKDJH '1$ ,VRODWLRQ f 6PDOO VFDOH N EDFWHULRSKDJH '1$ SUHSDUDWLRQV ZHUH FDUULHG RXW DV GHVFULEHG E\ 6FKZHL]HU f $ KLJK WLWHU O\VDWH RI N EDFWHULRSKDJH PO ZDV WUDQVIHUUHG WR D PO PLFURFHQWULIXJH WXEH FRQWDLQLQJ , b 6'6 ZYf 6XEVHTXHQWO\ cLO '(3& ZDV DGGHG DQG WKH WXEH PL[HG E\ LQYHUVLRQ 1H[W MXO RI 0 7ULV} &O S+ f DQG , 0 ('7$ ZHUH DGGHG WKH WXEH PL[HG DJDLQ E\ LQYHUVLRQ DQG LQFXEDWHG IRU PLQXWHV DW r& $IWHU WKH LQFXEDWLRQ , 0 VRGLXP DFHWDWH S+ f ZDV DGGHG DQG WKH LQFXEDWLRQ FRQWLQXHG IRU PLQXWHV RQ ZHW LFH 7KH VROXWLRQ ZDV WKHQ FHQWULIXJHG DW ; J IRU PLQXWHV DW r& )ROORZLQJ FHQWULIXJDWLRQ WKH VXSHUQDWDQW ZDV GHFDQWHG WR D IUHVK PLFURFHQWULIXJH WXEH 7KH WXEH ZDV ILOOHG ZLWK b HWKDQRO r&f PL[HG E\ LQYHUVLRQ DQG FHQWULIXJHG DW ; J IRU PLQXWHV DW URRP WHPSHUDWXUH

PAGE 38

7KH VXSHUQDWDQW ZDV GLVFDUGHG DQG WKH SHOOHW ZDVKHG ZLWK PO b HWKDQRO DW URRP WHPSHUDWXUH 7KH SHOOHW ZDV GULHG EULHIO\ XQGHU YDFXXP DQG UHVXVSHQGHG LQ O 7(6 EXIIHU P0 7ULV}&O S+ f P0 1D&O P0 ('7$f 7KH '1$ ZDV DSSOLHG WR D PO FROXPQ RI %LR*HO $P DJDURVH EHDGV DQG HOXWHG ZLWK 7(6 EXIIHU )UDFWLRQV ZHUH FROOHFWHG DQG DQDO\]HG RQ D b DJDURVH SODWH FRQWDLQLQJ QTPO HWKLGLXP EURPLGH )UDFWLRQV FRQWDLQLQJ $ EDFWHULRSKDJH '1$ ZHUH SRROHG DQG SUHFLSLWDWHG ZLWK 0 1D&O DQG YROXPHV RI b HWKDQRO r&f PL[HG E\ LQYHUVLRQ DQG FHQWULIXJHG DW ; J IRU PLQXWHV DW URRP WHPSHUDWXUH 7KH VXSHUQDWDQW ZDV GLVFDUGHG DQG WKH SHOOHW ZDVKHG ZLWK PO b HWKDQRO DW URRP WHPSHUDWXUH 7KH SHOOHW ZDV GULHG EULHIO\ XQGHU YDFXXP UHVXVSHQGHG LQ , P0 7ULV} &O S+ f P0 ('7$ FRQWDLQLQJ '1DVH IUHH 51DVH LJPOf DQG LQFXEDWHG IRU PLQXWHV DW r& )ROORZLQJ LQFXEDWLRQ WKH '1$ ZDV VWRUHG DW r& 6LQJOH6WUDQGHG 0 %DFWHULRSKDJH 7HPSODWH ,VRODWLRQ 6LQJOHVWUDQGHG 0 %DFWHULRSKDJH WHPSODWHV IRU VHTXHQFLQJ FORQHG LQVHUWV ZHUH SUHSDUHG DV GHVFULEHG E\ 6DQJHU HW DO f DQG 0HVVLQJ f $Q H[SRQHQWLDOO\ JURZLQJ FXOWXUH RI KRVW FHOOV ;/O%OXH f VHH DSSHQGL[ &f ZDV GLOXWHG LQ PO RI <71 PHGLXP LQIHFWHG ZLWK D SXULILHG 0 SODTXH DQG LQFXEDWHG IRU KRXUV DW r& ZLWK YLJRURXV VKDNLQJ )ROORZLQJ WKH

PAGE 39

LQFXEDWLRQ PO RI WKH FXOWXUH ZDV WUDQVIHUUHG WR D PLFURFHQWULIXJH WXEH DQG VSXQ LQ DQ (SSHQGRUI FHQWULIXJH DW ; J IRU PLQXWHV DW r& 7KH VXSHUQDWDQW POf ZDV WUDQVIHUUHG WR D PLFURFHQWULIXJH WXEH FRQWDLQLQJ LOO RI b 3(* 0 1D&O DQG LQFXEDWHG DW URRP WHPSHUDWXUH IRU PLQXWHV 7KH UHPDLQLQJ VXSHUQDWDQW ZDV SODFHG LQWR D FOHDQ PLFURFHQWULIXJH WXEH DQG VWRUHG DW r& DV D SKDJH VWRFN $IWHU WKH LQFXEDWLRQ WKH SKDJH VXVSHQVLRQ ZDV FHQWULIXJHG DW ; J IRU PLQXWHV DW URRP WHPSHUDWXUH DQG WKH VXSHUQDWDQW ZDV FDUHIXOO\ UHPRYHG DQG GLVFDUGHG 7KH SKDJH SHOOHW ZDV UHVXVSHQGHG LQ , 7(6 EXIIHU P0 7ULV}&O S+ f P0 1D&O P0 ('7$f E\ YRUWH[LQJ IRU VHFRQGV (TXLOLEUDWHG SKHQRO c[Of ZDV DGGHG WR WKH YLUXV VXVSHQVLRQ YRUWH[HG IRU WZR VHFRQGV LQFXEDWHG DW URRP WHPSHUDWXUH IRU PLQXWHV YRUWH[HG IRU DQ DGGLWLRQDO VHFRQGV DQG FHQWULIXJHG DW ; J IRU PLQXWHV DW URRP WHPSHUDWXUH 6LQJOHVWUDQGHG WHPSODWH '1$ ZDV SUHFLSLWDWHG IURP c[O RI DTXHRXV SKDVH ZLWK ILO 0 VRGLXP DFHWDWH , JO\FRJHQ PJPOf DQG , b HWKDQRO RQ FUXVKHG GU\ LFH IRU PLQXWHV DQG SHOOHWHG E\ FHQWULIXJDWLRQ DW ; J IRU PLQXWHV DW r& 7KH VXSHUQDWDQW ZDV GHFDQWHG DQG WKH SHOOHW ZDVKHG ZLWK , b HWKDQRO DW URRP WHPSHUDWXUH 7KH '1$ SHOOHW ZDV GULHG XQGHU YDFXXP UHVXVSHQGHG LQ c[O 7(6 EXIIHU DQG VWRUHG DW r&

PAGE 40

5HFRPELQDQW 3ODVPLG '1$V '1$ IUDJPHQWV FRQWDLQLQJ +, +$ +% + DQG + JHQHV IURP UHFRPELQDQW $ &KDURQ $ SKDJH f ZHUH VXEFORQHG LQWR S%5 RU S8& f DQG XQSXEOLVKHG GDWDf 7KH SODVPLG 5*$3'+ f ZDV JHQHURXVO\ SURYLGHG E\ 'U 3K -HDQWHXU DQG WKH SODVPLG *P f E\ 'U .HQQHWK 6RSUDQR DIWHU SUHSDUDWLRQ IURP D ES 3VW IUDJPHQW RI KXPDQ PLFURJOREXOLQ NLQGO\ SURYLGHG E\ 'U ,WDNXUD +DUYDUG 8QLYHUVLW\ ,VRODWLRQ DQG 3XULILFDWLRQ RI 3ODVPLG '1$ %DFWHULD FRQWDLQLQJ SODVPLGV ZHUH JURZQ LQ <71 PHGLXP FRQWDLQLQJ ]JPO DPSLFLOOLQ DQG DPSOLILFDWLRQ ZDV LQGXFHG LQ WKH SUHVHQFH RI JPO FKORUDPSKHQLFRO 3ODVPLG '1$ ZDV LVRODWHG DQG SXULILHG XVLQJ WKH IROORZLQJ PHWKRGV 5DSLG 6PDOO 6FDOH 3ODVPLG '1$ ,VRODWLRQ f 6PDOO VFDOH SODVPLG LVRODWLRQ ZDV FDUULHG RXW XVLQJ WKH UDSLG ERLOLQJ PHWKRG RI +ROPHV DQG 4XLJOH\ f $Q RYHUQLJKW FXOWXUH POf RI SODVPLGFRQWDLQLQJ EDFWHULD ZDV WUDQVIHUUHG WR D PLFURFHQWULIXJH WXEH DQG FHQWULIXJHG LQ DQ (SSHQGRUI FHQWULIXJH DW ; J IRU PLQXWHV DW URRP WHPSHUDWXUH 7KH VXSHUQDWDQW ZDV GHFDQWHG DQG WKH SHOOHW YRUWH[HG IRU VHFRQGV DQG UHVXVSHQGHG LQ Q 67(7 EXIIHU b VXFURVH b WULWRQ ; P0 ('7$ P0 7ULV}&O S+ ff $IWHU UHVXVSHQVLRQ [ RI IUHVKO\ SUHSDUHG

PAGE 41

O\VR]\PH PJPOf ZDV DGGHG WR WKH FHOO VXVSHQVLRQ WKH PL[WXUH ERLOHG IRU VHFRQGV DQG FHQWULIXJHG DW ; J IRU PLQXWHV DW URRP WHPSHUDWXUH 7KH SHOOHW ZDV UHPRYHG IURP WKH PLFURFHQWULIXJH WXEH XVLQJ D WRRWKSLFN DQG WKH '1$ SUHFLSLWDWHG IURP WKH VXSHUQDWDQW ZLWK , LVRSURSDQRO DW URRP WHPSHUDWXUH IRU PLQXWHV )ROORZLQJ SUHFLSLWDWLRQ WKH '1$ ZDV SHOOHWHG E\ FHQWULIXJDWLRQ DW ; J IRU PLQXWHV DW URRP WHPSHUDWXUH WKH VXSHUQDWDQW GHFDQWHG DQG WKH SHOOHW ZDVKHG ZLWK cM b HWKDQRO DW URRP WHPSHUDWXUH 7KH '1$ SHOOHW ZDV GULHG XQGHU YDFXXP UHVXVSHQGHG LQ LO P0 7ULV} &O P0 ('7$ S+ f FRQWDLQLQJ '1DVHIUHH 51DVH QJPOf DQG LQFXEDWHG IRU PLQXWHV DW r& )ROORZLQJ LQFXEDWLRQ WKH '1$ ZDV VWRUHG DW r& /DUJH 6FDOH 3ODVPLG '1$ ,VRODWLRQ /DUJH VFDOH LVRODWLRQ RI SODVPLG '1$ ZDV FDUULHG RXW XVLQJ WKH DONDOLQH O\VLV PHWKRG GHVFULEHG LQ %LUQERLP DQG 'RO\ f DQG PRGLILHG E\ ,VK+RURZLF] f $ PO FXOWXUH RI SODVPLGFRQWDLQLQJ EDFWHULD ZDV FHQWULIXJHG DW USP LQ D %HFNPDQ -$ URWRU ; Jf IRU PLQXWHV DW r& 7KH VXSHUQDWDQW ZDV GHFDQWHG DQG WKH FHOO SHOOHW UHVXVSHQGHG LQ PO DONDOLQH O\VLV VROXWLRQ VHH DSSHQGL[ %f FRQWDLQLQJ PJ O\VR]\PH DQG LQFXEDWHG IRU PLQXWHV DW URRP WHPSHUDWXUH $IWHU WKH LQFXEDWLRQ PO RI DONDOLQH O\VLV VROXWLRQ r VHH DSSHQGL[ %f ZDV DGGHG

PAGE 42

WKH VROXWLRQ ZDV PL[HG JHQWO\ E\ LQYHUWLQJ XQWLO QR FOXPSV UHPDLQHG DQG IXUWKHU LQFXEDWHG IRU PLQXWHV DW r& IROORZHG E\ WKH DGGLWLRQ RI PO LFH FROG 0 SRWDVVLXP DFHWDWH S+ f DQG LQFXEDWLRQ DW r& IRU DQ DGGLWLRQDO PLQXWHV 8SRQ FRPSOHWLRQ RI WKH LQFXEDWLRQ WKH VROXWLRQ ZDV WUDQVIHUUHG WR D PO SRO\FDUERQDWH FHQWULIXJH WXEH DQG FHQWULIXJHG DW USP LQ D %HFNPDQ -$ URWRU ; Jf IRU PLQXWHV DW r& 7KH VXSHUQDWDQW ZDV WUDQVIHUUHG WR D PO FRUH[ FHQWULIXJH ERWWOH FRQWDLQLQJ PO LVRSURSDQRO LQFXEDWHG DW URRP WHPSHUDWXUH IRU PLQXWHV DQG WKH '1$ FROOHFWHG E\ FHQWULIXJDWLRQ DW USP LQ D %HFNPDQ -$ URWRU ; Jf IRU PLQXWHV DW r& $IWHU WKH FHQWULIXJDWLRQ WKH VXSHUQDWDQW ZDV GHFDQWHG DQG WKH SHOOHW ZDVKHG ZLWK PO b HWKDQRO DW URRP WHPSHUDWXUH 7KH '1$ SHOOHW ZDV GULHG XQGHU YDFXXP DQG UHVXVSHQGHG LQ PO P0 7ULV}&O P0 ('7$ S+ f 7KH '1$ FROOHFWHG DIWHU LVRSURSDQRO SUHFLSLWDWLRQ ZDV SXULILHG E\ &V&O EXR\DQW GHQVLW\ JUDGLHQW FHQWULIXJDWLRQ DV GHVFULEHG E\ &OHZHOO DQG +HOLQVNL f 6ROLG &V&O J ZDV DGGHG WR WKH UHVXVSHQGHG '1$ POf DQG WKH VROXWLRQ ZDV PL[HG JHQWO\ XQWLO DOO RI WKH VDOW ZDV GLVVROYHG $ PJPO VWRFN RI HWKLGLXP EURPLGH PO IRU HYHU\ PO RI '1$ VROXWLRQ ZDV DGGHG DQG WKH ILQDO VROXWLRQ WUDQVIHUUHG WR %HFNPDQ TXLFN VHDO XOWUDFHQWULIXJH WXEHV 7KH SODVPLG '1$ ZDV EDQGHG E\ FHQWULIXJDWLRQ DW USP LQ D %HFNPDQ 7 XOWUDFHQWULIXJH URWRU ; Jf IRU KRXUV DW

PAGE 43

r& 7KH EDQG RI VXSHUFRLOHG SODVPLG '1$ ZDV FROOHFWHG XVLQJ DQ JDXJH QHHGOH DQG WKH '1$ SDVVHG RYHU D FDWLRQ H[FKDQJH FROXPQ RI 'RZH[ :; IROORZHG E\ FKURPDWRJUDSK\ WKURXJK D ; FP %LR*HO $P FROXPQ GHYHORSHG ZLWK P0 7ULV}&O P0 ('7$ S+ f )UDFWLRQV FRQWDLQLQJ SODVPLG '1$ 9Rf ZHUH SRROHG DQG HWKDQRO SUHFLSLWDWHG 7KH SXULILHG '1$ ZDV TXDQWLWDWHG DQG VWRUHG DW HLWKHU r& RU r& $OWHUQDWLYHO\ DIWHU FROOHFWLRQ RI WKH VXSHUFRLOHG SODVPLG '1$ EDQG WKH '1$ VROXWLRQ ZDV WUDQVIHUUHG WR D GLDO\VLV VDF DQG GLDO\]HG VHTXHQWLDOO\ IRU KRXUV DW r& DJDLQVW ; YROXPHV RI P0 7ULV}&O P0 ('7$ S+ f 7KH SXULILHG '1$ ZDV WKHQ TXDQWLWDWHG DQG VWRUHG DW HLWKHU r& RU r& ,VRODWLRQ DQG 3XULILFDWLRQ RI (XNDU\RWLF *HQRPLF '1$ (XNDU\RWLF JHQRPLF '1$ ZDV LVRODWHG DQG SXULILHG HVVHQWLDOO\ DV GHVFULEHG E\ *URVV%HOODUG HW DO f DQG PRGLILHG E\ %OLQ DQG 6WDIIRUG f $SSUR[LPDWHO\ ; HXNDU\RWLF FHOOV ZHUH FROOHFWHG E\ FHQWULIXJDWLRQ DW USP LQ DQ ,(& URWRU ; Jf IRU PLQXWHV DW r& 7KH VXSHUQDWDQW ZDV GHFDQWHG DQG WKH FHOOV JHQWO\ UHVXVSHQGHG LQ FROG 3%6 DQG UHFROOHFWHG E\ FHQWULIXJDWLRQ DW USP LQ DQ ,(& URWRU ; Jf IRU PLQXWHV DW r& $IWHU FHQWULIXJDWLRQ WKH VXSHUQDWDQW ZDV GHFDQWHG DQG WKH SHOOHW ZDV YRUWH[HG JHQWO\ UHVXVSHQGHG LQ PO '1$ O\VLV VROXWLRQ ; 66& ; 66& 0 1D&O 0 VRGLXP

PAGE 44

FLWUDWH S+ f b 6'6 ZYf MXJPO SURWHLQDVH .f DQG LQFXEDWHG IRU KRXUV DW URRP WHPSHUDWXUH ZLWK YHU\ JHQWO\ VKDNLQJ 8SRQ FRPSOHWLRQ RI WKH LQFXEDWLRQ WKH VROXWLRQ ZDV H[WUDFWHG ZLWK DQ HTXDO YROXPH RI HTXLOLEUDWHG SKHQRO DQG FHQWULIXJHG DW USP LQ DQ ,(& URWRU ; Jf IRU PLQXWHV DW URRP WHPSHUDWXUH 1H[W WKH VROXWLRQ ZDV H[WUDFWHG ZLWK DQ HTXDO YROXPH RI &+&,$$ YYf DQG DQ HTXDO YROXPH RI HTXLOLEUDWHG SKHQRO DQG FHQWULIXJHG DW USP LQ DQ ,(& URWRU ; Jf IRU PLQXWHV DW URRP WHPSHUDWXUH 7KLV H[WUDFWLRQ ZDV UHSHDWHG XQWLO D FOHDU LQWHUSKDVH ZDV YLVLEOH 7KH ILQDO H[WUDFWLRQ ZDV ZLWK DQ HTXDO YROXPH RI &+&,$$ YYf IROORZHG E\ FHQWULIXJDWLRQ DW USP LQ DQ ,(& URWRU ; Jf IRU PLQXWHV DW URRP WHPSHUDWXUH 8SRQ FRPSOHWLRQ RI WKH H[WUDFWLRQV WKH DTXHRXV SKDVH ZDV VSOLW LQWR WZR IUDFWLRQV RI HTXDO YROXPH DQG WUDQVIHUUHG WR PO FRUH[ FHQWULIXJH WXEHV 7KH '1$ ZDV SUHFLSLWDWHG ZLWK PO 0 VRGLXP DFHWDWH DQG PO b HWKDQRO RYHUQLJKW DW r& 7KH SUHFLSLWDWHG JHQRPLF '1$ ZDV KRRNHG RXW ZLWK D EHQW JODVV URG UHVXVSHQGHG LQ P0 7ULV}&O P0 ('7$ S+ f DQG VWRUHG DW r& ,Q DGGLWLRQ FHOOXODU '1$V XVHG IRU FKURPRVRPH ORFDOL]DWLRQ ZHUH LVRODWHG DV GHVFULEHG f ,VRODWLRQ DQG 3XULILFDWLRQ RI 0DPPDOLDQ 51$ f ,VRODWLRQ RI WRWDO FHOOXODU PDPPDOLDQ 51$ ZDV FDUULHG RXW DV SUHYLRXVO\ GHVFULEHG f &HOOV JURZLQJ LQ

PAGE 45

VXVSHQVLRQ FXOWXUH ZHUH KDUYHVWHG E\ FHQWULIXJDWLRQ DW USP LQ DQ ,(& URWRU ; Jf IRU PLQXWHV DW r& DQG ZDVKHG WZLFH ZLWK FROG ; VSLQQHU VDOW VROXWLRQ *,%&2f 7KH FHOOV ZHUH O\VHG LQ D VROXWLRQ ; FHOOVPO O\VLV VROXWLRQf FRQWDLQLQJ P0 7ULVm&O S+ f P0 ('7$ [JPO 396 b 6'6 ZYf DQG JPO SURWHLQDVH IRU PLQXWHV DW URRP WHPSHUDWXUH )ROORZLQJ WKH LQFXEDWLRQ 1D&O ZDV DGGHG WR 0 DQG WKH DTXHRXV SKDVH ZDV H[WUDFWHG RQFH ZLWK DQ HTXDO YROXPH RI D PL[WXUH YYf RI HTXLOLEUDWHG SKHQRO DQG &+&,$$ YYf DQG WZLFH ZLWK DQ HTXDO YROXPH RI &+&,$$ YYf 8SRQ FRPSOHWLRQ RI WKH H[WUDFWLRQV WKH WRWDO QXFOHLF DFLG ZDV SUHFLSLWDWHG XVLQJ ILO 0 VRGLXP DFHWDWH S+ f DQG WKUHH YROXPHV RI b HWKDQRO RYHUQLJKW DW r& 7KH WRWDO QXFOHLF DFLG ZDV FROOHFWHG E\ FHQWULIXJDWLRQ DW USP LQ D %HFNPDQ -$ URWRU ; Jf IRU PLQXWHV DW r& $IWHU WKH FHQWULIXJDWLRQ WKH VXSHUQDWDQW ZDV GHFDQWHG DQG WKH SHOOHW ZDVKHG ZLWK PO b HWKDQRO DW URRP WHPSHUDWXUH 7KH SHOOHW ZDV GULHG XQGHU YDFXXP UHVXVSHQGHG LQ '1DVH GLJHVWLRQ EXIIHU P0 7ULV}&O S+ f P0 &D&Of DQG LQFXEDWHG DW r& IRU PLQXWHV ZLWK PJPO '1DVH HOHFWURSKRUHWLFDOO\ SXUHf ZKLFK KDG EHHQ SUHWUHDWHG ZLWK SURWHLQDVH IRU KRXUV f )ROORZLQJ WKH LQFXEDWLRQ 1D&O ZDV DGGHG WR 0 6'6 WR b ZYf DQG H[WUDFWLRQV ZHUH FDUULHG RXW DV GHVFULEHG DERYH 8SRQ FRPSOHWLRQ RI WKH H[WUDFWLRQV WKH 51$ ZDV SUHFLSLWDWHG XVLQJ

PAGE 46

D ILQDO FRQFHQWUDWLRQ RI P0 VRGLXP DFHWDWH DQG WKUHH YROXPHV RI b HWKDQRO RYHUQLJKW DW r& 7KH WRWDO 51$ ZDV FROOHFWHG E\ FHQWULIXJDWLRQ DW USP LQ D %HFNPDQ -$ URWRU ; Jf IRU PLQXWHV DW r& )ROORZLQJ WKH FHQWULIXJDWLRQ WKH VXSHUQDWDQW ZDV GHFDQWHG DQG WKH SHOOHW ZDVKHG ZLWK PO b HWKDQRO DW URRP WHPSHUDWXUH 7KH SHOOHW ZDV GULHG XQGHU YDFXXP UHVXVSHQGHG LQ GG+ TXDQWLWDWHG DQG VWRUHG DW r& 6HOHFWLRQ RI 3RO\ $r 51$ 3RO\ $ 51$ ZDV VHOHFWHG XVLQJ D VLQJOH SDVV RYHU ROLJR G7FHOOXORVH DV GHVFULEHG E\ 0DQLDWLV HW DO f $ VLQJOH ROLJR G7FHOOXORVH VHOHFWLRQ JUHDWO\ HQULFKHV IRU SRO\ $ 51$ EXW GRHV QRW WRWDOO\ UHPRYH DOO RI WKH SRO\ $n 51$ 7RWDO FHOOXODU +H/D 51$ LQ ORDGLQJ EXIIHU P0 7ULV+&O S+ f P0 /L&O P0 ('7$ b 6'6 ZYff ZDV KHDWHG WR r& IRU PLQXWHV DQG DSSOLHG WR SUHZDVKHG ROLJR G7FHOOXORVH FROXPQV 7KH FROXPQV KDG EHHQ SUHZDVKHG ZLWK Lf WKUHH FROXPQ YROXPHV GG+ LLf WKUHH FROXPQ YROXPHV 1 1D2+ P0 ('7$ DQG LLLf WKUHH FROXPQ YROXPHV ORDGLQJ EXIIHU $IWHU DGGLWLRQ RI WKH VDPSOH WR WKH FROXPQ WKH IORZWKURXJK ZDV FROOHFWHG KHDWHG WR r& IRU PLQXWHV FRROHG DQG UHDSSOLHG WR WKH FROXPQ 7KH FROXPQ ZDV WKHQ ZDVKHG ZLWK WR FROXPQ YROXPHV RI ORDGLQJ EXIIHU IROORZHG E\ FROXPQ YROXPHV RI ORDGLQJ EXIIHU FRQWDLQLQJ P0 1D&O 7KH SRO\ $ 51$ ZDV HOXWHG ZLWK WR FROXPQ

PAGE 47

YROXPHV RI HOXWLRQ EXIIHU P0 7ULV}&O S+ f P0 ('7$ b 6'6 ZYff DQG SUHFLSLWDWHG ZLWK P0 VRGLXP DFHWDWH S+ f DQG YROXPHV b HWKDQRO RYHUQLJKW DW r& 51$ ZDV FROOHFWHG E\ FHQWULIXJDWLRQ DW USP LQ D %HFNPDQ -$ URWRU ; Jf IRU PLQXWHV DW r& )ROORZLQJ WKH FHQWULIXJDWLRQ WKH VXSHUQDWDQW ZDV GHFDQWHG DQG WKH SHOOHW ZDVKHG ZLWK PO b HWKDQRO DW URRP WHPSHUDWXUH 7KH SHOOHW ZDV GULHG XQGHU YDFXXP UHVXVSHQGHG LQ GG+ TXDQWLWDWHG DQG VWRUHG DW r& 6SHFWURSKRWRPHWULF 4XDQWLWDWLRQ RI '1$ DQG 51$ 1XFOHLF DFLGV ERWK '1$ DQG 51$ ZHUH URXWLQHO\ TXDQWLWDWHG E\ 2' DW QP $Q 2' DW QP 2'f RI FRUUHVSRQGV WR DSSUR[LPDWHO\ QJPO IRU GRXEOHVWUDQGHG '1$ [JPO IRU VLQJOHVWUDQGHG '1$ DQG 51$ DQG cMOJPO IRU ROLJRQXFOHRWLGHV f 7KH SXULW\ RI WKH QXFOHLF DFLG ZDV RIWHQ DVVHVVHG E\ FDOFXODWLQJ WKH QP QP RSWLFDO GHQVLW\ UDWLR 2'2'Jf 3XUH SUHSDUDWLRQV RI '1$ DQG 51$ KDYH DQ 2'2' RI DQG UHVSHFWLYHO\ 3UHSDUDWLRQ RI 5DGLRODEHOHG '1$ 5DQGRP 2OLJRQXFOHRWLGH 3ULPHG /DEHOLQJ $ UDQGRP ROLJRQXFOHRWLGH SULPLQJ WHFKQLTXH f VLPSOLILHG E\ 5REHUWV DQG :LOVRQ f HPSOR\LQJ 7 '1$ SRO\PHUDVH f ZDV FDUULHG RXW XVLQJ D UDQGRP SULPHU NLW 7KH YROXPH RI D VDPSOH FRQWDLQLQJ QJ RI '1$

PAGE 48

WHPSODWH OLQHDUL]HG SODVPLG '1$ RU DQ LVRODWHG '1$ IUDJPHQWf WR EH ODEHOHG ZDV DGMXVWHG WR LOO ZLWK GG+ 7KH VROXWLRQ ZDV WKHQ KHDWHG LQ D ERLOLQJ + EDWK IRU PLQXWHV DQG FHQWULIXJHG EULHIO\ WR FROOHFW WKH FRQGHQVDWLRQ $IWHU KHDW GHQDWXUDWLRQ , RI ; SULPHU EXIIHU VXSSOLHG E\ WKH PDQXIDFWXUHUf LO ODEHOHG QXFOHRWLGH a [FLf >D3@G&73 a &LPPROHf f DQG LOO GLOXWHG 7 '1$ SRO\PHUDVH XQLWVLO LQ HQ]\PH GLOXWLRQ EXIIHU VXSSOLHG E\ WKH PDQXIDFWXUHUf ZHUH DGGHG WR WKH WHPSODWH '1$ DQG WKH PL[WXUH ZDV LQFXEDWHG DW r& IRU PLQXWHV 8SRQ FRPSOHWLRQ RI WKH LQFXEDWLRQ SHULRG WKH UHDFWLRQ ZDV TXHQFKHG E\ WKH DGGLWLRQ RI LOO RI VWRS PL[WXUH 7KH YROXPH RI WKH UHDFWLRQ PL[WXUH ZDV DGMXVWHG WR PO ZLWK (OXWLSG ORZ VDOW EXIIHU P0 1D&O P0 7ULV}&O S+ f P0 ('7$f DQG WKH ODEHOHG '1$ SXULILHG XVLQJ D 6FKOHLFKHU t 6FKXHOO (OXWLSG FROXPQ DV GHVFULEHG E\ WKH PDQXIDFWXUHU 7KH ODEHOHG '1$ ZDV VWRUHG DW r& XQWLO XVH /DEHOLQJ WKH 7HUPLQL RI '1$ f '1$ K\EULGL]DWLRQ SUREHV ODEHOHG DW WKH WHUPLQL ZHUH SUHSDUHG DV GHVFULEHG E\ 2n)DUUHOO f 7KH YROXPH RI WKH VROXWLRQ FRQWDLQLQJ WKH '1$ IUDJPHQW WR EH ODEHOHG ZDV DGMXVWHG WR LO ZLWK GG+ %6$ LOO RI D PJPO VWRFN ZDV DGGHG WR WKH '1$ VROXWLRQ DORQJ ZLWK LOO RI %RHKULQJHU 0DQQKHLP UHVWULFWLRQ HQGRQXFOHDVH ; GLJHVWLRQ

PAGE 49

EXIIHU / DQG P 7 '1$ SRO\PHUDVH XQLW0Of DQG WKH VROXWLRQ LQFXEDWHG IRU PLQXWHV DW r& ,PPHGLDWHO\ IROORZLQJ WKH LQFXEDWLRQ 0O RI HDFK RI WKH IROORZLQJ ZDV DGGHG P0 G*73 P0 G$73 P0 773 DQG  cMOFLf RI >D3@G&73 &LPPROHf DQG LQFXEDWLRQ ZDV FRQWLQXHG IRU PLQXWH DW r& 8SRQ FRPSOHWLRQ RI WKH LQFXEDWLRQ 0O RI P0 G&73 ZDV DGGHG DQG WKH LQFXEDWLRQ FRQWLQXHG IRU PLQXWHV DW r& 7KH UHDFWLRQ ZDV VWRSSHG E\ KHDWLQJ WKH VROXWLRQ IRU PLQXWHV DW r& IROORZHG E\ WKH DGGLWLRQ RI 0O RI P0 ('7$ 7KH YROXPH RI WKH TXHQFKHG UHDFWLRQ PL[WXUH ZDV EURXJKW WR PO ZLWK (OXWLSG ORZ VDOW EXIIHU DQG WKH ODEHOHG '1$ SXULILHG XVLQJ D 6FKOHLFKHU t 6FKXHOO (OXWLSG FROXPQ DV GHVFULEHG E\ WKH PDQXIDFWXUHU 7KH ODEHOHG '1$ ZDV VWRUHG DW r& XQWLO XVH /DEHOLQJ WKH n 7HUPLQL RI '1$ '1$V WR EH ODEHOHG DW WKH WHUPLQL DQG XVHG DV K\EULGL]DWLRQ SUREHV ZHUH WUHDWHG ZLWK &,3 DV GHVFULEHG E\ &KDFRQDV DQG YDQ GH 6DQGH f DQG PRGLILHG E\ 0DQLDWLV HW DO f IROORZHG E\ n HQG ODEHOLQJ ZLWK 7 SRO\QXFOHRWLGH NLQDVH DQG >\3@$73 &LPPROHf DV GHVFULEHG E\ 0D[DP DQG *LOEHUW f DQG PRGLILHG E\ 0DQLDWLV HW DO f 7KH YROXPH RI WKH VROXWLRQ FRQWDLQLQJ WKH '1$ IUDJPHQW WR EH ODEHOHG ZDV DGMXVWHG WR 0O ZLWK GG+ &,3 ; EXIIHU P0 7ULV}&O S+ f P0 0J&O P0 =Q&O P0 VSHUPLGLQHf 0O DQG QO

PAGE 50

RI &,3 XQLWV0O IRU '1$ IUDJPHQWV ZLWK SURWUXGLQJ n WHUPLQL RU XQLW0O IRU '1$ IUDJPHQWV ZLWK EOXQW RU UHFHVVHG n WHUPLQLf ZHUH DGGHG WR WKH '1$ VROXWLRQ DQG LQFXEDWHG IRU PLQXWHV DW r& IROORZHG E\ LQFXEDWLRQ IRU PLQXWHV DW r& 8SRQ FRPSOHWLRQ RI WKH r& LQFXEDWLRQ D VHFRQG DOLTXRW RI &,3 ZDV DGGHG WR WKH UHDFWLRQ PL[WXUH DQG WKH VXFFHVVLYH LQFXEDWLRQV ZHUH UHSHDWHG 7KH UHDFWLRQ ZDV TXHQFKHG E\ WKH DGGLWLRQ RI 0O GG+ , ; 67( EXIIHU P0 7ULVr&O S+ f 0 1D&O P0 ('7$f DQG [ b 6'6 ZYf DQG WKH &,3 ZDV KHDW LQDFWLYDWHG IRU PLQXWHV DW r& 8SRQ FRPSOHWLRQ RI WKH LQFXEDWLRQ WKH UHDFWLRQ PL[WXUH ZDV H[WUDFWHG WZLFH ZLWK DQ HTXDO YROXPH RI HTXLOLEUDWHG SKHQRO DQG WZLFH ZLWK DQ HTXDO YROXPH RI &+&OM,$$ YYf )ROORZLQJ WKH DGGLWLRQ RI LO 0 1D&O 0O JO\FRJHQ PJPOf DQG 0O b HWKDQRO WKH '1$ ZDV SUHFLSLWDWHG RQ FUXVKHG GU\ LFH IRU PLQXWHV DQG SHOOHWHG E\ FHQWULIXJDWLRQ DW ; J IRU PLQXWHV DW r& $IWHU FHQWULIXJDWLRQ WKH VXSHUQDWDQW ZDV GHFDQWHG DQG WKH SHOOHW ZDVKHG ZLWK cLO b HWKDQRO DW URRP WHPSHUDWXUH 7KH SHOOHW RI '1$ ZDV GULHG XQGHU YDFXXP UHVXVSHQGHG LQ 0O P0 7ULVr&O P0 ('7$ S+ f DQG VWRUHG DW r& XQWLO WKH n HQG ODEHOLQJ UHDFWLRQ ZDV FDUULHG RXW 6SHUPLGLQH 0O RI D P0 VWRFN DQG 0O ; NLQDVH EXIIHU P0 7ULVr&O S+ f P0 0J&O P0 PHUFDSWRHWKDQROf ZHUH DGGHG WR WKH VROXWLRQ RI &,3

