CA2394841A1

CA2394841A1 - Nucleic acids, proteins, and antibodies

Info

Publication number: CA2394841A1
Application number: CA002394841A
Authority: CA
Inventors: Steven C. Barash; Steven M. Ruben; Craig A. Rosen
Original assignee: Steven C. Barash; Steven M. Ruben; Human Genome Sciences, Inc.; Craig A. Rosen
Current assignee: Human Genome Sciences Inc
Priority date: 2000-01-31
Filing date: 2001-01-17
Publication date: 2001-08-02
Also published as: WO2001055322A8; CA2392398A1; WO2001055324A3; WO2001059064A2; WO2001055304A2; WO2001055315A2; WO2001055350A8; CA2395671A1; WO2001055322A2; WO2001055303A2; CA2393652A1; WO2001055200A1; WO2001055302A3; WO2001055323A3; WO2001055325A2; WO2001055368A1; WO2001054473A2; WO2001055303A8; WO2001055323A2; CA2395403A1

Abstract

The present invention relates to novel connective tissue related polynucleotides and the polypeptides encoded by these polynucleotides herein collectively known as "connective tissue antigens", and the use of such connective tissue antigens for detecting disorders of connective tissue(s), particularly the presence of cancer and cancer metastases. More specifically, isolated connective tissue associated nucleic acid molecules are provided encoding novel connective tissue associated polypeptides. Novel connective tissue polypeptides and antibodies that bind to these polypeptides are provided. Also provided are vectors, host cells, and recombinant and synthetic methods for producing human connective tissue associated polynucleotides and/or polypeptides. The invention further relates to diagnostic and therapeutic methods useful for diagnosing, treating, preventing and/or prognosing disorders related to connective tissue(s), including cancer, and therapeutic methods for treating such disorders. The invention further relates to screening methods for identifying agonists and antagonsits of polynucleotides and polypeptides of the invention. The present invention further relates to methods and/or compositions for inhibiting the production and function of the polypeptides of the present invention.

Description

DEMANDE OU BREVET VOLUMINEUX
LA PRESENTE PARTIE DE CETTE DEMANDE OU CE BREVET COMPREND
PLUS D'UN TOME.

~~ TTENANT LES PAGES 213 A 340 NOTE : Pour les tomes additionels, veuillez contacter 1e Bureau canadien des brevets JUMBO APPLICATIONS/PATENTS
THIS SECTION OF THE APPLICATION/PATENT CONTAINS MORE THAN ONE
VOLUME

NOTE: For additional volumes, please contact the Canadian Patent Office NOM DU FICHIER / FILE NAME
NOTE POUR LE TOME / VOLUME NOTE:

Nucleic Acids, Proteins, and Antibodies [001] This application refers to a "Sequence Listing" that is provided only on electronic media in computer readable form pursuant to Administrative Instructions Section 801(a)(i). The Sequence Listing forms a part of this description pursuant to Rule 5.2 and Administrative Instructions Sections 801 to 806, and is hereby incorporated in its entirety.

[002] The Sequence Listing is provided as an electronic file (PC009PCT
seqList.txt, 4,080,491 bytes in size, created on January 12, 2001) on four identical compact discs (CD-R), labeled "COPY 1," "COPY 2," "COPY 3," and "CRF." The Sequence Listing complies with Annex C of the Administrative Instructions, and may be viewed, for example, on an IBM-PC machine running the MS-Windows operating system by using the V viewer software, version 2000 (see World Wide Web URL:
http://www.fileviewer.com).
Field of the Invention

[003] The present invention relates to novel connective tissue related polynucleotides, the polypeptides encoded by these polynucleotides herein collectively referred to as "connective tissue antigens," and antibodies that immunospecifically bind these polypeptides, and the use of such connective tissue polynucleotides, antigens, and antibodies for detecting, treating, preventing and/or prognosing disorders of connective tissue(s), including, but not limited to, the presence of cancer and- cancer metastases.
More specifically, isolated connective tissue nucleic acid molecules are provided encoding novel connective tissue polypeptides. Novel connective tissue polypeptides and antibodies that bind to these polypeptides are provided. Also provided are vectors, host cells, and recombinant and synthetic methods for producing human connective tissue polynucleotides, polypeptides, and/or antibodies. The invention further relates to diagnostic and therapeutic methods useful for diagnosing, treating, preventing and/or prognosing disorders related to connective tissue(s), including cancer, and therapeutic methods for treating such disorders. The invention further relates to screening methods for identifying agonists and antagonists of polynucleotides and polypeptides of the invention. The invention further relates to methods and/or compositions for inhibiting or promoting the production and/or function of the polypeptides of the invention.
Background of the Invention

[004] Connective tissues include those tissues that support and connect the various parts of the body. These tissues originate primarily in the cells of the mesodermal (middle tissue) layer of the embryo, and comprise the fibrous and elastic connective tissues, the adipose (fatty) tissues, and cartilage and bone. Connective tissues are composed of a variable structure of cells and fibers surrounded by an intercellular matrix that may be a fluid, solid, or gel, depending on the function of the particular connective tissue. White fibrous connective tissue forms most of the tendons and ligaments. Yellow elastic connective tissue forms such structures as the pads between the vertebrae and the elastic elements of the arterial walls and the trachea. Among other types of connective tissue, cartilage takes part in the formation of joints and the development of bone, and fat tissue provides a cushion for the support of such vital organs as the kidneys and stores excess food for use when needed. Lymphatic tissue and blood are also related in embryonic development to the connective tissues.

[005] Disorders that affect joints and their components--muscles, bones, cartilage, and tendons--are considered connective tissue diseases because these structures contain large amounts of connective tissue. However, many connective tissue diseases are also a type of autoimmune disease, involving immune reactions in which something triggers the immune system to react against the body's own tissues and to produce abnormal antibodies that attack these tissues (autoantibodies).

[006] Diseases of connective tissue may be due to genetic inheritance (such as, e.g., Marfan syndrome, and Ehlers-Danlos syndrome); or alternatively may not have inheritance which can be defined by gene abnormalities (such as, e.g., systemic lupus erythematosus, rheumatoid arthritis, scleroderma, polymyositis, and dermatomyositis).

[007] A connective tissue disorder developing in about 1 percent of the population, Rheumatoid arthritis, is an autoimmune disease in which joints, usually including those of the hands and feet, are symmetrically inflamed, resulting in swelling, pain, and often the eventual destruction of the joint's interior. Its exact cause isn't known, but many different factors, including genetic predisposition, may influence the autoimmune reaction. In this disease, the immune system attacks the tissue that lines and cushions joints.
Eventually, the cartilage, bone, and ligaments of the joint erode, causing scars to form within the joint. The joints deteriorate at a highly variable rate. Rheumatoid arthritis may produce a low-grade fever and occasionally an inflammation of blood vessels (vasculitis) that causes nerve damage or leg sores (ulcers). Inflammation of the membranes around the lungs (pleurisy) or the sac surrounding the heart (pericarditis) or inflammation and scarring of the lungs can lead to chest pain, difficulty in breathing, and abnormal heart function. Some people develop swollen lymph nodes, Sjogren's syndrome, or an eye inflammation. Still's disease is a variation of rheumatoid arthritis in which high fever and other generalized symptoms develop first. Many people with rheumatoid arthritis have distinctive antibodies in their blood, for example, rheumatoid factor.
Usually, the higher the level of rheumatoid factor in the blood, the more severe the rheumatoid arthritis and the poorer the prognosis.

[008] Psoriatic arthritis is a form of arthritis that occurs in people who have psoriasis of the skin or nails. The disease resembles rheumatoid arthritis but doesn't produce the antibodies characteristic of rheumatoid arthritis. Psoriasis (a skin condition causing flare-ups of red, scaly rashes and thickened, pitted nails) may precede or follow the joint inflammation. The arthritis usually affects joints of the fingers and toes, although other joints, including the hips and spine, are often affected as well. The joints may become swollen and deformed when inflammation is chronic. The skin and joint symptoms may appear and disappear together. The prognosis for psoriatic arthritis is usually better than that for rheumatoid arthritis because fewer joints are affected. Nonetheless, the joints can be severely damaged.

[009] A chronic, recurnng disorder of unknown cause, discoid lupus erythematosus, is characterized by clearly defined round, red patches on the skin. The disorder is more common in females, most often women in their 30s. The characteristic rash may persist or may come and go for years and the appearance of the patches changes over time.
Mouth sores are very common. If the disorder isn't treated, each patch gradually spreads outward.
The central area degenerates, leaving a scar. In particularly scaly areas, the plugged hair follicles dilate, leaving pits shaped like carpet tacks. Scarnng can cause widespread hair loss. The rash may be accompanied by achy joints and a decreased number of white blood cells but is only infrequently accompanied by the more severe symptoms of systemic lupus erythematosus.

[010] Systemic lupus erythematosus (lupus) is an autoimmune disease that results in episodes of inflammation in joints, tendons, and other connective tissues and organs.
Different tissues and organs become inflamed in different people, and the severity of the disease ranges from mild to debilitating, depending on the number and variety of antibodies that appear and the organs affected. About 90 percent of the people who have lupus are young women in their late teens to 30s, but children, mostly girls, and older men and women can also be affected. Occasionally, certain heart drugs (hydralazine, procainamide, and beta-blockers) can cause a lupus-like syndrome that disappears after the drug is discontinued. The number and variety of antibodies that can appear in lupus are greater than those in any other disease, and they, along with other unknown factors, determine which symptoms develop. Lupus can be quite mild, or it can be devastating, disabling, or fatal. Because symptoms vary greatly, lupus may resemble many other diseases. For example, the connective tissue of joints is commonly affected in lupus, and the arthritis that results may resemble rheumatoid arthritis. Lupus may resemble epilepsy or some psychologic disorders when the brain is affected. Although lupus can be chronic and ongoing, it usually flares up intermittently. What triggers a flare-up of lupus in people who are predisposed to it often isn't known, although sunlight seems to be one factor.
About 90 percent of people with lupus have joint inflammation, which ranges from intermittent mild aches to severe arthritis in several joints. Years of joint symptoms may precede other symptoms. Long-standing joint inflammation can lead to deformity and permanent damage to the joint and surrounding tissue, but the bone doesn't erode as it does in rheumatoid arthritis.

[011] A chronic disease of unknown cause, Scleroderma (systemic scleroisis), is characterized by degenerative changes and scarnng in the skin, joints, and internal organs and by blood vessel abnormalities. Fortunately, scleroderma is relatively rare.
Approximately 300,000 people in the United States have the condition, which is more common in women, and those between the ages of 20 and 40. This disease is not contagious, nor is it inherited. Scleroderma results from an overproduction of collagen, the main supportive (connective tissue) protein in the body. While several theories exist as to why this occurs, no definitive cause has been established. Scleroderma may occur as part of mixed connective tissue disease. The usual initial symptoms are thickening and swelling of the ends of the fingers. Raynaud's phenomenon, in which the fingers suddenly become very pale and tingle or become numb in response to cold or emotional upset, is also common. Aches and pains in several joints often accompany early symptoms.
Heartburn, difficulty in swallowing, and shortness of breath are occasionally the first symptoms of scleroderma, but usually they appear later, if the esophagus, heart, and lungs become damaged.

[012] The CREST syndrome, also called limited cutaneous sclerosis (scleroderma), is usually a less severe form of the disease that's less likely to cause serious internal organ damage. The acronym CREST applies to the following manifestations: calcinosis (calcification in the skin); Raynaud's phenomenon (a sequence of color changes in the skin in response to cold); esophageal dysfunction (such as reflux or difficulty in swallowing); sclerodactyly (hardening of the skin of the fingers or toes); and telangiectasia (dilatation of tiny blood vessels, particularly of the skin).
Skin damage is limited to the fingers. People who have the CREST syndrome can develop pulmonary hypertension, which can cause heart and respiratory failure. The course of scleroderma is variable and unpredictable. Sometimes scleroderma worsens rapidly and becomes fatal.
At other times, it affects only the skin for decades before affecting internal organs, although some damage to internal organs such as the esophagus is almost inevitable, even in the CREST syndrome. The prognosis is worst for those who have early symptoms of heart, lung, or kidney damage. No drug can stop the progression of scleroderma.

[013] A chronic inflammatory disorder characterized by excessive dryness of the eyes, mouth, and other mucous membranes, is Sjogren's syndrome. This syndrome is often associated with other symptoms more characteristic of rheumatoid arthritis or systemic lupus erythematosus (lupus). Sjogren's syndrome is thought to be an autoimmune disease, but its cause isn't known. It's less common than rheumatoid arthritis and more prevalent in women than in men. Lymphoma, a cancer of the lymphatic system, is 44 times more common in people who have Sjogren's syndrome than in the general population.
The prognosis depends on the potential of the antibodies to damage vital organs.
Rarely, pneumonia, kidney failure, or lymphoma is fatal. No cure for Sjogren's syndrome is available, but symptoms can be relieved.

[014] Polymyositis is a chronic connective tissue disease characterized by painfizl inflammation and degeneration of the muscles; dermatomyositis is polyrnyositis accompanied by skin inflammation. These diseases result in disabling muscle weakness and deterioration. The weakness typically occurs in the shoulders and hips but can affect muscles symmetrically throughout the body. Polymyositis and dermatomyositis usually occur in adults from ages 40 to 60 or in children from ages S to 15 years.
Women are twice as likely as men to develop either disease. In adults, these diseases may occur alone or as part of other connective tissue diseases. The cause is unknown. Viruses or autoimmune reactions may play a role. Cancer may also trigger the diseases-an autoimmune reaction against cancer may be directed against a substance in the muscles as well. Symptoms, which may begin during or just after an infection, include muscle weakness (particularly in the upper arms, hips, and thighs), muscle and joint pain, Raynaud's phenomenon, a rash, difficulty in swallowing, a fever, fatigue, and weight loss.
In dermatomyositis, rashes tend to appear at the same time as periods of muscle weakness and other symptoms.

[015] Mixed connective tissue disease is a collection of symptoms similar to those of several connective tissue diseases: systemic lupus erythematosus, scleroderma, polymyositis, and dermatomyositis. About 80 percent of the people who have this disease are women. It affects people from ages 5 to 80. Its cause is unknown, though an autoimmune reaction is likely. The typical symptoms are Raynaud's phenomenon (hands and feet that become white in spots and painful when chilled), joint aches or arthritis, swollen hands, muscle weakness, difficulty in swallowing, heartburn, and shortness of breath. Raynaud's phenomenon may precede other symptoms by many years.
Regardless of how this disease starts, it tends to worsen, and symptoms spread to several parts of the body. Mixed connective tissue disease damages the muscle fibers, so the muscles may feel weak and sore, especially in the shoulders and hips. Although the esophagus is usually affected, it seldom causes difficulty in swallowing and isn't painful.
Fluid may collect in or around the lungs. In some people, lung dysfunction is the most serious problem, causing shortness of breath during exertion and heart strain.
Sjogren's syndrome may develop. Over time, most people develop symptoms that are more typical of lupus or scleroderma.

[016] Relapsing polychondritis is an uncommon disorder characterized by episodes of painful, destructive inflammation of the cartilage and other connective tissues in the ears, joints, nose, voice box (larynx), windpipe (trachea), bronchi, eyes, heart valves, kidneys, and blood vessels: This disorder affects men and women equally, usually in middle age.

[017] Vasculitis is an inflammation of blood vessels. Vasculitis is not a disease but rather a disease process that occurs in a number of autoimmune connective tissue diseases, such as rheumatoid arthritis and systemic lupus erythematosus.
Vasculitis can also occur without connective tissue involvement. No one knows what triggers vasculitis in most people, but in some, hepatitis viruses are involved. Cells of the immune system, which cause inflammation, surround and infiltrate the affected blood vessels, destroying them and possibly damaging the tissues they supply. The blood vessels can become leaky or clogged; either condition disrupts blood flow to nerves, organs, and other parts of the body. Symptoms may result from direct damage to the blood vessels or damage to tissues whose blood supply is impaired. Vasculitis may be limited to veins, large arteries, small arteries, or capillaries, or it may be limited to vessels in one part of the body, such as the head, leg, or kidney. Disorders such as the Henoch-Schonlein syndrome, erythema nodosum, polyarteritis nodosa, temporal (giant cell) arteritis, and Takayasu's arteritis are characterized by vasculitis limited to blood vessels of a particular size or depth.

[018] Polyarteritis nodosa is a disease in which segments of medium-sized arteries become inflamed and damaged, reducing the blood supply to the organs they supply. This disease is often fatal if not treated adequately. It usually develops at 40 to 50 years of age but can occur at any age. Men are three times more likely than women to develop it. Its cause is unknown, but reactions to some drugs and vaccines may cause it. Viral and bacterial infections sometimes appear to trigger the inflammation, but most often no triggering event or substance can be found. The disease can be mild at first but fatal within several months, or it can develop subtly as a chronic debilitating disease.

[019] Connective tissue disorders father include, but are not limited to, Wegener's granulomatosis, an uncommon disease that often begins with an inflammation of the lining of the nose, sinuses, throat, or lungs and may progress to an inflammation of blood vessels throughout the body (generalized vasculitis) or fatal kidney disease;
Reiter's syndrome, an inflammation of the joints and tendon attachments at the joints, often accompanied by an inflammation of the eye's conjunctiva and the mucous membranes, such as those of the mouth, urinary tract, vagina, and penis, and by a distinctive rash;
Beh~et's syndrome, a chronic, relapsing inflammatory disease that can produce recurring, painful mouth sores, skin blisters, genital sores, and swollen joints; and Ankylosing spondylitis, a disease characterized by an inflammation of the spine and large joints, resulting in stiffness and pain.