PAGE 51

WUHDWHG '1$ DQG LQFXEDWHG IRU PLQXWHV DW r& $IWHU WKH LQFXEDWLRQ WKH VROXWLRQ ZDV VQDS IUR]HQ LQ FUXVKHG GU\ LFH 7KH IUR]HQ VROXWLRQ ZDV TXLFNO\ WKDZHG DGGHG WR , 0FLf RI >\f§3@ $73 a &LPPROHf DQG LQFXEDWHG IRU PLQXWHV DW r& 7KH UHDFWLRQ ZDV VWRSSHG E\ WKH DGGLWLRQ RI [O P0 ('7$ DQG PO (OXWLSG ORZ VDOW EXIIHU DQG WKH ODEHOHG '1$ ZDV SXULILHG XVLQJ D 6FKOHLFKHU t 6FKXHOO (OXWLSG FROXPQ DV GHVFULEHG E\ WKH PDQXIDFWXUHU 7KH ODEHOHG '1$ ZDV VWRUHG DW r& XQWLO XVH $QDO\VLV RI 5HFRPELQDQW '1$ &ORQHV 0DSSLQJ E\ 6LQJOH DQG 0XOWLSOH 5HVWULFWLRQ (QGRQXFOHDVH 'LJHVWLRQV 6LQJOH DQG GRXEOH GLJHVWV ZHUH FDUULHG RXW RQ SODVPLG DQG $ EDFWHULRSKDJH '1$V XVLQJ UHVWULFWLRQ HQGRQXFOHDVHV DQG DQDO\]HG HOHFWURSKRUHWLFDOO\ RQ KRUL]RQWDO DJDURVH JHOV f ,QGLUHFW (QG/DEHOHG 0DSSLQJ 5HVWULFWLRQ PDSSLQJ RI UHFRPELQDQW $ SKDJH E\ LQGLUHFW HQGODEHOOLQJ ZDV FDUULHG RXW DV GHVFULEHG E\ 5DFNZLW] HW DO f XVLQJ K\EULGL]DWLRQ FRQGLWLRQV GHILQHG E\ /LWWOH DQG &URVV f 2OLJRQXFOHRWLGHV FRPSOHPHQWDU\ WR WKH FRV VHTXHQFH RI $ SKDJH ZHUH HQG ODEHOHG XVLQJ 7 SRO\QXFOHRWLGH NLQDVH 2OLJRQXFOHRWLGHV QJf LQ , ZHUH FRPELQHG ZLWK LO ; NLQDVH EXIIHU cMO, a QFLf RI >\3@$73 &LPPROHf cMO, P0 VSHUPLGLQH

PAGE 52

, GG+ DQG LOO RI XQLW[O 7 SRO\QXFOHRWLGH NLQDVH DQG WKH UHDFWLRQ PL[WXUH ZDV LQFXEDWHG IRU PLQXWHV DW r& 7KH UHDFWLRQ ZDV VWRSSHG E\ DGGLQJ LOO P0 ('7$ DQG LOO P0 7ULV} &O P0 ('7$ S+ f DQG KHDWLQJ IRU PLQXWHV DW r& 7KH UHDFWLRQ PL[WXUH ZDV H[WUDFWHG ZLWK DQ HTXDO YROXPH RI D PL[WXUH YYf RI HTXLOLEUDWHG SKHQRO DQG &+&,$$ YYf )ROORZLQJ WKH DGGLWLRQ RI MXO 0 1D&O LOO JO\FRJHQ PJPOf DQG LOO b HWKDQRO WKH ROLJRQXFOHRWLGHV ZHUH SUHFLSLWDWHG RQ FUXVKHG GU\ LFH IRU PLQXWHV DQG SHOOHWHG E\ FHQWULIXJDWLRQ DW ; J IRU PLQXWHV DW r& $IWHU FHQWULIXJDWLRQ WKH VXSHUQDWDQW ZDV GHFDQWHG DQG WKH SHOOHW ZDVKHG ZLWK LOO b HWKDQRO DW URRP WHPSHUDWXUH 7KH SHOOHW ZDV GULHG XQGHU YDFXXP UHVXVSHQGHG LQ cLO P0 7ULV}&O P0 ('7$ S+ f DQG VWRUHG DW r& XQWLO WKH K\EULGL]DWLRQ ZDV FDUULHG RXW 7R QO RI SDUWLDOO\ UHVWULFWHG $ SKDJH '1$ LTf ZDV DGGHG [O RI D VROXWLRQ FRQWDLQLQJ b 6'6 ZYf P0 ('7$ LLTLLO SURWHLQDVH N DQG QJ RI n HQG ODEHOHG ROLJRQXFOHRWLGHV DQG WKH PL[WXUH ZDV LQFXEDWHG IRU PLQXWHV DW r& ,PPHGLDWHO\ IROORZLQJ WKH LQFXEDWLRQ WKH PL[WXUH ZDV WUDQVIHUUHG WR r& DQG K\EULGL]HG IRU PLQXWHV )ROORZLQJ WKH K\EULGL]DWLRQ WKH VDPSOHV ZHUH SODFHG RQ ZHW LFH DQG [O 73( WUDFNLQJ G\H P0 7ULV}&O S+ f P0 1D+3 PRQREDVLFf P0 ('7$ b EURPRSKHQRO EOXH DQG b [\OHQH F\DQRO

PAGE 53

b JO\FHUROf ZDV DGGHG 7KH VDPSOHV ZHUH WKHQ ORDGHG RQWR D b DJDURVH 73( P0 7ULV}&O S+ f P0 1D+3 PRQREDVLFf P0 ('7$f JHO DQG HOHFWURSKRUHVHG IRU KRXUV DW ZDWWV ZLWK EXIIHU UHFLUFXODWLRQ )ROORZLQJ HOHFWURSKRUHVLV WKH JHO ZDV GULHG XQGHU YDFXXP IRU KRXU DW r& $XWRUDGLRJUDSK\ ZDV SHUIRUPHG XVLQJ SUHIODVKHG .RGDN ;$5 ;UD\ ILOP DQG D &URQH[ /LJKWQLQJ 3OXV VFUHHQ DW & ZRXOG OLNH WR DFNQRZOHGJH 'U 3DXO / 5RPDLQ DQG 6X]LH 3LODSLO IRU WKHLU ZRUN RQ WKH PDSSLQJ DQDO\VLV RI WKH $++*( JHQRPLF FORQH 6RXWKHUQ %ORW $QDO\VLV 5HVWULFWLRQ HQGRQXFOHDVH GLJHVWHG '1$V ZHUH VHSDUDWHG E\ HOHFWURSKRUHVLV WKURXJK DJDURVH JHOV DQG WUDQVIHUUHG WR =HWD3UREH EORWWLQJ PHPEUDQH DV GHVFULEHG E\ 5HHG DQG 0DQQ f 7KH DJDURVH JHO ZDV VRDNHG LQ P0 +& IRU PLQXWHV DW URRP WHPSHUDWXUH ULQVHG EULHIO\ LQ GG+ DQG WKH '1$ ZDV WUDQVIHUUHG ZLWK 1 1D2+ RYHUQLJKW DW URRP WHPSHUDWXUH )ROORZLQJ WUDQVIHU WKH PHPEUDQH ZDV EDNHG LQ D YDFXXP RYHQ DW & IRU KRXU SUHZDVKHG LQ b 6'6 ZYf ; 66& IRU RQH KRXU DW & DQG WKHQ SUHK\EULGL]HG DQG K\EULGL]HG XVLQJ D PRGLILFDWLRQ RI WKH PHWKRG GHVFULEHG E\ :DKO HW DO f 3UHK\EULGL]DWLRQ ZDV FDUULHG RXW IRU WZR KRXUV LQ P0 VRGLXP SKRVSKDWH S+ f b 6'6 ZYf b IRUPDPLGH b GH[WUDQ VXOIDWH DQG [JPO GRXEOH VWUDQGHG (A FROL '1$ $IWHU SUHK\EULGL]DWLRQ

PAGE 54

3ODEHOHG SUREH ZDV DGGHG WR WKH PL[ WR D FRQFHQWUDWLRQ RI ; GSPPO DQG K\EULGL]HG IRU KRXUV DW r& 7KH K\EULGL]HG PHPEUDQH ZDV WKHQ ZDVKHG WKUHH WLPHV DV IROORZV f LQ PO P0 SRWDVVLXP SKRVSKDWH S+ f IRU PLQXWHV DW URRP WHPSHUDWXUH f LQ PO P0 SRWDVVLXP SKRVSKDWH S+ f IRU PLQXWHV DW r& f LQ PO ; 66& DQG b 6'6 ZYf IRU PLQXWHV DW r& $XWRUDGLRJUDSK\ ZDV SHUIRUPHG XVLQJ SUHIODVKHG .RGDN ;$5 ;UD\ ILOP DQG D &URQH[ /LJKWQLQJ 3OXV VFUHHQ DW r& 3ODEHOHG SUREHV XVHG IRU 6RXWKHUQ DQDO\VLV RI WKH JHQRPLF +% FORQH $++*( ZHUH DV IROORZV +, QW 3VW ,(FR 5, IUDJPHQW IURP S)1& f +$ QW 6VW IUDJPHQW IURP S)) f +% QW (FR 5,;KR IUDJPHQW IURP $++& )LJXUH f + QW (FR 5, IUDJPHQW IURP S67 f + QW +LQG ,,,(FR 5, IUDJPHQW IURP S)$ f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

PAGE 55

UHSHDWHG XQWLO QR DGGLWLRQDO OLTXLG ZDV VTXHH]HG RXW RI WKH SXUHH 7KH '1$ IUDJPHQW ZDV SXULILHG XVLQJ D 6FKOHLFKHU t 6FKXHOO (OXWLSG FROXPQ DV GHVFULEHG E\ WKH PDQXIDFWXUHU DQG ZDV VWRUHG DW r& $QDO\VLV RI 0DPPDOLDQ 51$ $JDURVH)RUPDOGHK\GH 'HQDWXULQJ *HO (OHFWURSKRUHVLV 51$ ZDV VL]H IUDFWLRQDWHG LQ D b DJDURVH ; 0236 P0 0236 S+ f P0 VRGLXP DFHWDWH P0 ('7$f b IRUPDOGHK\GH JHO XVLQJ D ; 0236 b YYf IRUPDOGHK\GH UXQQLQJ EXIIHU D PRGLILFDWLRQ RI WKH PHWKRG XVHG E\ /HKUDFK HW DO f 1RUWKHUQ $QDO\VLV 1RUWKHUQ EORW DQDO\VLV ZDV FDUULHG RXW DV IROORZV WKH 51$ ZDV VL]H IUDFWLRQDWHG DV GHVFULEHG DERYH DQG WUDQVIHUUHG DV GHVFULEHG E\ 7KRPDV f WR =HWD3UREH EORWWLQJ PHPEUDQH XVLQJ ; 66& DV WKH WUDQVIHU EXIIHU )ROORZLQJ WUDQVIHU WKH PHPEUDQH ZDV EDNHG LQ D YDFXXP RYHQ DW r& IRU KRXU SUHZDVKHG LQ b 6'6 ZYf ; 66& IRU RQH KRXU DW r& DQG WKHQ SUHK\EULGL]HG DQG K\EULGL]HG XVLQJ D PRGLILFDWLRQ RI WKH PHWKRG GHVFULEHG E\ :DKO HW DO f 3UHK\EULGL]DWLRQ ZDV FDUULHG RXW IRU WZR KRXUV LQ P0 VRGLXP SKRVSKDWH S+ f b 6'6 ZYf b IRUPDPLGH b GH[WUDQ VXOIDWH DQG JPO GRXEOH VWUDQGHG (BcB FROL '1$ $IWHU SUHK\EULGL]DWLRQ 3ODEHOHG SUREH ZDV DGGHG WR

PAGE 56

WKH PL[ WR D FRQFHQWUDWLRQ RI ; GSPPO DQG K\EULGL]HG IRU KRXUV WKH WHPSHUDWXUH RI WKH K\EULGL]DWLRQ ZDV GHSHQGHQW XSRQ SUREH VL]H DQG SHUFHQW *& FRQWHQW DQG UDQJHG IURP r&r&f 7KH K\EULGL]HG =HWD3UREH PHPEUDQH ZDV WKHQ ZDVKHG ILYH WLPHV DV IROORZV f PLQXWHV DW URRP WHPSHUDWXUH ZLWK ; 66& ; 'HQKDUGWnV f PLQXWHV DW r& ZLWK ; 66& ; 'HQKDUGWnV f PLQXWHV DW & ZLWK ; 66& b 6'6 ZYf f PLQXWHV DW r& ZLWK ; 66& b 6'6 ZYf f PLQXWHV DW r& ZLWK ; 66& b 6'6 ZYf $XWRUDGLRJUDSK\ ZDV SHUIRUPHG XVLQJ SUHIODVKHG .RGDN ;$5 ;UD\ ILOP DQG D &URQH[ /LJKWQLQJ 3OXV VFUHHQ DW r& 7KH GHYHORSHG ;UD\ ILOPV ZHUH DQDO\]HG E\ ODVHU GHQVLWRPHWU\ XVLQJ D /.% *HO6FDQ ;/ GHQVLWRPHWHU SDUWLFOH DQDO\VLV RI K\EULGL]HG PHPEUDQHV ZDV FDUULHG RXW XVLQJ D HWDJHQ :DOWKDP 0$f %HWDVFRSH EORW DQDO\]HU 6, 1XFOHDVH 3URWHFWLRQ $QDO\VLV 6, DQDO\VLV ZDV FDUULHG RXW DFFRUGLQJ WR %HUN DQG 6KDUS f DV PRGLILHG E\ +DHJHPDQ HW DO f '1$ IUDJPHQWV XVHG DV SUREHV SUHSDUDWLRQ GHVFULEHG DERYHf ZHUH WUHDWHG ZLWK &,3 DV GHVFULEHG E\ &KDFRQDV DQG YDQ GH 6DQGH f DQG PRGLILHG E\ 0DQLDWLV HW DO f IROORZHG E\ n HQG ODEHOLQJ ZLWK 7 SRO\QXFOHRWLGH NLQDVH DQG >\3@$73 &LPPROHf DV GHVFULEHG E\ 0D[DP DQG *LOEHUW f DQG PRGLILHG E\ 0DQLDWLV HW DO f $OWHUQDWLYHO\ '1$

PAGE 57

IUDJPHQWV ZHUH n HQG ODEHOHG ZLWK 7 '1$ SRO\PHUDVH DQG >D3@G&73 &LPPROHf DV GHVFULEHG E\ 2n)DUUHOO f 7KH 51$ WR EH DQDO\]HG ZDV FRSUHFLSLWDWHG ZLWK ODEHOHG '1$ SUREH XVLQJ 0 1D&O LO JO\FRJHQ PJPOf DQG YROXPHV RI b HWKDQRO IRU PLQXWHV RQ FUXVKHG GU\ LFH DQG SHOOHWHG E\ FHQWULIXJDWLRQ DW ; J IRU PLQXWHV DW r& $IWHU FHQWULIXJDWLRQ WKH VXSHUQDWDQW ZDV GHFDQWHG DQG WKH SHOOHW ZDVKHG ZLWK L b HWKDQRO DW URRP WHPSHUDWXUH 7KH SHOOHW ZDV GULHG XQGHU YDFXXP UHVXVSHQGHG LQ _LO ; 6, K\EULGL]DWLRQ EXIIHU ; 0 1D&O P0 3LSHV S+ f P0 ('7$f DQG LO GHLRQL]HG UHFU\VWDOOL]HG IRUPDPLGH +\EULGL]DWLRQ PL[WXUHV LO ZHUH KHDWHG DW r& IRU PLQXWHV DQG WKHQ DQQHDOHG DW r& IRU DW OHDVW WKUHH KUV $QQHDOLQJ UHDFWLRQV ZHUH JXHQFKHG ZLWK HLJKW YROXPHV LO 6, QXFOHDVH EXIIHU P0 1D&O P0 VRGLXP DFHWDWH S+ f P0 =Q6f IROORZHG E\ LQFXEDWLRQ DW r& IRU PLQXWHV ZLWK XQLWV RI 6, QXFOHDVH LQ WKH SUHVHQFH RI LJ VDOPRQ VSHUP '1$ $IWHU 6, GLJHVWLRQ WKH VDPSOHV ZHUH H[WUDFWHG ZLWK DQ HTXDO YROXPH RI D PL[WXUH RI HTXLOLEUDWHG SKHQRO DQG &+&O,$$ YYf DQG HWKDQRO SUHFLSLWDWHG XVLQJ 0 1D&O LO JO\FRJHQ PJPOf DQG YROXPHV RI b HWKDQRO RYHUQLJKW DW r& 7KH SURWHFWHG IUDJPHQW ZDV SHOOHWHG E\ FHQWULIXJDWLRQ DW ; J IRU PLQXWHV DW r& )ROORZLQJ FHQWULIXJDWLRQ WKH VXSHUQDWDQW ZDV GHFDQWHG DQG WKH SHOOHWV ZHUH ZDVKHG

PAGE 58

ZLWK LOO b HWKDQRO DW URRP WHPSHUDWXUH 7KH SHOOHWV ZHUH GULHG XQGHU YDFXXP UHVXVSHQGHG LQ QO ORDGLQJ EXIIHU b IRUPDPLGH P0 ('7$ b EURPRSKHQRO EOXH DQG b [\OHQH F\DQROf DQG WKH VDPSOHV ZHUH KHDW GHQDWXUHG DQG ORDGHG RQWR D b SRO\DFU\ODPLGH VHTXHQFLQJ JHO FRQWDLQLQJ 0 XUHD f $IWHU HOHFWURSKRUHVLV WKH JHO ZDV VRDNHG LQ b DFHWLF DFLG b PHWKDQRO IRU PLQXWHV WR UHPRYH XUHD GULHG XQGHU YDFXXP DW r& DQG SODFHG DJDLQVW SUHIODVKHG .RGDN ;$5 ;UD\ ILOP DQG D &URQH[ /LJKWQLQJ 3OXV VFUHHQ DW r& ,Q 9LWUR 1XFOHDU 5XQRQ 7UDQVFULSWLRQ $QDO\VLV 7KH LQ YLWUR QXFOHDU UXQRQ WUDQVFULSWLRQ DVVD\V ZHUH SHUIRUPHG XVLQJ D PRGLILFDWLRQ f RI WKH PHWKRG XVHG E\ )OLQW HW DO f &HOOV ZHUH KDUYHVWHG E\ FHQWULIXJDWLRQ DQG WKH FHOO SHOOHWV ZDVKHG WZLFH LQ FROG LVRWRQLF EXIIHU P0 SRWDVVLXP SKRVSKDWH P0 7ULV+&O S+ f P0 0J&O P0 PHUFDSWRHWKDQROf &HOOV ZHUH GLVUXSWHG E\ KRPRJHQL]DWLRQ VWURNHVf ZLWK D 'RXQFH KRPRJHQL]HU :KHDWRQ W\SH $ SHVWOH $IWHU !b RI WKH FHOOV KDG EHHQ O\VHG QXFOHL ZHUH SHOOHWHG E\ FHQWULIXJDWLRQ DW USP IRU PLQXWHV LQ DQ ,(& FHQWULIXJH DW r& DQG UHVXVSHQGHG LQ QXFOHL VWRUDJH EXIIHU P0 7ULV+&O S+ f P0 0J&O P0 ('7$f FRQWDLQLQJ b JO\FHURO 1XFOHL ZHUH DOLTXRWHG DQG XVHG IUHVK LQ WKH LQ YLWUR WUDQVFULSWLRQ UHDFWLRQV 5HDFWLRQV W\SLFDOO\ FRQWDLQHG QXFOHL

PAGE 59

Q&L D3873 &LPPROHf P0 $73 P0 *73 DQG P0 &73 LQ D ILQDO YROXPH RI cMO DQG ZHUH LQFXEDWHG IRU PLQXWHV ZLWK LQWHUPLWWHQW VKDNLQJ DW r& 5DGLRODEHOHG 51$V ZHUH LVRODWHG E\ WUHDWPHQW RI QXFOHL ZLWK '1DVH [JPOf LQ WKH SUHVHQFH RI 0 1D&O P0 7ULV+&O S+ f P0 0J&O IRU PLQXWHV DW URRP WHPSHUDWXUH 7KH PL[WXUH ZDV WKHQ LQFXEDWHG ZLWK SURWHLQDVH JPOf IRU PLQXWHV DW r& LQ WKH SUHVHQFH RI P0 1D&O P0 ('7$ P0 7ULV+&O S+ f DQG P0 0J&O 6RGLXP DFHWDWH S+ f ZDV DGGHG WR P0 DQG QXFOHLF DFLGV ZHUH H[WUDFWHG VHYHUDO WLPHV E\ WKH KRW SKHQRO PHWKRG f 7R WKH DTXHRXV VROXWLRQ RI 3ODEHOHG 51$V QT RI \HDVW 51$ DQG YROXPHV RI b HWKDQRO ZHUH DGGHG 3UHFLSLWDWLRQ ZDV RYHUQLJKW DW r& 5DGLRODEHOHG WUDQVFULSWV ZHUH UHVXVSHQGHG LQ PO RI P0 7ULV+&O S+ f P0 ('7$ DQG DQ DOLTXRW RI HDFK VDPSOH ZDV SUHFLSLWDWHG ZLWK [J \HDVW 51$ DQG FROG b 7&$ 7&$SUHFLSLWDEOH FRXQWV ZHUH GHWHUPLQHG E\ OLTXLG VFLQWLOODWLRQ VSHFWURPHWU\ )ROORZLQJ SUHSDUDWLRQ RI 6RXWKHUQ EORWV f RI HOHFWURSKRUHWLFDOO\VHSDUDWHG UHVWULFWLRQ HQGRQXFOHDVH GLJHVWHG SODVPLG '1$V RU VORW EORWV SUHSDUHG XVLQJ D VORW EORW DSSDUDWXV ZLWK FRQGLWLRQV DV GHVFULEHG E\ 6FKOHLFKHU DQG 6FKXHOO ,QFf RI OLQHDUL]HG SODVPLG '1$V '1$ H[FHVV K\EULGL]DWLRQV ZHUH FDUULHG RXW 6RXWKHUQ RU VORW EORWV RQ QLWURFHOOXORVH ZHUH SUHK\EULGL]HG LQ 0 1D&O P0

PAGE 60

7ULV+&O S+ f P0 ('7$ b 6'6 ZYf ; 'HQKDUGWnV MXJPO GHQDWXUHG (B FROL '1$ DQG P0 VRGLXP S\URSKRVSKDWH DW r& IRU DW OHDVW KRXUV +\EULGL]DWLRQV ZHUH FRQGXFWHG DW r& IRU KRXUV LQ 0 1D&O P0 7ULV+&O S+ f P0 ('7$ b 6'6 ZYf ; 'HQKDUGWnV JPO (BcB FROL '1$ ZLWK 3ODEHOHG WUDQVFULSWV DW ; ; 7&$SUHFLSLWDEOH GSPPO RI K\EULGL]DWLRQ VROXWLRQ %ORWV ZHUH ZDVKHG DW r& IRU PLQXWHV LQ IUHVK SUHK\EULGL]DWLRQ VROXWLRQ KRXU LQ ; 66& b 6'6 ZYf RYHUQLJKW LQ ; 66& b 6'6 ZYf DQG KRXU LQ ; 66& b 6'6 ZYf $XWRUDGLRJUDSK\ ZDV SHUIRUPHG RQ DLU GULHG ILOWHUV ZLWK SUHn IODVKHG ;$5 RU &URQH[ ILOP DQG &URQH[ /LJKWQLQJ 3OXV VFUHHQV DW r& 7KH GHYHORSHG ;UD\ ILOPV ZHUH DQDO\]HG E\ ODVHU GHQVLWRPHWU\ XVLQJ D /.% *HO6FDQ ;/ GHQVLWRPHWHU ZRXOG OLNH WR DFNQRZOHGJH $QQD 5DPVH\(ZLQJ IRU KHU ZRUN RQ WKH WUDQVFULSWLRQDO DQDO\VLV RI WKH +%*/ JHQH /LEUDU\ 6FUHHQLQJ /DPEGD TWOO F'1$ /LEUDU\ 6FUHHQLQJ $ $JWOO KXPDQ OLYHU F'1$ OLEUDU\ PDGH IURP WRWDO FHOOXODU SRO\ $ 51$ DQG JHQHURXVO\ SURYLGHG E\ 'U 6 / & :RR %D\ORU &ROOHJH RI 0HGLFLQH 'HSDUWPHQW RI &HOO %LRORJ\f ZDV VFUHHQHG ZLWK WKH 3ODEHOHG KXPDQ KLVWRQH SUREHV LQGLFDWHG LQ )LJXUH 7KH OLEUDU\ VFUHHQLQJ ZDV

PAGE 61

FDUULHG RXW DV LQGLFDWHG LQ )LJXUH DFFRUGLQJ WR WKH SURFHGXUH GHVFULEHG E\ %HQWRQ DQG 'DYLV f ZLWK WKH IROORZLQJ PRGLILFDWLRQV LQ WKH K\EULGL]DWLRQ FRQGLWLRQV 7KH ILOWHUV ZHUH SUHK\EULGL]HG DW r& IRU WZR KRXUV LQ D VROXWLRQ FRQWDLQLQJ ; 'HQKDUGWnV ; b ILFROO b SRO\YLQ\OS\UUROLGRQHf ; 663( ; 0 VRGLXP FKORULGH P0 VRGLXP SKRVSKDWH P0 ('7$ S+ f b 6'6 ZYf b ZY %6$ DQG QTPO GRXEOH VWUDQGHG ( FROL '1$ +\EULGL]DWLRQ ZDV FDUULHG RXW DW r& IRU KRXUV LQ D VROXWLRQ FRQWDLQLQJ ; 'HQKDUGWnV ; 663( b 6'6 ZYf b ZY %6$ QJPO (/ FROL '1$ DQG ; GSPPO KHDWGHQDWXUHG SUREH '1$ IUDJPHQW +\EULGL]HG ILOWHUV ZHUH WKHQ ZDVKHG DW r& IRU PLQXWHV LQ P0 SRWDVVLXP SKRVSKDWH S+ f IROORZHG E\ WZR PLQXWH ZDVKHV DW r& LQ ; 663( b 6'6 ZYf $XWRUDGLRJUDSK\ ZDV SHUIRUPHG XVLQJ .RGDN ;$5 ;UD\ ILOP DQG D &URQH[ /LJKWQLQJ 3OXV VFUHHQ DW r& 3ODTXH GRW DQDO\VLV ZDV FDUULHG RXW DFFRUGLQJ WR WKH SURFHGXUH GHVFULEHG E\ 3RZHOO HW DO A f $ LO DOLTXRW RI SODWLQJ EDFWHULD ZDV SODFHG LQWR D WXEH DQG PO RI 1=&<0 WRS DJDU ZDV DGGHG WR WKH WXEH YRUWH[HG JHQWO\ DQG SRXUHG RQWR D 1=&<0 ERWWRP DJDU SODWH 8SRQ VROLGLILFDWLRQ

PAGE 62

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f IURP D ; &KDURQ $ JHQRPLF '1$ OLEUDU\

PAGE 63

+ + A Ws ;++* (FR 5O / f§r +LQG ,,, ;++* ;++* D?

PAGE 64

)LJXUH 2XWOLQH RI WKH VFUHHQLQJ VFKHPH IRU WKH $FUWOO SRO\ $ F'1$ OLEUDU\ $ $JWOO KXPDQ OLYHU F'1$ OLEUDU\ ZDV VFUHHQHG DV GHVFULEHG E\ %HQWRQ DQG 'DYLV f 7KH $JWOO F'1$ OLEUDU\ ZDV JURZQ RQ PP GLDPHWHU SHWUL GLVKHV )ROORZLQJ RYHUQLJKW LQFXEDWLRQ GXSOLFDWH QLWURFHOOXORVH ILOWHUV ZHUH OLIWHG IURP HDFK SODWH DQG SUHSDUHG DV GHVFULEHG f 2QH VHW RI ILOWHUV ZDV K\EULGL]HG WR WKH + DQG + SUREH DW r& DQG WKH RWKHU VHW WR WKH +$ +% DQG +, SUREHV DW r& DV GHVFULEHG E\ /DZQ HW DO f +\EULGL]HG ILOWHUV ZHUH WKHQ ZDVKHG DQG DXWRUDGLRJUDSK\ SHUIRUPHG 7KH SRVLWLYH SODTXHV RU SODTXH DUHDVf ZHUH SLFNHG DQG VFUHHQHG D VHFRQG WLPH 6LQJOH ZHOO LVRODWHG SRVLWLYH SODTXHV ZHUH SLFNHG DQG VFUHHQHG D WKLUG WLPH 8SRQ FRPSOHWLRQ RI WKH WKLUG URXQG RI VFUHHQLQJ ++ SRVLWLYHV DQG +$+%+ SRVLWLYH SODTXHV KDG EHHQ REWDLQHG 3RVLWLYH SODTXHV ZHUH VHSDUDWHG DFFRUGLQJ WR LQGLYLGXDO KLVWRQH FODVV XVLQJ WKH KXPDQ KLVWRQH SUREHV GHVFULEHG LQ )LJXUH E\ SODTXH GRW DQDO\VLV

PAGE 65

[JW 3RO\ $ F'1$ /LEUDU\ +XPDQf 3ODWH 3RVLWLYHV DQG A 5HSHDW 7ZR 7LPHV L 3LFN 3RVLWLYHV &KHFN )RU 3RVLWLYHV 8SRQ &RPSOHWLRQ RI 7KLUG 5RXQG RI 6FUHHQLQJ + +$ 3RVLWLYHV ,// +$ +% 3RVLWLYHV 6HSDUDWH 3ODTXHV

PAGE 66

RI WKH WRS DJDU , DOLTXRWV RI $ EDFWHULRSKDJH GLOXWLRQV ZHUH VSRWWHG RQWR WKH SODWH DQG DOORZHG WR GU\ 7KH SODWH ZDV LQYHUWHG DQG LQFXEDWHG DW r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f $ FRPSOHWH HTXLYDOHQW RI WKLV $(0%/ KXPDQ JHQRPLF '1$ OLEUDU\ ; SKDJHf ZDV JURZQ RQ PP GLDPHWHU SHWUL GLVKHV DQG VFUHHQHG ZLWK D 3ODEHOHG +% n QRQWUDQVODWHG SUREH QW 6DF IUDJPHQW LVRODWHG IURP $++&f DFFRUGLQJ WR WKH SURFHGXUH GHVFULEHG E\ %HQWRQ DQG 'DYLV f /( f SODWLQJ EDFWHULD PO ZHUH PL[HG ZLWK SIX RI UHFRPELQDQW SKDJH DQG LQFXEDWHG IRU PLQXWHV DW r& 1H[W PO PHOWHG WRS DJDURVH r&f ZDV DGGHG DQG WKH PL[WXUH SRXUHG RQWR D PP 1=&<0 ERWWRP DJDU SODWH 7KH WRS DJDURVH ZDV DOORZHG WR VROLGLI\ DW URRP WHPSHUDWXUH DQG WKHQ LQFXEDWHG KRXUV 2U XQWLO WKH SODTXHV UHDFK D GLDPHWHU RI DSSUR[LPDWHO\ PPf DW r&

PAGE 67

)ROORZLQJ RYHUQLJKW LQFXEDWLRQ WKH SODWHV ZHUH FKLOOHG IRU PLQXWHV DW r& )RU ILOWHU OLIWLQJ SODWHV ZHUH UHPRYHG IURP WKH FROG ER[ DQG QLWURFHOOXORVH ILOWHUV FDUHIXOO\ ODLG RQ WRS RI WKH DJDURVH IRU PLQXWHV 7KH RULHQWDWLRQ RI WKH ILOWHUV ZDV PDUNHG LQ WKUHH DV\PPHWULF SRVLWLRQV XVLQJ D QLWURFHOOXORVH PDUNLQJ SHQ 7KH ILOWHUV ZHUH WKHQ UHPRYHG DQG VRDNHG LQ GHQDWXULQJ VROXWLRQ 0 1D&O 1 1D2+f IRU VHFRQGV DQG WUDQVIHUUHG LQWR QHXWUDOL]LQJ VROXWLRQ 0 1D&O 0 7ULV}&O S+ ff IRU PLQXWHV 6XEVHJXHQWO\ WKH ILOWHUV ZHUH ULQVHG LQ ; 663( EORWWHG GU\ RQ :KDWPDQ 00 SDSHU DQG EDNHG IRU KRXUV DW r& LQ D YDFXXP RYHQ +\EULGL]DWLRQ RI WKHVH ILOWHUV ZDV FDUULHG RXW HVVHQWLDOO\ DV GHVFULEHG E\ /DZQ HW DO f ZLWK WKH IROORZLQJ PRGLILFDWLRQV LQ WKH K\EULGL]DWLRQ FRQGLWLRQV 7KH ILOWHUV ZHUH SUHK\EULGL]HG DW r& IRU WZR KRXUV LQ D VROXWLRQ FRQWDLQLQJ ; 'HQKDUGWnV ; 663( b 6'6 ZYf b ZY %6$ DQG JPO GRXEOH VWUDQGHG (BcB FROL '1$ +\EULGL]DWLRQ ZDV FDUULHG RXW DW r& IRU KRXUV LQ D VROXWLRQ FRQWDLQLQJ ; 'HQKDUGWnV ; 663( b 6'6 ZYf b %6$ JPO (/ FROL '1$ DQG ; GSPPO KHDWGHQDWXUHG SUREH '1$ IUDJPHQW +\EULGL]HG ILOWHUV ZHUH WKHQ ZDVKHG PO ZDVK VROXWLRQILOWHUf DW r& IRU KRXU LQ P0 SRWDVVLXP SKRVSKDWH S+ f IROORZHG E\ WZR KRXU ZDVKHV DW r& LQ ; 663( b 6'6 ZYf $XWRUDGLRJUDSK\ ZDV SHUIRUPHG XVLQJ .RGDN ;$5 ;UD\ ILOP DQG D &URQH[ /LJKWQLQJ 3OXV VFUHHQ DW r& )ROORZLQJ