[020] Connective disorders can also involve the skin. For example, cellulitis, is an acute noncontagious inflammation of the connective tissue of the skin, resulting from Staphylococcus, Streptococcus, or other bacterial infection. Keloids develop from an overgrowth of scar tissue at the site of a skin injury.

[021] Ehler Danlos syndrome is one of the inheritable connective tissues disorders along with: Marfan syndrome, pseudoxantoma elasticum, osteogenese imperfecta, chondrodysplasias, epidermolysis bullosa and Alport syndrome. It comprises a group of ten different subtypes. The main clinical manifestations are skin fragility, abnormal scar formation, excessive bruising, joint laxity and sometimes rupture of viscera and arteries.
The basic defect is in the synthesis of collagen type I and III, leading to low tensile strength of skin and artery wall.

[022] Cutis laxa is a rare, inherited or acquired connective tissue disorder in which skin becomes inelastic and hangs loosely in folds. Clinical presentation and the mode of inheritance show considerable heterogeneity; and autosomal dominant, autosomal recessive, and X-linked recessive patterns have been noted in inherited forms.
The precise cause is unknown, but may be due to abnormal elastin metabolism resulting in markedly reduced dermal elastin content.

[023] The discovery of new human connective tissue associated polynucleotides, the polypeptides encoded by them, and antibodies that immunospecifically bind these polypeptides, satisfies a need in the art by providing new compositions which are useful in the diagnosis, treatment, prevention and/or prognosis of disorders of connective tissues, including, but not limited to, rheumatoid arthritis, psoriatic arthritis, discoid lupus erythematosus, systemic lupus erythematosus, scleroderma, CREST syndrome, Sjogren's syndrome, polymyositis, dermatomyositis, mixed connective tissue disease, relapsing polychondritis, vasculitis, Henoch-Schonlein syndrome, erythema nodosum, polyarteritis nodosa, temporal (giant cell) arteritis, Takayasu's arteritis, Wegener's granulomatosis, Reiter's syndrome, Behcet's syndrome, ankylosing spondylitis, cellulitis, keloids, Ehler Danlos syndrome, Marfan syndrome, pseudoxantoma elasticum, osteogenese imperfecta, chondrodysplasias, epidermolysis bullosa, Alport syndrome, cubs laxa, genetic disorders affecting skeleton, skin and muscles; formation of excessive scar tissue;
deposition of pathological amounts of connective tissue in body organs, including kidney, intestines and heart, and in liver by liver cirrhosis, in skin by scleroderma, in lung by lung fibrosis, in bone marrow by leukemia, in blood vessels by atherosclerosis, and in joints by rheumatic diseases.
Summary of the Invention

[024] The present invention relates to novel connective tissue related polynucleotides, the polypeptides encoded by these polynucleotides herein collectively referred to as "connective tissue antigens," and antibodies that immunospecifically bind these polypeptides, and the use of such connective tissue polynucleotides, antigens, and antibodies for detecting, treating, preventing and/or prognosing disorders of the connective tissue, including, but not limited to, the presence of cancer and cancer metastases. More specifically, isolated connective tissue nucleic acid molecules are provided encoding novel connective tissue polypeptides. Novel connective tissue polypeptides and antibodies that bind to these polypeptides are provided. Also provided are vectors, host cells, and recombinant and synthetic methods for producing human connective tissue polynucleotides, polypeptides, and/or antibodies. The invention further relates to diagnostic and therapeutic methods useful for diagnosing, treating, preventing and/or prognosing disorders related to connective tissue, including cancer, and therapeutic methods for treating such disorders. The invention further relates to screening methods for identifying agonists and antagonists of polynucleotides and polypeptides of the invention. The invention further relates to methods and/or compositions for inhibiting or promoting the production and/or function of the polypeptides of the invention.
Detailed Description Tables

[025] Table 1A summarizes some of the polynucleotides encompassed by the invention (including cDNA clones related to the sequences (Clone >D NO:Z), contig sequences (contig identifier (Contig >D:) and contig nucleotide sequence identifier (SEQ
m NO:X)) and further summarizes certain characteristics of these polynucleotides and the polypeptides encoded thereby. The first column provides a unique clone identifier, "Clone m NO:Z", for a cDNA plasmid related to each connective tissue associated contig sequence disclosed in Table 1 A. The second column provides a unique contig identifier, "Contig )D:" for each of the contig sequences disclosed in Table 1A. The third column provides the sequence identifier, "SEQ >D NO:X", for each of the contig polynucleotide sequences disclosed in Table 1A. The fourth column, "ORF (From-To)", provides the location (i.e., nucleotide position numbers) within the polynucleotide sequence of SEQ
ID NO:X that delineate the preferred open reading frame (ORF) shown in the sequence listing and referenced in Table 1A as SEQ )D NO:Y (column 5). Column 6 lists residues comprising predicted epitopes contained in the polypeptides encoded by each of the preferred ORFs (SEQ ID NO:Y). Identification of potential immunogenic regions was performed according to the method of Jameson and Wolf (CABIOS, 4:181-186 (1988));
specifically, the Genetics Computer Group (GCG) implementation of this algorithm, embodied in the program PEPT1Z7ESTRUCTURE (Wisconsin Package v10.0, Genetics Computer Group (GCG), Madison, Wisc.). This method returns a measure of the probability that a given residue is found on the surface of the protein.
Regions where the antigenic index score is greater than 0.9 over at least 6 amino acids are indicated in Table 1A as "Predicted Epitopes." In particular embodiments, connective tissue associated polypeptides of the invention comprise, or alternatively consist of, one, two, three, four, five or more of the predicted epitopes described in Table 1 A. It will be appreciated that depending on the analytical criteria used to predict antigenic determinants, the exact address of the determinant may vary slightly. Column 7, "Tissue Distribution"
shows the expression profile of tissue, cells, and/or cell line libraries which express the polynucleotides of the invention. The first number in column 7 (preceding the colon), represents the tissue/cell source identifier code corresponding to the code and description provided in Table 4. Expression of these polynucleotides was not observed in the other tissues and/or cell libraries tested. For those identifier codes in which the first two letters are not "AR", the second number in column 7 (following the colon), represents the number of times a sequence corresponding to the reference polynucleotide sequence (e.g., SEQ )D NO:X) was identified in the tissue/cell source. Those tissue/cell source identifier codes in which the first two letters are "AR" designate information generated using DNA
array technology. Utilizing this technology, cDNAs were amplified by PCR and then transferred, in duplicate, onto the array. Gene expression was assayed through hybridization of first strand cDNA probes to the DNA array. cDNA probes were generated from total RNA extracted from a variety of different tissues and cell lines.
Probe synthesis was performed in the presence of 33P dCTP, using oligo(dT) to prime reverse transcription. After hybridization, high stringency washing conditions were employed to remove non-specific hybrids from the array. The remaining signal, emanating from each gene target, was measured using a Phosphorimager. Gene expression was reported as Phosphor Stimulating Luminescence (PSL) which reflects the level of phosphor signal generated from the probe hybridized to each of the gene targets represented on the array. A local background signal subtraction was performed before the total signal generated from each array was used to normalize gene expression between the different hybridizations. The value presented after "[array code]:" represents the mean of the duplicate values, following background subtraction and probe normalization. One of skill in the art could routinely use this information to identify normal and/or diseased tissues) which show a predominant expression pattern of the corresponding polynucleotide of the invention or to identify polynucleotides which show predominant and/or specific tissue and/or cell expression. Column 8, "Cytologic Band,"
provides the chromosomal location of polynucleotides corresponding to SEQ D7 NO:X.
Chromosomal location was determined by finding exact matches to EST and cDNA sequences contained in the NCBI (National Center for Biotechnology Information) UniGene database.
Given a presumptive chromosomal location, disease locus association was determined by comparison with the Morbid Map, derived from Online Mendelian Inheritance in Man (Online Mendelian Inheritance in Man, OMIMTM. McKusick-Nathans Institute for Genetic Medicine, Johns Hopkins University (Baltimore, MD) and National Center for Biotechnology Information, National Library of Medicine (Bethesda, MD) 2000.
World Wide Web URL: http://www.ncbi.nlm.nih.gov/omim~. If the putative chromosomal location of the Query overlapped with the chromosomal location of a Morbid Map entry, an OMIM identification number is provided in Table 1A, column 9 labeled "OMIM
Disease References)". A key to the OMIM reference identification numbers is provided in Table 5.

[026] Table 2 summarizes homology and features of some of the polypeptides of the invention. The first column provides a unique clone identifier, "Clone ID
NO:Z", corresponding to a cDNA disclosed in Table 1A. The second column provides the unique contig identifier, "Contig 117:" corresponding to contigs in Table 1A and allowing for correlation with the information in Table 1 A. The third column provides the sequence identifier, "SEQ ~ NO:X", for the contig polynucleotide sequences. The fourth column provides the analysis method by which the homology/identity disclosed in the row was determined. Comparisons were made between polypeptides encoded by the polynucleotides of the invention and either a non-redundant protein database (herein referred to as "NR"), or a database of protein families (herein referred to as "PFAM") as further described below. The fifth column provides a description of PFAM/NR
hits having significant matches to a polypeptide of the invention. Column six provides the accession number of the PFAM/NR hit disclosed in the fifth column. Column seven, "Score/Percent Identity", provides a quality score or the percent identity, of the hit disclosed in column five. Columns 8 and 9, "NT From" and "NT To" respectively, delineate the polynucleotides in "SEQ 117 NO:X" that encode a polypeptide having a significant match to the PFAM/NR database as disclosed in the fifth column. In specific embodiments, polypeptides of the invention comprise, or alternatively consist of, an amino acid sequence encoded by the polynucleotides in SEQ ID NO:X as delineated in columns 8 and 9, or fragments or variants thereof.

[027] Table 3 provides polynucleotide sequences that may be disclaimed according to certain embodiments of the invention. The first column provides a unique clone identifier, "Clone >D NO:Z", for a cDNA clone related to connective tissue associated contig sequences disclosed in Table 1A. The second column provides the sequence identifier, "SEQ ID NO:X", for contig polynucleotide sequences disclosed in Table 1A.
The third column provides the unique contig identifier, "Contig ID", for contigs disclosed in Table 1A. The fourth column provides a unique integer 'a' where 'a' is any integer between 1 and the final nucleotide minus 15 of SEQ ID NO:X, represented as "Range of a", and the fifth column provides a unique integer 'b' where 'b' is any integer between 15 and the final nucleotide of SEQ ID NO:X, represented as "Range of b", where both a and b correspond to the positions of nucleotide residues shown in SEQ >D NO:X, and where b is greater than or equal to a + 14. For each of the polynucleotides shown as SEQ ID
NO:X, the uniquely defined integers can be substituted into the general formula of a-b, and used to describe polynucleotides which may be preferably excluded from the invention. In certain embodiments, preferably excluded from the polynucleotides of the invention (including polynucleotide fragments and variants as described herein and diagnostic and/or therapeutic uses based on these polynucleotides) are at least one, two, three, four, five, ten, or more of the polynucleotide sequences) having the accession numbers) disclosed in the sixth column of this Table (including for example, published sequence in connection with a particular BAC clone). In further embodiments, preferably excluded from the invention are the specific polynucleotide sequences) contained in the clones corresponding to at least one, two, three, four, five, ten, or more of the available material having the accession numbers identified in the sixth column of this Table (including for example, the actual sequence contained in an identified BAC
clone).

[028] Table 4 provides a key to the tissue/cell source identifier code disclosed in Table 1A, column 7. Column 1 provides the key to the tissue/cell source identifier code disclosed in Table 1A, Column 7. Columns 2-5 provide a description of the tissue or cell source. Codes corresponding to diseased tissues are indicated in column 6 with the word "disease". The use of the word "disease" in column 6 is non-limiting. The tissue or cell source may be specific (e.g. a neoplasm), or may be disease-associated (e.g., a tissue sample from a normal portion of a diseased organ). Furthermore, tissues and/or cells lacking the "disease" designation may still be derived from sources directly or indirectly involved in a disease state or disorder, and therefore may have a further utility in that disease state or disorder. In numerous cases where the tissue/cell source is a library, column 7 identifies the vector used to generate the library.

[029] Table 5 provides a key to the OM1MTM reference identification numbers disclosed in Table 1A, column 9. OMIM reference identification numbers (Column 1) were derived from Online Mendelian Inheritance in Man (Online Mendelian Inheritance in Man, OMIMTM. McKusick-Nathans Institute for Genetic Medicine, Johns Hopkins University (Baltimore, MD) and National Center for Biotechnology Information, National Library of Medicine, (Bethesda, MD) 2000. World Wide Web URL:
http://www.ncbi.nlm.nih.gov/omim~. Column 2 provides diseases associated with the cytologic band disclosed in Table 1A, column 8, as determined from the Morbid Map database.

[030] Table 6 summarizes ATCC Deposits, Deposit dates, and ATCC designation numbers of deposits made with the ATCC in connection with the present application.

[031] Table 7 shows the cDNA libraries sequenced, tissue source description, vector information and ATCC designation numbers relating to these cDNA libraries.

[032] Table 8 provides a physical characterization of clones encompassed by the invention. The first column provides the unique clone identifier, "Clone ID
NO:Z", for certain cDNA clones of the invention, as described in Table 1A. The second column provides the size of the cDNA insert contained in the corresponding cDNA
clone.

Definitions

[033] The following definitions are provided to facilitate understanding of certain terms used throughout this specification.

[034] In the present invention, "isolated" refers to material removed from its original environment (e.g., the natural environment if it is naturally occurring), and thus is altered "by the hand of man" from its natural state. For example, an isolated polynucleotide could be part of a vector or a composition of matter, or could be contained within a cell, and still be "isolated" because that vector, composition of matter, or particular cell is not the original environment of the polynucleotide. The term "isolated" does not refer to genomic or cDNA libraries, whole cell total or mRNA preparations, genomic DNA
preparations (including those separated by electrophoresis and transferred onto blots), sheared whole cell genomic DNA preparations or other compositions where the art demonstrates no distinguishing features of the polynucleotide sequences of the present invention.

[035] As used herein, a "polynucleotide" refers to a molecule having a nucleic acid sequence encoding SEQ ID NO:Y or a fragment or variant thereof, a nucleic acid sequence contained in SEQ ID NO:X (as described in column 3 of Table 1A) or the complement thereof, a cDNA sequence contained in Clone ID NO:Z (as described in column 1 of Table 1A and contained within a library deposited with the ATCC);
a nucleotide sequence encoding the polypeptide encoded by a nucleotide sequence in SEQ
)D NO:B as defined in column 6 of Table 1B or a fragment or variant thereof;
or a nucleotide coding sequence in SEQ ID NO:B as defined in column 6 of Table 1B
or the complement thereof. For example, the polynucleotide can contain the nucleotide sequence of the full length cDNA sequence, including the 5' and 3' untranslated sequences, the coding region, as well as fragments, epitopes, domains, and variants of the nucleic acid sequence. Moreover, as used herein, a "polypeptide" refers to a molecule having an amino acid sequence encoded by a polynucleotide of the invention as broadly defined (obviously excluding poly-Phenylalanine or poly-Lysine peptide sequences which result from translation of a polyA tail of a sequence corresponding to a cDNA).

[036] As used herein, a "connective tissue antigen" refers collectively to any polynucleotide disclosed herein (e.g., a nucleic acid sequence contained in SEQ ID NO:X
or the complement therof, or cDNA sequence contained in Clone >D NO:Z, or a nucleotide sequence encoding the polypeptide encoded by a nucleotide sequence in SEQ

>D NO:B as defined in column 6 of Table 1 B, or a nucleotide coding sequence in SEQ )D
NO:B as defined in column 6 of Table 1B or the complement thereof and fragments or variants thereof as described herein) or any polypeptide disclosed herein (e.g., an amino acid sequence contained in SEQ )D NO:Y, an amino acid sequence encoded by SEQ
>D
NO:X, or the complement thereof, an amino acid sequence encoded by the cDNA
sequence contained in Clone >D NO:Z, an amino acid sequence encoded by SEQ )D
NO:B, or the complement thereof, and fragments or variants thereof as described herein).
These connective tissue antigens have been determined to be predominantly expressed in connective tissues, including normal or diseased tissues (as shown in Table 1A
column 7 and Table 4).