PAGE 68

K\EULGL]DWLRQ WKH SRVLWLYH SODTXHV RU SODTXH DUHDVf ZHUH SLFNHG DQG SODFHG LQWR PO RI 60 FRQWDLQLQJ D GURS RI FKORURIRUP 7KHVH SRVLWLYH SODTXHV ZHUH VFUHHQHG D VHFRQG WLPH E\ K\EULGL]DWLRQ 6LQJOH ZHOO LVRODWHG SRVLWLYH SODTXHV ZHUH SLFNHG DQG VFUHHQHG D WKLUG WLPH &ORQLQJ DQG &RQVWUXFW 3UHSDUDWLRQ 6XEFORQLQJ LQWR 3ODVPLG 9HFWRUV '1$ IUDJPHQWV ZHUH LVRODWHG IURP DJDURVH JHOV DQG OLJDWLRQV LQWR WKH DSSURSULDWH YHFWRUV ZHUH FDUULHG RXW XVLQJ 7 '1$ OLJDVH DV GHVFULEHG E\ WKH PDQXIDFWXUHU 7KH OLJDWLRQ SURGXFWV ZHUH XVHG WR WUDQVIHFW FRPSHWHQW (Br FROL '+D f VHH DSSHQGL[ %f EDFWHULD 7KH QW 3VW IUDJPHQW QW (FR 5, IUDJPHQW DQG QW (FR 5, IUDJPHQW IURP $++*( )LJXUH f ZHUH LVRODWHG DQG FORQHG LQWR S8& DQG WKH UHVXOWLQJ VXEFORQHV WHUPHG S*/ S*/,2O DQG S*/ UHVSHFWLYHO\ &RQVWUXFW 3UHSDUDWLRQ &RQVWUXFW S*/69f LV WKH FRQWURO FRQVWUXFW FRQWDLQLQJ WKH HQWLUH $++*( +% JHQH SURPRWHU DQG QW RI n XQWUDQVODWHG UHJLRQ )LJXUH f 7KH QW 3VW IUDJPHQW IURP $++*( ZDV LVRODWHG DQG OLJDWHG XVLQJ 7 '1$ OLJDVH LQWR WKH 3VW VLWH RI S8& WR REWDLQ WKH LQWHUPHGLDWH S*/f &RQVWUXFW ZDV REWDLQHG E\ LVRODWLQJ WKH (FR 5,(FR 5, 69 HQKDQFHU HOHPHQW IURP

PAGE 69

S69($ f EOXQWHQGLQJ LWV WHUPLQL ZLWK WKH .OHQRZ IUDJPHQW RI '1$ SRO\PHUDVH DQG OLJDWLQJ LW LQWR WKH %DP +, VLWH RI S*/ ZKLFK KDG SUHYLRXVO\ EHHQ EOXQWHQGHG ZLWK WKH .OHQRZ IUDJPHQW &RQVWUXFW ,, S*/+269f LV VLPLODU WR FRQVWUXFW EXW KDV WKH n (FR 5,3VW IUDJPHQW FRQWDLQLQJ WKH SRO\$f DGGLWLRQ VHJXHQFHV GHOHWHG )LJXUH f 7KH QW (FR 5, IUDJPHQW IURP $++*( ZDV LVRODWHG EOXQWHQGHG XVLQJ WKH .OHQRZ IUDJPHQW DQG OLJDWHG XVLQJ 7 '1$ OLJDVH LQWR WKH +LH ,, VLWH RI S8&69( ZKLFK FRQWDLQV WKH 69 HQKDQFHU HOHPHQW IURP S69($ f UHVXOWLQJ LQ S*/+269 &RQVWUXFW ,,, S*/69f LV VLPLODU WR FRQVWUXFW EXW KDV WKH QW ;KR IUDJPHQW FRQWDLQLQJ WKH KLVWRQH n KDLUSLQ PRWLI GHOHWHG )LJXUH f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

PAGE 70

)LJXUH 2XWOLQH RI WKH FORQLQJ VFKHPH IRU WKH FRQVWUXFWLRQ RI WKH +% FRQVWUXFW 3*/69 &RQVWUXFW S*/69f LV WKH FRQWURO FRQVWUXFW FRQWDLQLQJ WKH HQWLUH $++*( +% JHQH SURPRWHU DQG QW RI n XQWUDQVODWHG UHJLRQ )LJXUH f 7KH QW 3VW IUDJPHQW IURP $++*( ZDV LVRODWHG DQG OLJDWHG XVLQJ 7 '1$ OLJDVH LQWR WKH 3VW VLWH RI S8& WR REWDLQ WKH LQWHUPHGLDWH S*/f &RQVWUXFW ZDV REWDLQHG E\ LVRODWLQJ WKH (FR 5,(FR 5, 69 HQKDQFHU HOHPHQW IURP S69($ f EOXQWHQGLQJ LWV WHUPLQL ZLWK WKH .OHQRZ IUDJPHQW RI '1$ SRO\PHUDVH DQG OLJDWLQJ LW LQWR WKH %DP +, VLWH RI S*/ ZKLFK KDG SUHYLRXVO\ EHHQ EOXQWHQGHG ZLWK WKH .OHQRZ IUDJPHQW 5HVWULFWLRQ HQGRQXFOHDVH VLWHV DUH DV IROORZV $ $VS % %DP +, ( (FR 5, + +LQG ,,, 3 3VW 6 6DF 6n 6SK ; ;KR D OHWWHU FRQWDLQLQJ D VHPLFLUFOH KDW LQGLFDWHV D UHVWULFWLRQ HQGRQXFOHDVH VLWH ZKLFK KDV EHHQ GHVWUR\HG GXH WR D EOXQWHQGLQJ UHDFWLRQ $++*( LV D $(0%/ UHFRPELQDQW EDFWHULRSKDJH ZKLFK ZDV LVRODWHG IURP D $(0%/ JHQRPLF '1$ OLEUDU\ VHH FKDSWHU f

PAGE 71

.E ( 2QW ( f 22QW 36f ; ;+ ;; E 363 $ /, , LPFG ,_/ ,f§8 22QW % .E ( 36f ; ;+ L 0 R ;; ( Vr 3+ B S*/ ( QW ( $ ++*( (3+ S8& .E ( (A n636n ; ;+ ;; L 3+ ( ( 69 (1+$1&(5 S*/ 69 8 r

PAGE 72

)LJXUH 2XWOLQH RI WKH FORQLQJ VFKHPH IRU WKH FRQVWUXFWLRQ RI WKH +% GHOHWLRQ FRQVWUXFW S*/+269 &RQVWUXFW ,, S*/+269f LV VLPLODU WR FRQVWUXFW EXW KDV WKH n (FR 5,3VW IUDJPHQW FRQWDLQLQJ WKH SRO\$f DGGLWLRQ VHTXHQFHV GHOHWHG )LJXUH f 7KH QW (FR 5, IUDJPHQW IURP ƒ++*( ZDV LVRODWHG EOXQWHQGHG XVLQJ WKH .OHQRZ IUDJPHQW DQG OLJDWHG XVLQJ 7 '1$ OLJDVH LQWR WKH +LH ,, VLWH RI S8&69( ZKLFK FRQWDLQV WKH 69 HQKDQFHU HOHPHQW IURP S69($ f UHVXOWLQJ LQ S*/+269 5HVWULFWLRQ HQGRQXFOHDVH VLWHV DUH DV IROORZV $ $VS % %DP +, ( (FR 5, + +LQG ,,, 3 3VW 6 6DF 6n 6SK ; ;KR D OHWWHU FRQWDLQLQJ D VHPLFLUFOH KDW LQGLFDWHV D UHVWULFWLRQ HQGRQXFOHDVH VLWH ZKLFK KDV EHHQ GHVWUR\HG GXH WR D EOXQWHQGLQJ UHDFWLRQ $++*( LV D $(0%/ UHFRPELQDQW EDFWHULRSKDJH ZKLFK ZDV LVRODWHG IURP D $(0%/ JHQRPLF '1$ OLEUDU\ VHH FKDSWHU f

PAGE 73

.E ( 22QW ( V 22QW ( 22QW ( QW ( 36f ; ;+A;; 363 $ LLL L % L $ L

PAGE 74

)LJXUH 2XWOLQH RI WKH FORQLQJ VFKHPH IRU WKH FRQVWUXFWLRQ RI WKH +% KDLUSLQ PRWLI PLQXV FRQVWUXFW 3*/69 &RQVWUXFW ,,, S*/69f LV VLPLODU WR FRQVWUXFW EXW KDV WKH QW ;KR IUDJPHQW FRQWDLQLQJ WKH KLVWRQH n KDLUSLQ PRWLI GHOHWHG )LJXUH f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n 6SK ; ;KR D OHWWHU FRQWDLQLQJ D VHPLFLUFOH KDW LQGLFDWHV D UHVWULFWLRQ HQGRQXFOHDVH VLWH ZKLFK KDV EHHQ GHVWUR\HG GXH WR D EOXQWHQGLQJ UHDFWLRQ 3ODVPLG */69 LV GHVFULEHG LQ )LJXUH

PAGE 75

F .E (IW IW36n ; ;+ A /0--f§, L LOO O/ S*/ 469 3+ ZB F & .E (3+ 3+ 8 *\FMRff§L .E (IW IW36 ; ;+ 0DL\ .E (3+ 3+ U8B 3*/4 F O .E (3+ ; n 3+ S* / F .E (IW-36f ; -! + ; .E 3+ G.E (IW IW36f ; ;+ ; 8 , 3+ S*/469

PAGE 76

6XEFORQLQJ LQWR %DFWHULRSKDJH 0 9HFWRUV '1$ IUDJPHQWV WR EH VHTXHQFHG ZHUH LVRODWHG IURP DJDURVH JHOV DQG OLJDWHG LQWR 0PSO DQGRU PSO UHSOLFDWLYH IRUP f XVLQJ 7 '1$ OLJDVH DV GHVFULEHG E\ WKH PDQXIDFWXUHU 7KH OLJDWLRQ SURGXFWV ZHUH XVHG WR WUDQVIHFW FRPSHWHQW ;/O%OXH f VHH DSSHQGL[ &f EDFWHULD 7UDQVIHFWLRQ RI '1$ LQWR %DFWHULDO &HOOV 3UHSDUDWLRQ RI &RPSHWHQW ( FROL &HOOV IRU 7UDQVIHFWLRQ &RPSHWHQW (BL FROL FHOOV ZHUH SUHSDUHG XVLQJ D PRGLILFDWLRQ RI WKH PHWKRG GHVFULEHG E\ 0DQGHO DQG +LJD f $Q RYHUQLJKW FXOWXUH RI (AB FROL ZDV GLOXWHG LQWR PO <71 PHGLXP DQG JURZQ DW r& ZLWK YLJRURXV VKDNLQJ XQWLO WKH 2' DW QP UHDFKHG DSSUR[LPDWHO\ 7KH FHOO FXOWXUH ZDV FKLOOHG RQ ZHW LFH IRU PLQXWHV VSOLW LQWR WZR PO WXEHV DQG WKHQ FHQWULIXJHG DW USP LQ DQ ,(& URWRU ; Jf IRU PLQXWHV DW r& )ROORZLQJ FHQWULIXJDWLRQ WKH VXSHUQDWDQW ZDV GLVFDUGHG DQG WKH FHOO SHOOHWV YRUWH[HG JHQWO\ UHVXVSHQGHG LQ PO FROG VWHULOH WUDQVIHFWLRQ EXIIHU P0 &D&O P0 3LSHV S+ f b JO\FHUROf DQG FHQWULIXJHG DW USP LQ DQ ,(& URWRU ; Jf IRU PLQXWHV DW r& 1H[W WKH VXSHUQDWDQW ZDV GLVFDUGHG DQG WKH FHOO SHOOHWV ZHUH JHQWO\ UHVXVSHQGHG LQ PO FROG VWHULOH WUDQVIHFWLRQ EXIIHU DQG LQFXEDWHG IRU PLQXWHV RQ ZHW LFH 8SRQ FRPSOHWLRQ RI WKH LQFXEDWLRQ WKH FHOOV ZHUH FHQWULIXJHG DW USP LQ DQ ,(& URWRU

PAGE 77

; Jf IRU PLQXWHV DW r& WKH VXSHUQDWDQW ZDV GLVFDUGHG DQG WKH FHOO SHOOHWV ZHUH JHQWO\ UHVXVSHQGHG LQ PO FROG VWHULOH WUDQVIHFWLRQ EXIIHU 7KH FRPSHWHQW FHOOV ZHUH VWRUHG DW r& DQG XVHG ZLWKLQ KRXUV RI SUHSDUDWLRQ 7UDQVIHFWLRQ RI 5HFRPELQDQW 3ODVPLG '1$ 7UDQVIHFWLRQ RI UHFRPELQDQW SODVPLG '1$ ZDV FDUULHG RXW XVLQJ D PRGLILFDWLRQ RI WKH PHWKRG GHVFULEHG E\ 0DQGHO DQG +LJD f $ IUDFWLRQ RI D OLJDWLRQ PL[WXUH f DQG LOO FRPSHWHQW (K FROL '+D f FHOOV ZHUH FRPELQHG LQ D PO PLFURFHQWULIXJH WXEH LQFXEDWHG IRU PLQXWHV RQ ZHW LFH DQG WKHQ LQFXEDWHG IRU PLQXWHV DW r& )ROORZLQJ WKH LQFXEDWLRQ WKH FHOOV ZHUH GLOXWHG ZLWK PO SUHZDUPHG <71 PHGLXP DQG LQFXEDWHG ZLWK VKDNLQJ IRU KRXU DW r& $Q DOLTXRW RI WKH WUDQVIHFWLRQ PL[WXUH ZDV SODWHG RQWR WKH DSSURSULDWH ERWWRP DJDU SODWH DQG LQFXEDWHG LQYHUWHG IRU KRXUV DW r& 7UDQVIHFWLRQ RI 5HFRPELQDQW %DFWHULRSKDJH 0 '1$ 7UDQVIHFWLRQ RI UHFRPELQDQW EDFWHULRSKDJH 0 '1$ ZDV FDUULHG RXW DV GHVFULEHG E\ 0HVVLQJ f $ IUDFWLRQ RI D OLJDWLRQ PL[WXUH f DQG ILO FRPSHWHQW (K FROL ;/O%OXH f FHOOV ZHUH FRPELQHG LQ D PP ; PP JODVV FXOWXUH WXEH LQFXEDWHG IRU PLQXWHV RQ ZHW LFH DQG WKHQ KHDWVKRFNHG IRU PLQXWHV DW r& ,PPHGLDWHO\ IROORZLQJ

PAGE 78

WKH LQFXEDWLRQ PO RI <71 WRS DJDU r&f ZDV DGGHG WR WKH WXEH YRUWH[HG JHQWO\ DQG SRXUHG RQWR D 1=&<0 ERWWRP DJDU SODWH 8SRQ VROLGLILFDWLRQ RI WKH WRS DJDU WKH SODWH ZDV LQFXEDWHG LQYHUWHG IRU KRXUV DW r& WR DOORZ IRU SODTXH IRUPDWLRQ '1$ 6HTXHQFLQJ 6DQJHU 'LGHR[Y0HGLDWHG &KDLQ 7HUPLQDWLRQ 0HWKRG 'LGHR[\ VHTXHQFLQJ f ZDV FDUULHG RXW XVLQJ D 6HTXHQDVH '1$ VHTXHQFLQJ NLW DFFRUGLQJ WR FRQGLWLRQV GHVFULEHG E\ WKH PDQXIDFWXUHU $IWHU WKH VHTXHQFLQJ UHDFWLRQV ZHUH FRPSOHWHG ORDGLQJ EXIIHU b IRUPDPLGH P0 ('7$ b EURPRSKHQRO EOXH DQG b [\OHQH F\DQROf ZDV DGGHG DQG WKH VDPSOHV ZHUH KHDW GHQDWXUHG DW r& IRU PLQXWHV DQG ORDGHG RQWR D b SRO\DFU\ODPLGH VHTXHQFLQJ JHO FRQWDLQLQJ 0 XUHD f )ROORZLQJ HOHFWURSKRUHVLV WKH JHO ZDV VRDNHG LQ b DFHWLF DFLG b PHWKDQRO IRU PLQXWHV WR UHPRYH XUHD GULHG XQGHU YDFXXP DW & XVLQJ VKHHWV RI :KDWPDQ 00 SDSHU DV EDFNLQJ DQG SODFHG DJDLQVW SUHIODVKHG .RGDN ;$5 RU &URQH[ ;UD\ ILOP DW URRP WHPSHUDWXUH &KURPRVRPDO LQ VLWX +\EULGL]DWLRQ &KURPRVRPDO LQ VLWX K\EULGL]DWLRQ ZDV SHUIRUPHG DV GHVFULEHG f 3UREH '1$ QW (FR 5,6DF IUDJPHQW LVRODWHG IURP $++& ff ZDV ODEHOHG ZLWK + WR D

PAGE 79

VSHFLILF DFWLYLW\ RI ; GSPQJ 6OLGHV FRQWDLQLQJ PHWDSKDVH FKURPRVRPHV IURP QRUPDO PDOH ;+@WK\PLGLQH LQWR 7&$SUHFLSLWDEOH PDWHULDO LQ D PLQXWH SXOVH f '1$ V\QWKHVLV ZDV LQKLELWHG E\ WUHDWLQJ WKH

PAGE 80

FHOO FXOWXUH ZLWK P0 K\GUR[\XUHD IRU KRXU EHJLQQLQJ IRXU KRXUV DIWHU UHOHDVH IURP WKH VHFRQG WK\PLGLQH EORFN &HOOV K\GUR[\XUHD WUHDWHG RU XQWUHDWHGf ZHUH KDUYHVWHG DW YDULRXV WLPHV DIWHU UHOHDVH IURP WKH VHFRQG WK\PLGLQH EORFN +/ &HOO &XOWXUH DQG 'LIIHUHQWLDWLRQ +XPDQ SURP\HORF\WLF OHXNHPLD FHOOV +/ FHOOVf ZHUH FXOWXUHG LQ OLTXLG VXVSHQVLRQ LQ 530, PHGLXP VXSSOHPHQWHG ZLWK b IHWDO FDOI VHUXP +/ FHOOV ZHUH SODWHG DW D FHOO GHQVLW\ RI ; PO JURZQ IRU KRXUV WR D FHOO GHQVLW\ RI ; PO DQG KDUYHVWHG E\ FHQWULIXJDWLRQ )RU GLIIHUHQWLDWLRQ RI +/ FHOOV FXOWXUHV DW D GHQVLW\ RI ; PO ZHUH WUHDWHG ZLWK 73$ DW D ILQDO FRQFHQWUDWLRQ RI Q0 DQG KDUYHVWHG DIWHU SHULRGV RI DQG GD\V 7/ &HOO &XOWXUH 0RXVH 7/ FHOOV ZHUH PDLQWDLQHG DV VXEFRQIOXHQW FXOWXUHV LQ '0(0 FRQWDLQLQJ b FDOI VHUXP LQ D PRLVW b & r& LQFXEDWRU 7UDQVIHFWLRQ DQG 7UDQVLHQW ([SUHVVLRQ RI 5HFRPELQDQW '1$ LQ 7/ &HOOV 7UDQVIHFWLRQ RI 7/ FHOOV ZDV FDUULHG RXW DV GHVFULEHG E\ *RUPDQ HW DO f )RXU KRXUV SULRU WR WUDQVIHFWLRQ 7/ FHOOV LQ PRQROD\HU DW b FRQIOXHQF\ ZHUH UHIHG ZLWK IUHVK PHGLXP 7KH FDOFLXP SKRVSKDWH'1$

PAGE 81

FRPSOH[ ZDV SUHSDUHG DV GHVFULEHG E\ *UDKDP DQG 9DQ GHU (E f XVLQJ ]J RI SODVPLG '1$ DQG ]J RI VDOPRQ VSHUP '1$ DV FDUULHU )RXU KRXUV DIWHU WKH DGGLWLRQ RI WKH FDOFLXP SKRVSKDWH'1$ FRPSOH[ WKH PHGLXP ZDV UHPRYHG DQG WKH FHOOV VXEMHFWHG WR D RQH PLQXWH VKRFN ZLWK b JO\FHURO YYf LQ PHGLXP FRQWDLQLQJ b IHWDO FDOI VHUXP ,QFXEDWLRQ ZDV FRQWLQXHG LQ IUHVK PHGLXP IRU KRXUV )ROORZLQJ LQFXEDWLRQ WKH FHOOV ZHUH KDUYHVWHG DQG WRWDO FHOOXODU 51$ ZDV LVRODWHG DQG DQDO\]HG E\ 6, QXFOHDVH SURWHFWLRQ DQDO\VLV 6HOHFWLRQ RI 6WDEOH 3RO\FORQDO 7/ &HOO /LQHV 7/ FHOO PRQROD\HUV ZHUH WUDQVIHFWHG DV DERYH XVLQJ PRODU TXDQWLWLHV RI FRQVWUXFW DQG S69QHR f ZLWKRXW XVLQJ FDUULHU '1$ ]J WRWDO '1$ TXDQWLW\f 6HOHFWLRQ RI FHOOV ZDV FDUULHG RXW DV GHVFULEHG E\ 6RXWKHUQ DQG %HUJ f XVLQJ *HQHWLFLQ VXOIDWHf 7KH PHGLXP SOXV GUXJ ZDV FKDQJHG HYHU\ WKUHH WR IRXU GD\V XQWLO UHVLVWDQW FRORQLHV JUHZ WR DSSUR[LPDWHO\ FP LQ GLDPHWHU 3RO\FORQDO FHOO OLQHV ZHUH HVWDEOLVKHG E\ FRPELQLQJ UHVLVWDQW FRORQLHV DV IROORZV WKH PHGLXP IURP HDFK SODWH FRQWDLQLQJ IURP UHVLVWDQW FRORQLHVf ZDV UHPRYHG DQG WKH SODWHV ZHUH WUHDWHG ZLWK PO WU\SVLQ b LQ EXIIHUHG VDOLQHf IRU PLQXWHV 7KH FHOOV ZHUH ULQVHG IURP WKH VXUIDFH RI WKH SODWH DQG D PO DOLTXRW RI WKH KHWHURJHQHRXV FHOO VXVSHQVLRQ ZDV WUDQVIHUUHG WR D SODWH FRQWDLQLQJ IUHVK

PAGE 82

PHGLXP DQG ZDV PDLQWDLQHG DV D VXEFRQIOXHQW FXOWXUH LQ '0(0 FRQWDLQLQJ b FDOI VHUXP LQ D PRLVW b & r& LQFXEDWRU 7HQ SRO\FORQDO FHOO OLQHV ZHUH HVWDEOLVKHG IURP WKH WUDQVIHFWLRQ RI HDFK RI WKH FRQVWUXFWV GHVFULEHG DERYH DQG DUH QXPEHUHG 3, WKURXJK 3 &HOO OLQHV 3 3 DQG 3 ZHUH FKRVHQ IRU IXUWKHU DQDO\VLV VHH FKDSWHU f EDVHG XSRQ WKHLU DELOLW\ WR GLIIHUHQWLDWH WR DGLSRF\WHV ,QGXFWLRQ DQG 'LIIHUHQWLDWLRQ RI 7/ &HOOV ,QGXFWLRQ RI 7/ FHOOV WR DGLSRF\WHV ZDV FDUULHG RXW DV GHVFULEHG E\ %HUQORKU HW DO f 7/ FHOOV ZHUH JURZQ WR FRQIOXHQFH DQG GD\V SRVW FRQIOXHQFH ZHUH JLYHQ IUHVK '0(0 FRQWDLQLQJ b IHWDO FDOI VHUXP Q0 LQVXOLQ Q0 GH[DPHWKDVRQH DQG P0 LVREXW\OOPHWK\O [DQWKLQH )RUW\HLJKW KRXUV ODWHU WKH PHGLXP ZDV UHSODFHG ZLWK IUHVK '0(0 VXSSOHPHQWHG ZLWK b IHWDO FDOI VHUXP $GLSRF\WH FRQYHUVLRQ ZDV GHWHFWHG ZLWKLQ GD\V RI LQGXFWLRQ &HOO &XOWXUH IRU &KURPRVRPH /RFDOL]DWLRQ ,VRODWLRQ SURSDJDWLRQ DQG FKDUDFWHUL]DWLRQ RI SDUHQWDO KXPDQ DQG PXULQH FHOOV DV ZHOO DV WKH KXPDQPRXVH VRPDWLF FHOO K\EULGV ZDV FDUULHG RXW DV SUHYLRXVO\ GHVFULEHG f LQ 'U &DUOR &URFHnV ODERUDWRU\ DW 7HPSOH 8QLYHUVLW\ 6FKRRO RI 0HGLFLQH )HOV 5HVHDUFK ,QVWLWXWH 3KLODGHOSKLD 3$ 7KH KXPDQ FKURPRVRPHV HLWKHU SDUWLDO RU

PAGE 83

FRPSOHWH FKURPRVRPHVf UHWDLQHG LQ WKH URGHQWKXPDQ K\EULGV XVHG LQ GHWHUPLQLQJ WKH FKURPRVRPDO ORFDWLRQ RI WKH KXPDQ +% KLVWRQH JHQH DUH VFKHPDWLFDOO\ LOOXVWUDWHG LQ )LJXUH % ZRXOG OLNH WR DFNQRZOHGJH 6KLUZLQ 3RFNZLQVH 'U .D\ +XHEQHU 'U /LQGD $ &DQQL]]DUR DQG 'U &DUOR &URFH IRU WKHLU ZRUN RQ WKH FKURPRVRPDO ORFDOL]DWLRQ RI WKH +%*/ JHQH 7+3 &HOO FXOWXUH $FXWH PRQRF\WLF OHXNHPLD FHOOV 7+3 f ZHUH PDLQWDLQHG LQ VXVSHQVLRQ FXOWXUHV LQ 530, PHGLXP VXSSOHPHQWHG ZLWK b IHWDO FDOI VHUXP DQG NDQDP\FLQ 0JPOf

PAGE 84

&+$37(5 ,62/$7,21 $1' &+$5$&7(5,=$7,21 2) $ F'1$ )520 $ +80$1 +,6721( +% *(1( :+,&+ ,6 5(&,352&$//< (;35(66(' ,1 5(/$7,21 72 5(3/,&$7,21'(3(1'(17 +% +,6721( *(1(6 '85,1* +/ &(// ',))(5(17,$7,21 ,QWURGXFWLRQ 5HSOLFDWLRQGHSHQGHQW KLVWRQH JHQHV GR QRW FRQWDLQ LQWURQV f DQG WKHLU P51$V DUH VWUXFWXUDOO\ VLPSOH WKH\ DUH QRW SRO\DGHQ\ODWHG DQG KDYH VKRUW n OHDGHU DQG n WUDLOHU VHTXHQFHV f +RZHYHU UHSOLFDWLRQGHSHQGHQW KLVWRQH P51$V GR KDYH D FKDUDFWHULVWLF VWHPORRS PRWLI f 5HSOLFDWLRQGHSHQGHQW KLVWRQH JHQHV DUH FRRUGLQDWHO\ H[SUHVVHG GXULQJ WKH FHOO F\FOH DQG WKHLU H[SUHVVLRQ LV FRXSOHG ZLWK '1$ V\QWKHVLV f 7KH DEXQGDQFH RI WKH UHSOLFDWLRQGHSHQGHQW KXPDQ KLVWRQH P51$V LV UHJXODWHG DW ERWK WKH WUDQVFULSWLRQDO DQG SRVW WUDQVFULSWLRQDO OHYHOV UHYLHZHG LQ f ,Q FRQWUDVW UHSODFHPHQW KLVWRQH JHQHV PD\ FRQWDLQ LQWURQV f DQG WKHLU P51$V ZKLFK DUH SRO\DGHQ\ODWHG DQG IUHTXHQWO\ FRQWDLQ ORQJ n OHDGHUV DQG n WUDLOHUV DUH VWUXFWXUDOO\ PRUH FRPSOH[ WKDQ WKHLU UHSOLFDWLRQGHSHQGHQW FRXQWHUSDUWV f $OWKRXJK WKH SURFHVVHV JRYHUQLQJ UHSODFHPHQW KLVWRQH P51$ OHYHOV DUH QRW ZHOO XQGHUVWRRG LW LV FOHDU WKDW WKHLU P51$V

PAGE 85

f DV ZHOO DV WLVVXHVSHFLILF KLVWRQH P51$V f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f 7ZR +, FORQHV +$ FORQHV +% FORQHV + FORQHV DQG + FORQHV ZHUH LGHQWLILHG 1R FORQH JDYH D SRVLWLYH VLJQDO ZLWK PRUH WKDQ RQH KLVWRQH SUREH )LJXUH f

PAGE 86

)LJXUH 3ODTXH GRW K\EULGL]DWLRQ DQDO\VLV RI SXWDWLYH SRVLWLYH FORQHV IURP WKH $JWOO SRO\ $ F'1$ OLEUDU\ VFUHHQLQJ 8SRQ FRPSOHWLRQ RI WKH WKLUG URXQG RI $JWOO OLEUDU\ VFUHHQLQJ VHH )LJXUH f ++ SRVLWLYHV DQG +$+%+ SRVLWLYH SODJXHV ZHUH VHSDUDWHG DFFRUGLQJ WR LQGLYLGXDO KLVWRQH FODVV E\ SODTXH GRW DQDO\VLV 6SRW DVVD\ DQDO\VLV ZDV FDUULHG RXW XVLQJ QO DOLTXRWV RI $ EDFWHULRSKDJH GLOXWLRQV a SIX 8SRQ SODTXH IRUPDWLRQ ILOWHU OLIWV ZHUH SUHSDUHG IURP WKH ++ SODWH DQG IURP WKH +$+%+ SODWH DQG HDFK K\EULGL]HG WR DQ LQGLYLGXDO KLVWRQH 3ODEHOHG SUREH VHH )LJXUH f DV LQGLFDWHG DERYH HDFK SDQHO 7KH ILOWHUV ZHUH WKHQ ZDVKHG DQG DXWRUDGLRJUDSK\ SHUIRUPHG XVLQJ .RGDN ;$5 ILOP

PAGE 88

5HVWULFWLRQ (QGRQXFOHDVH 0DSSLQJ RI 3RVLWLYH &ORQHV ,VRODWHG )URP WKH ,JWOO 3RO\ $ F'1$ /LEUDU\ 6FUHHQLQJ 7ZHOYH FORQHV VXVSHFWHG RI FRQWDLQLQJ KXPDQ KLVWRQH JHQHV ZHUH SLFNHG IRU IXUWKHU DQDO\VLV WZR IURP HDFK KLVWRQH FODVV DV ZHOO DV WZR UDQGRPO\ SLFNHG DGGLWLRQDO FORQHVf DQG PDSSHG XVLQJ D FRPELQDWLRQ RI UHVWULFWLRQ HQGRQXFOHDVH GRXEOH GLJHVWLRQV DQG VRXWKHUQ EORW DQDO\VLV XVLQJ WKH SUREHV LQ )LJXUH f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f ZHUH IRXQG WR FRQWDLQ KLVWRQH SURWHLQ FRGLQJ VHTXHQFH

PAGE 89

)LJXUH 5HVWULFWLRQ HQGRQXFOHDVH 6RXWKHUQ PDSSLQJ RI $++& (DFK ODQH RI D b DJDURVH JHO ZDV ORDGHG ZLWK J RI $++& '1$ UHVWULFWHG ZLWK ( (FR 5, ; ;KR 6 6DF + +LQG ,,, (+ (FR 5,+LQG ,,, ;+ ;KR ,+LQG ,,, 6+ 6DF ,+LQG ,,, (% (FR 5,%DP +, /DQH 8 ZDV ORDGHG ZLWK QT XQFXW $++& '1$ DQG ODQH 0 ZLWK nODEHOHG $ '1$ GLJHVWHG ZLWK (FR 5, DQG +LQG ,,, DQG ZLWK nODEHOHG S%5 '1$ GLJHVWHG ZLWK +LQI )ROORZLQJ VHSDUDWLRQ RI WKH '1$ IUDJPHQWV HOHFWURSKRUHWLFDOO\ WKH '1$ ZDV WUDQVIHUUHG WR =HWDSUREH Q\ORQ PHPEUDQH DQG K\EULGL]HG WR D 3ODEHOHG +% SUREH VHH )LJXUH f $ SKRWRJUDSK RI WKH DERYH JHO VWDLQHG ZLWK HWKLGLXP EURPLGH LV VKRZQ WR WKH ULJKW RI WKH ILJXUH IRU UHIHUHQFH $ UHVWULFWLRQ PDS RI $++& LV LOOXVWUDWHG DW WKH WRS RI WKH ILJXUH $JWOO DUPV DUH UHSUHVHQWHG E\ FORVHG ER[HV DW WKH WHUPLQL RI WKH UHVWULFWLRQ PDS 5HVWULFWLRQ HQ]\PH DEEUHYLDWLRQV DUH DV IROORZV ( (FR 5, ; ;KR 6 6VW ,

PAGE 90

ESO 8 ( ; 6 + (+ 0 ;+ 6+ (% $K+& 0$33,1*

PAGE 91

)LJXUH 5HVWULFWLRQ HQGRQXFOHDVH PDSV RI SRVLWLYH FORQHV LVRODWHG IURP WKH NDWOO SRO\ $ F'1$ OLEUDU\ VFUHHQLQJ 5HVWULFWLRQ HQGRQXFOHDVH PDSV RI SRVLWLYH FORQHV LVRODWHG IURP WKH $JWOO SRO\ $ F'1$ OLEUDU\ VFUHHQLQJ DUH LOOXVWUDWHG 5HVWULFWLRQ HQGRQXFOHDVH VLWHV DUH DV GHSLFWHG LQ WKH WRS ULJKW FRUQHU RI WKH ILJXUH $JWOO DUPV DUH UHSUHVHQWHG E\ FORVHG ER[HV DW WKH WHUPLQL RI WKH HDFK UHVWULFWLRQ PDS $ GHQRWHV D IUDJPHQW JLYLQJ D SRVLWLYH K\EULGL]DWLRQ VLJQDO ZLWK D KLVWRQH FRGLQJ SUREH

PAGE 92

; '1$ )/$1.,1* (&2 5O ,16(576 ;KKF + , ESU (&2 5O 7 +,1' ,,, I ;+2 W 667 cW %$0 +, } .31 I + +$9 ,

PAGE 93

)LJXUH 6HTXHQFLQJ VWUDWHJ\ RQ WKH SRVLWLYH FORQHV LVRODWHG IURP WKH $TWOO SRO\ $ F'1$ OLEUDU\ VFUHHQLQJ $UURZV UHSUHVHQW UHJLRQV RI WKH ,JWOO FORQHV ZKLFK ZHUH VHJXHQFHG E\ 6DQJHU GLGHR[\ VHTXHQFLQJ $ F EHORZ D PDS LQGLFDWHV D UHVWULFWLRQ IUDJPHQW ZKLFK K\EULGL]HG WR D KLVWRQH SURWHLQ FRGLQJ SUREH VHH )LJXUH f DQG D & WR WKH OHIW RI D PDS PDUNV WKH FORQHV LGHQWLILHG E\ VHTXHQFH DQDO\VLV f WR FRQWDLQ KLVWRQH SURWHLQ FRGLQJ VHTXHQFHV $Q $ WR WKH OHIW RI D PDS PDUNV WKH FORQHV LGHQWLILHG E\ VHTXHQFH DQDO\VLV WR FRQWDLQ SRO\$f VWUHWFKHV DW WKH HQG RI WKH LQVHUW 5HVWULFWLRQ HQGRQXFOHDVH VLWHV DUH DV GHSLFWHG DW WKH WRS RI WKH ILJXUH $JWOO DUPV DUH UHSUHVHQWHG E\ FORVHG ER[HV DW WKH WHUPLQL RI WKH HDFK UHVWULFWLRQ PDS

PAGE 94

(&2 5O '1$ )/$1.,1* (&2 5O ,16(576 +,1' ,,, W ;+2 g ESU 667 A .31 +% &$ s s 6/ + &$ +$ -/ K + +

PAGE 95

DQG FORQHV $++& $++& DQG $++&f WR FRQWDLQ D VWUHWFK RI SRO\$f DW WKH WHUPLQXV RI WKHLU LQVHUWV )LJXUH f 6HTXHQFLQJ RI D 3RO\ $ +% F'1$ 2I WKH F'1$ FORQHV FRQWDLQLQJ +% KLVWRQH VHTXHQFHV REWDLQHG IURP WKH LQLWLDO VFUHHQLQJ RI WKH ODPEGD OLEUDU\ IRXU ZHUH PDSSHG XVLQJ D FRPELQDWLRQ RI UHVWULFWLRQ HQGRQXFOHDVH GRXEOH GLJHVWLRQV DQG VRXWKHUQ EORW DQDO\VLV DQG WKH FORQH ZLWK WKH ODUJHVW LQVHUW )LJXUH $++& QWf WKDW DOVR FRQWDLQHG D SRO\$f WUDFW )LJXUH f ZDV WKHQ VHOHFWHG IRU GHWDLOHG VHTXHQFH DQDO\VLV 5HJLRQV RI WKH KXPDQ +% KLVWRQH F'1$ $++& ZKLFK ZHUH VHTXHQFHG DUH LQGLFDWHG E\ DUURZV LQ )LJXUH DQG WKH QXFOHRWLGH VHTXHQFH LV SUHVHQWHG LQ )LJXUH $f 7KH $++& F'1$ HQFRGHV +% DPLQR DFLGV DQG WKH WUDQVODWLRQ VWRS FRGRQ $OVR FRQWDLQHG LQ WKH F'1$ VHTXHQFH LV WKH SRO\$f DGGLWLRQ VHTXHQFH $$7$$$ ORFDWHG QW GRZQVWUHDP IURP WKH WUDQVODWLRQ VWRS FRGRQ DQG D SRO\$f VWUHWFK ORFDWHG QW GRZQVWUHDP RI WKH SRO\$f DGGLWLRQ VHTXHQFH 7KH SURWHLQ FRGLQJ UHJLRQ LV *& ULFK bf ZKHUHDV WKH n XQn WUDQVODWHG WUDLOHU LV UHODWLYHO\ *& SRRU bf 6KRZQ LQ )LJXUH %f DUH SHUWLQHQW UHVWULFWLRQ VLWHV DQG WKH $++& IUDJPHQWV XVHG DV SUREHV IRU QRUWKHUQ EORW DQDO\VLV

PAGE 96

)LJXUH 6HTXHQFLQJ VWUDWHJ\ IRU WKH KXPDQ +% KLVWRQH F'1$ $++& $UURZV LQGLFDWH UHJLRQV RI WKH KXPDQ +% KLVWRQH F'1$ ++& ZKLFK ZHUH VHTXHQFHG E\ 6DQJHU GLGHR[\ VHTXHQFH DQDO\VLV f 7KH +% SURWHLQ FRGLQJ UHJLRQ LV LOOXVWUDWHG E\ D KDWFKHG ER[ 5HVWULFWLRQ HQ]\PH DEEUHYLDWLRQV DUH DV IROORZV ( (FR 5, ; ;KR 6 6VW $JWOO DUPV DUH QRW UHSUHVHQWHG LQ WKLV LOOXVWUDWLRQ

PAGE 97

QW

PAGE 98

)LJXUH $f 1XFOHRWLGH DQG GHGXFHG DPLQR DFLG VHTXHQFH RI WKH KXPDQ +% KLVWRQH F'1$ $++& %f 5HVWULFWLRQ PDS RI WKH F'1$ $++& $f 7KH ++& F'1$ HQFRGHV +% DPLQR DFLGV DQG WKH WUDQVODWLRQ VWRS FRGRQ $OVR FRQWDLQHG LQ WKH $++& F'1$ VHTXHQFH LV WKH SRO\ $ DGGLWLRQ VHTXHQFH $$7$$$ ORFDWHG QW GRZQVWUHDP IURP WKH SURWHLQ VWRS FRGRQ DQG D SRO\ $ VWUHWFK ORFDWHG QW GRZQVWUHDP IURP WKH ODVW QXFOHRWLGH RI WKH SRO\ $ DGGLWLRQ VHTXHQFH 6ROLG GRXEOH XQGHUOLQHG f VHTXHQFHV UHSUHVHQW SRWHQWLDO VSOLFH VLWHV %f 7KH +% SURWHLQ FRGLQJ UHJLRQ LV LOOXVWUDWHG E\ D FORVHG ER[ DQG WKH $JWOO DUPV DUH UHSUHVHQWHG E\ WKH RSHQ ER[HV 6ROLG EDUV EHORZ WKH UHVWULFWLRQ PDS LOOXVWUDWHV $++& SUREHV XVHG IRU QRUWKHUQ DQDO\VLV 5HVWULFWLRQ HQ]\PH DEEUHYLDWLRQV DUH DV IROORZV ( (FR 5, ; ;KR 6 6VW ,

PAGE 99

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f 352%( QW f 352%(

PAGE 100

&RPSDULVRQ RI WKH $++& 3UHGLFWHG $PLQR $FLG 6HTXHQFH :LWK 2WKHU +% $PLQR $FLG 6HTXHQFHV ,Q )LJXUH $f LV VKRZQ D FRPSDULVRQ RI VHOHFWHG DPLQR DFLGV IURP WKH SUHGLFWHG KLVWRQH +% VHTXHQFH RI $++& ZLWK D PXOWLVSHFLHV FRQVHQVXV +% SURWHLQ VHTXHQFH f D KXPDQ JHQRPLF VHTXHQFH f DQG VHYHUDO FKLFNHQ JHQRPLF VHTXHQFHV f 7KH DPLQR DFLGV VKRZQ YDU\ DW WKH LQGLFDWHG SRVLWLRQV DPRQJ WKH JHQHV UHSRUWHG LQ )LJXUH $f 7KHVH DPLQR DFLGV DUH HLWKHU VSHFLHVVSHFLILF WKDW LV WKH\ DUH FRQVHUYHG DPRQJ NQRZQ FKLFNHQ +% JHQHV EXW PD\ GLIIHU EHWZHHQ WKH FKLFNHQ DQG RWKHU VSHFLHV UHSRUWHG LQ )LJXUH $f DPLQR DFLGV DQG f RU VXEW\SH VSHFLILF ZKLFK PD\ YDU\ DPRQJ +% SURWHLQ YDULDQWV ZLWKLQ D VSHFLHV RU EHWZHHQ VSHFLHV DPLQR DFLGV DQG f 7KH DPLQR DFLG VHTXHQFH GDWD IRU KXPDQ +% KLVWRQHV DUH WRR LQFRPSOHWH WR SHUPLW FODVVLILFDWLRQ RI WKH ++& +% SURWHLQ DV D UHSOLFDWLRQGHSHQGHQW UHSODFHPHQW RU WLVVXHVSHFLILF YDULDQW +RZHYHU WKH SUHGLFWHG SURWHLQ LV b VLPLODU WR WKH UHSOLFDWLRQ GHSHQGHQW ++* +% SURWHLQ f ZLWK DPLQR DFLG GLIIHUHQFHV DW SRVLWLRQV DQG )LJXUH $ff DQG LGHQWLFDO WR WKH PXOWLVSHFLHV FRQVHQVXV +% VHTXHQFH FRPSLOHG E\ :HOOV f 7KH ++& SUHGLFWHG DPLQR DFLG VHTXHQFH LV DOVR VLPLODU WR WKH S+K$S+K& +% VHTXHQFH UHSRUWHG E\ =KRQJ HW DO f H[FHSW LQ WKH UHJLRQ RI DPLQR DFLGV WKURXJK

PAGE 101

)LJXUH $f &RPSDULVRQ RI WKH $++& KLVWRQH +% DPLQR DFLG VHTXHQFH ZLWK WKH VHTXHQFHV RI RWKHU +%V %f '1$ VHTXHQFH FRPSDULVRQ RI WKH XQWUDQVODWHG UHJLRQ RI VHOHFWHG KLVWRQH VHTXHQFHV $f $ $++& KXPDQ F'1$ % PXOWLVSHFLHV FRQVHQVXV +% SURWHLQ VHTXHQFH f & ƒ++* KXPDQ JHQRPLF VHTXHQFH f S%%$ FKLFNHQ JHQRPLF VHTXHQFH f ( S33G FKLFNHQ JHQRPLF VHTXHQFH f ) S.5OD S33G S55F S55H DQG S%5$ FKLFNHQ JHQRPLF VHTXHQFHV f $&+ DQG $&+$&+ FKLFNHQ JHQRPLF VHTXHQFHV f + S6W+% FKLFNHQ JHQRPLF VHTXHQFH f 'DVKHG OLQHV LQGLFDWH XQGHWHUPLQHG DPLQR DFLGV GXH WR LQFRPSOHWH QXFOHRWLGH VHTXHQFH LQIRUPDWLRQ DQG DQ DVWHULVN LQGLFDWHV DQ DPLQR DFLG ZKLFK LV LGHQWLFDO WR WKH $++& SUHGLFWHG DPLQR DFLG LQ WKH VDPH SRVLWLRQ 7KH DPLQR DFLG SRVLWLRQ LV LQGLFDWHG E\ WKH EROG QXPEHU DW WKH WRS RI HDFK FROXPQ DQG WKH RULJLQ RI HDFK VHTXHQFH LV LQGLFDWHG LQ WKH ULJKW KDQG FROXPQ $PLQR DFLGV ZKLFK GR QRW YDU\ DPRQJ WKH JHQHV UHSRUWHG DERYH DUH QRW LOOXVWUDWHG %f $++& KXPDQ +% F'1$ S.5OD FKLFNHQ +% JHQRPLF VHTXHQFH f S%%$ FKLFNHQ +% JHQRPLF VHTXHQFH f $++* KXPDQ +% JHQRPLF VHTXHQFH f S+K$S+K& KXPDQ +% JHQRPLF VHTXHQFH f S+K* KXPDQ +$ JHQRPLF VHTXHQFH f S67 KXPDQ + JHQRPLF VHTXHQFH f KXPDQ + JHQRPLF VHTXHQFH f KXPDQ +$= F'1$ VHTXHQFH f S6W+% FKLFNHQ JHQRPLF VHTXHQFH f 1XPEHUV IODQNHG E\ GRWV LQGLFDWH WKH QXPEHU RI EDVHV FRQWDLQHG EHWZHHQ WKH WZR DGMDFHQW UHSRUWHG VHTXHQFHV $ EODQN VSDFH LV D JDS FUHDWHG WR IDFLOLWDWH DOLJQPHQW $ SOXV VLJQ V\PEROL]HV WKH SUHVHQFH RI D SRO\ $r WDLO LQ WKH P51$ DQG D TXHVWLRQ PDUN LQGLFDWHV WKH ODFN RI GDWD FRQFHUQLQJ SRO\DGHQ\ODWLRQ RU WKH QXPEHU RI EDVHV FRQWDLQHG EHWZHHQ WZR DGMDFHQW UHSRUWHG VHTXHQFHV $UURZV LQGLFDWH WKH FRQVHUYHG n VWHPORRS PRWLI VHTXHQFH 7KH FRQVHUYHG VHTXHQFHV DUH GLVSOD\HG LQ WKH WRS OLQH LQ EROG SULQW

PAGE 102

$ $ f§ f§ $VS *O\ /\V $UJ 6HU OLH *O\ OLH 7KU 6HU +XPDQ % /HX $OD r r r r r r r r r r 0XOWL & 9DO $OD r r r r r 9DO r 9DO r /\V +XPDQ 9DO 7KU *O\ $VS r r $OD r 6HU r r r &KLFNHQ ( f§ f§ f§ f§ f§ f§ f§ r r r OLH fN &KLFNHQ ) 9DO 7KU *O\ $VS $UJ /\V r r r r r r &KLFNHQ 9DO 7KU *O\ $VS r /\V r r r r r r &KLFNHQ + 9DO 7KU *O\ $VS $UJ r 7KU r r r r r &KLFNHQ % 72&$ &&&$$$ **&7&7777&$*$*&&$&& *$*&7 $$7$$$ 7*$ 7$* 7$$ 77$ 7$$ 7*$ 7$$ 7$$ 7$$ 7$* 7*&$ 7*&* 77&7 7*&$ 7*&$ 7*&$ 7*7$ 77&$ ff7**$ 7*&*f 7&&$$$ **&7&7777&$*$*&&$&&*$*&7$$7$$$ &&&$$$ **&7&7777&**$*&&$&& ***&7"""" &&&$$$&**&7&7777&**$*&&&$$ ***&7"""" &&&$$$ **&7&7777&$*$*&&$&7*$*&7$7$7$&7*$&$7*7*$$*$7 &&&7$$&**&7&7777$$*$*&&$&&*$*&7**7*&77*7$777&&7&&7& &$*&$$ **&7&7777&$*$*&&$&&*$*&7*77*7*&777**$77$7*&& &&&$$$ **&7&7777&$*$*&&$&&7$*&7*7*$7$$777777*77*7&7 77&7&$$$$$$$$$$$$$$$$777&7&77&77&&7*77$7$77$$$ 77&&77*77$7&7&$**$&7&7$$$7$&7&7$$&$*&7*7$$7$$$ &&&$$$ **&7&7777&$*$*&&$&& *$*7*"""" RR XL

PAGE 103

&RPSDULVRQ RI 1RQWUDQVODWHG 6HTXHQFHV )URP $++& DQG 2WKHU +LVWRQH F'1$V DQG *HQHV 7KH n QRQWUDQVODWHG VHTXHQFHV IURP $++& DQG RWKHU KLVWRQH F'1$V DQG JHQHV DUH LOOXVWUDWHG LQ )LJXUH %f 7KH KLVWRQH n VWHPORRS HOHPHQW f LV SUHVHQW LQ DOO RI WKH SRO\ $n KLVWRQH VHTXHQFHV f ,QFOXGHG LQ WKLV FRPSDULVRQ DUH WKUHH SRO\ $ KXPDQ KLVWRQH VHTXHQFHV f DQG RQH SRO\ $ FKLFNHQ W+% VHTXHQFH f RI WKHVH RQO\ $++& DQG WKH FKLFNHQ W+% VHTXHQFH FRQWDLQ D n VWHPORRS PRWLI 7KH $++& +% DQG WKH +$= VHTXHQFHV f FRQWDLQ WKH $$7$$$ SRO\$f DGGLWLRQ HOHPHQW f ZKLOH WKH + VHTXHQFH f FRQWDLQV DQ $77$$$ HOHPHQW DOVR LPSOLFDWHG LQ SRO\DGHQ\ODWLRQ f VHH )LJXUH %ff +RZHYHU WKH W+% VHTXHQFH GRHV QRW DSSHDU WR FRQWDLQ HLWKHU SRO\DGHQ\ODWLRQ HOHPHQW f ,Q DGGLWLRQ WR FRQWDLQLQJ D VWHPORRS PRWLI $++& LV GLVWLQFW IURP RWKHU NQRZQ KXPDQ SRO\ $ KLVWRQH VHTXHQFHV DV LW FRQWDLQV WKH ORQJHVW n WUDLOHUf§ QW DV FRPSDUHG WR IRU + f DQG IRU +$= f ++& P51$ LV ([SUHVVHG 7KURXJKRXW WKH +H/D &HOO &\FOH DQG 'XULQJ ,QKLELWLRQ RI '1$ 6\QWKHVLV 7R GHWHUPLQH WKH H[WHQW RI FHOO F\FOH UHJXODWLRQ RI WKH ++& +% KLVWRQH JHQH ZH ILUVW PHDVXUHG WKH OHYHO RI ++& P51$ WKURXJKRXW WKH +H/D FHOO F\FOH 3UHVHQWHG LQ )LJXUHV DQG DUH WKH UHVXOWV RI QRUWKHUQ EORW DQDO\VLV RI SRO\ $ 51$ LVRODWHG IURP V\QFKURQL]HG +H/D

PAGE 104

)LJXUH ++& P51$ OHYHOV GXULQJ WKH +H/D FHOO F\FOH 3RO\ $ 51$ ZDV LVRODWHG IURP V\QFKURQL]HG +H/D FHOOV SULRU WR f DQG DIWHU DQG KRXUVf UHOHDVH IURP D GRXEOH WK\PLGLQH EORFN DV ZHOO DV IURP XQV\QFKURQL]HG +H/D FHOOV LQ ORJDULWKPLF /,f JURZWK ,Q D VHSDUDWH H[SHULPHQW SRO\ $ 51$ ZDV LVRODWHG IURP XQV\QFKURQL]HG +H/D FHOOV LQ ORJDULWKPLF JURZWK ZKLFK KDG EHHQ WUHDWHG ZLWK WKH '1$ V\QWKHVLV LQKLELWRU K\GUR[\XUHD +8f DV ZHOO DV IURP FHOOV ZKLFK ZHUH QRW WUHDWHG /f MXJ RI HDFK 51$ VDPSOH ZDV VL]H IUDFWLRQDWHG SDVVLYHO\ WUDQVIHUUHG WR =HWD3UREH Q\ORQ PHPEUDQH SUHZDVKHG SUHK\EULGL]HG DQG K\EULGL]HG WR WKH IROORZLQJ 3ODEHOHG SUREHV QW (FR 5,6VW n IUDJPHQW IURP $++& SDQHO $f SODVPLG 5*$3'+ f FRQWDLQLQJ *$3'+ VHTXHQFHV IURP UDW SDQHO %f 7KH ILOWHUV ZHUH WKHQ ZDVKHG DQG DXWRUDGLRJUDSK\ SHUIRUPHG XVLQJ .RGDN ;$5 ILOP /DQH 0 FRQWDLQV 51$ PDUNHUV SXUFKDVHG IURP %HWKHVGD 5HVHDUFK /DEV 7KH VL]HV RI WKH YDULRXV EDQGV DUH LQGLFDWHG LQ .E E\ WKH QXPEHUV RQ WKH ULJKW VLGH RI WKH ILJXUH

PAGE 106

)LJXUH +% P51$ OHYHOV GXULQJ WKH +H/D FHOO F\FOH 1RUWKHUQ EORW DQDO\VLV ZDV FDUULHG RXW RQ WKH VDPH +H/D VDPSOHV XVHG LQ )LJXUH ZLWK WKH 3ODEHOHG QW (FR 5,;KR IUDJPHQW IURP $++& DV SUREH SDQHO $f 3DQHO % FRQWDLQV D VKRUWHU H[SRVXUH RI WKH .E EDQGV IURP SDQHO $ 7KH VL]HV RI WKH YDULRXV EDQGV DUH LQGLFDWHG LQ .E RQ WKH ULJKW VLGH RI WKH ILJXUH DUH

PAGE 108

FHOOV SULRU WR DQG DIWHU UHOHDVH IURP D GRXEOH WK\PLGLQH EORFN DV ZHOO DV IURP XQV\QFKURQL]HG +H/D FHOOV LQ ORJDULWKPLF JURZWK DQG DIWHU WUHDWPHQW ZLWK WKH '1$ V\QWKHVLV LQKLELWRU K\GUR[\XUHD 7KH SHDN RI 6 SKDVH DV GHWHUPLQHG E\ +WK\PLGLQH LQFRUSRUDWLRQ RFFXUUHG DW KRXUV DIWHU UHOHDVH IURP WKH VHFRQG WK\PLGLQH EORFN GDWD QRW VKRZQf 7R IROORZ WKH P51$ OHYHOV RI DOO +% VSHFLHV WZR GLIIHUHQW SUREHV IURP $++& ZHUH XVHG 2QH SUREH ZDV WKH QW (FR 5,;KR IUDJPHQW )LJXUH %ff ZKLFK VSDQV WKH +% SURWHLQ FRGLQJ UHJLRQ DQG KDV VWURQJ VHTXHQFH VLPLODULW\ ZLWK RWKHU UHSRUWHG +% VHTXHQFHV 7KH VHFRQG SUREH ZDV WKH QW (FR 5,6VW IUDJPHQW )LJXUH %ff WKDW VSDQV WKH GLVWDO KDOI RI WKH n QRQWUDQVODWHG WUDLOHU 1RUWKHUQ EORW DQDO\VLV XVLQJ WKH n VSHFLILF SUREH UHYHDOHG D QW EDQG WKDW ZDV SUHVHQW WKURXJKRXW WKH +H/D FHOO F\FOH )LJXUH $ff $V DQ LQWHUQDO FRQWURO IRU WKH DPRXQW RI P51$ SHU ODQH WKLV EORW ZDV UHSUREHG ZLWK WKH 3ODEHOHG SODVPLG 5*$3'+ f ZKLFK FRQWDLQV UDW F'1$ VHTXHQFHV FRGLQJ IRU *$3'+ )LJXUH %ff *$3'+ P51$ OHYHOV KDYH EHHQ VKRZQ LQ RXU ODE WR UHPDLQ FRQVWDQW WKURXJKRXW WKH +H/D FHOO F\FOH ,Q FHOOV LQ ZKLFK '1$ V\QWKHVLV ZDV LQKLELWHG ZLWK K\GUR[\XUHD WKH OHYHO RI WKH QW ++& P51$ VSHFLHV GHFUHDVHG E\ DSSUR[LPDWHO\ SHUFHQW )LJXUH $ff DV LQGLFDWHG E\ GHQVLWRPHWU\ DQG QRUPDOL]DWLRQ WR *$3'+ P51$ OHYHOV )LJXUH %ff

PAGE 109

:KHQ QRUWKHUQ EORW DQDO\VLV ZDV FDUULHG RXW XVLQJ WKH SURWHLQFRGLQJ IUDJPHQW IURP ++& )LJXUHV $f DQG %ff +% P51$V ZHUH REVHUYHG DW QW DQG QW LQ DGGLWLRQ WR ORZ PROHFXODU ZHLJKW +% VSHFLHV PLJUDWLQJ DW DSSUR[LPDWHO\ QW ,W VKRXOG EH QRWHG WKDW D VLQJOH URXQG RI VHOHFWLRQ ZLWK ROLJR G7FHOOXORVH JUHDWO\ HQULFKHV IRU SRO\ $ 51$ EXW GRHV QRW WRWDOO\ UHPRYH DOO WKH SRO\ $n 51$ 7KH QW +% P51$V ZHUH GHWHFWHG WKURXJKRXW WKH +H/D FHOO F\FOH EXW WKHLU OHYHOV FOHDUO\ IOXFWXDWHG DV D IXQFWLRQ RI WKH FHOO F\FOH )LJXUHV $f DQG f§%ff /HYHOV RI WKH QW DQG QW P51$V UHPDLQHG UHODWLYHO\ FRQVWDQW WKURXJKRXW WKH FHOO F\FOH LQ FRPSDULVRQ WR WKH QW UHSOLFDWLRQGHSHQGHQW P51$V )LJXUH $ff 'HQVLWRPHWULF DQDO\VLV UHYHDOHG WKDW DIWHU WUHDWPHQW RI SUROLIHUDWLQJ +H/D FHOOV ZLWK K\GUR[\XUHD WKH OHYHOV RI WKH ++& P51$ QWf DV ZHOO DV WKH QW P51$ GURSSHG WR DSSUR[LPDWHO\ SHUFHQW RI WKHLU QRQLQKLELWHG OHYHOV ZKHUHDV WKH QW UHSOLFDWLRQGHSHQGHQW P51$V GHFUHDVHG E\ b )LJXUH $ff $Q DGGLWLRQDO +% FORQH ZLWK D GLIIHUHQW UHVWULFWLRQ SDWWHUQ IURP WKDW RI $++& KDV EHHQ LVRODWHG IURP D SRO\ $ F'1$ OLEUDU\ VHH FORQH $++& LQ )LJXUH f ZKLFK VXJJHVW WKDW WKH QW UHSOLFDWLRQ LQGHSHQGHQW +% P51$ VSHFLHV LV QRW WKH WUDQVFULSWLRQ SURGXFW RI ++& JHQH EXW SRVVLEO\ WKDW RI WKH SXWDWLYH ++& JHQH

PAGE 110

7UDQVFULSWLRQ RI WKH ++& +LVWRQH *HQH 3XULQD WKH +H/D &HOO &\FOH 7R GHWHUPLQH WKH H[WHQW WR ZKLFK WUDQVFULSWLRQ FRQWULEXWHV WR WKH FHOO F\FOH UHJXODWLRQ RI ++& JHQH H[SUHVVLRQ ZH FDUULHG RXW LQ YLWUR UXQRQ WUDQVFULSWLRQ DQDO\VLV RI QXFOHL LVRODWHG IURP V\QFKURQL]HG +H/D FHOOV DIWHU UHOHDVH IURP D GRXEOH WK\PLGLQH EORFN )LJXUH f 'XULQJ 6 SKDVH KRXUVf WUDQVFULSWLRQ IURP WKH ++& JHQH ZDV WZR IROG KLJKHU WKDQ LWV OHYHO RXWVLGH RI 6 SKDVH KRXUVf ,Q FRPSDULVRQ ZH REVHUYHG WKDW GXULQJ 6 SKDVH WRWDO + KLVWRQH DQG +&$7 IXVLRQ JHQH WUDQVFULSWLRQ LQFUHDVHG WR IROG RYHU WKH OHYHOV RXWVLGH RI 6 SKDVH )LJXUH f ++& P51$ LV ([SUHVVHG LQ D 5HFLSURFDO 5HODWLRQVKLS :LWK 5HSOLFDWLRQ'HSHQGHQW +% P51$V 'XULQJ WKH 2QVHW RI +/ &HOO 'LIIHUHQWLDWLRQ :H H[DPLQHG WKH OHYHO RI ++& P51$ IROORZLQJ LQGXFWLRQ RI +/ SURP\HORF\WLF OHXNHPLD FHOOV DORQJ WKH PRQRF\WLF SDWKZD\ DV SUROLIHUDWLRQ FHDVHV DQG PRQRF\WH VSHFLILF JHQH H[SUHVVLRQ LV LQLWLDWHG 51$ ZDV LVRODWHG IURP SUROLIHUDWLQJ +/ FHOOV DQG IURP FHOOV DW YDULRXV WLPHV DIWHU LQGXFWLRQ RI GLIIHUHQWLDWLRQ ZLWK 73$ 1RUWKHUQ EORW DQDO\VHV XVLQJ WKH n VSHFLILF SUREH )LJXUH $ff DQG WKH SURWHLQFRGLQJ IUDJPHQW )LJXUH f UHYHDOHG WKDW WKH QW DQG QW +% KLVWRQH P51$V SUHVHQW LQ YHU\ ORZ OHYHOV LQ WKH SUROLIHUDWLQJ +/ VDPSOH ZHUH LQGXFHG XSRQ 73$ WUHDWPHQW %DVHG RQ GHQVLWRPHWULF DQDO\VLV WKH

PAGE 111

)LJXUH 7UDQVFULSWLRQ RI WKH ++& KLVWRQH JHQH GXULQJ WKH +H/D FHOO F\FOH 7KH SRO\FORQDO +H/D FHOO OLQH SO ZKLFK FRQWDLQV DQ + KLVWRQH&$7 IXVLRQ JHQH ZDV V\QFKURQL]HG YLD GRXEOH WK\PLGLQH EORFN &HOOV ZHUH KDUYHVWHG DW WKH *O6 ERUGHU 35(f LQ PLG 6SKDVH KRXUVf DQG LQ *0 KRXUVf 5DGLRODEHOHG WUDQVFULSWV GSPf IURP LQ YLWUR QXFOHDU UXQRQ WUDQVFULSWLRQ ZHUH K\EULGL]HG WR VORW EORWV FRQWDLQLQJ OLQHDUL]HG SODVPLG '1$V + UHSOLFDWLRQ GHSHQGHQW KXPDQ + KLVWRQH JHQH f +% QW n (FR 5, IUDJPHQW IURP WKH $++& UHSOLFDWLRQLQGHSHQGHQW +% F'1$ DQG &$7 + SURPRWHU&$7 IXVLRQ JHQH 7KH K\EULGL]HG ILOWHUV ZHUH WKHQ ZDVKHG DQG DXWRUDGLRJUDSK\ SHUIRUPHG XVLQJ ;$5 RU &URQH[ ILOP DQG &URQH[ /LJKWQLQJ 3OXV VFUHHQV DW r& 'HQVLWRPHWULF DQDO\VLV ZDV SHUIRUPHG DQG WKH UHVXOWDQW YDOXHV IRU WUDQVFULSWLRQ RI WKH KLVWRQH JHQHV DQG +&$7 IXVLRQ JHQH ZHUH QRUPDOL]HG WR WKRVH RI WKH 6 ULERVRPDO 51$ JHQH QRW VKRZQf 7KH UHVXOWV REWDLQHG LQ VL[ VLPLODU H[SHULPHQWV DUH UHSUHVHQWHG LQ WKH WDEOH

PAGE 112

35( +5 +5 + +% &$7 5$7,2 2) 75$16&5,37,21 $7 7+5(( +2856 5(/$7,9( 72 7(1 +2856 WWO2 $%62/87( s Q 1250$/,=(' s Q

PAGE 113

)LJXUH ++& P51$ OHYHOV GXULQJ SUROLIHUDWLRQ DQG GLIIHUHQWLDWLRQ RI +/ FHOOV 3RO\ $ 51$ AT IURP SUROLIHUDWLQJ 3f DQG GLIIHUHQWLDWHG 'LIIf +/ FHOOV ZDV XVHG IRU 1RUWKHUQ EORW DQDO\VLV 51$ ZDV LVRODWHG IURP GLIIHUHQWLDWHG +/ FHOOV DW ,Gf Gf DQG GD\V Gf DIWHU WUHDWPHQW ZLWK 73$ 7KH 3ODEHOHG SUREHV ZHUH WKH QW (FR 5,6VW n QRQFRGLQJ IUDJPHQW IURP $++& SDQHO $f DQG WKH QW 3VW LQVHUW IURP S*P f SDQHO %f

PAGE 115

)LJXUH +% P51$ OHYHOV GXULQJ SUROLIHUDWLRQ DQG GLIIHUHQWLDWLRQ RI +/ FHOOV 3RO\ $ 51$ QJ IURP SUROLIHUDWLQJ 3f DQG GLIIHUHQWLDWHG 'LIIf +/ FHOOV ZDV XVHG IRU 1RUWKHUQ EORW DQDO\VLV ZLWK WKH 3ODEHOHG (FR 5,;KR +% FRGLQJUHJLRQ IUDJPHQW RI $++& DV SUREH 51$ ZDV LVRODWHG IURP GLIIHUHQWLDWHG +/ FHOOV DW ,Gf Gf Gf Gf DQG GD\V Gf DIWHU WUHDWPHQW ZLWK 73$ 7KH VL]HV RI WKH YDULRXV EDQGV DUH LQGLFDWHG LQ .E RQ WKH ULJKW VLGH RI WKH ILJXUH

PAGE 117

QW ++& P51$ ZDV SUHVHQW DW D IROG KLJKHU OHYHO E\ GD\ WKDQ LQ WKH SUROLIHUDWLQJ VDPSOH )LJXUH f 7KH QW P51$ SUHVHQW DW D YHU\ ORZ OHYHO LQ WKH SUROLIHUDWLQJ +/ FHOOV DOVR LQFUHDVHG XQWLO GD\ )LJXUH f 7KH QW +% P51$V ZHUH GHWHFWHG DW D KLJK OHYHO LQ WKH SUROLIHUDWLQJ +/ VDPSOH EXW UDSLGO\ GHFOLQHG XQWLO DW GD\ WKH\ KDG GHFUHDVHG DSSUR[LPDWHO\ IROG IURP WKHLU OHYHO LQ WKH SUROLIHUDWLQJ VDPSOH $V LOOXVWUDWHG LQ )LJXUH D UHFLSURFDO UHODWLRQVKLS H[LVWV GXULQJ +/ GLIIHUHQWLDWLRQ EHWZHHQ WKH KLJK PROHFXODU ZHLJKW +% P51$V ZKLFK LQFUHDVH IROG DQG WKH QW +% P51$V ZKLFK GHFUHDVH WR OHVV WKDQ b RI WKHLU OHYHO LQ SUROLIHUDWLQJ +/ FHOOV 'LVFXVVLRQ 6WUXFWXUDO $QDO\VLV RI WKH $++& +% F'1$ $OWKRXJK FRPSDULVRQ RI WKH QXFOHRWLGH VHTXHQFH RI $++& ZLWK VHYHUDO RWKHU +% KLVWRQH VHTXHQFHV UHYHDOHG WKDW WKH SURWHLQ FRGLQJ UHJLRQ LV KLJKO\ FRQVHUYHG WKH VWULNLQJ GLIIHUHQFHV LQ WKH n WUDLOLQJ VHTXHQFHV VXJJHVW WKDW WKH $++& F'1$ LV XQLTXH DPRQJ +% KLVWRQH VHTXHQFHV GHVFULEHG WKXV IDU 7KH $++& +% F'1$ LV WKH RQO\ +% KXPDQ KLVWRQH VHTXHQFH UHSRUWHG ZKLFK FRQWDLQV ERWK WKH KLJKO\ FRQVHUYHG KLVWRQH n VWHPORRS VWUXFWXUH f DQG D SRO\$f DGGLWLRQ VHTXHQFH f IROORZHG E\ D SRO\$f VWUHWFK DOWKRXJK 0DQQLURQL HW DO f KDYH UHSRUWHG DQ

PAGE 118

)LJXUH 'HQVLWRPHWULF DQDO\VLV RI WKH QW UHSOLFDWLRQGHSHQGHQW +% P51$ DQG WKH QW ++& P51$ OHYHOV GXULQJ GLIIHUHQWLDWLRQ RI +/ FHOOV 'HQVLWRPHWULF DQDO\VLV RI WKH QW UHSOLFDWLRQ GHSHQGHQW +% P51$ 2GDVKHG OLQHf DQG WKH QW ++& P51$ VROLG OLQHf OHYHOV GXULQJ GLIIHUHQWLDWLRQ RI +/ FHOOV 7KH P51$ OHYHOV DUH SUHVHQWHG DV D IUDFWLRQ RI WKH PD[LPXP IRU HDFK P51$ YHUWLFDO D[LVf