[037] In the present invention, "SEQ >D NO:X" was often generated by overlapping sequences contained in multiple clones (contig analysis). A representative clone containing all or most of the sequence for SEQ >D NO:X is deposited at Human Genome Sciences, Inc. (HGS) in a catalogued and archived library. As shown, for example, in column 1 of Table 1A, each clone is identified by a cDNA Clone m (identifier generally referred to herein as Clone )D NO:Z). Each Clone >D is unique to an individual clone and the Clone )D is all the information needed to retrieve a given clone from the HGS library.
Furthermore, certain clones disclosed in this application have been deposited with the ATCC on October 5, 2000, having the ATCC designation numbers PTA 2574 and PTA
2575; and on January S, 2001, having the depositor reference numbers TS-l, TS-2, AC-l, and AC-2. In addition to the individual cDNA clone deposits, most of the cDNA
libraries from which the clones were derived were deposited at the American Type Culture Collection (hereinafter "ATCC"). Table 7 provides a list of the deposited cDNA
libraries. One can use the Clone >D NO:Z to determine the library source by reference to Tables 6 and 7. Table 7 lists the deposited cDNA libraries by name and links each library to an ATCC Deposit. Library names contain four characters, for example, "HTWE." The name of a cDNA clone (Clone >D NO:Z) isolated from that library begins with the same four characters, for example "HTWEP07". As mentioned below, Table 1A
correlates the Clone >D NO:Z names with SEQ >D NO:X. Thus, starting with an SEQ >I7 NO:X, one can use Tables 1A, 6 and 7 to determine the corresponding Clone >D NO:Z, which library it came from and which ATCC deposit the library is contained in. Furthermore, it is possible to retrieve a given cDNA clone from the source library by techniques known in the art and described elsewhere herein. The ATCC is located at 10801 University Boulevard, Manassas, Virginia 20110-2209, USA. The ATCC deposits were made pursuant to the terms of the Budapest Treaty on the international recognition of the deposit of microorganisms for the purposes of patent procedure.

[038] In specific embodiments, the polynucleotides of the invention are at least 15, at least 30, at least 50, at least 100, at least 125, at least 500, or at least 1000 continuous nucleotides but are less than or equal to 300 kb, 200 kb, 100 kb, 50 kb, 15 kb, 10 kb, 7.5 kb, 5 kb, 2.5 kb, 2.0 kb, or 1 kb, in length. In a further embodiment, polynucleotides of the invention comprise a portion of the coding sequences, as disclosed herein, but do not comprise all or a portion of any intron. In another embodiment, the polynucleotides comprising coding sequences do not contain coding sequences of a genomic flanking gene (i.e., 5' or 3' to the gene of interest in the genome). In other embodiments, the polynucleotides of the invention do not contain the coding sequence of more than 1000, 500, 250, 100, 50, 25, 20, 15, 10, 5, 4, 3, 2, or 1 genomic flanking gene(s).

[039] A "polynucleotide" of the present invention also includes those polynucleotides capable of hybridizing, under stringent hybridization conditions, to sequences contained in SEQ >D NO:X, or the complement thereof (e.g., the complement of any one, two, three, four, or more of the polynucleotide fragments described herein), the polynucleotide sequence delineated in columns 8 and 9 of Table 2 or the complement thereof, and/or cDNA sequences contained in Clone >D NO:Z (e.g., the complement of any one, two, three, four, or more of the polynucleotide fragments, or the cDNA clone within the pool of cDNA clones deposited with the ATCC, described herein) and/or the polynucleotide sequence delineated in column 6 of Table 1B or the complement thereof.
"Stringent hybridization conditions" refers to an overnight incubation at 42 degree C in a solution comprising 50% formamide, 5x SSC (750 mM NaCI, 75 mM trisodium citrate), 50 mM
sodium phosphate (pH 7.6), 5x Denhardt's solution, 10% dextran sulfate, and 20 ~g/ml denatured, sheared salmon sperm DNA, followed by washing the filters in O.lx SSC at about 65 degree C.

[040] Also contemplated are nucleic acid molecules that hybridize to the polynucleotides of the present invention at lower stringency hybridization conditions.
Changes in the stringency of hybridization and signal detection are primarily accomplished through the manipulation of formamide concentration (lower percentages of formamide result in lowered stringency), salt conditions, or temperature.
For example, lower stringency conditions include an overnight incubation at 37 degree C in a solution comprising 6X SSPE (20X SSPE = 3M NaCI; 0.2M NaH2P04; 0.02M EDTA, pH 7.4), 0.5% SDS, 30% formamide, 100 ug/ml salmon sperm blocking DNA; followed by washes at 50 degree C with 1XSSPE, 0.1% SDS. In addition, to achieve even lower stringency, washes performed following stringent hybridization can be done at higher salt concentrations (e.g. 5X SSC).

[041] Note that variations in the above conditions may be accomplished through the inclusion and/or substitution of alternate blocking reagents used to suppress background in hybridization experiments. Typical blocking reagents include Denhardt's reagent, BLOTTO, heparin, denatured salmon sperm DNA, and commercially available proprietary formulations. The inclusion of specific blocking reagents may require modification of the hybridization conditions described above, due to problems with compatibility.

[042] Of course, a polynucleotide which hybridizes only to polyA+ sequences (such as any 3' terminal polyA+ tract of a cDNA shown in the sequence listing), or to a complementary stretch of T (or U) residues, would not be included in the definition of "polynucleotide," since such a polynucleotide would hybridize to any nucleic acid molecule containing a poly (A) stretch or the complement thereof (e.g., practically any double-stranded cDNA clone generated using oligo dT as a primer).

[043] The polynucleotide of the present invention can be composed of any polyribonucleotide or polydeoxribonucleotide, which may be unmodified RNA or DNA
or modified RNA or DNA. For example, polynucleotides can be composed of single-and double-stranded DNA, DNA that is a mixture of single- and double-stranded regions, single- and double-stranded RNA, and RNA that is mixture of single- and double-stranded regions, hybrid molecules comprising DNA and RNA that may be single-stranded or, more typically, double-stranded or a mixture of single- and double-stranded regions. In addition, the polynucleotide can be composed of triple-stranded regions comprising RNA or DNA or both RNA and DNA. A polynucleotide may also contain one or more modified bases or DNA or RNA backbones modified for stability or for other reasons. "Modified" bases include, for example, tritylated bases and unusual bases such as inosine. A variety of modifications can be made to DNA and RNA; thus, "polynucleotide" embraces chemically, enzymatically, or metabolically modified forms.

[044] The polypeptide of the present invention can be composed of amino acids joined to each other by peptide bonds or modified peptide bonds, i.e., peptide isosteres, and may contain amino acids other than the 20 gene-encoded amino acids. The polypeptides may be modified by either natural processes, such as posttranslational processing, or by chemical modification techniques which are well known in the art. Such modifications are well described in basic texts and in more detailed monographs, as well as in a voluminous research literature. Modifications can occur anywhere in a polypeptide, including the peptide backbone, the amino acid side-chains and the amino or carboxyl termini. It will be appreciated that the same type of modification may be present in the same or varying degrees at several sites in a given polypeptide. Also, a given polypeptide may contain many types of modifications. Polypeptides may be branched, for example, as a result of ubiquitination, and they may be cyclic, with or without branching. Cyclic, branched, and branched cyclic polypeptides may result from posttranslation natural processes or may be made by synthetic methods. Modifications include acetylation, acylation, ADP-ribosylation, amidation, covalent attachment of flavin, covalent attachment of a heme moiety, covalent attachment of a nucleotide or nucleotide derivative, covalent attachment of a lipid or lipid derivative, covalent attachment of phosphotidylinositol, cross-linking, cyclization, disulfide bond formation, demethylation, formation of covalent cross-links, formation of cysteine, formation of pyroglutamate, formylation, gamma-carboxylation, glycosylation, GPI anchor formation, hydroxylation, iodination, methylation, myristoylation, oxidation, pegylation, proteolytic processing, phosphorylation, prenylation, racemization, selenoylation, sulfation, transfer-RNA
mediated addition of amino acids to proteins such as arginylation, and ubiquitination.
(See, for instance, PROTEINS - STRUCTURE AND MOLECULAR PROPERTIES, 2nd Ed., T. E. Creighton, W. H. Freeman and Company, New York (1993);
POSTTRANSLATIONAL COVALENT MODIFICATION OF PROTEINS, B. C.
Johnson, Ed., Academic Press, New York, pgs. 1-12 (1983); Seifter et al., Meth.
Enzymol. 182:626-646 (1990); Rattan et al., Ann. N.Y. Acad. Sci. 663:48-62 (1992).)

[045] "SEQ ID NO:X" refers to a polynucleotide sequence described, for example, in Tables 1A or 2, while "SEQ ID NO:Y" refers to a polypeptide sequence described in column 5 of Table 1A. SEQ ID NO:X is identified by an integer specified in column 3 of Table 1A. The polypeptide sequence SEQ ID NO:Y is a translated open reading frame (ORF) encoded by polynucleotide SEQ ID NO:X. "Clone ID NO:Z" refers to a cDNA
clone described in column 1 of Table 1A.

[046] "A polypeptide having biological activity" refers to a polypeptide exhibiting activity similar to, but not necessarily identical to, an activity of a polypeptide of the present invention, including mature forms, as measured in a particular biological assay, with or without dose dependency. In the case where dose dependency does exist, it need not be identical to that of the polypeptide, but rather substantially similar to the dose-dependence in a given activity as compared to the polypeptide of the present invention (i.e., the candidate polypeptide will exhibit greater activity or not more than about 25-fold less and, preferably, not more than about tenfold less activity, and most preferably, not more than about three-fold less activity relative to the polypeptide of the present invention).