PAGE 119

&-L E

PAGE 120

+$ KLVWRQH LVRSURWHLQ +$;f ZKLFK LV HQFRGHG E\ D SRO\ $ P51$ FRQWDLQLQJ WKH n VWHPORRS VWUXFWXUH DQG D SRO\$f DGGLWLRQ VHTXHQFH $++& LV DOVR GLVWLQFW IURP WKH RWKHU SRO\ $ KLVWRQH VHTXHQFHV UHSRUWHG LQ WKDW LW FRQWDLQV WKH ORQJHVW n WUDLOHU 7KH VWUXFWXUDO HOHPHQWV GHVFULEHG IRU $++& WKH KLJKO\ FRQVHUYHG KLVWRQH n VWHPORRS VWUXFWXUH DQG D SRO\$f DGGLWLRQ VHTXHQFH IROORZHG E\ D SRO\$f VWUHWFKf PD\ SOD\ D UROH LQ FRQIHUULQJ ERWK UHSOLFDWLRQ GHSHQGHQW DQG UHSOLFDWLRQLQGHSHQGHQW SURSHUWLHV &HOO &\FOH 5HJXODWLRQ RI WKH ++& +% +LVWRQH *HQH :KHQ +% P51$ OHYHOV ZHUH PHDVXUHG WKURXJKRXW WKH +H/D FHOO F\FOH WKH QW DQG QW P51$V GLG QRW XQGHUJR FHOOF\FOH IOXFWXDWLRQV DV GLG WKH UHSOLFDWLRQGHSHQGHQW KLVWRQH P51$V f ,Q DGGLWLRQ WKH OHYHO RI WKH QW ++& P51$ VSHFLHV LQ SUROLIHUDWLQJ +H/D FHOOV GURSSHG RQO\ b DIWHU WUHDWPHQW ZLWK K\GUR[\XUHD VXJJHVWLQJ LQFRPSOHWH FRXSOLQJ WR '1$ V\QWKHVLV 7KLV LV FRQVLVWHQW ZLWK UHVXOWV RQ WKH UDW WHVWLVVSHFLILF + DQG VHD XUFKLQ DKLVWRQH + P51$V FRQWDLQLQJ WKH n VWHPORRS PRWLI ZKLFK KDYH EHHQ REVHUYHG WR EH VWDEOH LQ WKH SUHVHQFH RI K\GUR[\XUHD f 7KH VWHDG\ VWDWH OHYHO RI WKH SRO\ $ QW P51$ LV PXFK ORZHU WKDQ WKH VWHDG\ VWDWH OHYHO RI FHOO F\FOH GHSHQGHQW +% P51$V LQ SUROLIHUDWLQJ FHOOV OHVV WKDQ bf )LJXUH DQG GDWD QRW VKRZQf KRZHYHU TXDQWLWDWLYH FRPSDULVRQV DUH GLIILFXOW GXH WR WKH IDFW WKDW

PAGE 121

VHYHUDO +% JHQHV FRQWULEXWH WR WKH FHOO F\FOH GHSHQGHQW +% P51$ SRRO ,Q YLWUR UXQRQ WUDQVFULSWLRQ DQDO\VLV RI QXFOHL LVRODWHG IURP +H/D FHOOV UHYHDOHG WKDW WUDQVFULSWLRQ IURP WKH ++& JHQH LV WZR IROG KLJKHU GXULQJ 6 SKDVH WKDQ RXWVLGH RI 6 SKDVH ZKHUHDV GXULQJ 6 SKDVH WRWDO + KLVWRQH DQG +&$7 IXVLRQ JHQH WUDQVFULSWLRQ LQFUHDVHG WR IROG RYHU WKH OHYHOV RXWVLGH RI 6 SKDVH )LJXUH f 7KHVH UHVXOWV VXJJHVW WKDW WUDQVFULSWLRQDO UHJXODWLRQ RI ++& H[SUHVVLRQ LV VLPLODU WR WKDW RI UHSOLFDWLRQGHSHQGHQW KLVWRQH JHQHV 2XU +H/D FHOO F\FOH GDWD UDLVH WKH TXHVWLRQ 'RHV WKH LQWHUQDO n VWHPORRS PRWLI LQIOXHQFH ++& P51$ SURFHVVLQJ DQGRU VWDELOLW\ RU DUH WKHVH FRQWUROOHG SULPDULO\ E\ RWKHU n WUDLOHU HOHPHQWV VXFK DV WKH ODUJH VL]H RI WKH n WUDLOHU VHTXHQFH DV ZHOO DV LWV SRO\$f WDLO" ,W LV DOVR SRVVLEOH WKDW QXFOHRWLGH VHTXHQFHV LQ SURWHLQ FRGLQJ UHJLRQV DUH LQYROYHG LQ KLVWRQH P51$ GHJUDGDWLRQ f 7KH OHYHO RI WKH QW ++& P51$ VSHFLHV LV YHU\ ORZ LQ SUROLIHUDWLQJ +H/D DQG +/ FHOOV WKLV REVHUYDWLRQ UDLVHV WKH SRVVLELOLW\ RI D GXDO SURFHVVLQJ SDWKZD\ LQ ZKLFK WKH PDMRULW\ RI WKH ++& WUDQVFULSWV DUH FOHDYHG WR WKH LQWHUQDO n VWHPORRS PRWLI OHDYLQJ RQO\ D PLQRULW\ LQ WKH QW IRUP 7KLV LV FRQVLVWHQW ZLWK UHSRUWV RI KLVWRQH JHQHV ZLWK DOWHUQDWLYH SDWKZD\V RI P51$ n HQG SURFHVVLQJ WKDW SURGXFH LQGHSHQGHQWO\ UHJXODWHG P51$V IURP D VLQJOH JHQH f 7KH SRVVLELOLW\ RI D GXDO SURFHVVLQJ SDWKZD\ IRU WKH ++&

PAGE 122

LWL51$ LV IXUWKHU VXSSRUWHG E\ WKH SUHVHQFH RI WKH VHTXHQFH $$$*7$$*7 SRVLWLRQ LQ )LJXUH $ff ORFDWHG QW n WR WKH VWHPORRS PRWLI ZKLFK LV QHDUO\ LGHQWLFDO WR WKH n VSOLFH VLWH FRQVHQVXV f 7KH $++& VHTXHQFH DOVR FRQWDLQV D SRWHQWLDO n VSOLFH VLWH f 7*7*7$7$*$7 SRVLWLRQ f WKDW LI VSOLFHG WR WKH $$$*7$$*7 n VSOLFH VLWH ZRXOG UHVXOW LQ D SRO\ $ P51$ VLPLODU LQ VL]H WR WKH EXON RI +% P51$ REVHUYHG LQ F\FOLQJ FHOOV 7KLV SRWHQWLDO VSOLFH MXQFWLRQ FRQIRUPV WR WKH *7$* UXOH f DQG LV LPPHGLDWHO\ SUHFHGHG E\ D S\ULPLGLQHULFK UHJLRQ DQG D EUDQFKSRLQW VHTXHQFH f 7&77$$& QW XSVWUHDP 6WDXEHU HW DO f REVHUYHG SRVVLEOH SURFHVVLQJ RI DQ + P51$ DW D VLPLODU n VSOLFH VLWH FRQVHQVXV ORFDWHG QW n WR WKH VWHPORRS PRWLI WR DQ XQGHILQHG n VSOLFH VLWH IXUWKHU GRZQVWUHDP 7KH QW +% P51$ REVHUYHG ZKHQ QRUWKHUQ EORW DQDO\VLV ZDV FDUULHG RXW XVLQJ WKH SURWHLQn FRGLQJ IUDJPHQW IURP $++& )LJXUH $ff FRXOG UHVXOW IURP H[SUHVVLRQ IURP DQRWKHU +% JHQH RU PD\ UHSUHVHQW GXDO SURFHVVLQJ RI WUDQVFULSWV IURP WKH ++& JHQH 5HJXODWLRQ RI ++& P51$ 'XULQJ 'LIIHUHQWLDWLRQ LQ +/ FHOOV 1RUWKHUQ EORW DQDO\VLV RI SRO\ $ 51$ LVRODWHG IURP SUROLIHUDWLQJ DQG GLIIHUHQWLDWHG +/ FHOOV GHPRQVWUDWHG WKDW WKH ++& P51$ LV UHJXODWHG LQ D YHU\ GLIIHUHQW PDQQHU WKDQ WKH P51$V IRU PRVW RWKHU +% KLVWRQHV 7KH QW ++& P51$ DQG UHSOLFDWLRQGHSHQGHQW +% KLVWRQH P51$

PAGE 123

OHYHOV DSSHDU WR EH UHJXODWHG LQ D UHFLSURFDO IDVKLRQ ZKHQ +/ FHOOV DUH WHUPLQDOO\ GLIIHUHQWLDWHG $ SRUWLRQ RI WKH QW UHSOLFDWLRQGHSHQGHQW +% KLVWRQH P51$ GHWHFWHG PD\ DULVH IURP WKH ++& JHQH GXH WR XWLOL]DWLRQ RI WKH n VWHPORRS VWUXFWXUH RU E\ D VSOLFLQJ HYHQW LQ WKH n WUDLOHU VHH FKDSWHU f 7KHVH UHVXOWV FRQILUP DQG H[WHQG RXU SUHYLRXV VWXGLHV GHPRQVWUDWLQJ WKH GRZQUHJXODWLRQ RI UHSOLFDWLRQGHSHQGHQW KLVWRQHV DQG WKHLU UHFLSURFDO UHODWLRQVKLS ZLWK D UHSOLFDWLRQLQGHSHQGHQW +% JHQH GXULQJ WKH RQVHW RI +/ FHOO GLIIHUHQWLDWLRQ f 7KH LQFUHDVH LQ QW ++& P51$ OHYHOV GXULQJ GLIIHUHQWLDWLRQ PD\ SURYLGH +% KLVWRQH SURWHLQV UHTXLUHG IRU WKH UHRUJDQL]DWLRQ DQG FRQGHQVDWLRQ RI FKURPDWLQ WDNLQJ SODFH GXULQJ GLIIHUHQWLDWLRQ $ VWULNLQJ VLPLODULW\ H[LVWV EHWZHHQ WKH XSUHJXODWLRQ RI WKH QW ++& +% P51$ GXULQJ GLIIHUHQWLDWLRQ LQ +/ FHOOV DQG WKH XSUHJXODWLRQ RI WKH FKLFNHQ W+% P51$ LQ SDFK\WHQH VSHUPDWRF\WHV f %RWK WKH ++& P51$ DQG WKH W+% P51$ FRQWDLQ WKH n VWHPORRS PRWLI LQ DGGLWLRQ WR D SRO\$f WDLO f 5HSOLFDWLRQGHSHQGHQW +% P51$V DUH DOVR WUDQVFULEHG IURP WKH VDPH JHQH DV WKH W+% P51$ EXW DUH SURFHVVHG E\ DQ DOWHUQDWH SDWKZD\ RI P51$ n HQG IRUPDWLRQ f 7KH H[LVWHQFH RI DOWHUQDWH n SURFHVVLQJ SDWKZD\V IRU WKLV FKLFNHQ +% JHQH UDLVHV WKH SRVVLELOLW\ WKDW D VLPLODU GXDO SURFHVVLQJ PD\ EH RSHUDWLYH IRU WKH ++& JHQH ZKLFK ZRXOG EH FRQVLVWHQW ZLWK WKH SUHVHQFH RI SRWHQWLDO VSOLFH

PAGE 124

VLWHV LQ WKH $++& VHTXHQFH )LJXUH f +RZHYHU WKH SUHVHQFH RI WKH n VWHPORRS PRWLI VXJJHVWV WKDW WKH ++& JHQH PD\ XWLOL]H WKH 8 PHGLDWHG KLVWRQH SURFHVVLQJ SDWKZD\ VHH FKDSWHU f LQ DGGLWLRQ WR WKH SRO\ $f DGGLWLRQ SDWKZD\ :H VSHFXODWH WKDW WKH ++& +% KLVWRQH JHQH HQFRGHV +% SURWHLQV ZKLFK DUH XVHG LQ WKUHH VLWXDWLRQV Lf GXULQJ '1$ V\QWKHVLV ZKHQ GHSRVLWLRQ RI QHZ KLVWRQH SURWHLQV LV UHTXLUHG IRU SDFNDJLQJ '1$ LQWR FKURPDWLQ LLf IRU UHSODFHPHQW RI ROG KLVWRQHV RQ DFWLYH JHQHV RU RI GDPDJHG KLVWRQHV LQ WKH JHQRPH LLLf GXULQJ GLIIHUHQWLDWLRQ ZKHQ WKH FHOOnV FKURPDWLQ LV EHLQJ UHRUJDQL]HG RU FRQGHQVHG DQG ZKHQ GHSRVLWLRQ RI QHZ KLVWRQH SURWHLQV PD\ EH UHTXLUHG :H VXJJHVW WKDW WKH SULPDU\ UROH IRU YDULDQW KLVWRQH JHQHV VXFK DV ++& LV WR SURYLGH KLVWRQHV IRU UHSODFHPHQW DQG UHRUJDQL]DWLRQ RU FRQGHQVDWLRQ ZKHQ WKH EXON RI UHSOLFDWLRQ GHSHQGHQW KLVWRQH P51$V KDYH EHHQ GHJUDGHG DQG WKH PDMRULW\ RI KLVWRQH SURWHLQ V\QWKHVLV KDV EHHQ VKXW GRZQ $OWKRXJK WKH ++& P51$ KDV YDULRXV VWUXFWXUDO IHDWXUHV WKDW PDNH LW XQLTXH DPRQJ KXPDQ +% KLVWRQH P51$V VXFK DV D ODUJH n IODQNLQJ UHJLRQ DQG D SRO\DGHQ\ODWLRQ VHTXHQFH LW DOVR VKDUHV HOHPHQWV ZLWK UHSOLFDWLRQGHSHQGHQW KLVWRQH P51$V VXFK DV WKH n VWHPORRS PRWLI 7KHVH HOHPHQWV PD\ SURYLGH WKH PHDQV IRU DOWHUQDWLYH P51$ SURFHVVLQJ DV WKH PHWDEROLF UHTXLUHPHQWV RI WKH FHOO DUH DOWHUHG +RZHYHU LW LV DOVR SRVVLEOH WKDW WKH GXDO SURFHVVLQJ SDWKZD\V DUH FRQVWLWXWLYH

PAGE 125

DQG WKDW WKH [Q51$V DUH GLIIHUHQWLDOO\ RU VHOHFWLYHO\ GHJUDGHG

PAGE 126

&+$37(5 $/7(51$7,9( n (1' 352&(66,1* ,6 ,192/9(' ,1 02'8/$7,1* (;35(66,21 2) $ +80$1 +,6721( +% 9$5,$17 *(1( /2&$7(' 21 &+5202620( ,1 5(63216( 72 &+$1*(6 ,1 7+( 352/,)(5$7,9( 67$7( 2) 7+( &(// ,QWURGXFWLRQ 7KH H[SUHVVLRQ RI UHSOLFDWLRQGHSHQGHQW KLVWRQH JHQHV LV FRXSOHG ZLWK '1$ V\QWKHVLV DQG WKH DEXQGDQFH RI WKHVH P51$V LV UHJXODWHG DW ERWK WKH WUDQVFULSWLRQDO DQG SRVW WUDQVFULSWLRQDO OHYHOV DQG UHYLHZHG LQ f 7UDQVFULSWLRQDO UHJXODWLRQ LQYROYHV D WR IROG LQFUHDVH LQ WKH UDWH RI WUDQVFULSWLRQ DW WKH *O6 SKDVH ERXQGDU\ ZLWK D UHWXUQ WR EDVDO OHYHO E\ PLG WR ODWH 6 SKDVH DQG UHYLHZHG LQ f 3RVW WUDQVFULSWLRQDO UHJXODWLRQ RI WKHVH KLVWRQH JHQHV LQYROYHV D UDSLG DQG VHOHFWLYH GHVWDELOL]DWLRQ RI KLVWRQH P51$V WRZDUG WKH HQG RI 6 SKDVH RU LQ UHVSRQVH WR LQKLELWLRQ RI '1$ V\QWKHVLV f ,Q FRQWUDVW WKH P51$ OHYHOV RI UHSOLFDWLRQLQGHSHQGHQW KLVWRQH JHQHV DUH QRW UHJXODWHG LQ WKH VDPH FHOO F\FOH GHSHQGHQW PDQQHU DV WKHLU UHSOLFDWLRQ GHSHQGHQW FRXQWHUSDUWV f $OWKRXJK VHYHUDO UHSOLFDWLRQGHSHQGHQW KXPDQ KLVWRQH +% JHQHV DQG RQH SVHXGRJHQH KDYH EHHQ GHVFULEHG f QR UHSOLFDWLRQLQGHSHQGHQW KXPDQ

PAGE 127

+% JHQHV KDYH EHHQ UHSRUWHG +RZHYHU ZH KDYH LVRODWHG DQG FKDUDFWHUL]HG D KXPDQ +% KLVWRQH F'1$ ZKLFK LV UHFLSURFDOO\ H[SUHVVHG LQ UHODWLRQ WR UHSOLFDWLRQGHSHQGHQW +% KLVWRQH JHQHV GXULQJ +/ FHOO GLIIHUHQWLDWLRQ f VHH FKDSWHU f +HUH ZH GHVFULEH WKH FORQLQJ DQG FKDUDFWHUL]DWLRQ RI WKLV YDULDQW KXPDQ +% KLVWRQH JHQH */f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f WR SUREH D $ KXPDQ JHQRPLF OLEUDU\ 2QH FRPSOHWH HTXLYDOHQW RI D $(0%/ KXPDQ JHQRPLF '1$ OLEUDU\ ; SKDJHf ZDV VFUHHQHG ZLWK D 3ODEHOHG +% n QRQWUDQVODWHG SUREH QW (FR 5,6DF IUDJPHQW LVRODWHG IURP $++& VHH )LJXUH ff 8SRQ FRPSOHWLRQ RI WKH WKLUG URXQG RI

PAGE 128

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f FRUUHVSRQGLQJ WR WKH ++& F'1$ ZDV VHOHFWHG IRU IXUWKHU DQDO\VLV 7KH DUURZ LQ WKH XSSHU ULJKW FRUQHU RI )LJXUH SRLQWV RXW FORQH $++*( GXULQJ WKH WKLUG URXQG RI WKH JHQRPLF OLEUDU\ VFUHHQLQJ 7KH QW (FR 5, IUDJPHQW FURVVK\EULGL]HG ZLWK WKH 3ODEHOHG QW (FR 5O;KR SURWHLQ FRGLQJ IUDJPHQW IURP $++& DQG WKH QW (FR 5, IUDJPHQW FURVVK\EULGL]HG ZLWK WKH 3ODEHOHG +% n QRQ WUDQVODWHG SUREH QW (FR 5,6DF IUDJPHQW LVRODWHG IURP ++& VHH )LJXUH ff 7KH QW DQG QW (FR 5, IUDJPHQWV IURP $++*( )LJXUH f ZHUH LVRODWHG DQG

PAGE 129

)LJXUH 7KLUG URXQG VFUHHQLQJ RI D $(0%/ KXPDQ DGXOW O\PSKRF\WH '1$ OLEUDU\ ZLWK D $++& QRQ WUDQVODWHG WUDLOHU SUREH 2QH FRPSOHWH HTXLYDOHQW RI D $(0%/ KXPDQ JHQRPLF '1$ OLEUDU\ ; SKDJHf ZDV JURZQ RQ PP GLDPHWHU SHWUL GLVKHV DQG VFUHHQHG ZLWK D 3ODEHOHG +% n QRQ WUDQVODWHG SUREH QW (FR 5,6DF IUDJPHQW LVRODWHG IURP $++& VHH )LJXUH ff ,OOXVWUDWHG LV D UHSUHVHQWDWLYH VHFWLRQ RI D ILOP REWDLQHG GXULQJ WKH WKLUG URXQG RI OLEUDU\ VFUHHQLQJ 7KH DUURZ LQ WKH XSSHU ULJKW FRUQHU RI WKH ILJXUH LQGLFDWHV FORQH $++*( ZKLFK ZDV RQH RI WKH FORQHV FKRVHQ IRU IXUWKHU DQDO\VLV

PAGE 131

)LJXUH ,QGLUHFW HQGODEHOHG PDSSLQJ RI WKH SXWDWLYH JHQRPLF +% FORQH ++*( ; SKDJH '1$ $++*(f ZDV SDUWLDOO\ UHVWULFWHG ZLWK ( (FR 5, % %DP +, ; ;KR 6 6DF RU $ $VS 2OLJRQXFOHRWLGHV FRPSOHPHQWDU\ WR WKH FRV VHTXHQFH RI ; SKDJH ZHUH n HQG ODEHOHG XVLQJ 7 SRO\QXFOHRWLGH NLQDVH DQG >\3@$73 DQG K\EULGL]HG WR J RI WKH SDUWLDOO\ UHVWULFWHG ; SKDJH '1$ )ROORZLQJ WKH K\EULGL]DWLRQ WKH VDPSOHV ZHUH ORDGHG RQWR D b DJDURVH 73( JHO DQG HOHFWURSKRUHVHG IRU KRXUV DW ZDWWV ZLWK EXIIHU UHFLUFXODWLRQ )ROORZLQJ HOHFWURSKRUHVLV WKH JHO ZDV GULHG XQGHU YDFXXP IRU KRXU DW r& $XWRUDGLRJUDSK\ ZDV SHUIRUPHG XVLQJ SUHIODVKHG .RGDN ;$5 ;UD\ ILOP DQG D &URQH[ /LJKWQLQJ 3OXV VFUHHQ DW r& /DQH 0O ZDV ORDGHG ZLWK nODEHOHG ; (FR 5, PDUNHU '1$ DQG ODQH 0 ZLWK nODEHOHG ; 6DO PDUNHU '1$ 7KH VL]HV RI WKH PDUNHU IUDJPHQWV DUH LQGLFDWHG RQ WKH OHIW VLGH RI WKH ILJXUH LQ .E

PAGE 133

)LJXUH 5HVWULFWLRQ PDS RI SXWDWLYH JHQRPLF +% FORQHV 5HVWULFWLRQ HQGRQXFOHDVH PDSV RI SRVLWLYH FORQHV LVRODWHG IURP WKH $(0%/ JHQRPLF '1$ OLEUDU\ VFUHHQLQJ DUH LOOXVWUDWHG 7KH (FR 5, UHVWULFWLRQ HQGRQXFOHDVH VLWHV (f DQG WKH VL]H RI HDFK (FR 5, IUDJPHQW DUH VKRZQ DERYH HDFK PDS 7KH +% SURWHLQ FRGLQJ UHJLRQ RI FORQH $++*( ZKLFK ZDV FKRVHQ IRU VHTXHQFH DQDO\VLV LV LOOXVWUDWHG E\ D FURVVKDWFKHG ER[ 7KH $(0%/ DUPV DUH QRW UHSUHVHQWHG LQ WKHVH PDSV

PAGE 134

+% 3527(,1 &2',1* .E ( ( ++*( O OXPLLLLP / ( ( ( ( ( ++*( 8 ( ( ( ( ( LFRQ TL RRRL ( ( ++*) / ( ( __ ( ( ( ( ( ++*( _ ( ( / (

PAGE 135

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f 7KH QW 3VW IUDJPHQW LOOXVWUDWHG LQ )LJXUH DQG FRQWDLQLQJ WKH HQWLUH +% JHQH IURP $++*( ZDV FORQHG LQWR S8& DQG LV WHUPHG S*/ 7KH QXFOHRWLGH DQG GHGXFHG DPLQR DFLG VHTXHQFHV RI WKH KXPDQ KLVWRQH +%*/ DQG D SRUWLRQ RI WKH DGMDFHQW +$ KLVWRQH */ DUH SUHVHQWHG LQ )LJXUH 7KH $++*( +% JHQH HQFRGHV D SURWHLQ LGHQWLFDO WR WKH VRPDWLF +% PRXVH SURWHLQ LQ NH\ DPLQR DFLG SRVLWLRQV 6HU $OD $VS /\V *O\ $VS *O\ *OXf DV UHYLHZHG E\ =ZHLGOHU f $ UHVWULFWLRQ PDS RI WKH JHQRPLF FORQH $++*( GHPRQVWUDWLQJ WKH ORFDWLRQ RI WKH +% SURWHLQ FRGLQJ UHJLRQ LV VKRZQ LQ )LJXUH 7KH $++*( +$+% JHQH SDLU VKDUH DQ LQWHUJHQLF UHJLRQ ZLWK D VSDFLQJ RI QW EHWZHHQ WKHLU SURWHLQ FRGLQJ UHJLRQV DQG QW EHWZHHQ WKHLU FDS VLWHV )LJXUH f 7KH +$ DQG

PAGE 136

)LJXUH 6RXWKHUQ EORW DQDO\VLV RI WKH JHQRPLF +% VXEFORQH 3*/5 (DFK ODQH RI D b DJDURVH JHO ZDV ORDGHG ZLWK QJ RI S*/5 '1$ UHVWULFWHG ZLWK ( (FR 5, (6 (FR 5,6DF (3 (FR 5,3VW ($ (FR 5,$VS $ $VS $6 $VS 6DF 3 3VW 6 6DF /DQHV 0 ZHUH ORDGHG ZLWK PDUNHU '1$V ZKLFK ZHUH XVHG WR VL]H WKH HWKLGLXP EURPLGH VWDLQHG '1$ IUDJPHQWV GDWD QRW VKRZQf )ROORZLQJ VHSDUDWLRQ RI WKH '1$ IUDJPHQWV HOHFWURSKRUHWLFDOO\ WKH '1$ ZDV WUDQVIHUUHG WR =HWDSUREH Q\ORQ PHPEUDQH DQG K\EULGL]HG WR D 3ODEHOHG SUREH QW (FR 5,6DF IUDJPHQW LVRODWHG IURP $++& VHH )LJXUH ff

PAGE 137

S*/ 5 ( (6 (3 ($ 0 0 $ $6 3 6 } } } }

PAGE 138

)LJXUH 6HTXHQFLQJ VWUDWHJ\ IRU WKH KXPDQ KLVWRQH JHQH +%*/ /LQHV KDYLQJ ILOOHG DUURZKHDGV LQGLFDWH UHJLRQV RI WKH KXPDQ +% KLVWRQH JHQRPLF FORQH $++*( ZKLFK ZHUH VHTXHQFHG E\ 6DQJHU GLGHR[\ VHTXHQFH DQDO\VLV f DQG OLQHV FRQWDLQLQJ KROORZ DUURZKHDGV LQGLFDWH UHJLRQV VHTXHQFHG XVLQJ WKH (;20(7+ '1$ VHTXHQFLQJ NLW SXUFKDVHG IURP 6WUDWDJHQH 7KH GRWWHG OLQH ZLWK WZR ILOOHG DUURZKHDGV UHSUHVHQW WKH UHJLRQ RI ++& ZKLFK KDG SUHYLRXVO\ EHHQ VHTXHQFHG LQ ERWK GLUHFWLRQV E\ 6DQJHU GLGHR[\ VHTXHQFH DQDO\VLV f VHH )LJXUH f 7KH +% SURWHLQ FRGLQJ UHJLRQ LV LOOXVWUDWHG E\ D KDWFKHG ER[ 5HVWULFWLRQ HQ]\PH DEEUHYLDWLRQV DUH DV IROORZV ( (FR 5, ; ;KR 6 6VW 3 3VW 6n 6RK + +LQG ,,, $(0%/ DUPV DUH QRW UHSUHVHQWHG LQ WKLV LOOXVWUDWLRQ

PAGE 139

3527(,1 &2',1*r1 f VWHP ORRS PRWLI $ $ $70 .E 6

PAGE 140

)LJXUH 1XFOHRWLGH DQG GHGXFHG DPLQR DFLG VHTXHQFHV RI WKH KXPDQ +% DQG +$ KLVWRQH JHQHV IURP WKH JHQRPLF FORQH $++*( 1XFOHRWLGHV DUH QXPEHUHG FRQWLQXRXVO\ IURP *OX RI WKH +$ JHQH WKURXJK WR WKH n WUDLOHU UHJLRQ RI WKH +% JHQH 7KH ORFDWLRQV RI WKH +$ DQG +% FDS VLWHV DUH PDUNHG E\ ILOOHG WULDQJOHV DW QW DQG QW UHVSHFWLYHO\ 7KH 7$7$ER[HV DQG &&$$7ER[HV IRU WKH +$ DQG +% JHQHV DV ZHOO DV DQ 27) VLWH DUH GLVSOD\HG LQ XQGHUOLQHG EROG IRQW 7KH n XQWUDQVODWHG UHJLRQ RI WKH +% JHQH FRQWDLQV WKH FRQVHUYHG KLVWRQH n VWHPORRS PRWLILQGLFDWHG E\ DUURZV DQG GLVSOD\HG LQ XQGHUOLQHG EROG IRQW DV ZHOO DV D SRO\$f DGGLWLRQ VHTXHQFH ORFDWHG QW GRZQVWUHDP IURP WKH SURWHLQ VWRS FRGRQ DOVR GLVSOD\HG LQ XQGHUOLQHG EROG IRQW 7KH n WHUPLQXV RI WKH QW FHOO F\FOH GHSHQGHQW +% P51$ LV LQGLFDWHG E\ DQ HPSW\ WULDQJOH DW QW DQG WKH n WHUPLQXV UHJLRQ RI WKH QW FHOO F\FOH LQGHSHQGHQW +% P51$ LV LQGLFDWHG E\ DUURZV DW QW QW &RQVHQVXV VSOLFH VLWH VHTXHQFHV DW QW DQG QW DUH LOOXVWUDWHG E\ XQGHUOLQHG EROG IRQW

PAGE 141

&7& *$* *$& &*& &*& &$7 *7$ *$& *** &*& *&& **& &&& &$& &&* &7& &*& *7$ *77 *&& *OX /HX 9DO $OD $OD 0HW 7\U 9DO 3UR $OD *O\ $OD *O\ 9DO $UJ *OX $OD 7\U $VQ *O\ 777 *&* &$* &$$ *&* *7* &$& 7&* &&& 7$& &** *$$ &7* *$* *&& $*& *&* **$ &*$ *&* /\V $UJ /HX /HX $UJ 0LV 9DO $UJ *O\ 9DO 3UR 3KH *LQ /HX *O\ $OD $UJ 6HU 6HU $UJ &*$ &77 **& &77 **& *&* **& &77 *&& 7&& 77* 777 *&& $&* $&& $*$ &$7 *$&7*$$$7 6HU /\V $OD /\V $OD $UJ $OD /\V *O\ *O\ *LQ /\V *O\ $UJ *O\ 6HU 0HW f§7$7$ER[ &$$**77$$$ &$&$*$&$$& *&7&$*$$&& 7&*&$$&$$$ $7$$*$77*$ $$*$*$$&&$ $*$**$$$*& &777$7$$*& f§ &&$$7ER[ &&$$7ER[ f§ 777&&7$*** *7****7&$$ *$$&*$*777 77&$77**7& &7$$7$7**& 77&$*$$&&* *&&$$7&$$* 7&$$*$*7&* 27) *7*77*77$& *7$&*&$77* &7*$$$$&*& $$**7&&*&$ &$&&*$$7** $$$7*$$&*$ &777&**$*& &&7$$777*& 7$7$ ER[f§ 0$$**7**7 WDWDDDDJDD 7&$**&&*&& $77&7&77$& 77&7777&77 **&7$$*&&* &*777*7$&7 *7*7&77$&& $7* &&7 *$$ &&* *&$ $$$ 7&& *&7 &&* *&* &&7 $$$ $$* **& 7&& $$* $$$ *&& *7& $&& 0HW 3UR *OX 3UR $OD /\V 6HU $OD 3UR $OD 3UR /\V /\V *O\ 6HU /\V /\V $OD 9DO 7KU $$$ *&& &$* $$* $$$ *$& **& $$* $$* &*& $$* &*& $*& &*& $$$ *$* $*& 7$& 7&& $7& /\V $OD *LQ /\V /\V $VS *O\ /\V /\V $UJ /\V $UJ 6HU $UJ /\V *OX 6HU 7\U 6HU OH 7$& *7* 7$& $$* *7* &7* $$* &$* *7& &$& &&& *$& $&& **& $7& 7&* 7&& $$* *&& $7* 7\U 9DO 7\U /\V 9DO /HX /\V *LQ 9DO +LV 3UR $VS 7KU *O\ OH 6HU 6HU /\V $OD 0HW **& $7& $7* $$& 7&& 77& *7& $$& *$& $7& 77& *$* &*& $7& *&* **$ *$* *&7 7&& &*& *O\ OH 0HW $VQ 6HU 3KH 9DO $VQ $VS OH 3KH *OX $UJ OH $OD *O\ *OX $OD 6HU $UJ &7* *&* &$& 7$& $$& $$* &*& 7&& $&& $7& $&$ 7&& &*& *$* $7& &$* $&* *&& *7* &*& /HX $OD +LV 7\U $VQ /\V $UJ 6HU 7KU OLH 7KU 6HU $UJ *OX OLH *LQ 7KU $OD 9DO $UJ &7* &7* &7* &&& **& *$* &7* *&& $$* &$& *&& *7* 7&& *$* **& $&& $$* *&* *7& $&& /HX /HX /HX 3UR *O\ *OX /HX $OD /\V +LV $OD 9DO 6HU *OX *O\ 7KU /\V $OD 9DO 7KU $$* 7$& $&& $*& 7&& $$* 7*$ *7&&&7*&& ***&$&7**& *&7&*&7&*& &*$*7&*&& **&7*&,,*$ /\V 7\U 7KU 6HU 6HU /\V 6WRS &7&&$$$**& 7&7777&$*$ *&&$&&&$&& 7$$7&$&7$* $$$$*$*&77 *77&$&77$7 7&&&77$*77 7&7777&$7$ $$*7$$*77$ 7777$*7*7* $$**7&$7** *$$$7**&$7 $&*7$*&777 77$$&7$777 **$$&7&*$* *7&&&&$*7* &*7&$77**$ 777*&7777* $$7&7$*$*& *7*7&777$& 7&$77*7*&7 *&77$*&&77 &&&$**$*7& **77&7&$$7 7$**&7*77* **$$7&&*&& 7&777$&&&* &&&&&$&7&& &*&&&&$&$& *&*&&&7**7 **&7&&77** *777*777&$ 77&7$$$$&* $$*7**&7*$ *77&**&7*7 &$777$$*$* $$&7&&$**$ &$&$$77&$* &&&***77&& *&$$$&$&7*

PAGE 142

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77&&77& 7&&,,,,&77 338 $&7&7&&&&7 &$7**7*77& &&&7&77$$$ **$**$**$* &7777$$777 $&$&77$&&$ &&7&$77,*& 7777&,**$* *&&$7*&$$7 $7$**&***$ &7$&$*$*77 $$7&7&&777 77$&$$$7*$ **&&$$*$*$ $*&&7&$77* *77&$&$*7& $7*&$*&7&$ 7$&7*7&&$& &&77*7$77& 7&$*$7*&$* *$&$$77*&$ 777-$*7777 $7777*7**$ **7 *&$*$$7 $777$&7&77 7&7*7&&$$& &&77*$77&7 *&&*$**$$* $&$&7*$7** 777*$7*$*7 *$77&$*&-* 77777**&7$ $***&7777* *$*&7*$7** &$****777* $7*$$7&&$$ $7*$*&7&7 $ *$&$77$7&$ &$*$&7*$$7 L $*$7&77$$& 7*7&7&&7$& $7*7*7*777 7 &$$$7*7*7 $7$*$7*&7$ 77*,,$77$$ 7$$$*77$&& $$,,$$,77$ O $$**&77*& ,**$77&777 ,$7&7$&$*7 *7$7777&7& $$7*$$7*$$ $77$$&$*$7 $&$$&77&&$ &-*7*7&7*7 $*$$$$*&*7 ***7*&&7&7 *$7&7***** $7*$&-$777 $7 $$*&&$&& $*&&$$$&7* &7 *&&&&$$$ **7*&$&7** **&,*$$$7$ *7&7*7-$&$ $**&&$&7*7 7$$7*&77$* 7$$77*$$$$ 7$7$7*$7&$ &$$7$*$&$* &$*7$&$$7$ *$7**$*$$, $$*7, &$$, $$*$7&*$$ &$&&$&,$*$ *&$& )LJXUH FRQWLQXHGf

PAGE 143

+% JHQHV HDFK FRQWDLQ FRQVHQVXV 7$7$ f DQG &&$$7 f ER[HV ZLWK WKH +% SURPRWHU DOVR LQFOXGLQJ DQ 27) HOHPHQW f )LJXUH f 7KH +%*/ JHQH LQFOXGHV D n QRQWUDQVODWHG UHJLRQ FRQWDLQLQJ WKH FRQVHUYHG KLVWRQH n VWHPORRS PRWLI DV ZHOO DV D SRO\$f DGGLWLRQ VHJXHQFH ORFDWHG QW GRZQVWUHDP IURP WKH SURWHLQ VWRS FRGRQ DQG FRQVHQVXV VSOLFH VLWH VHTXHQFHV DW QW DQG QW )LJXUH DQG f 7KXV WKLV +% KLVWRQH JHQH FRQWDLQV VWUXFWXUDO HOHPHQWV FRPPRQO\ IRXQG LQ FHOO F\FOH GHSHQGHQW KLVWRQH JHQHV DV ZHOO DV WKRVH UHSRUWHG IRU FRQVWLWXWLYHO\ H[SUHVVHG KLVWRQH JHQHV 7KH +%*/ JHQH LV FRPSOHWHO\ LGHQWLFDO WR WKH $++& F'1$ f DQG DSSDUHQWO\ FRQWDLQV QR LQWHUYHQLQJ VHTXHQFHV *HQRPLF 2UJDQL]DWLRQ RI $++*( 7R GHWHUPLQH LI DSDUW IURP WKH +$+% SDLU DGGLWLRQDO KLVWRQH JHQHV DUH ORFDWHG ZLWKLQ WKH $++*( JHQRPLF VHJPHQW ZH FDUULHG RXW 6RXWKHUQ EORW DQDO\VLV RI (FR 5, GLJHVWHG $++*( '1$ )LJXUH f XVLQJ 3ODEHOHG SUREHV IRU KLVWRQHV +, +$ +% + DQG + 6HH 0DWHULDOV DQG 0HWKRGV VHFWLRQ RQ 6RXWKHUQ EORW DQDO\VLV IRU D GHWDLOHG GHVFULSWLRQ RI WKHVH KLVWRQH SUREHVf 7KH UHVXOWV FRQILUPHG WKH SUHVHQFH RI DQ +$ DQG DQ +% KLVWRQH JHQH ZLWKLQ WKH QW (FR 5, IUDJPHQW RI $++*( EXW QR DGGLWLRQDO KLVWRQH JHQHV ZHUH GHWHFWHG )LJXUH f +RZHYHU WKLV GRHV QRW H[FOXGH WKH SRVVLELOLW\ WKDW DGGLWLRQDO KLVWRQH JHQHV PD\ EH

PAGE 144

)LJXUH 5HVWULFWLRQ GDWD IRU WKH JHQRPLF FORQH ++*( 7KH ORFDWLRQV RI WKH +$ DQG +% SURWHLQ FRGLQJ UHJLRQV ZLWKLQ WKH ++*( LQVHUW DUH LOOXVWUDWHG E\ KDWFKHG ER[HV ORFDWHG ZLWKLQ WKH QW (FR 5, IUDJPHQW 7KH QW 3VW IUDJPHQW FRQWDLQLQJ WKH HQWLUH +% P51$ FRGLQJ UHJLRQ DQG WKH FRQVHUYHG KLVWRQH n VWHPORRS PRWLI LV H[SDQGHG EHORZ WKH $++*( LQVHUW 3UREH $ XVHG IRU 6, QXFOHDVH SURWHFWLRQ DQDO\VLV LV UHSUHVHQWHG EHORZ WKH 3VW IUDJPHQW 5HVWULFWLRQ HQGRQXFOHDVH VLWHV DUH DV IROORZV $ $VS % %DP +, ( (FR 5, + +LQG ,,, 1 1HR 3 3VW 6 6DF 6n 6SK ; ;KR ,

PAGE 145

.E ( 2QW ( 22QW 36f ; M+A& V 363 $ 22QW ( QW ( % L $ +$ +% e W SnVf [ [KQ[N L V 6 3UREH $ ‘ .E f§ 3527(,1 &2',1* n 3 f67(0 /223 027,) 36f ; ; + 1 ;; A n n + .\L .E 3 6 3UREH %

PAGE 146

)LJXUH 6RXWKHUQ EORW DQDO\VLV RI WKH JHQRPLF FORQH $++*( ZLWK SUREHV IRU KLVWRQHV +, +$ +% + DQG + )LYH ODQHV RI D b DJDURVH JHO ZHUH ORDGHG ZLWK /WJ (FR 5, UHVWULFWHG $++*( '1$ HDFK DQG WKH IUDJPHQWV VHSDUDWHG HOHFWURSKRUHWLFDOO\ 7KH '1$ ZDV WUDQVIHUUHG WR =HWDSUREH Q\ORQ PHPEUDQH DQG HDFK ODQH LQGLYLGXDOO\ K\EULGL]HG WR HLWKHU D KLVWRQH +$ +% +, + RU + 3ODEHOHG '1$ SUREH DV LQGLFDWHG DW WKH WRS RI WKH ILJXUH VHH 0DWHULDOV DQG 0HWKRGV VHFWLRQ RQ 6RXWKHUQ EORW DQDO\VLV IRU D GHWDLOHG GHVFULSWLRQ RI WKH SUREHVf 7KH ILOOHG WULDQJOH LQGLFDWHV WKH SRVLWLRQ RI WKH QW $++*( IUDJPHQW FRQWDLQLQJ WKH +$ DQG +% JHQHV

PAGE 148

ZLWKLQ SUR[LPLW\ EXW RXWVLGH WKH ERXQGDULHV RI WKH $++*( JHQRPLF VHJPHQW 7R HYDOXDWH LI WKH +%*/ JHQH LV SUHVHQW DV D VLQJOH FRS\ ZLWKLQ WKH JHQRPH ZH FDUULHG RXW 6RXWKHUQ EORW DQDO\VLV RI +H/D JHQRPLF '1$ UHVWULFWHG ZLWK HLWKHU (FR 5, +LQG ,,, RU 3VW )LJXUH f XVLQJ D 3ODEHOHG SUREH QW (FR 5,6DF IUDJPHQW LVRODWHG IURP $++& )LJXUH f WKDW VSDQV WKH GLVWDO KDOI RI WKH +%*/ n QRQ WUDQVODWHG WUDLOHU )LJXUH QW QW f $V FDQ EH VHHQ LQ )LJXUH WKH n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

PAGE 149

)LJXUH 6RXWKHUQ EORW DQDO\VLV RI +H/D JHQRPLF '1$ ZLWK D +%*/ SUREH (DFK ODQH RI D b DJDURVH JHO ZDV ORDGHG ZLWK QT RI +H/D JHQRPLF '1$ UHVWULFWHG ZLWK ( (FR 5, + +LQG ,,, 3 3VW /DQH 0 ZDV ORDGHG ZLWK ODEHOHG ; (FR 5,+LQG ,,, PDUNHU '1$ )ROORZLQJ VHSDUDWLRQ RI WKH '1$ IUDJPHQWV HOHFWURSKRUHWLFDOO\ WKH '1$ ZDV WUDQVIHUUHG WR =HWDSUREH Q\ORQ PHPEUDQH DQG K\EULGL]HG WR D 3ODEHOHG SUREH QW (FR 5,6DF IUDJPHQW LVRODWHG IURP $++& VHH )LJXUH ff WKDW VSDQV WKH GLVWDO KDOI RI WKH +%*/ n QRQWUDQVODWHG WUDLOHU VHH )LJXUH QW QW f 7KH VL]HV RI WKH PDUNHU IUDJPHQWV DUH LQGLFDWHG WR WKH OHIW RI WKH DXWRUDGLRJUDP LQ .E

PAGE 151

)LJXUH 6RXWKHUQ EORW DQDO\VLV RI '1$V IURP PRXVH KXPDQ VRPDWLF FHOO K\EULGV SUREHG ZLWK D 3ODEHOHG '1$ IUDJPHQW IURP WKH KXPDQ KLVWRQH JHQH +%*/ $Q b DJDURVH JHO ZDV ORDGHG ZLWK [JODQH '1$ IURP PRXVH ODQH f KXPDQ ODQH f K\EULG F ODQH f K\EULG 3% ODQH f K\EULG 6 ODQH f K\EULG FO ODQH f K\EULG FO ODQH f K\EULG ODQH f K\EULG FOO ODQH f K\EULG 3KO ODQH f K\EULG $$ ODQH f K\EULG D ODQH f K\EULG 1 ODQH f K\EULG 6 ODQH f K\EULG $% ODQH f K\EULG & ODQH f K\EULG ODQH f DQG K\EULG %' ODQH f FOHDYHG ZLWK 3VW VHSDUDWHG HOHFWURSKRUHWLFDOO\ WUDQVIHUUHG Q\ORQ PHPEUDQH DQG K\EULGL]HG WR D 3ODEHOHG SUREH QW (FR 5,6DF IUDJPHQW LVRODWHG IURP ++& VHH )LJXUH ff WKDW VSDQV WKH GLVWDO KDOI RI WKH +%*/ n QRQ WUDQVODWHG WUDLOHU )LJXUH QW QW f &KURPRVRPHV UHWDLQHG E\ WKH K\EULGV DUH GHSLFWHG LQ )LJXUH

PAGE 152

NES

PAGE 153

)LJXUH 3UHVHQFH RI WKH KXPDQ KLVWRQH JHQH +%*/ LQ D SDQHO RI URGHQW KXPDQ K\EULGV $ VKDGHG ER[ LQGLFDWHV WKDW WKH K\EULG QDPHG LQ WKH OHIW FROXPQ FRQWDLQV WKH FKURPRVRPH LQGLFDWHG LQ WKH XSSHU URZ D VKDGHG WULDQJOH LQ WKH ORZHU ULJKW FRUQHU RI D ER[ LQGLFDWHV WKH SUHVHQFH RI WKH ORQJ DUP RI WKH FKURPRVRPH RU SDUW RI WKH ORQJ DUP UHSUHVHQWHG E\ DQ HYHQ VPDOOHU IUDFWLRQ RI VKDGLQJf D VKDGHG WULDQJOH LQ WKH XSSHU OHIW FRUQHU RI D ER[ LQGLFDWHV WKH SUHVHQFH RI WKH VKRUW DUP RU SDUWLDO VKRUW DUPf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

PAGE 154

+XPDQ &KURPRVRPHV +\EULG ; < HO 6 ‘ B HO 1 */ D F FO $$ %' & 6 3KO ‘ FO $% 3% 3% U $ 8! GLII +E

PAGE 155

FKURPRVRPH VHH )LJXUH IRU WKH FKURPRVRPDO FRQWHQW RI WKH K\EULGVf 7KXV WKH SUHVHQFH RI WKH +%*/ KLVWRQH JHQH LQ K\EULG FHOOV FRUUHODWHV ZLWK WKH SUHVHQFH RI KXPDQ FKURPRVRPH DV VXPPDUL]HG LQ )LJXUH $GGLWLRQDOO\ WKH +%*/ ORFXV ZDV DEVHQW LQ D K\EULG 3% ZKLFK UHWDLQV PRVW RI WKH VKRUW DUP f RI FKURPRVRPH DQG ZDV SUHVHQW LQ D K\EULG ZKLFK UHWDLQV SRUWLRQV RI WKH ORQJ DUP RI FKURPRVRPH EXW QRW WKH VKRUW DUP GDWD QRW VKRZQ EXW VXPPDUL]HG LQ )LJXUH f 7KXV VRPDWLF FHOO K\EULG DQDO\VHV VXJJHVW ORFDOL]DWLRQ RI +%*/ RQ WKH ORQJ DUP RI FKURPRVRPH 7R FRQILUP DQG UHILQH WKH ORFDWLRQ RI WKH +%*/ JHQH RQ FKURPRVRPH LQ VLWX K\EULGL]DWLRQ ZDV FDUULHG RXW RQ PHWDSKDVH FKURPRVRPHV IURP D QRUPDO PDOH VXEMHFW ;
PAGE 156

)LJXUH 5HJLRQDO ORFDOL]DWLRQ RI WKH KXPDQ KLVWRQH JHQH +%*/ RQ KXPDQ FKURPRVRPH 7KH LQ VLWX K\EULGL]DWLRQ RI WKH +%*/ SUREH WR FKURPRVRPH LV LOOXVWUDWHG GLDJUDPPDWLFDOO\ 7KH UHVXOWV LQGLFDWH WKDW ,TOT LV WKH ORFDWLRQ RI WKLV JHQH

PAGE 157

ff ffff f f f f f ff f f ff f f fffff f f f f f fff ‘ ZP &2 ,7:&2&0 &0 &2 &0 &0 7&0&2AQR 7 &0 nU &0 &2/,n &2 &2&2&2&2 &2 &0 &0 WW W &0 &0 &0 &0 &0 &2 &2 7I f0n

PAGE 158

JHQHV 6HYHUDO KLVWRQHV LQFOXGLQJ +, + DQG + DOVR PDS WR WKH OT VXEUHJLRQ f +%*/ P51$ /HYHOV LQ 3UROLIHUDWLQJ 7+3 &HOOV :H KDYH SUHYLRXVO\ GHPRQVWUDWHG WKDW WKH +%*/ JHQH H[SUHVVHV D QW FHOO F\FOH LQGHSHQGHQW SRO\ $ P51$ FRQWDLQLQJ FRQVHQVXV VSOLFH VLWH VHTXHQFHV FKDSWHU f 7KHVH VSOLFH VLWH VHTXHQFHV SURYLGH WKH FHOO ZLWK D SRWHQWLDO PHFKDQLVP E\ ZKLFK DOWHUQDWLYH P51$V PD\ EH H[SUHVVHG IURP D VLQJOH JHQH $OWHUQDWLYHO\ WKH FRQVHUYHG KLVWRQH n VWHPORRS PRWLI SUHVHQW LQ WKH n QRQWUDQVODWHG UHJLRQ RI WKH +%*/ JHQH )LJXUH $ DQG ,%f SURYLGHV DQ DGGLWLRQDO DYHQXH IRU WKH SURGXFWLRQ RI DOWHUQDWLYH P51$V 7R GHWHUPLQH LI WKH +%*/ JHQH H[SUHVVHV WZR P51$ VSHFLHV ZH IROORZHG WKH P51$ OHYHOV RI DOO +% VSHFLHV LQ SUROLIHUDWLQJ 7+3 FHOOV E\ QRUWKHUQ EORW DQDO\VLV XVLQJ WZR GLIIHUHQW +% 3ODEHOHG SUREHV 2QH SUREH ZDV DQ +% SURWHLQ FRGLQJ IUDJPHQW IURP $++& )LJXUH ODQH $f DQG WKH VHFRQG ZDV D QW IUDJPHQW LVRODWHG IURP WKH +%*/ n QRQWUDQVODWHG OHDGHU )LJXUH ODQH %f 7KH SURWHLQ FRGLQJ SUREH FURVV K\EULGL]HV ZLWK DOO WKH +% P51$V LQ WKH FHOO DQG GHWHFWV +% P51$ VSHFLHV ZLWK VL]HV RI QW EDQG f QW QRW ZHOO UHVROYHG IURP WKH VWURQJ QW VLJQDOf DQG QW EDQG f )LJXUH ODQH $f 7KH n QRQWUDQVODWHG SUREH LV VSHFLILF IRU P51$ WUDQVFULEHG IURP WKH +%*/ JHQH DQG GHWHFWV QW EDQG f DQG

PAGE 159

)LJXUH 1RUWKHUQ EORW DQDO\VLV RI +%*/ P51$ OHYHOV LQ SUROLIHUDWLQJ 7+3 FHOOV 7RWDO FHOOXODU 7+3 51$ J ZDV VL]H IUDFWLRQDWHG SDVVLYHO\ WUDQVIHUUHG WR =HWDSUREH Q\ORQ PHPEUDQH DQG K\EULGL]HG WR WKH 3ODEHOHG QW (FR 5O;KS +% SURWHLQ FRGLQJ IUDJPHQW IURP $++& )LJXUH f DV SUREH ODQH $f $Q LGHQWLFDO QRUWKHUQ DQDO\VLV ZDV FDUULHG RXW XVLQJ WKH 3ODEHOHG QW +DH ,,,1OD ,,, IUDJPHQW )LJXUH QW QW f LVRODWHG IURP WKH +%*/ n QRQ WUDQVODWHG OHDGHU ODQH %f /DQHV $ DQG % FRQWDLQ UHVXOWV IURP DXWRUDGLRJUDSK\ DQG ODQH & FRQWDLQV WKH UHVXOWV IURP D MSDUWLFOH DQDO\VLV XVLQJ D -HWDJHQ %HWD6FRSH EORW DQDO\]HUf RI WKH ODQH % Q\ORQ PHPEUDQH ,Q DGGLWLRQ WKH SDQHO VKRZQ RQ WKH ULJKW VLGH RI WKH ILJXUH FRQWDLQV WKH QXPHULFDO UHVXOWV RI TXDQWLWDWLYH SDUWLFOH DQDO\VLV RI WKH K\EULGL]HG PHPEUDQH LQ ODQH % ZLWK UHDGLQJ EHLQJ RI WKH QW EDQG UHDGLQJ EHLQJ RI WKH QW EDQG DQG UHDGLQJV DQG EHLQJ RI EDFNJURXQG UHJLRQV RI WKH Q\ORQ PHPEUDQH

PAGE 160

$ &RXQWV

PAGE 161

QW EDQG f +% P51$ VSHFLHV )LJXUH ODQHV % DQG &f 7KH SDQHO VKRZQ RQ WKH ULJKW VLGH RI )LJXUH FRQWDLQV WKH QXPHULFDO UHVXOWV RI TXDQWLWDWLYH SDUWLFOH DQDO\VLV RI WKH QW EDQG WKH QW EDQG DQG EDFNJURXQG UHJLRQV RI WKH Q\ORQ PHPEUDQH IURP WKH QRUWKHUQ EORW VKRZQ LQ ODQH & 7KH LGHQWLW\ RI WKH QW +% P51$ VSHFLHV ZDV YHULILHG E\ K\EULGL]DWLRQ RI WKLV QRUWKHUQ EORW ZLWK D XQLTXH VHTXHQFH SUREH VSHFLILF IRU WKH n QRQ WUDQVODWHG UHJLRQ RI WKH +%*/ JHQH 7KH UHVXOWV LQ )LJXUH GHPRQVWUDWH WKDW WKH +%*/ JHQH HQFRGHV D QW +% P51$ LQ DGGLWLRQ WR WKH QW P51$ SUHYLRXVO\ GHVFULEHG FKDSWHU f ZLWK WKH QW P51$ EHLQJ PRUH DEXQGDQW WKDQ WKH QW VSHFLHV LQ SUROLIHUDWLQJ 7+3 FHOOV +%*/ P51$ /HYHOV LQ 6\QFKURQL]HG +H/D &HOOV 7R HYDOXDWH WKH FHOO F\FOH GHSHQGHQW QDWXUH RI +%*/ P51$V ZH FDUULHG RXW QRUWKHUQ EORW DQDO\VLV RQ WRWDO FHOOXODU 51$ LVRODWHG IURP V\QFKURQL]HG +H/D FHOOV DIWHU UHOHDVH IURP D GRXEOH WK\PLGLQH EORFN DQG RQ VDPSOHV HQULFKHG IRU SRO\ $ 51$ XVLQJ D VLQJOH SDVV RYHU ROLJR G7FHOOXORVH $ VLQJOH ROLJR G7FHOOXORVH VHOHFWLRQ JUHDWO\ HQULFKHV IRU SRO\ $ 51$ EXW GRHV QRW WRWDOO\ UHPRYH DOO RI WKH SRO\ $n 51$ +\EULGL]DWLRQ ZDV FDUULHG RXW XVLQJ D QW 3ODEHOHG IUDJPHQW LVRODWHG IURP WKH +%*/ n QRQWUDQVODWHG OHDGHU WKDW LV VSHFLILF IRU +%*/ P51$V

PAGE 162

7KH QW +%*/ P51$ VSHFLHV ZDV GHWHFWHG GXULQJ WKH +H/D FHOO F\FOH )LJXUH f ZLWK LWV OHYHO UHDFKLQJ D PD[LPXP GXULQJ WKH SHDN RI '1$ V\QWKHVLV KRXUVf 'HVSLWH KDYLQJ HQULFKHG IRU SRO\ $ 51$ ZH ZHUH XQDEOH WR GHWHFW WKH QW +%*/ P51$ GXH WR WKH VPDOO VL]H RI WKH K\EULGL]DWLRQ SUREH DQG WKH ORZ DEXQGDQFH RI WKH QW +%*/ P51$ VHH FKDSWHU DQG )LJXUH f 7KHVH UHVXOWV HVWDEOLVK WKDW WKH QW +%*/ P51$ VSHFLHV LV UHJXODWHG LQ D FHOO F\FOH GHSHQGHQW PDQQHU XQOLNH WKH QW VSHFLHV ZKLFK ZH KDYH SUHYLRXVO\ GHPRQVWUDWHG WR EH FRQVWLWXWLYHO\ H[SUHVVHG GXULQJ WKH +H/D FHOO F\FOH FKDSWHU f 0DSSLQJ RI WKH +%*/ P51$ 6WDUW 6LWH 7R GHWHUPLQH WKH +%*/ P51$ VWDUW VLWH 6, QXFOHDVH SURWHFWLRQ DQDO\VLV ZDV FDUULHG RXW RQ WRWDO FHOOXODU DQG SRO\ $ 51$ XVLQJ D QW +LQG ,,,1FR SUREH GHULYHG IURP WKH +%*/ JHQH )LJXUH QW QW f DQG n HQG ODEHOHG DW WKH 1HR VLWH )LJXUH f $ QW +% SURWHFWHG IUDJPHQW ZDV GHWHFWHG LQGLFDWLQJ WKH FDS VLWH IRU WKH +%*/ P51$V LV ORFDWHG QW XSVWUHDP RI WKH $7* WUDQVODWLRQDO VWDUW FRGRQ )LJXUH f 7KH +% IUDJPHQW ZDV GHWHFWHG SULPDULO\ DW WKH SHDN RI 6 SKDVH KRXUVf LQ WKH WRWDO FHOOXODU 51$ VDPSOH )LJXUH f KRZHYHU RQ ORQJHU H[SRVXUH D SURWHFWHG IUDJPHQW RI WKH VDPH VL]H ZDV REVHUYHG RXWVLGH RI 6 SKDVH DQG LQ WKH SRO\ $ 51$ VDPSOHV

PAGE 163

)LJXUH 1RUWKHUQ EORW DQDO\VLV RI +%*/ P51$ OHYHOV LQ V\QFKURQL]HG +H/D FHOOV 7RWDO FHOOXODU 51$ ZDV LVRODWHG IURP V\QFKURQL]HG FHOOV DIWHU DQG KRXUVf UHOHDVH IURP D GRXEOH WK\PLGLQH EORFN DQG HQULFKHG IRU SRO\ $ 51$ XVLQJ D VLQJOH SDVV RYHU ROLJR G7FHOOXORVH DV GHVFULEHG E\ 0DQLDWLV HW DO f $ VLQJOH ROLJR G7FHOOXORVH VHOHFWLRQ JUHDWO\ HQULFKHV IRU SRO\ $ 51$ EXW GRHV QRW WRWDOO\ UHPRYH DOO RI WKH SRO\ $n 51$ 7KH VHOHFWHG 51$ J ZDV VL]H IUDFWLRQDWHG SDVVLYHO\ WUDQVIHUUHG WR =HWDSUREH Q\ORQ PHPEUDQH DQG K\EULGL]HG WR WKH 3ODEHOHG QW +DH ,,,1OD ,,, IUDJPHQW )LJXUH QW QW f LVRODWHG IURP WKH +%*/ n QRQWUDQVODWHG OHDGHU 7KH DUURZKHDG DW WKH OHIW RI WKH DXWRUDGLRJUDP LQGLFDWHV WKH GHWHFWHG +%*/ EDQG

PAGE 165

)LJXUH 6, QXFOHDVH PDSSLQJ RI WKH +%*/ P51$ VWDUW VLWH 7RWDO FHOOXODU DQG SRO\ $ 51$V ZHUH LVRODWHG IURP V\QFKURQL]HG +H/D FHOOV SULRU WR f DQG DIWHU DQG KRXUVf UHOHDVH IURP D GRXEOH WK\PLGLQH EORFN 7RWDO FHOOXODU 51$ 7RWf [J DQG [J SRO\ $ 51$ $f ZHUH XVHG IRU 6, DQDO\VLV 6, DQDO\VLV ZDV FDUULHG RXW XVLQJ D QW +LQG ,,,1FR SUREH GHULYHG IURP WKH +%*/ JHQH )LJXUH QW QW f DQG n HQG ODEHOHG DW WKH 1HR VLWH /DQHV 0 DQG 0r ZHUH ORDGHG ZLWK nODEHOHG S%5 +LQI DQG S%5 +SD ,, PDUNHUV UHVSHFWLYHO\ 7KH VL]HV RI YDULRXV PDUNHU EDQGV DUH JLYHQ RQ WKH OHIW VLGH RI WKH ILJXUH 7KH QW +% SURWHFWHG IUDJPHQW LV LQGLFDWHG E\ DQ DUURZKHDG LQ WKH FHQWHU RI WKH DXWRUDGLRJUDP

PAGE 167

GHPRQVWUDWLQJ WKDW WKH FDS VLWHV IRU ERWK +%*/ P51$V DUH WKH VDPH ,Q DGGLWLRQ 6, QXFOHDVH SURWHFWLRQ DQDO\VLV FDUULHG RXW RQ +H/D 51$ XVLQJ D QW ;KR ,+LQG ,,, SUREH GHULYHG IURP WKH +$*/ JHQH )LJXUH QW QW f DQG n HQG ODEHOHG DW WKH ;KR VLWH SURGXFHG D QW +$ SURWHFWHG IUDJPHQW LQGLFDWLQJ WKDW WKH FDS VLWH IRU WKH +$*/ P51$V LV ORFDWHG QW XSVWUHDP RI WKH $7* WUDQVODWLRQDO VWDUW FRGRQ )LJXUH f 7KH 6, QXFOHDVH SURWHFWLRQ DQDO\VLV FOHDUO\ GLVWLQJXLVKHG EHWZHHQ FRUUHFWO\ LQLWLDWHG +$*/ WUDQVFULSWV DQG WUDQVFULSWV IURP RWKHU QRQLGHQWLFDO +$ KLVWRQH JHQHV )LJXUH f 7KH +$ SURWHFWHG IUDJPHQW ZDV GHWHFWHG SULPDULO\ DW WKH SHDN RI 6 SKDVH KRXUVf 0DSSLQJ RI WKH +%*/ P51$ 7HUPLQL 7R LQYHVWLJDWH ZKHWKHU WKH n WHUPLQXV RI WKH QW +%*/ P51$ LV W\SLFDO RI FHOO F\FOH GHSHQGHQW KLVWRQH P51$V 6, QXFOHDVH SURWHFWLRQ DQDO\VLV ZDV FDUULHG RXW RQ WRWDO FHOOXODU )LJXUH ODQH f DQG SRO\ $ )LJXUH ODQH f 51$ LVRODWHG IURP SUROLIHUDWLQJ +H/D FHOOV XVLQJ D n HQG ODEHOHG SUREH GHULYHG IURP WKH ++*( +% JHQH )LJXUH 6, SUREH $f $ QW SURWHFWHG IUDJPHQW PDSSLQJ DSSUR[LPDWHO\ WR WKH +%*/ VWRS FRGRQ DQG D QW SURWHFWHG IUDJPHQW PDSSLQJ WR WKH FRQVHQVXV KLVWRQH P51$ WHUPLQXV f ZHUH GHWHFWHG LQ WKH WRWDO

PAGE 168

)LJXUH 6, PDSSLQJ RI WKH +$*/ n P51$ VWDUW VLWH 7RWDO FHOOXODU DQG SRO\ $ 51$V ZHUH LVRODWHG IURP V\QFKURQL]HG +H/D FHOOV SULRU WR f DQG DIWHU DQG KRXUVf UHOHDVH IURP D GRXEOH WK\PLGLQH EORFN 7RWDO FHOOXODU 51$ 7RWf MXJ DQG QT SRO\ $ 51$ $f ZHUH XVHG IRU 6, DQDO\VLV 6, DQDO\VLV ZDV FDUULHG RXW XVLQJ D QW ;KR ,+LQG ,,, SUREH GHULYHG IURP WKH +$*/ JHQH )LJXUH QW QW f DQG n HQG ODEHOHG DW WKH ;KR VLWH /DQHV 0 DQG 0r ZHUH ORDGHG ZLWK nODEHOHG S%5 +LQI DQG S%5 +RD ,, PDUNHUV UHVSHFWLYHO\ 7KH VL]HV RI YDULRXV PDUNHU EDQGV DUH JLYHQ RQ WKH OHIW VLGH RI WKH ILJXUH /DQHV $ & DQG 7 ZHUH ORDGHG ZLWK WKH SURGXFWV RI GLGHR[\ VHTXHQFLQJ UHDFWLRQV RQ 0 VLQJOH VWUDQGHG WHPSODWH '1$ FORQH f XWLOL]LQJ D SULPHU n *7777&&&$*7&$&*$&nf 7KH QW +$ SURWHFWHG IUDJPHQW LV LQGLFDWHG E\ DQ DUURZKHDG DW WKH ULJKW RI WKH DXWRUDGLRJUDP

PAGE 170

)LJXUH 6, QXFOHDVH SURWHFWLRQ DQDO\VLV RI WKH WHUPLQL RI WKH +%*/ P51$ 7RWDO FHOOXODU ODQH f DQG SRO\ $ ODQH f 51$V ZHUH LVRODWHG IURP SUROLIHUDWLQJ +H/D FHOOV 7RWDO FHOOXODU 51$ QT DQG cLT SRO\ $ 51$ ZHUH XVHG IRU 6, DQDO\VLV 6, DQDO\VLV ZDV FDUULHG RXW XVLQJ D QW 1HR ,(FR 5, SUREH GHULYHG IURP WKH +%*/ JHQH )LJXUH QW QW )LJXUH 6, SUREH $f DQG n HQG ODEHOHG /DQH 0 ZDV ORDGHG ZLWK nODEHOHG S%5 +SD ,, PDUNHUV 7KH VL]HV RI YDULRXV PDUNHU EDQGV DUH JLYHQ RQ WKH OHIW VLGH RI ODQH 0 /DQHV $ & DQG 7 ZHUH ORDGHG ZLWK WKH SURGXFWV RI GLGHR[\ VHTXHQFLQJ UHDFWLRQV RQ 0PSO VLQJOH VWUDQGHG WHPSODWH '1$ XWLOL]LQJ D SULPHU *7777&&&$*7&$&*$& nf 7KH QW +%*/ SURWHFWHG IUDJPHQW LV LQGLFDWHG E\ D ILOOHG WULDQJOH DQG WKH QW +% VWRS FRGRQ SURWHFWHG IUDJPHQW LV LQGLFDWHG E\ DQ HPSW\ WULDQJOH 7KH DUURZKHDG DW WKH WRS RI WKH DXWRUDGLRJUDP LQGLFDWHV SURWHFWLRQ WR WKH HQG RI 6, SUREH $ f

PAGE 172

FHOOXODU 51$ VDPSOH )LJXUH ODQH f ,Q WKH SRO\ $51$ VDPSOH ZH REVHUYHG D SURWHFWHG IUDJPHQW FRUUHVSRQGLQJ WR WKH QW SUREH LQGLFDWLQJ SURWHFWLRQ WR WKH HQG RI 6, SUREH $ )LJXUH ODQH f ZKLFK ZRXOG EH FRQVLVWHQW ZLWK WKH SUHVHQFH RI D QW SRO\ $ P51$ SURGXFHG IURP WKH +%*/ JHQH VHH )LJXUH DQG FKDSWHU f 7DNHQ WRJHWKHU WKHVH UHVXOWV YHULI\ WKH SUHVHQFH RI WZR +%*/ P51$ VSHFLHV DQG GHPRQVWUDWH WKDW WKH n WHUPLQXV RI WKH QW +%*/ P51$ PDSV WR WKH FRQVHQVXV KLVWRQH P51$ WHUPLQXV ZKLFK LV FKDUDFWHULVWLF RI FHOO F\FOH GHSHQGHQW KLVWRQH P51$V 'LVFXVVLRQ 6WUXFWXUH DQG 2UJDQL]DWLRQ RI WKH +%*/ *HQH :H KDYH LVRODWHG DQG VHTXHQFHG D YDULDQW KXPDQ +% KLVWRQH JHQH */f ZKLFK H[SUHVVHV DOWHUQDWLYH P51$V UHJXODWHG GLIIHUHQWLDOO\ GXULQJ WKH +H/D FHOO F\FOH 2XU SUHYLRXV VWXGLHV XWLOL]LQJ WKH +%*/ F'1$ $++&f GHPRQVWUDWHG WKDW WKH QW +%*/ P51$ VSHFLHV LV UHFLSURFDOO\ H[SUHVVHG LQ UHODWLRQ WR UHSOLFDWLRQGHSHQGHQW +% KLVWRQH JHQHV GXULQJ +/ FHOO GLIIHUHQWLDWLRQ FKDSWHU f 7KH QXFOHRWLGH DQG GHGXFHG DPLQR DFLG VHTXHQFHV UHYHDO WKDW WKH KXPDQ +%*/ JHQH HQFRGHV D SURWHLQ LGHQWLFDO WR WKH VRPDWLF +% PRXVH SURWHLQ LQ NH\ DPLQR DFLG SRVLWLRQV f 7KH +% KLVWRQH YDULDQW LV SUHVHQW LQ VPDOO DPRXQWV LQ UDSLGO\ JURZLQJ WLVVXHV f