[047] Table 1 A summarizes some of the polynucleotides encompassed by the invention (including contig sequences (SEQ ID NO:X) and clones (Clone m NO:Z) and further summarizes certain characteristics of these polynucleotides and the polypeptides encoded thereby.
Polynucleotides and Polypeptides T A Rr Ti' 1 A

H
a~

a L

w U
.
, .
bD '~

o a o ~e ''~
~4 U

:

o w o0 ~ ~' ~

N M O
o ;? ~poa x V

o = '~ ~ b -d ..
~n~' N N N M N N M N ~ N
j N ~

~

~ cG ap~ O M N O O O ~
y ~ .. ' m s. ~ ~ a1 N o0 00 o0N N N N ~p N
y v~ ~
~ ~

M M O N N N
O
D

C/~~ x C/~~ C/~ C/~x x x x r-~ x .

M
O ~p O M N ~ ~ '-. oo ~O Q1 ~ ~"' M .~ M ~ M ~ ~ M i~ N
N V7 ~ M
~

i l~ V7 ~
t; ~

~ ~ ~~ ~ d ~ o H ~ , , ~
v~ (~ H ~ o fs, ~-1 d per C7 ts ~ d , .., '-~

o 0 0 0., o o ~ o 0 0 0 o o 0 ~

v ~ ~ ~, ,~ ~ ~ ~ p ~, ~. 0 .r N ~ ~-. y.., ~ O N ~,,~ O N O
.-, O o0 ~.-~ v~ ~D
,~ ~ l' .~

N .~ ,-, N ~ N M Q\ r' N
N M pp N M
~ .-~ M

~ , _>, ,., , s.,. ~,N y..,bA ~ v~
~ >, cn ~'w p ~
s.. ~ ~ ~

~ N N , s..
C7 C7 ~n C7 v~, C7~ N ~ ~., U ~
C7 v~ ,-, p... d U
~l v~
a O' ~ O .~ .~N M ~r ~n~ ~ o00, N O
A

O O ooO O O O O O O O o0 o ~

~~ ~ ~

..
y D ~ M O N N ~ ~ o0~ N c~~

N ~' ~ O V' ~.,~~,~M M ~ V7 , M N
,~M N N i ' ' ~ ' ' i ~ ~D

i ~ i i _ ,~M ~ M ~!1 ~ ~ \O~ M l~ ~ 00 G~ N ,~,~ .-. .-.

'r .--~N N M ~ ~ ~DI~00 O~O M ~ N

~ N ~ N N

~z Gp t~vW D ~tv~ O ~h~OO M ~O ~ t~ N

~ l~ N ~ O~a1 ~ d' (~ N I~

V1M M 0000 I~ ~ ~ ~ t~00 A N ~O ~Ov~N ~n v~,-,~ ~nO~ M ~t I~

V7N N l~N ~ ~ M O~ O O o0 t~
U ~ ~

~ o, a,~nt 00 0ot ~ ~ ~ o0 ~'M pp~ ~ O M ~ M M N
N

N I~ ~ d E-H 'J'.' r~
~ ~ ~ ~ U U

0 ~ V U ~ ~ U

z d d d d x x x x x x x x x x x O N

d' O O

x x b b N N N N N N N N -~ N N N N N N M N

N N N N N N N N o0 N N N N N N N N

~ V' d'~ ~ ~ ~ ~ ~n ~ ~ d' ~' Wit'~t ~''~t O O O O O O O O O O O O O O O O O

x x x x x x x x x x x x x x x x x . .

M
N ~ I~ ~ M ,-. D ~ ~ M ,-.~~ v'i.~
.~ ,-, Ov 00 _ ' ~
~

~, ~ ~ '~ ~ ~' pp v~ ~ ~ ~ ~
~, '~ ~ a ~ ~ ~ d-0 c~
~ ' ~~ ; ~
c~~ ~ a o~

> c~ ~ ~ , ~ _ ~
~

o E'~, o p o p U C7 t 0 r o ~ o o o o o o o 0 .~ o o N , ~ p ,~01 pp~,N p ~ p p v~
~ N o M ~p00 ~' ~' ~''' ~... ~ 00 ~

M

N ;' ~ Q\ M ~p ,~~ N M ,~ ~ ~ .~ .~M
~O i ,~ .~ ~ i i N N , N
i ~ ~ i b0 N i ~ v~ by i i i ~ i ~ , by i ~ by b-0 p ~ ~ ~ N O ~ ~ cnw ~ ~ p ~ ~
a d > ~ ~ ~ ~ H U a ~
a x ~ w C7 ~
a C7 ~ , H H

N M ~ ~WO t~ 00Q1O .--~N M ~ ~n'O l~00 ,~r,~--~.-n .~,~N N N N N N N N N

~nv~ ~n~n~n ~n v~~n~ v~~nv~ ~n ~nv~ ~n~n 00 N N M N ~ ~O M O
N M

~ l~~O o0 I~N M o0~ v~ 00 I~t~ O ~O

N .-r M ~ V7 d' N M ~ ~ M ~O M ~ 00 N M

--~v~ ~ ~ ~ ~ ~ ~ ~ ~ m -' V7M ~ i i i ~ ,~00 M ~

O~~ N N ~ ~Ooo O M ~ N O o0 ~ ~n M .~ .~ M

M ~ ~n~Ol~ 00 01O ~ N M d' v7 ~Ol~ 00Q\

N N N N N N N M M M M M M M M M M

01~O l~~n~ O O ~ oo ~ N ~ N ~D~O ~nO

v7O 01O ~O ~O ~ ~nN O V1l~ O ~ M ~OM

v7M ~ l~v7 M 00(~O M v700 O M N ~ 00 ~pV1 I~~ O ~D O~~ l~ I~M ~O O~ M l~ O ~

~ \O ~ M l~ ~O ~ O v7 00l~00 ~ M N l~l~

l~00 ~ l~01 ~D I~l~I~ l~I~I~ 00 t~I~ Q1l0 00 ~ l~l~ 00 ~ 00V p 00Ov ~ 'O00 M ,~ ~OM l~ l0(~00 N V7V7 M M

o o o o a x ~ x r~ U w w a x c~ x x U U U U U U U U A ~1f.~~1 ~ w r~.~ w w x x x x x x x x x x x x x x x x x ~

~O N N o ,~ ~' x a x x -d .b ~ -d Mo o .

a x r..
.-.

N .~ ~ M N N N N N N ~ N N N N N ~
M

I~ p O l~M N N N N N N N N N N N N
~

N ~p O N ~ 01O~d\O~ 0101 d101 O1 01 d1d1 O O

p .o ~ ~ ~nv,~n~n ~n~n ~n~n ~n ~n ~n~n a a 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 ,b .~

x H x x x x x x x x x x x x x x x ~

~ ~ ~; ~

c~j ij '~ ~ pp(~
N ~

., c ~ pp '~ N ,'~~
~' ~ ~, ~ ~p ~ ~ ~, N ~ ~ c N

, ~ _ ' d v~
p ~ ~' Cl~ ~ ' ci~
G~
p 'U H
~ , ~ d P O
H ~ a O

p p., O p a O O -i O p p O ~ p p ~

p ~, p p p p p ~..., ~, N ,~, ~' '-' ,._,l~
~ 00 M O 00 ,-, pp Q

N ~ ~ ~ .~ 00 N Wit' ~ N ~ 00 M '!~ ~O
~ I~ ,-, ~ ' ~ ciW N , , ' ~, ~
' ~

v w w C7 d W d ~'1 a ci~~
i a~ ~ d ' ~ o ~ ~. ., r a 01 O ~ N M d'~ ~Ol~ oo~ O ~ N M ~t~n N M M M M M M M M M M d'~ ~ ~ ~t<f' N N ~ ~ ~ ~ ~ ~ ~ N N ~ ~ N

M ~ ~OO N V~.~01 ~n~ ~ N O~ ~ ~DM

N 00 ~O d'M N ~O~ ~ ~ Qv o0~ M 00 N N

.-rN , , ~'N , , ~ ~ , N , ~O
' ' , , , O ~ , , ~ N , , r-, , M , M ~ .-i O ~ ~ M M N ~ M ~ M

N M N ~

O ~ N M '~t~ ~Ol~00 01O ~ N M d' V1~O

~t ~ ~ ~ ~'~ ~ ~ ~ ~ V~ v~~n ~n v7 v~V~

lp ,~ 00 ~ ~ ~O~ M V7 (~M 01O~ ~ M M <h ~O ~O l~N Ovo0~Ov1 ~ I~ v~I~ l~ ~D ~ O

l~ I~ I~ O~O~01N I~N N N ~O~ ~ O~ ~ \O

v1 01 ~ 00.~tnM ~ V1 M M N M V7 O~ O <Y

O N ~ ~ d'O M ~ O ~ M ~nN -~ ~n -,~O

I~ t~ 00t~t~O~t~t~ t~a1 t~a1 a1 I~ t~00 M N M ~O~OO ~ ~ M ~ M I~O O M ~ 0 ~ ~ ~ ~ ~ ~ ~' w v~ rn x ~C U

C C C7 C ~
.7 7 7 x x x x x x x x x x x x x x x x x N

M

O N

,_, v~ v~ ,~ O

o~ x x p x O ~O "~ .b h O ~

a N N N N ~ .~ .-, N N ~ ~ N N N N
i ~
;

N N N p . ,- d' ~ M M M M V7 ~ ~ V7 ,- ~ ' ' l~

01 01Oyp o0 Q1 01 01 ~ Q\ 01 ~ ~ ctd ~ ~
p V'1V1~ O ~ O ~ ~ V7 V7V~ V~ M M M M
y~

O O O O O p O O O O O O O O O O
~ O

x x x H xpp x x x x x x x x x x . . .