PAGE 173

+RZHYHU LQ WKRVH WLVVXHV ZLWK ORZ JURZWK UDWHV LQ WKH DGXOW +% LQFUHDVHV GUDPDWLFDOO\ LQ UHODWLYH DPRXQW GXULQJ DQG HVSHFLDOO\ MXVW DIWHU WKH SRVWQDWDO JURZWK SKDVH WR UHDFK D SODWHDX DW DERXW PRQWKV f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f 3UHYLRXVO\ QXFOHDU UXQRQ WUDQVFULSWLRQ DQDO\VLV UHYHDOHG D WZR IROG LQFUHDVH LQ WUDQVFULSWLRQ RI WKH +%*/ JHQH GXULQJ 6 SKDVH FKDSWHU f 6HTXHQFH DQDO\VLV VKRZV WKDW WKH +%*/ JHQH FRQWDLQV DSDUW IURP D FRQVHQVXV 7$7$ER[ D KLVWRQH SURPRWHU VSHFLILF &&$$7 ER[ f DV ZHOO DV DQ 27) HOHPHQW f ZKLFK DUH DOO SURPRWHU HOHPHQWV W\SLFDO RI FHOO F\FOH GHSHQGHQW +% KLVWRQH JHQHV 7KH FHOO F\FOH WUDQVFULSWLRQ SDWWHUQ RI WKH +%*/ JHQH DQG WKH DSSDUHQW VLPLODULWLHV RI LWV SURPRWHU HOHPHQWV WR WKRVH RI FHOO F\FOH GHSHQGHQW +% JHQHV VXJJHVW WKDW WUDQVFULSWLRQDO UHJXODWLRQ LV QRW WKH VROH PRGXODWRU LQ FRQWUROOLQJ +% YDULDQW UDWLRV

PAGE 174

3RVW7UDQVFULSWLRQDO 5HJXODWLRQ RI WKH +%*/ *HQH :H KDYH REVHUYHG WKDW WKH +%*/ JHQH H[SUHVVHV D QW FHOO F\FOH GHSHQGHQW P51$ LQ DGGLWLRQ WR WKH QW FHOO F\FOH LQGHSHQGHQW P51$ SUHYLRXVO\ GHVFULEHG FKDSWHU f 7KH n HQG RI WKH FHOO F\FOH GHSHQGHQW P51$ WHUPLQDWHV LPPHGLDWHO\ IROORZLQJ WKH UHJLRQ RI K\SKHQDWHG G\DG V\PPHWU\ ZKHUHDV WKH FHOO F\FOH LQGHSHQGHQW P51$ KDV D QW QRQWUDQVODWHG WUDLOHU WKDW FRQWDLQV WKH UHJLRQ RI K\SKHQDWHG G\DG V\PPHWU\ DQG D SRO\ $f DGGLWLRQ VHTXHQFH IROORZHG E\ D SRO\ $f WDLO 7KH DELOLW\ RI WKH FHOO WR XWLOL]H WZR PHFKDQLVPV IRU P51$ n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f +RZHYHU WKH OHYHO RI WKH QW +%*/ VSHFLHV LQFUHDVHV DSSUR[LPDWHO\ IROG GXULQJ +/ FHOO GLIIHUHQWLDWLRQ ZKHUHDV WKH OHYHOV RI FHOO F\FOH GHSHQGHQW +% P51$V GHFUHDVH WR OHVV WKDQ b RI WKRVH LQ

PAGE 175

SUROLIHUDWLQJ FHOOV FKDSWHU f 7KLV UHVXOW ZRXOG VXJJHVW WKDW DW WKH RQVHW RI GLIIHUHQWLDWLRQ WKH 8 PHGLDWHG f KLVWRQH PDWXUDWLRQ SURFHVV LV VORZHG RU VKXW GRZQ DOORZLQJ DGGLWLRQDO WLPH IRU SRO\DGHQ\ODWLRQ RI WKH KLVWRQH SUHP51$ DQG UHVXOWLQJ LQ DQ LQFUHDVH RI WKH QW +%*/ P51$ OHYHO ,Q VXSSRUW RI WKLV VXJJHVWLRQ +RIIPDQQ DQG %LUQVWLHO KDYH REVHUYHG WKDW n VHTXHQFHV RI WKH 8 VQ513 ZKLFK K\EULGL]H ZLWK WKH KLVWRQH GRZQVWUHDP VSDFHU PRWLI GXULQJ n SURFHVVLQJ f DUH RFFOXGHG LQ WKH *2 VWDJH RI WKH FHOO F\FOH f DQG 9DVVHURW HW DO KDYH REVHUYHG WKDW GXULQJ WKH WUDQVLWLRQ IURP D SUROLIHUDWLYH WR D QRQGLYLGLQJ VWDWH ZKLOH UDW P\REODVWV GLIIHUHQWLDWH WR P\RWXEHV WKH n HQGV RI 8 VQ513V DUH SURWHFWHG DJDLQVW PLFURFRFFDO QXFOHDVH DWWDFN f DQG SUHVXPDEO\ XQDYDLODEOH IRU KLVWRQH n HQG SURFHVVLQJ ,Q DGGLWLRQ /VFKHU DQG 6FKPSHUOL KDYH VKRZQ D KLVWRQH n SURFHVVLQJ IDFWRU EHFRPHV OLPLWLQJ LQ *ODUUHVWHG FHOOV f 6WDXEHU DQG 6FKPSHUOL KDYH GHPRQVWUDWHG n SURFHVVLQJ RI SUHP51$ SOD\V D PDMRU UROH LQ SUROLIHUDWLRQGHSHQGHQW UHJXODWLRQ RI KLVWRQH P51$ OHYHOV f DQG /LX HW DO KDYH VKRZQ WKDW WKH HIILFLHQF\ RI n HQG IRUPDWLRQ FRQWULEXWHV WR WKH UHODWLYH OHYHOV RI GLIIHUHQW KLVWRQH P51$V f 7KLV PHFKDQLVP PD\ EH RSHUDWLYH IRU RWKHU KLVWRQH JHQHV ZKLFK KDYH EHHQ REVHUYHG WR SURGXFH PXOWLSOH VSHFLHV RI P51$ E\ DOWHUQDWLYH n HQG SURFHVVLQJ f

PAGE 176

$OWHUQDWLYHO\ WKH QW P51$ PD\ UHVXOW IURP SUH P51$ WUDQVFULSWV EHLQJ SRO\DGHQ\ODWHG GXH WR WKH SUR[LPLW\ RI D SRO\ $ DGGLWLRQ VHTXHQFH WR WKH VWHPORRS VWUXFWXUH EHIRUH WKH\ DUH SURFHVVHG E\ WKH KLVWRQH VSHFLILF n HQGRQXFOHRO\WLF FOHDYDJH UHDFWLRQ 6XFK P51$ VOLSSDJH ZRXOG LQFUHDVH DV D FHOO HQWHUV D QRQSUROLIHUDWLYH VWDWH ZKHUH IDFWRUV VXFK DV WKH 8 VQ513 DQG WKH KHDW ODELOH IDFWRU HVVHQWLDO IRU FHOO F\FOH GHSHQGHQW KLVWRQH P51$ n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n HQG SURFHVVLQJ RI WKH SUHP51$V 7KHVH UHVXOWV VXJJHVW WKDW n SURFHVVLQJ RI KLVWRQH +% P51$ SUHFXUVRUV PD\ SOD\ DQ LPSRUWDQW UROH LQ WKH PRGXODWLRQ RI +% SURWHLQ OHYHOV GXULQJ WKH WUDQVLWLRQ IURP D SUROLIHUDWLYH WR D QRQGLYLGLQJ VWDWH

PAGE 177

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f ZKLFK VRPHWLPHV FRQWDLQHG LQWURQV f ,Q DGGLWLRQ %RUXQ HW DO REVHUYHG WKDW D IUDFWLRQ RI KXPDQ KLVWRQH P51$V FRQWDLQHG VKRUW WUDFWV RI SRO\$f f (DUO\ LQ WKH FRXUVH RI WKLV SURMHFW :HOOV DQG .HGHV UHSRUWHG WKH FORQLQJ RI D F'1$ IRU D KXPDQ + SRO\DGHQ\ODWHG P51$ KDYLQJ LQWHUYHQLQJ VHTXHQFHV f %DVHG RQ WKHVH REVHUYDWLRQV DQG RXU K\SRWKHVLV WKDW EHFDXVH FRQVWLWXWLYHO\ H[SUHVVHG KLVWRQH SURWHLQV DUH V\QWKHVL]HG WKURXJKRXW WKH FHOO F\FOH f WKHLU P51$V PD\ EH VWDEOH LQ WKH DEVHQFH RI '1$ V\QWKHVLV ZH SURSRVHG

PAGE 178

WKDW VWUXFWXUDO IHDWXUHV RI FRQVWLWXWLYHO\ H[SUHVVHG KLVWRQH P51$V PD\ DOORZ WKHP WR HVFDSH WKH PHFKDQLVPV ZKLFK UDSLGO\ DQG VHOHFWLYHO\ GHVWDELOL]H UHSOLFDWLRQGHSHQGHQW KLVWRQH P51$V WRZDUG WKH HQG RI 6 SKDVH RU LQ UHVSRQVH WR LQKLELWLRQ RI '1$ V\QWKHVLV f $OWHUQDWLYHO\ WKH DEVHQFH RI VWUXFWXUDO HOHPHQWV FRPPRQO\ IRXQG LQ UHSOLFDWLRQ GHSHQGHQW KLVWRQH P51$V FRXOG KDYH WKH VDPH UHVXOW 7KHUHIRUH ZH IRFXVHG RXU DWWHQWLRQ RQ WKH FORQLQJ RI D YDULDQW KXPDQ KLVWRQH JHQH DQG FKDUDFWHUL]DWLRQ RI LWV VWUXFWXUDO IHDWXUHV DQG WKHLU HIIHFW RQ H[SUHVVLRQ LQ UHVSRQVH WR FKDQJHV LQ WKH JURZWK VWDWH RI WKH FHOO ,Q RUGHU WR LVRODWH D YDULDQW KXPDQ KLVWRQH JHQH ZH ILUVW FORQHG DQG FKDUDFWHUL]HG D YDULDQW KXPDQ KLVWRQH F'1$ $++& f IURP D SRO\ $ F'1$ OLEUDU\ FKDSWHU f $++& FRQWDLQV DQ +% SURWHLQ FRGLQJ UHJLRQ WKDW LV IODQNHG DW WKH r HQG E\ D QW QRQWUDQVODWHG WUDLOHU WKDW LQFOXGHV D UHJLRQ RI K\SKHQDWHG G\DG V\PPHWU\ DQG D SRO\$f DGGLWLRQ VHTXHQFH IROORZHG E\ D SRO\$f WDLO :H GHPRQVWUDWHG WKDW WKH ++& P51$ LV UHJXODWHG LQ D FRPSOH[ PDQQHU ZLWK UHVSHFW WR WKH +H/D FHOO F\FOH DQG +/ FHOO GLIIHUHQWLDWLRQ 1RUWKHUQ EORW DQDO\VLV LQGLFDWHG WKDW WKH OHYHOV RI WKH QW ++& P51$ VSHFLHV GLG QRW YDU\ VLJQLILFDQWO\ GXULQJ WKH +H/D FHOO F\FOH LQ FRPSDULVRQ WR UHSOLFDWLRQGHSHQGHQW +% P51$V ZKLFK DUH HOHYDWHG IROG KLJKHU GXULQJ 6 SKDVH WKDQ GXULQJ SKDVH 1RUWKHUQ EORW DQDO\VLV DOVR UHYHDOHG D UHFLSURFDO UHODWLRQVKLS GXULQJ WKH

PAGE 179

RQVHW RI +/ GLIIHUHQWLDWLRQ EHWZHHQ WKH H[SUHVVLRQ RI WKH ++& +% JHQH DQG WKH UHSOLFDWLRQGHSHQGHQW +% JHQHV 7KH OHYHOV RI WKH QW ++& +% VSHFLHV LQFUHDVHG DSSUR[LPDWHO\ IROG GXULQJ +/ FHOO GLIIHUHQWLDWLRQ ZKHUHDV WKH OHYHOV RI FHOO F\FOH GHSHQGHQW +% P51$V GHFUHDVHG WR OHVV WKDQ b RI WKRVH LQ SUROLIHUDWLQJ FHOOV $OWKRXJK ZH QRWLFHG WKH VWHDG\ VWDWH OHYHO RI ++& P51$ ZDV UHJXODWHG GLIIHUHQWLDOO\ WR WKDW RI FHOO F\FOH GHSHQGHQW +% KLVWRQH P51$V ZKLFK ZRXOG EH FRQVLVWHQW ZLWK WKH GLVVLPLODU n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f IURP D KXPDQ JHQRPLF '1$ OLEUDU\ :H GHPRQVWUDWHG WKDW +%*/ H[SUHVVHV DOWHUQDWLYH P51$V UHJXODWHG GLIIHUHQWLDOO\ GXULQJ WKH +H/D FHOO F\FOH ZKLFK LV FRQVLVWHQW ZLWK WKH SUHVHQFH RI n VWUXFWXUDO HOHPHQWV FRPPRQO\ IRXQG LQ FHOO F\FOH GHSHQGHQW KLVWRQH JHQHV DV ZHOO DV WKRVH HQGRJHQRXV WR FRQVWLWXWLYHO\ H[SUHVVHG KLVWRQH

PAGE 180

JHQHV 7KH n HQG RI WKH FHOO F\FOH UHJXODWHG P51$ WHUPLQDWHV LPPHGLDWHO\ IROORZLQJ WKH UHJLRQ RI K\SKHQDWHG G\DG V\PPHWU\ W\SLFDO RI PRVW KLVWRQH P51$V ZKHUHDV WKH FRQVWLWXWLYHO\ H[SUHVVHG P51$ ++&f KDV D QW QRQ WUDQVODWHG WUDLOHU WKDW FRQWDLQV WKH VDPH UHJLRQ RI K\SKHQDWHG G\DG V\PPHWU\ EXW LV SRO\DGHQ\ODWHG 7KHVH UHVXOWV GHPRQVWUDWH WKDW DOWHUQDWLYH n HQG SURFHVVLQJ IXUWKHU LQFUHDVHV WKH YHUVDWLOLW\ E\ ZKLFK FHOOV FDQ PRGXODWH WKH V\QWKHVLV RI UHSOLFDWLRQGHSHQGHQW DV ZHOO DV YDULDQW KLVWRQH SURWHLQV GXULQJ WKH FHOO F\FOH DQG DW WKH RQVHW RI GLIIHUHQWLDWLRQ 7R EHJLQ WR H[SORUH ZKDW UROHV WKH n VWHPORRS PRWLI DQG WKH SRO\$f DGGLWLRQ VHTXHQFH SOD\ LQ WKH UHJXODWLRQ RI +%*/ P51$ OHYHOV DQG WR GHWHUPLQH LI WKH\ DUH ERWK HVVHQWLDO IRU H[SUHVVLRQ RI WKH +%*/ JHQH ZH FDUULHG RXW D SUHOLPLQDU\ H[SHULPHQW H[DPLQLQJ WKH H[SUHVVLRQ RI YDULRXV +%*/ GHOHWLRQ PXWDQWV LQ 7/ FHOOV )LJXUH f :H DQDO\]HG WRWDO FHOOXODU 51$ IURP FRQIOXHQW DQG GLIIHUHQWLDWHG 7/ FHOOV DQG FHOO OLQHV HVWDEOLVKHG DV GHVFULEHG LQ PDWHULDOV DQG PHWKRGV 6, DQDO\VLV ZDV FDUULHG RXW RQ WRWDO FHOOXODU 51$ LVRODWHG IURP FRQIOXHQW 7/ FHOO OLQHV 3 3 3 DQG FRQIOXHQW 7/ FHOOV DV ZHOO DV GLIIHUHQWLDWHG 7/ FHOO OLQHV 3 3 3 DQG GLIIHUHQWLDWHG 7/ FHOOV ,Q DGGLWLRQ 6, DQDO\VLV ZDV FDUULHG RXW RQ WRWDO FHOOXODU 51$ DQG SRO\ $ 51$ LVRODWHG IURP SUROLIHUDWLQJ +H/D FHOOV &HOO OLQH

PAGE 181

)LJXUH 6, DQDO\VLV RI WKH n WHUPLQL RI +% P51$ H[SUHVVHG LQ 7/ FHOO OLQHV 7RWDO FHOOXODU ODQH f DQG SRO\ $ ODQH f 51$V ZHUH LVRODWHG IURP SUROLIHUDWLQJ +H/D FHOOV ,Q DGGLWLRQ WRWDO FHOOXODU 51$ ZDV LVRODWHG IURP FRQIOXHQW 7/ FHOO OLQHV 3 ODQH f 3 ODQH f 3 ODQH f DQG FRQIOXHQW 7/ FHOOV ODQH f DQG GLIIHUHQWLDWHG 7/ FHOO OLQHV 3 ODQH f 3 ODQH f 3 ODQH f DQG GLIIHUHQWLDWHG 7/ FHOOV ODQH f 7RWDO FHOOXODU 51$ QJ DQG J SRO\ $ 51$ ZHUH XVHG IRU 6, DQ£O\VLV 6, DQDO\VLV RQ DOO VDPSOHV ZLWK WKH H[FHSWLRQ RI 51$ VDPSOHV LVRODWHG IURP WKH 7/ FHOO OLQH 3 ZDV FDUULHG RXW XVLQJ D QW 1HR ,(FR 5, SUREH GHULYHG IURP WKH +%*/ JHQH )LJXUH QW QW )LJXUH 6, SUREH $f DQG n HQG ODEHOHG DQG WKHVH VDPSOHV ORDGHG LQ ODQHV 6, DQDO\VLV RQ 51$ VDPSOHV LVRODWHG IURP WKH 7/ FHOO OLQH 3 ZDV FDUULHG RXW XVLQJ D QW 1FR ,(FR 5, SUREH GHULYHG IURP WKH FRQVWUXFW */69 VHH )LJXUH f ZKLFK LV VLPLODU WR 6, SUREH $ EXW ZLWK WKH QW ;KR IUDJPHQW GHOHWHG )LJXUH 6, SUREH %f 6, SUREH % ZDV n HQG ODEHOHG DQG WKH 3 VDPSOHV ZHUH ORDGHG LQ ODQHV DQG /DQH 0 ZDV ORDGHG ZLWK nODEHOHG S%5 +SD ,, PDUNHUV 7KH VL]HV RI YDULRXV PDUNHU EDQGV DUH JLYHQ RQ WKH OHIW VLGH RI ODQH 0 /DQHV $ & DQG 7 ZHUH ORDGHG ZLWK WKH SURGXFWV RI GLGHR[\ VHTXHQFLQJ UHDFWLRQV RQ 0PSO VLQJOH VWUDQGHG WHPSODWH '1$ XWLOL]LQJ D SULPHU n*7777&&&$*7&$&*$&nf 7KH QW +%*/ SURWHFWHG IUDJPHQW LV LQGLFDWHG E\ D ILOOHG WULDQJOH DQG WKH QW +% VWRS FRGRQ SURWHFWHG IUDJPHQW LV LQGLFDWHG E\ DQ HPSW\ WULDQJOH

PAGE 183

3 LV D FRQWURO FHOO OLQH FRQWDLQLQJ WKH HQWLUH +%*/ JHQH )LJXUH f DQG FHOO OLQHV 3 )LJXUH f DQG 3 )LJXUH f FRQWDLQ GHOHWLRQV RI WKH n VWHPORRS PRWLI DQG SRO\$f DGGLWLRQ VHTXHQFH UHVSHFWLYHO\ $ QW SURWHFWHG IUDJPHQW PDSSLQJ DSSUR[LPDWHO\ WR WKH +%*/ VWRS FRGRQ DQG D QW SURWHFWHG IUDJPHQW PDSSLQJ WR WKH FRQVHQVXV KLVWRQH P51$ WHUPLQXV f ZHUH GHWHFWHG LQ WKH WRWDO FHOOXODU 51$ VDPSOHV IRU +H/D DQG 7/ FHOO OLQH 3 )LJXUH ODQHV DQG f +RZHYHU RQO\ WKH QW SURWHFWHG IUDJPHQW DQ +% VWRS FRGRQ SURWHFWLRQf ZDV GHWHFWHG IRU WKH FHOO OLQH 3 DQG WKH 7/ FRQWURO VDPSOH )LJXUH ODQHV DQG f LQGLFDWLQJ WKH SUHVHQFH RI +% KLVWRQH P51$V LQ WKHVH VDPSOHV EXW QRQH VSHFLILF IRU WKH */69 FRQVWUXFW )LJXUH f 7KH 6, DQDO\VLV RI WKH 51$ IURP WKH 3 FHOO OLQH ZDV XQLQWHUSUHWDEOH )LJXUH ODQHV DQG f D FRQWLQXRXV VLJQDO ZDV GHWHFWHG WKURXJKRXW WKH OHQJWK RI WKH JHO HYHQ ZLWK D VKRUWHU H[SRVXUH 6LPLODU UHVXOWV ZHUH REWDLQHG LQ WUDQVLHQW H[SUHVVLRQ DVVD\V XVLQJ 51$ LVRODWHG KRXUV DIWHU WUDQVIHFWLRQ RI 7/ FHOOV ZLWK WKH */69 FRQVWUXFW GDWD QRW VKRZQf 7KHVH GDWD LQGLFDWH WKDW WKH 3 FHOO OLQH H[SUHVVHV WKH +%*/ P51$ ZKHUHDV WKH 3 P51$ GRHV QRW DFFXPXODWH WR GHWHFWDEOH OHYHOV LQ WKH 3 FHOO OLQH 7KH UHVXOWV IRU WKH 3 FHOO OLQH DUH XQLQWHUSUHWDEOH $OWKRXJK WKHVH UHVXOWV DUH SUHOLPLQDU\ WKH\ GR VXJJHVW WKDW GHOHWLQJ D SRUWLRQ RI WKH n WUDLOHU RI

PAGE 184

WKH +%*/ JHQH FRQWDLQLQJ WKH SRO\$f DGGLWLRQ VHTXHQFH FRQVWUXFW */69 )LJXUH f DOWHUV WKH H[SUHVVLRQ RI RU n SURFHVVLQJ RI WKH +%*/69 LQ51$ UHVXOWLQJ LQ QR GHWHFWDEOH DFFXPXODWLRQ RI +%*/69 P51$ $GGLWLRQDO UHVHDUFK PD\ OHDG WR D EHWWHU XQGHUVWDQGLQJ RI WKH UROH WKDW WKH n VWHPORRS PRWLI DQG WKH SRO\$f DGGLWLRQ VHTXHQFH SOD\ LQ WKH UHJXODWLRQ RI +%*/ P51$ $ V\VWHPDWLF DQDO\VLV RI FRQVWUXFWV FRQWDLQLQJ GHOHWLRQV WKURXJKRXW WKH n WUDLOLQJ UHJLRQ RI WKH +%*/ JHQH PD\ SURYLGH LQVLJKW LQWR ZKDW VHTXHQFHV DUH HVVHQWLDO IRU H[SUHVVLRQ RI VWDEOH UHSOLFDWLRQ GHSHQGHQW DQG FRQVWLWXWLYHO\ H[SUHVVHG +%*/ P51$V ,Q DGGLWLRQ VXFK DQDO\VHV PD\ UHYHDO ZKHWKHU VHTXHQFHV LQ FORVH SUR[LPLW\ WR WKH VWHPORRS PRWLI DUH VXIILFLHQW IRU H[SUHVVLRQ RI D UHSOLFDWLRQ GHSHQGHQW +%*/ P51$ RU ZKHWKHU DGGLWLRQDO VHTXHQFHV IXUWKHU GRZQVWUHDP DUH UHTXLUHG ,Q VXPPDU\ WKH ZRUN SUHVHQWHG LQ WKLV GLVVHUWDWLRQ GHPRQVWUDWHV WKDW WKH YDULDQW +%*/ JHQH H[SUHVVHV DOWHUQDWLYH P51$V ZKRVH SURFHVVLQJ LV GHSHQGHQW XSRQ VWUXFWXUDO HOHPHQWV SUHVHQW LQ WKH n WUDLOHU VXFK DV WKH r VWHPORRS PRWLI DQG SRO\ $f DGGLWLRQ VHTXHQFHf 7KH +%*/ JHQH KDV n VWUXFWXUDO IHDWXUHV LQKHUHQW WR ERWK FHOO F\FOH GHSHQGHQW DQG FRQVWLWXWLYHO\ H[SUHVVHG +% JHQHV DQG WKXV DOORZV IRU WKH H[SUHVVLRQ RI LWV +% JHQH SURGXFW GXULQJ WKH FHOO F\FOH DV ZHOO DV DW WKH RQVHW RI GLIIHUHQWLDWLRQ 7KH H[SUHVVLRQ RI DOWHUQDWLYH P51$V E\ WKH

PAGE 185

+%*/ JHQH SURYLGHV WKH FHOO ZLWK WKH YHUVDWLOLW\ WR PRGXODWH WKH V\QWKHVLV RI UHSOLFDWLRQGHSHQGHQW +% SURWHLQV GXULQJ WKH WUDQVLWLRQ IURP D SUROLIHUDWLYH WR D QRQGLYLGLQJ VWDWH

PAGE 186

$33(1',; $ 5($*(176 )25 3523$*$7,21 $1' 0$,17(1$1&( 2) %$&7(5,$ $1' %$&7(5,23+$*( $PSLFLOOLQ PTPO 2QH JUDP RI WKH VRGLXP VDOW RI DPSLFLOOLQ ZDV GLVVROYHG LQ PO GG+ WKH S+ RI WKH VROXWLRQ DGMXVWHG WR E\ DGGLQJ 0 +& GURSZLVH DQG WKH YROXPH EURXJKW WR PO 7KH VROXWLRQ ZDV VWHULOL]HG E\ ILOWUDWLRQ DQG GLYLGHG LQWR PO DOLTXRWV ZKLFK ZHUH VWRUHG DW r& 7KH ZRUNLQJ FRQFHQWUDWLRQ RI DPSLFLOOLQ XVHG ZDV JPO $PSLFLOOLQ NLOOV RQO\ JURZLQJ EDFWHULD DQG IXQFWLRQV E\ LQKLELWLQJ FHOO ZDOO V\QWKHVLV E\ EORFNLQJ IRUPDWLRQ RI WKH SHSWLGRJO\FDQ FURVVOLQN UHYLHZHG E\ 'DYLHV DQG 6PLWK f :D[PDQ DQG 6WURPLQJHU ff &KORUDPSKHQLFRO PTPO 7ZR JUDPV RI FKORUDPSKHQLFRO ZHUH GLVVROYHG LQ PO b HWKDQRO DQG VWRUHG DW r& )RU DPSOLILFDWLRQ RI SODVPLGV WKH ZRUNLQJ FRQFHQWUDWLRQ RI FKORUDPSKHQLFRO XVHG ZDV [JPO &KORUDPSKHQLFRO LQKLELWV SURWHLQ V\QWKHVLV LQ EDFWHULD E\ ELQGLQJ WR WKH 6 ULERVRPDO VXEXQLW DQG LQKLELWLQJ WKH SHSWLG\OWUDQVIHUDVH UHDFWLRQ UHYLHZHG E\ 'DYLHV DQG 6PLWK ff

PAGE 187

*OXFRVH b ;f 7ZHQW\ JUDPV RI JOXFRVH ZHUH GLVVROYHG LQ GG+ DQG WKH YROXPH EURXJKW WR PO 7KH VROXWLRQ ZDV VWHULOL]HG E\ ILOWUDWLRQ DQG VWRUHG DW r& 0DOWRVH b ;f 7ZHQW\ JUDPV RI PDOWRVH ZHUH GLVVROYHG LQ GG+ DQG WKH YROXPH EURXJKW WR PO 7KH VROXWLRQ ZDV VWHULOL]HG E\ ILOWUDWLRQ DQG VWRUHG DW r& 1=&<0 0HGLXP &RPSRQHQW 3HU OLWHU 1= DPLQH J 1D&O J FDVDPLQR DFLGV J %DFWR\HDVW H[WUDFW J 0J6} + J &RPSRQHQWV ZHUH GLVVROYHG LQ GG+ EULQJLQJ WKH VROXWLRQ WR b ILQDO YROXPH DQG WKH S+ DGMXVWHG WR E\ DGGLQJ 1 1D2+ GURSZLVH 7KH YROXPH ZDV QH[W EURXJKW WR b RI WKH GHVLUHG ILQDO YROXPH DQG WKH PHGLXP VWHULOL]HG E\ DXWRFODYLQJ

PAGE 188

1=&<0 %RWWRP $JDU 3ODWHV &RPSRQHQW 3HU OLWHU %DFWRDJDU J 1=&<0 PHGLXP &RPSRQHQWV ZHUH DGGHG WR DQ HUOHQPH\HU IODVN DQG WKH VXVSHQVLRQ ZDV VWHULOL]HG E\ DXWRFODYLQJ $IWHU UHPRYDO IURP WKH DXWRFODYH WKH IODVN ZDV FDUHIXOO\ VZLUOHG DQG WKHQ FRROHG WR r& LQ D ZDWHU EDWK $IWHU FRROLQJ WKH ERWWRP DJDU ZDV SRXUHG LQWR GLVSRVDEOH SODVWLF SHWUL GLVKHV DQG VWRUHG DW r& 1=&<0 7RS $JDU &RPSRQHQW 3HU OLWHU %DFWRDJDU J 1=&<0 PHGLXP &RPSRQHQWV ZHUH DGGHG WR DQ HUOHQPH\HU IODVN DQG WKH VXVSHQVLRQ ZDV VWHULOL]HG E\ DXWRFODYLQJ $IWHU UHPRYDO IURP WKH DXWRFODYH WKH IODVN ZDV FDUHIXOO\ VZLUOHG DQG WKHQ FRROHG WR r& LQ D ZDWHU EDWK $IWHU FRROLQJ WKH WRS DJDU ZDV GLYLGHG LQWR PO DOLTXRWV LQ FXOWXUH WXEHV WKH FDSV VHDOHG ZLWK SDUDILOP WR SUHYHQW PRLVWXUH ORVV DQG VWRUHG DW URRP WHPSHUDWXUH 3ULRU WR XVH WKH WRS DJDU ZDV PHOWHG E\ SODFLQJ WKH WXEHV LQ D r& ZDWHU EDWK IRU PLQXWHV DQG WKHQ FRROHG WR r& EHIRUH XVH

PAGE 189

60 0HGLXP &RPSRQHQW 3HU OLWHU 1D&O J 0J6m + 0 7ULV}&O S+ f PO J b JHODWLQ PO &RPSRQHQWV ZHUH GLVVROYHG LQ GG+ EULQJLQJ WKH VROXWLRQ WR b ILQDO YROXPH DQG WKH S+ DGMXVWHG WR E\ DGGLQJ 1 1D2+ GURSZLVH 7KH YROXPH ZDV QH[W EURXJKW WR b RI WKH GHVLUHG ILQDO YROXPH WKH PHGLXP VWHULOL]HG E\ DQG DXWRFODYLQJ 7HWUDF\FOLQH PTPO 7HWUDF\FOLQH K\GURFKORULGH Jf ZDV GLVVROYHG LQ PO HWKDQROZDWHU b YYf DQG VWRUHG DW r& (WKDQROZDWHU b YYf ZDV SUHSDUHG E\ PL[LQJ PO b HWKDQRO DQG PO GG+ 7KH ZRUNLQJ FRQFHQWUDWLRQ RI WHWUDF\FOLQH XVHG ZDV JPO 7HWUDF\FOLQH LV OLJKW VHQVLWLYH VR VROXWLRQV DQG SODWHV FRQWDLQLQJ WKH DQWLELRWLF ZHUH ZUDSSHG LQ IRLO IRU VWRUDJH 0DJQHVLXP LRQV DUH DQWDJRQLVWV RI WHWUDF\FOLQH VR PHGLD FRQWDLQLQJ PDJQHVLXP VDOWV HJ 1=&<0f ZHUH QRW XVHG ZKHQ WHWUDF\FOLQH ZDV UHTXLUHG 7HWUDF\FOLQH LQKLELWV EDFWHULDO SURWHLQ V\QWKHVLV E\ ELQGLQJ WR WKH 6 VXEXQLW RI WKH ULERVRPH ZKLFK SUHYHQWV ELQGLQJ RI DPLQRDF\O W51$ WR WKH ULERVRPH $ VLWH WKHUHE\

PAGE 190

LQKLELWLQJ ULERVRPDO WUDQVORFDWLRQ UHYLHZHG E\ 'DYLHV DQG 6PLWK ff <71 0HGLXP &RPSRQHQW %DFWRWU\SWRQH %DFWR\HDVW H[WUDFW 1D&O 3HU OLWHU J J J &RPSRQHQWV ZHUH GLVVROYHG LQ GG+ EULQJLQJ WKH VROXWLRQ WR b ILQDO YROXPH DQG WKH S+ DGMXVWHG WR E\ DGGLQJ 1 1D2+ GURSZLVH 7KH YROXPH ZDV QH[W EURXJKW WR b RI WKH GHVLUHG ILQDO YROXPH DQG WKH PHGLXP VWHULOL]HG E\ DXWRFODYLQJ <71 %RWWRP $JDU 3ODWHV &RPSRQHQW 3HU OLWHU %DFWRDJDU J <71 PHGLXP &RPSRQHQWV ZHUH DGGHG WR DQ HUOHQPH\HU IODVN DQG WKH VXVSHQVLRQ ZDV VWHULOL]HG E\ DXWRFODYLQJ $IWHU UHPRYDO IURP WKH DXWRFODYH WKH IODVN ZDV FDUHIXOO\ VZLUOHG DQG WKHQ FRROHG WR r& LQ D ZDWHU EDWK $IWHU FRROLQJ WKH ERWWRP DJDU ZDV SRXUHG LQWR GLVSRVDEOH SODVWLF SHWUL GLVKHV DQG VWRUHG DW r& ,I QHFHVVDU\ DPSLFLOOLQ ZDV DGGHG WR WKH <71 DJDU WR D ILQDO FRQFHQWUDWLRQ RI JPO RU