M
N
~ ~

, ~ ~

~', ~ ~' , ~,' N r ~. ~ ~ bA N
~ i C/~ ~ _ ~ _ ~ ~' ~
'-' ~ ~ ~ ~ P (~
Vl H C7 a O . -~ ~

~ ~~~ ~ ~ ~~ O~ O

_O_ _O _p v~ _O _O _O
~

N .-~ r' N ~, 00 ~-.
r' d. ,~ ~O N

_>, a~ N ~ p _cd ~ v~
s: _~ by _~ _cii s~.
~

H H C7 x ~ H A, ~ x U ci~ C7 ~ d d v~

-~

l~0001O ~ N M ~ ~ ~D I~ 00 01O

~t~n v~ ~n ~n ~n~n ~n ~n v~ v~~o ~n ~n~n~n~n ~n ~n ~ v~v~ v~ ~n ~n ~ v~

0o a1 ~n O ,~ ~ ~n N O ~

~D N ~ O l~ 00 l~ -~ _,I~ ~ 00 N O t~

V7 M N O~M ~--~~ ~ , M , ~ ~ M M
~, ~,~

.-wm "'~ ~ y 1 ~ ~' ~ , ~ i i i i ~ ~ i ~ ~ ~ ~

N N M ~ M N O N
M N ~

M N M ~ N ~ N

l~ 00a1O -~ N M ~ ~WO I~ 00 Q\ O

~D

N I~M ~tO~ l~ t~ O N ~O Ov d' ~ o0a\

I~ I~~ N 01 N N N N l~ O N ~n o000 01 N O l~.~ 00 0o O O l~ ~O ~O

l~ ~ D 00I~ O M O~ 00O ~ M M I~M

00 N Q\O M M .-~ lp ~ M ~D l~ l~ N I~

~O l~~O~1v'~ 01 a1 a\ Q\a\ O~ O~ 01 01O~

N N ~ ~ ~ ,~ M N ~ M ~ N O

0 o O

~

x x x x x ~ x x x x x x N N N N N N N N N N N N N N N N N N N

Vo ~n Vo~n~n ~n ~n~O~O ~O,--~.--~.--~,-.-~~--~.-r'.-r,--~ .--s M M M M M M M O O O O O O O O O O O O

O O O O O O O O O O O O O O O O O O O

x x x x x x x x x x x x x x x x x x x N ~ M ~ N N N
' ~ M

~ I ~j n t~ ~
~ ' .~ ~ I~ M
~ ~
~

~' ' ~ v~ y_, _W_,~ ~ W_, ~
b0 ~ ~ ~ _~, ~.

~" ~" ~
[~ C/7 ~ ~ ~ O ""'~ '> ~' '~' ~ O x O O O O O

O O p p O p O O O ~ ~ O O O O
~' ' ~ N ~ N ~ i-. .N ~ N t'' N ~D M .~ 00 .~ .~-i ~'\O M ~" ~ ,~ t~ .--i ~
V7 ,~ ~ ~ ,~ N .~ N
M M

i i i i i i , i i i i i i N ~ i i i i ,~ J' i ~
~ ~ ~ ~ ~ N ~ ~ N
' ~ ~ ~

~a~ ~H a ~~ , ~wc~ a~ ~ ~ c~c~c7c~~
ac7 N M ~ vW O l~00Ov O ~ N M ~ v~~OI~00 a\

v~ ~n ~n~nv~ ~n V~v-~~n ~n~n ~ v~~ ~n~n~ ~n 00 ~ 0~0M O ~ N o0~ N ~ ~j M ~' O ~ O N
lp V7V'100 V7 M M M N ~ ~ M

N ~ , , ~ , i , , i ~, N N ~ M N M M M

~ N O ' O O Q\O_ ~ ~ ~ , ~ ' M N ~O
N ' ~ ~ ~ ~,N M ~ , r..
~

~ M N M N N ~ ~ l n ~ n n N M ~t~n~O l~ ooO~O ~ N M ~ ~n~Ol~ooOv O

00 00 ~Ol~M 01 V7.~Q~ lp~ ~ 00V7N 00~ M V'1 O ~O l~O~O~ O l~v~~ ~ ~n o0N ~ O ~ N M M

l~ ~D ~O~ -~ I~ N ~ ~ ~'~t ~ 00M ~ M ~ON v'1 M M ~'41 M N l~t~ I~V1 ~ N ~ ~ O O r1 t~ l~t~N l~ M M M M O O M N ~ M ~ I~ I~

01 ~ 01V7 01 l~~ V7 ~ l~ ~ Q\N 01N 01Q\ 01 ppM ~ M N N N o0l~ ~ N M t~
i N V1 [~01~ ~ ~ 01N 00M M O 'fit'O.~p ~ M
c7 a ~ d d ~ ~ d ~ d a p ~
d ~ w w w w w w w w w x x x x x x x x x x x x ~ x x x x x b 0o a1 c~i o ~

x x x b b -~ p ~ p .
a o x r' M N ~ N N ~-~ N N a ~j ~ N N
,~ ~O 00 00 ~O
00 00 00 00 d' ~ 00 01 Q1 ~p O v~ O 01 01 O O O ~n v~ ~n ~n ~ ~ o pp ~ x d' O O O O O O O O O pp ~ O .b O O
x x xx .xxx .xxpp a ~x x .--. ~ c~
I~ O~ wj _~ ~ ,-i N 01 ~ ~-~ ~!j N
p ~ N ~ ~ N M ~ tip ~ ~' M ,~_, N ~ ~ '' ~ ~ ~ ~, ~1 d '.~'' d U a; ~ ~' ~ ~ ~ ~ , j C7 ~ o ~ ~ H H
C7 ~ C7 d ~ ~ ~ H
O O ~ O O d1 O O ~ O O O ~ N ~D p 00 ,-, ~' 01 O ''-' .~ ~ l~ O ~-' r-' M ~ N O M O ''."' Y y ~p I~
\O "r ~ ~ N N M I~ .r .~ ~ M ~ .~ M 01 ~ '~ .~ r., M
s?, c~ ~ >, ~ a. ~ >, ~ ~ ~ ~ o on do ~ a, on p p _~
a> a~ ~ .~ v~ .~ ,~ ~ ~ .~ ~.~ a~ '~ '~' ~ H d ~ ~ a, C7 a v~ d d d t7 t7 d d C7 c7 a H ~ w O ~~ N M '~ ~n ~O l~ oo Ov O

~mn ~ ~n ~n ~n ~mn v~ ~n ~n ~n M ~ ~ ~ '~ OM ~ due' O d' ~ U
M V7 ,~ M
i i ~ ~ M i i ~ ~ i M ~n N '-' .--r N ~ ~ M
N N N ~
N M ~ 'n ~O l~ oo O~ O O O
Q\ 01 01 Ov 01 01 Q1 01 01 .~ .~ '-r M O M V~ ~O I~ M N N 00 ~O O
v0 I~ 00 00 N O v~ 01 l~ '~t ~n ~O
l~ N ~D O ~D ~-~ a1 ~ M oo N M
Q1 ~ WO v7 ~ ~p 01 N ~D O M
00 I~ M ~O N ~n ~ l~ O ~D ~n t~
I~ 01 O\ l~ ~ 00 01 00 l~ d' I~ O~
00 O O in lN0 N O o~0 N ~ ~'' ~ ~ M 00 01 p~q pwq p~q paq ~ ~ ~ w rx V U U
w ww w~~
x ~ ~~ x~x xx x 0 0 0 0 0 0 0 o v-i a; ,--~ .-r .-~ .~ o ~
V; .-~ 0 0 o wi a;