PAGE 191

WHWUDF\FOLQH WR QTPO LPPHGLDWHO\ SULRU WR SRXULQJ RI WKH SODWHV <71 7RS $JDU &RPSRQHQW 3HU OLWHU %DFWRDJDU J <71 PHGLXP &RPSRQHQWV ZHUH DGGHG WR DQ HUOHQPH\HU IODVN DQG WKH VXVSHQVLRQ ZDV VWHULOL]HG E\ DXWRFODYLQJ $IWHU UHPRYDO IURP WKH DXWRFODYH WKH IODVN ZDV FDUHIXOO\ VZLUOHG DQG WKHQ FRROHG WR r& LQ D ZDWHU EDWK $IWHU FRROLQJ WKH WRS DJDU ZDV GLYLGHG LQWR PO DOLTXRWV LQ FXOWXUH WXEHV WKH FDSV VHDOHG ZLWK SDUDILOP WR SUHYHQW PRLVWXUH ORVV DQG VWRUHG DW URRP WHPSHUDWXUH 3ULRU WR XVH WKH WRS DJDU ZDV PHOWHG E\ SODFLQJ WKH WXEHV LQ D r& ZDWHU EDWK IRU PLQXWHV DQG WKHQ FRROHG WR r& EHIRUH XVH <71 7RS $JDURVH <71 WRS DJDURVH ZDV SUHSDUHG HVVHQWLDOO\ DV GHVFULEHG IRU <71 WRS DJDU EXW ZLWK DJDURVH VXEVWLWXWLQJ IRU DJDU 7RS DJDURVH SURYLGHV D ILUPHU VXUIDFH IRU OLIWLQJ QLWURFHOOXORVH ILOWHUV IURP WKH SODWHV GXULQJ EDFWHULRSKDJH VFUHHQLQJ

PAGE 192

$SSHQGL[ % 5($*(176 )25 18&/(,& $&,' ,62/$7,21 $1' 385,),&$7,21 3KHQRO (TXLOLEUDWHG 3KHQRO ZDV UHGLVWLOOHG DW r& +\GUR[\TXLQROLQH DGGHG WR D ILQDO FRQFHQWUDWLRQ RI b DQG VWRUHG LQ PO DOLTXRWV DW r& XQGHU QLWURJHQ JDV $OLTXRWV RI SKHQRO ZHUH UHPRYHG IURP WKH IUHH]HU DQG PHOWHG LQ D r& ZDWHU EDWK 7KH PHOWHG SKHQRO ZDV WKHQ H[WUDFWHG RQFH ZLWK D b YROXPH RI 0 7ULV}&O S+ f IROORZHG E\ VHYHUDO HTXDO YROXPH H[WUDFWLRQV ZLWK P0 7ULV}&O S+ f P0 ('7$ XQWLO WKH S+ ZDV DSSUR[LPDWHO\ 7KH HTXLOLEUDWHG SKHQRO ZDV WKHQ VWRUHG DW r& XQGHU P0 7ULV} &O S+ f P0 ('7$ LQ D EURZQ JODVV ERWWOH $ONDOLQH /YVLV 6ROXWLRQ &RPSRQHQW 3HU OLWHU b D' *OXFRVH PO 0 7ULV}&O S+ f PO P0 ('7$ PO )LQDO FRQFHQWUDWLRQ b ZYf P0 P0 &RPSRQHQWV ZHUH FRPELQHG DQG WKH VROXWLRQ EURXJKW WR OLWHU ILQDO ZLWK WKH DGGLWLRQ RI PO GG+

PAGE 193

$ONDOLQH /YVLV 6ROXWLRQ &RPRRQHQW 3HU PO )LQDO FRQFHQWUDWLRQ b 6'6 ZYf PO b ZYf 0 1D2+ PO P1 7KH 0 1D2+ VWRFN PO ZDV DGGHG WR PO GG+ DQG PL[HG WKRURXJKO\ 1H[W WKH b 6'6 ZYf VWRFN PO ZDV DGGHG DQG WKH VROXWLRQ PL[HG $ONDOLQH O\VLV VROXWLRQ ZDV PDGH IUHVK IRU HDFK SODVPLG SUHSDUDWLRQ

PAGE 194

$SSHQGL[ & %$&7(5,$/ 675$,16 86(' 6WUDLQ '+D /( ;/O%OXH < 5HPDUNV 5HIHUHQFH 6XLWDEOH IRU JHQHUDWLQJ %5/r1 f F'1$ OLEUDULHV DQG IRU RWKHU SODVPLG FORQLQJ SURFHGXUHV /( ZDV SUHSDUHG E\ / (QTXLVW DQG LV D GHULYDWLYH RI VWUDLQ (' %RUFN HW DO f 0XUUD\ HW DO f (QTXLVW HW DOA f 6WDELOLW\ RI WKH )n %XOORFN HW DOA f HSLVRPH LV SURYLGHG E\ WKH WHWUDF\FOLQH UHVLVWDQW PDUNHU ORFDWHG RQ WKH )n HSLVRPH < ZDV GHULYHG IURP
PAGE 195

5()(5(1&(6 $OOIUH\ 9* 9& /LWWDX DQG $( 0LUVN\ 2Q WKH UROH RI KLVWRQHV LQ UHJXODWLQJ ULERQXFOHLF DFLG V\QWKHVLV LQ WKH FHOO QXFOHXV 3URF 1DWO $FDG 6FL 86$ $OWHUPDQ 5%0 & 6SUHFKHU 5 *UDYHV :3 0DU]OXII DQG $, 6NRXOWFKL 5HJXODWHG ([SUHVVLRQ RI D FKLPHULF KLVWRQH JHQH LQWURGXFHG LQWR PRXVH ILEUREODVWV 0RO &HOO %LRO %DXPEDFK // ) 0DUDVKL 0 3OXPE 6WHLQ DQG 6WHLQ ,QKLELWLRQ RI '1$ UHSOLFDWLRQ FRRUGLQDWHO\ UHGXFHV FHOOXODU OHYHOV RI FRUH DQG +, KLVWRQH P51$V UHTXLUHPHQW IRU SURWHLQ V\QWKHVLV %LRFKHPLVWU\ %DXPEDFK // *6 6WHLQ DQG -/ 6WHLQ 5HJXODWLRQ RI KXPDQ KLVWRQH JHQH H[SUHVVLRQ WUDQVFULSWLRQDO DQG SRVW WUDQVFULSWLRQDO FRQWURO LQ WKH FRXSOLQJ RI KLVWRQH PHVVHQJHU 51$ VWDELOLW\ ZLWK '1$ UHSOLFDWLRQ %LRFKHPLVWU\ %HQWRQ :' DQG 5: 'DYLV 6FUHHQLQJ ;JW UHFRPELQDQW FORQHV E\ K\EULGL]DWLRQ WR VLQJOH SODTXHV LQ VLWX 6FLHQFH %HUN $DQG 3$ 6KDUS 6SOLFHG HDUO\ P51$V RI VLPLDQ YLUXV 3URF 1DWO $FDG 6FL 86$ %HUQORKU '$ &: $QJXV 0' /DQH 0$ %RODQRZVNL DQG 7.HOO\-U ([SUHVVLRQ RI VSHFLILF P51$V GXULQJ DGLSRVH GLIIHUHQWLDWLRQ ,GHQWLILFDWLRQ RI DQ P51$ HQFRGLQJ D KRPRORJXH RI P\HOLQ 3 SURWHLQ 3URF 1DWO $FDG 6FL 86$ %HWKHVGD 5HVHDUFK /DERUDWRULHV &DWDORJXH t UHIHUHQFH JXLGH %HWKHVGD 5HVHDUFK /DERUDWRULHV /LIH 7HFKQRORJLHV ,QF *DLWKHUVEXUJ 0' %LUFKPLHU & 5 *URVVFKHGO DQG 0/ %LUQVWLHO *HQHUDWLRQ RI DXWKHQWLF n WHUPLQL RI DQ +$ LQ YLYR LV GHSHQGHQW RQ D VKRUW LQYHUWHG '1$ UHSHDW DQG RQ VSDFHU VHTXHQFHV &HOO

PAGE 196

%LUG 5& )$ -DFREV 6WHLQ 6WHLQ DQG %+ 6HOOV &RRUGLQDWH UHJXODWLRQ RI KLVWRQH P51$V GXULQJ JURZWK DQG GLIIHUHQWLDWLRQ RI UDW P\REODVWV %LRFKLP %LRSK\V $FWD %LUQERLP +& DQG 'RO\ $ UDSLG DONDOLQH H[WUDFWLRQ SURFHGXUH IRU VFUHHQLQJ UHFRPELQDQW SODVPLG '1$ 1XFOHLF $FLGV 5HV %LUQVWLHO 0/ 0 %XVVOLQJHU DQG 6WUXE 7UDQVFULSWLRQ WHUPLQDWLRQ DQG n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
PAGE 197

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n3 /DEHOLQJ RI 51$ DQG '1$ UHVWULFWLRQ IUDJPHQWV 0HWK (Q]\PRO &KDOORQHU 3% 6% 0RVV DQG 0 *URXGLQH ([SUHVVLRQ RI UHSOLFDWLRQGHSHQGHQW KLVWRQH JHQHV LQ DYLDQ VSHUPDWLGV LQYROYHV DQ DOWHUQDWH SDWKZD\ RI P51$ f HQG IRUPDWLRQ 0RO &HOO %LRO &KDXEHW 1 0 &KDERXWH % &OPHQW 0 (KOLQJ 3KLOLSSV DQG & *LJRW 7KH KLVWRQH + DQG + P51$V DUH SRO\DGHQ\O\ODWHG LQ PDL]H 1XFOHLF $FLGV 5HV &KHQJ $ 1DQGL 6 &OHUN DQG $, 6NRXOWFKL 'LIIHUHQW nHQG SURFHVVLQJ SURGXFHV WZR LQGHSHQGHQWO\ UHJXODWHG P51$V IURP D VLQJOH +, KLVWRQH JHQH 3URF 1DWO $FDG 6FL 86$ &KHQJ DQG $, 6NRXOWFKL 5DSLG LQGXFWLRQ RI SRO\DGHQ\ODWHG +, KLVWRQH P51$V LQ PRXVH HU\WKUROHXNHPLD FHOOV LV UHJXODWHG E\ FP\F 0RO &HOO %LRO &KRGFKR\ 1 %/HYLQH & 6SUHFKHU $, 6NRXOWFKL DQG :) 0DU]OXII ([SUHVVLRQ RI PRXVH KLVWRQH JHQHV WUDQVFULSWLRQ LQWR n LQWHUJHQLF '1$ DQG FU\SWLF SURFHVVLQJ VLWHV GRZQVWUHDP IURP WKH n HQG RI WKH + JHQH 0RO &HOO %LRO

PAGE 198

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f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n 51$ SURFHVVLQJ UHDFWLRQ (0%2 'DOWRQ 6 -5 &ROHPDQ DQG -5( :HOOV 7UDQVFULSWLRQ RI WKH KLVWRQH + JHQH LV QRW 6SKDVH UHJXODWHG 0RO &HOO %LRO 'DYLHV DQG ', 6PLWK 3ODVPLGGHWHUPLQHG UHVLVWDQFH WR DQWLPLFURELDO DJHQWV $QQX 5HY 0LFURELRO

PAGE 199

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
PAGE 200

)ULHGPDQ '/ DQG 0,PSHULDOH 51$ n HQG IRUPDWLRQ LQ WKH FRQWURO RI JHQH H[SUHVVLRQ $QQX 5HY *HQHW )ULVFKDXI $0 + /HKUDFK $ 3RXVWND DQG 1 0XUUD\ /DPEGD UHSODFHPHQW YHFWRUV FDUU\LQJ SRO\OLQNHU VHTXHQFHV 0RO %LRO *LFN 2 $ .UDPHU : .HOOHU DQG 0/ %LUQVWLHO *HQHUDWLRQ RI KLVWRQH P51$ n HQGV E\ HQGRQXFOHRO\WLF FOHDYDJH RI WKH SUHP51$ LQ D VQ513GHSHQGHQW LQ YLWUR UHDFWLRQ (0%2 *RUPDQ &0 /) 0RIIDW DQG %+ +RZDUG 5HFRPELQDQW JHQRPHV ZKLFK H[SUHVV FKORUDPSKHQLFRO DFHW\OWUDQVIHUDVH LQ PDPPDOLDQ FHOOV 0RO &HOO %LRO *RXOLDQ 0 6+ *RXOLDQ (( &RGG DQG $= %OXPHQILHOG 3URSHUWLHV RI ROLJRGHR[\QXFOHRWLGHV WKDW GHWHUPLQH SULPLQJ DFWLYLW\ ZLWK (VFKHULFKLD FROL GHR[\ULERQXFOHLF DFLG SRO\PHUDVH %LRFKHPLVWU\ *UDKDP )/ DQG $YDQ GHU (E $ QHZ WHFKQLTXH IRU WKH DVVD\ RI LQIHFWLYLW\ RI KXPDQ DGHQRYLUXV '1$ 9LURORJ\ *UDQG\ '. DQG -% 'RGJVRQ 6WUXFWXUH DQG RUJDQL]DWLRQ RI WKH FKLFNHQ +% KLVWRQH JHQH IDPLO\ 1XFOHLF $FLGV 5HV *UDQG\ '. -' (QJHO DQG -% 'RGJVRQ &RPSOHWH QXFOHRWLGH VHTXHQFH RI D FKLFNHQ +E KLVWRQH JHQH %LRO &KHP *UDYHV 5$ 1% 3DQGH\ 1 &KRGFKR\ DQG :) 0DU]OXII 7UDQVODWLRQ LV UHTXLUHG IRU UHJXODWLRQ RI KLVWRQH P51$ GHJUDGDWLRQ &HOO *UHHQ / 5 YDQ $QWZHUSHQ 6WHLQ 6WHLQ 3 7ULSSXWL % (PDQXHO 6HOGHQ DQG & &URFH $ PDMRU KXPDQ KLVWRQH JHQH FOXVWHU RQ WKH ORQJ DUP RI FKURPRVRPH 6FLHQFH *UHHQ 05 3UHP51$ VSOLFLQJ $QQX 5HY *HQHW *ULPHV 6 3 :HLV]&DUULQJWRQ + 'DXP ,,, 6PLWK / *UHHQ :ULJKW 6WHLQ DQG 6WHLQ $ UDW KLVWRQH + JHQH FORVHO\ DVVRFLDWHG ZLWK WKH WHVWLVVSHFLILF +LW JHQH ([S &HOO 5HV

PAGE 201

*URVV : 6FKDIIQHU 7HOIRUG DQG 0 %LUQVWLHO 0ROHFXODU DQDO\VLV RI WKH KLVWRQH JHQH FOXVWHU RI 3VDPPHFKLQXV PLOLDULV ,,, SRODULW\ DQG DV\PPHWU\ RI WKH KLVWRQHFRGLQJ VHTXHQFHV &HOO *URVV%HOODUG 0 3 2XGHW DQG 3 &KDPEQ ,VRODWLRQ RI KLJKPROHFXODUZHLJKW '1$ IURP PDPPDOLDQ FHOOV (XU %LRFKHP +DHJHPDQ ,VHUHQWDQW *KH\VHQ DQG : )LHUV &KDUDFWHUL]DWLRQ RI WKH PDMRU DOWHUHG OHDGHU VHTXHQFH RI ODWH P51$ LQGXFHG E\ 69 GHOHWLRQ PXWDQW GO 1XFOHLF $FLGV 5HV +DJHQEFKOH 2 5 %RYH\ DQG 5$
PAGE 202

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f KLVWRQH JHQH 0RO &HOO %LRO ,JR.HPHQHV 7 : +U] DQG +* =DFKDX &KURPDWLQ $QQX 5HY %LRFKHP ,VHQEHUJ +LVWRQHV $QQX 5HY %LRFKHP ,VK+RURZLF] DQG -) %XUNH 5DSLG DQG HIILFLHQW FRVPLG FORQLQJ 1XFOHLF $FLGV 5HV

PAGE 203

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n SURFHVVLQJ LQ WZR DYLDQ KLVWRQH +, JHQHV *HQHV t 'HYHORSPHQW .ULHJ 3$ DQG '$ 0HOWRQ )RUPDWLRQ RI WKH n HQG RI KLVWRQH P51$ E\ SRVWWUDQVODWLRQDO SURFHVVLQJ 1DWXUH .ULHJ 3$ $5RELQV $ &ROPDQ DQG -5( :HOOV &KLFNHQ KLVWRQH + P51$ WKH SRO\DGHQ\ODWHG 51$ ODFNV WKH FRQVHUYHG KLVWRQH n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

PAGE 204

/HYHQVRQ 5* DQG .% 0DUFX 2Q WKH H[LVWHQFH RI SRO\DGHQ\ODWHG KLVWRQH P51$ LQ ;HQRSXV ODHYLV RRF\WHV &HOO /HYLQH %1 &KRGFKR\ :) 0DU]OXII DQG $, 6NRXOWFKL &RXSOLQJ RI UHSOLFDWLRQ W\SH KLVWRQH [Q51$ OHYHOV WR '1$ V\QWKHVLV UHTXLUHV WKH VWHPORRS VHTXHQFH DW WKH n HQG RI WKH P51$ 3URF 1DWO $FDG 6FL 86$ /LIWRQ 53 0/ *ROGEHUJ 5: .DUS DQG '6 +RJQHVV 7KH RUJDQL]DWLRQ RI WKH KLVWRQH JHQHV LQ 'URVRSKLOD PHODQRTDVWHU IXQFWLRQDO DQG HYROXWLRQDU\ LPSOLFDWLRQV &ROG 6SULQJ +DUERU 6\PS 4XDQW %LRO /LWWOH 3)5 DQG 6+ &URVV $ FRVPLG YHFWRU WKDW IDFLOLWDWHV UHVWULFWLRQ HQ]\PH PDSSLQJ 3URF 1DWO $FDG 6FL 86$ /LX 7%/HYLQH $, 6NRXOWFKL DQG :) 0DU]OXII 7KH HIILFLHQF\ RI nHQG IRUPDWLRQ FRQWULEXWHV WR WKH UHODWLYH OHYHOV RI GLIIHUHQW KLVWRQH P51$V 0RO &HOO %LRO /VFKHU % DQG 6FKPSHUOL 51$ n SURFHVVLQJ UHJXODWHV KLVWRQH P51$ OHYHOV LQ D PDPPDOLDQ FHOO F\FOH PXWDQW $ SURFHVVLQJ IDFWRU EHFRPHV OLPLWLQJ LQ *ODUUHVWHG FHOOV (0%2 /VFKHU % & 6WDXEHU 5 6FKLQGOHU DQG 6FKPSHUOL )DLWKIXO FHOOF\FOH UHJXODWLRQ RI D UHFRPELQDQW PRXVH KLVWRQH + JHQH LV FRQWUROOHG E\ VHTXHQFHV LQ WKH nWHUPLQDO SDUW RI WKH JHQH 3URF 1DWO $FDG 6FL 86$ 0DQGHO 0 DQG $ +LJD &DOFLXPGHSHQGHQW EDFWHULRSKDJH '1$ LQIHFWLRQ 0RO %LRO 0DQLDWLV 7 () )ULWVFK DQG 6DPEURRN 0ROHFXODU FORQLQJ D ODERUDWRU\ PDQXDOf &ROG 6SULQJ +DUERU /DERUDWRU\ &ROG 6SULQJ +DUERU 1HZ
PAGE 205

0DQQLURQL & :0 %RQQHU DQG &/ +DWFK +$; D KLVWRQH LVRSURWHLQ ZLWK D FRQVHUYHG &WHUPLQDO VHTXHQFH LV HQFRGHG E\ D QRYHO P51$ ZLWK ERWK '1$ UHSOLFDWLRQ W\SH DQG SRO\$f n SURFHVVLQJ VLJQDOV 1XFOHLF $FLGV 5HV 0DUDVKL ) 6 +HOPV $ 6KLHOV 6 6LOYHUVWHLQ '6 *UHHQVSDQ 6WHLQ DQG 6WHLQ (QKDQFHUIDFLOLWDWHG H[SUHVVLRQ RI SURNDU\RWLF DQG HXNDU\RWLF JHQHV XVLQJ KXPDQ KLVWRQH JHQH n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n HQG IRUPDWLRQ LQ YLWUR 0RO &HOO %LRO 0RZU\ ./ DQG -$ 6WHLW] ,GHQWLILFDWLRQ RI WKH KXPDQ 8 VQ513 DV RQH RI VHYHUDO IDFWRUV LQYROYHG LQ WKH n HQG PDWXUDWLRQ RI KLVWRQH SUHPHVVHQJHU 51$nV 6FLHQFH 0RZU\ ./ DQG -$ 6WHLW] VQ513 PHGLDWRUV RI n HQG SURFHVVLQJ IXQFWLRQDO IRVVLOV" 7UHQGV %LRFKHP 6FL

PAGE 206

0XUUD\ 1( :%UDPPDU DQG 0XUUD\ /DPEGRLG SKDJHV WKDW VLPSOLI\ WKH UHFRYHU\ RI LQ YLWUR UHFRPELQDQWV 0ROHF JHQ *HQHW 1HYLQV -5 7KH SDWKZD\ RI HXNDU\RWLF P51$ IRUPDWLRQ $QQX 5HY %LRFKHP 1RUUDQGHU 7 .HPSH DQG 0HVVLQJ &RQVWUXFWLRQ RI LPSURYHG 0 YHFWRUV XVLQJ ROLJRGHR[\QXFOHRWLGHGLUHFWHG PXWDJHQHVLV *HQH 2n)DUUHOO 3 5HSODFHPHQW V\QWKHVLV PHWKRG RI ODEHOLQJ '1$ IUDJPHQWV )RFXV ff§ 3DQGH\ 1% DQG :) 0DU]OXII 7KH VWHPORRS VWUXFWXUH DW WKH n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

PAGE 207

3URNRSS .0 7KH QXFOHRWLGH VHTXHQFHV RI D KXPDQ +% KLVWRQH FRGLQJ UHJLRQ DQG D SRUWLRQ RI LWV IODQNLQJ UHJLRQV 06F WKHVLV 8QLYHUVLW\ RI )ORULGD *DLQHVYLOOH )/ 3URXGIRRW 1DQG ** %URZQOHH n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f 5RVV DQG .REV + KLVWRQH PHVVHQJHU 51$ GHFD\ LQ FHOOIUHH H[WUDFWV LQLWLDWHV DW RU QHDU WKH WHUPLQXV DQG SURFHHGV n WR n 0RO %LRO 5RVV .REV %UHZHU DQG 6: 3HOW] 3URSHUWLHV RI WKH H[RQXFOHDVH DFWLYLW\ WKDW GHJUDGHV + KLVWRQH P51$ %LRO &KHP 5RVV 6: 3HOW] .REV DQG %UHZHU +LVWRQH P51$ GHJUDGDWLRQ LQ YLYR WKH ILUVW GHWHFWDEOH VWHS RFFXUV DW RU QHDU WKH n WHUPLQXV 0RO &HOO %LRO 5XEHUWL 3 )UDJDSDQH 3 3LHUDQGUHL$PDOGL ( %HFFDUL ) $PDOGL DQG %R]]RQL &KDUDFWHUL]DWLRQ RI KLVWRQH JHQHV LVRODWHG IURP ;HQRSXV ODHYLV DQG ;HQRSXV WURSLFDOLV JHQRPLF OLEUDULHV 1XFOHLF $FLGV 5HV 5XGHUPDQ -9 DQG 0/ 3DUGXH $ SRUWLRQ RI DOO PDMRU FODVVHV RI KLVWRQH PHVVHQJHU 51$ LQ DPSKLELDQ RRF\WHV LV SRO\DGHQ\ODWHG %LRO &KHP 6DOVHU : *ORELQ P51$ VHTXHQFHV DQDO\VLV RI EDVH SDLULQJ DQG HYROXWLRQDU\ LPSOLFDWLRQV &ROG 6SULQJ +DUERU 6\PS 4XDQW %LRO

PAGE 208

6DQJHU ) 6 1LFNOHQ DQG $5 &RXOVRQ '1$ VHTXHQFLQJ ZLWK FKDLQWHUPLQDWLQJ LQKLELWRUV 3URF 1DWO $FDG 6FL 86$ 6FKDXIHOH ) *0 *LOPDUWLQ : %DQQZDUWK DQG 0/ %LUQVWLHO &RPSHQVDWRU\ PXWDWLRQV VXJJHVW WKDW EDVHSDLULQJ ZLWK D VPDOO QXFOHDU 51$ LV UHTXLUHG WR IRUP WKH n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n SURFHVVLQJ RI KLVWRQH SUHP51$ 0RO &HOO %LRO

PAGE 209

6RXWKHUQ (0 'HWHFWLRQ RI VSHFLILF VHTXHQFHV DPRQJ '1$ IUDJPHQWV VHSDUDWHG E\ JHO HOHFWURSKRUHVLV 0RO %LRO 6RXWKHUQ 3DQG 3 %HUJ 7UDQVIRUPDWLRQ RI PDPPDOLDQ FHOOV WR DQWLELRWLF UHVLVWDQFH ZLWK D EDFWHULDO JHQH XQGHU FRQWURO RI WKH 69 HDUO\ UHJLRQ SURPRWHU 0RO $SSO *HQHW 6WDXEHU & % /VFKHU 5 (FNQHU ( /GWVFKHU DQG 6FKPSHUOL $ VLJQDO UHJXODWLQJ PRXVH KLVWRQH + P51$ OHYHOV LQ D PDPPDOLDQ FHOO F\FOH PXWDQW DQG VHTXHQFHV FRQWUROOLQJ 51$ n SURFHVVLQJ DUH ERWK FRQWDLQHG ZLWKLQ WKH VDPH ES IUDJPHQW (0%2 6WDXEHU & DQG 6FKPSHUOL n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f +LVWRQH JHQHV -RKQ :LOH\ t 6RQV 1HZ
PAGE 210

7DERU 6 +( +XEHU DQG && 5LFKDUGVRQ (VFKHULFKLD FROL 7KLRUHGR[LQ FRQIHUV SURFHVVLYLW\ RQ WKH '1$ SRO\PHUDVH DFWLYLW\ RI WKH JHQH SURWHLQ RI EDFWHULRSKDJH 7 %LRO &KHP 7D\ORU -0 5 ,OOPHQVHH DQG 6XPPHUV (IILFLHQW WUDQVFULSWLRQ RI 51$ LQWR '1$ E\ DYLDQ VDUFRPD YLUXV SRO\PHUDVH %LRFKLP %LRSK\V $FWD 7KRPDV 36 +\EULGL]DWLRQ RI GHQDWXUHG 51$ DQG VPDOO '1$ IUDJPHQWV WUDQVIHUUHG WR QLWURFHOOXORVH 3URF 1DWO $FDG 6FL 86$ 7ULSSXWL 3 %6 (PDQXHO &+ &URFH /* *UHHQ *6 6WHLQ DQG -/ 6WHLQ +XPDQ KLVWRQH JHQHV PDS WR PXOWLSOH FKURPRVRPHV 3URF 1DWO $FDG 6FL 86$ 7VDL <+ DQG /6 +QLOLFD 7LVVXHVSHFLILF KLVWRQHV LQ WKH HU\WKURF\WHV RI FKLFNHQ DQG WXUWOH ([S &HOO 5HV 7VXFKL\D 6 < .RED\DVKL < *RWR + 2NXPXUD 6 1DNDH 7 .RQQR DQG 7DGD ,QGXFWLRQ RI PDWXUDWLRQ LQ FXOWXUHG KXPDQ PRQRF\WLF OHXNHPLD FHOOV E\ D SKRUERO GLHVWHU ,QW &DQFHU 7XOOLV 5+ DQG + 5XELQ &DOFLXP SURWHFWV '1DVH IURP SURWHLQDVH $ QHZ PHWKRG IRU WKH UHPRYDO RI FRQWDPLQDWLQJ 51DVH IURP '1DVH $QDO %LRFKHP 8UEDQ 0. 6* )UDQNOLQ DQG $ =ZHLGOHU ,VRODWLRQ DQG FKDUDFWHUL]DWLRQ RI WKH KLVWRQH YDULDQWV LQ FKLFNHQ HU\WKURF\WHV %LRFKHPLVWU\ YDQ :LMQHQ $5) 0DVVXQJ -/ 6WHLQ DQG *6 6WHLQ +XPDQ +, KLVWRQH JHQH SURPRWHU &&$$7 ER[ ELQGLQJ SURWHLQ +L1)% LV D PRVDLF IDFWRU %LRFKHPLVWU\ YDQ :LMQHQ $./ :ULJKW 5) 0DVVXQJ 0 *HUUHWVHQ -/ 6WHLQ DQG *6 6WHLQ 7ZR WDUJHW VLWHV IRU SURWHLQ ELQGLQJ LQ WKH SURPRWHU UHJLRQ RI D FHOO F\FOH UHJXODWHG KXPDQ +, KLVWRQH JHQH 1XFOHLF $FLGV 5HV 9DVVHURW $3 +RIIPDQQ 7DKLWL DQG 0/ %LUQVWLHO n 3URFHVVLQJ RI KLVWRQH 51$ SUHFXUVRUV 0RO %LR 5HSRUWV

PAGE 211

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

PAGE 212


PAGE 213

%,2*5$3+,&$/ 6.(7&+ 'DYLG *HRUJH &ROODUW ZDV ERUQ LQ :HLUWRQ :HVW 9LUJLQLD RQ 0D\ +H DWWHQGHG %OHVVHG 6DFUDPHQW 6FKRRO LQ :LQWHUVYLOOH 2KLR IURP WR DQG 6W 3HWHUnV VFKRRO LQ +XURQ 2KLR IURP WR +H DWWHQGHG (GLVRQ +LJK 6FKRRO 0LODQ 2KLRf DQG JUDGXDWHG LQ +H WKHQ HQWHUHG )LUHODQGV &ROOHJH RI %RZOLQJ *UHHQ 6WDWH 8QLYHUVLW\ +XURQ 2KLRf LQ WKH IDOO RI WUDQVIHUUHG WR WKH PDLQ FDPSXV DW %RZOLQJ *UHHQ 2KLR LQ WKH IDOO RI DQG JUDGXDWHG ZLWK D %DFKHORU RI 6FLHQFH GHJUHH LQ FKHPLVWU\ LQ WKH VXPPHU RI +H EHJDQ UHVHDUFK LQ WKH ODERUDWRU\ RI 'UV *DU\ DQG -DQHW 6WHLQ DQG LQ WKH IDOO RI KH HQWHUHG WKH JUDGXDWH SURJUDP LQ WKH 'HSDUWPHQW RI %LRFKHPLVWU\ DQG 0ROHFXODU %LRORJ\ DW WKH 8QLYHUVLW\ RI )ORULGD 2Q $XJXVW KH PDUULHG .DWKOHHQ /HDK 'XGHFN LQ *DLQHVYLOOH )ORULGD DQG RQ 2FWREHU *RG EOHVVHG WKHP ZLWK WKH ELUWK RI D VRQ 7LPRWK\ *HUDUG &ROODUW $IWHU JUDGXDWLRQ KH ZLOO SXUVXH UHVHDUFK DV D SRVWGRFWRUDO IHOORZ ZLWK 'U *UD\ &URXVH LQ WKH 'HSDUWPHQW RI %LRORJ\ DW (PRU\ 8QLYHUVLW\ LQ $WODQWD *HRUJLD

PAGE 214

, FHUWLI\ WKDW KDYH UHDG WKLV VWXG\ DQG WKDW LQ P\ RSLQLRQ LW FRQIRUPV WR DFFHSWDEOH VWDQGDUGV RI VFKRODUO\ SUHVHQWDWLRQ DQG LV IXOO\ DGHTXDWH LQ VFRSADQG TXDOLW\ DV D GLVVHUWDWLRQ IRU WKH GHJUHH RI 'RFWRIASII 3KLORVRSK\ ;tf§ U\ 6 6eJ=Q &KDLU 3URIHVVRU RI %LRFKHPLVWU\ DQG 0ROHFXODU %LRORJ\ FHUWLI\ WKDW KDYH UHDG WKLV VWXG\ DQG WKDW LQ P\ RSLQLRQ LW FRQIRUPV WR DFFHSWDEOH VWDQGDUGV RI VFKRODUO\ SUHVHQWDWLRQ DQG LV IXOO\ DGHTXDWH LQ VFRSH DQG TXDOLW\ DV D GLVVHUWDWLRQ IRU WKH GHJUHH RI 'RFWRU RI 3KLORVRSK\ -D£HW /6WHLQ&RFKDLU 3URIHVVRU RI ,PPXQRORJ\ DQG 0HGLFDO 0LFURELRORJ\ FHUWLI\ WKDW KDYH UHDG WKLV VWXG\ DQG WKDW LQ P\ RSLQLRQ LW FRQIRUPV WR DFFHSWDEOH VWDQGDUGV RI VFKRODUO\ SUHVHQWDWLRQ DQG LV IXOO\ DGHTXDWH LQ VFRSH DQG TXDOLW\ DV D GLVVHUWDWLRQ IRU WKH GHJUHH RI 'RFWRU RI 3KLORVRSK\ 3KLOLS O$OSLV 3URIHVVRUnRI %LRFKHPLVWU\ DQG 0ROHFXODU %LRORJ\ FHUWLI\ WKDW KDYH UHDG WKLV VWXG\ DQG WKDW LQ P\ RSLQLRQ LW FRQIRUPV WR DFFHSWDEOH VWDQGDUGV RI VFKRODUO\ SUHVHQWDWLRQ DQG LV IXOO\ DGHTXDWH LQ VFRSH DQG TXDOLW\ DV D GLVVHUWDWLRQ IRU WKH GHJUHH RI 'RFWRU RI 3KLORVRSK\ +DUU\ 61LFN $VVRFLDWH 3URIHVVRU RI %LRFKHPLVWU\ DQG 0ROHFXODU %LRORJ\

PAGE 215

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n 7URIHVVRU RI %LRORJ\ FHUWLI\ WKDW KDYH UHDG WKLV VWXG\ DQG WKDW LQ P\ RSLQLRQ LW FRQIRUPV WR DFFHSWDEOH VWDQGDUGV RI VFKRODUO\ SUHVHQWDWLRQ DQG LV IXOO\ DGHTXDWH LQ VFRSH DQG TXDOLW\ DV D GLVVHUWDWLRQ IRU WKH GHJUHH RI 'RFWRU RI 3KLORVRSK\ (LOHHQ +LFNH\ 3URIHVVRU RI %LRORJ\

PAGE 216

7KLV GLVVHUWDWLRQ ZDV VXEPLWWHG WR WKH *UDGXDWH )DFXOW\ RI WKH &ROOHJH RI 0HGLFLQH DQG WR WKH *UDGXDWH 6FKRRO DQG ZDV DFFHSWHG DV SDUWLDO IXOILOOPHQW RI WKH UHTXLUHPHQWV IRU WKH GHJUHH RI 'RFWRU RI 3KLORVRSK\ 0D\ 'HDQ &ROOHJH RI 0HGLFLQH 'HDQ *UDGXDWH 6FKRRO

PAGE 217

81,9(56,7< 2) )/25,'$


UNIVERSITY OF FLORIDA
3 1262 08554 6223