O O O O O o0 v1 O ~ O ~D ~O ~O N ~ 00 M O
O O l~

N ~ -a ~ ~- I~ O l~ O O ~ d' ~ l~ 00 ~ N 01 ~ l~ l~ O M

N vp .~ ,~ .~ M I~ M ~ ~ 00 00 0o O O ~ M
a\ N 01 01 O O

O O M M M M d' v1 ~O ~D ~D ~O \O 00 00 ~ Q\
O M ~ N O O

M

..r 'b O

a~

N M N

. d' r~

pp p N N N ~' O

N M I~ M I~
00 ~
~

i i i H

O O O ~ O O
O

N r"' ~ ~O 00 ~D
~ l~

M N ,-, N r.r ~O ~

~ ap ap d0 ~
~'' ~

N M

d1 d1 a1 m v~ ~n N

O N
M

M

O O O

M N I~

~n N o0 ~D O

N o0 N

~n O

U

N o0 OO
01 00 ~ ~t N " ~ ~ V1 ~ 41 ,~ ~ Qv ~ Q1 rr ~--y\ l~ I~ O O Wit' ~O O O
°O a a x x ~ox ~ x p ~ a 'C~ 'b '~ b p a 'd M ~b ~xa ~ ~ ~ ~ x'z' ~
N N ~--~ ~~ N r; ~ ~ N ~ ~' ,.~ N N N
_ _ V7 \O ~ M ~ ~ 00 pv 0~1 N ~j N 0~1 O O ~ 0~1 0~1 0~1 O ~ ~ O O O ~ p O O O
x~ x~x~;x~x ~ax~x~M~xxx O N ~ v'i N y0 Qv d' dwD M N d' M v~
s~ l ~ N O ~ ~ s' 00 ~ O ci~ N
a ~ v~ ~' ~ as ~~ C7 ~n v~ o o C7 0 ~ o ~ C7 ~' o ° ~' o ~' ° o 0 0 0 0 00 0, ~ ,~ ~O ~' oo ~ ~ ~ O ~n .-r .~ M M .-, M ~ ~' ~ M M ,~ N ~
~ s~' p ~ ~1. bA ~, ~, s~' p ~ ap C7 d C7 E-1 Ii. Q d ~ C7 C7 H p , C7 ate...
V7 M ~ l~ 00 01 O ~ N M ~ V~ lp l~
01 00 01 01 01 Q\ O O O O O O O O
U ~ N
N ~ O ~n l~ O oo N N '~ N O
V~ .~ ~ M ~ ~ M ~ v~ V~ M N I~ N
~ N M ~ N N M N ~ N
v7 O O1 ~ ~ ~ ~O
M M ~ '~ N M '~ 01 M M N d' '~t I~ o0 Q1 O ~-~ N M ~ ~ ~D t~ o0 O 41 O O O .-r .~ .~ .~ .-. .--i ~ .-w ~
.~ vmn .~ 00 M ~~ N O ~O O~ WO
01 01 M ~n Q1 O ~ ~ N l~ 00 ~O O
01 O N M ~O 00 M l~ l~ N M ~D
N N M O M O l~ O o ~n ~ ~n ~ l~ l~ M ~ l~ 00 00 ~n I~
O~ 01 l~ 00 00 ~D Q\ 01 l~ ~ 00 l~ 01 D\
ONp ~ O ~ N ~ ~ ~ ~ 0~0 M N
U ~ U
x x x x x x xx xxx b a ri c d O ~ '-' ~ ~' ~ r' .-i .

Q\

b O ~ p ~ O

x xo xpoo a a x a a N ~ .~ V7N M N M ,~ N N rrj N
, ri .-r (wj .--~
~ ~--i ~ ~ .-r .--s '--i 0 Npp ~ ~ 1 1 ~ O~OV ~ ~ ~I~oo~l~

~ N ~ 'ch~i'~t~ O d1 '~ ~'l~ ~ 01 ~ M 01 l~

O O O ~ O O O O O O ~ O O p ~ ~ ~ O
~ ~ ~

x~; x xpp x x x x x HOO x x aappp x N

N

O ~ N o0 N O wj ~ 01 o0 p~ ~ c N N ~ M M
I ~n op ~ M
O
t ~ N ~ ~ ~ ~
~

,~ s~ ~, ~, _J, v~ s~ Jw N
_~, O v' j, _~

~ C!~ ~ ~ ~ E-',~ C/~ (7 N
CJ ~ O a C7 CJ

O O ~ ~ ~ O ~ O O O
O N O

f' ~n ~p O~ ~ M ~ ~ M ~' m ~ '-' N v~
N ' I~ vp ~~ N r',~ ~ M oo ,~ M Wit ~D N
' ' ' , , ~ ~ ' , ~ ~ O
. ~ ' ~ ~ O ~
, ~ O ~-' O ~ ~
O
O

a v C7~ a dd C7 w>
a dC7~
a '~

, ~a ~
~ ., , 0o Qv O ~ N M ~ vWD l~ 00Q\ O

O O ,~ .~.~,-,,~,-V.--~ ,~ .~,~ cy N N oo l~

~i _ _ '~ ~ N ~

I~~n d'M ~t N ~

, .~, r'~ M M , , ' ' , , O , N , ~ , , M
~ M

N N ~O .-,o0 N ~n oo .~.-, O
~

N N .~ .~ N

Ov O ~ N M '~t~n~ I~ oo a1O .-r N N N N N N N N N N M M

C~ N O M l~~ l~N M M t~01 O

M N M -~00M 00M ~ O ~ l~ N

M M M ~nN N O ~n M N t~ N

M 01 I~ V1\OM N l~M -1 O~~ M

OW E V7 .-.M I~00N ~O \O t~01 I~

~ Q1~ ~ D\

N N N .~N p ~t~nO ~' N M op ~ ~ ~ ~ ~ w ~ z ~ z x ~ x x ~ ~ x x x x x cr a, M
N

N N N N N N M N N N N N M ~ N N N N N
N

O O O O O O O O O O O O r; O O O O O
O
O

~'~t'~~ d'Wit'~f'~t~ ~ ~ d'~ ~ ~ ~h O O O O O O O O O O O O p O O O O O O

O O O O O O O O O O O O ., O O O O O
O

~ 07G707 ~ ~
d' vp ~ ~ y D ~ j, ~, ~ ~ ~
~ ~

~ ~ ' V7~' N d N _ _ CJ ~ C/~ CJ _ CJ

, o v~a cn,~

0 0 ~, 0 0 0 0 0 0 0 0 o O ~ ~ ~ ~ N I~ N ~'~ ~ Qv N l~
~

N ,~ .~ ~D~n v~ M
o N ~O ~

J' ~ ~' cd~ ~ ~ ~ ~ ~ ~, ~ N
~ ~

C7 ~ C7 7 C7 . H ~ ~ , U , v i C7 a d d U a -~ ~
H

r , E

N M ~ V7~OI~00O1O ~ N M ~ ~ ~ ~Ol~00 N N N N N N N N N M M M M o0 M M M M M

~D~

M .~ N Ov N 0o ,~O

M O ~ v~ ~ V~N O ~nN 01 ~ ~ N ~ l~~ N M
' M M ~ ~ N M 00M ~ ~ V7 N V7 N N ~,N d ~ M , .-r, N .~ .~~ , N i O lD~ ~ ~ M ~ \p~ ~ ~ M l~ ~ ~ ~ 00 M ~ ~ N '-' ~

~ ,~M M Q1 .~ .~ I~ M ~p<Y
o M '~

N ,-~N

N M ~ VW O l~0001O ~ N M V' ~ ~ ~ l~00a\

M M M M M M M M ~ d'~t ~'~ ~ ~ ~'d'~t O ~ ~ ~ l~N l~~ M l~00 ~'.-~ I~ N 01~ N t~

N O N M l~~D~ ooI~O '~t I~~n ~n ~GooM ~ a\

~ oo~ O a\~ l~~ oo~ON o0N ~t I~O o0~nN

l~t~~ lp M N ~O01M ~ eh M I~ 00 ~ d'~Ol~O

N N M M t~o0N ~ I~N N I~M M O l~l~Q1et v~~ v~Ov W O ~nI~~nI~v~ v~00 0o I~l~~ ~ 00 ~ ~ ~ ~ ~

~--~l~ N O~~t~ N ~ M N M V7 s ~ U ~

H a a v~v~ ~ z C z r~ a H ~

0 0 0 0 0 0 x 0 0 0 0 x x x x x x x x x x x x x x x x x b N N N N N M N N N ~ N M N N N M M ~ N N N

O O O O N ~ 000000000000 0000 0000000000 0000 d' d'~ ~ ~ ~ ~O~O~ ~O~D~O ~D~O ~D~O~O~D~D ~O~

O O O O M O O O O O O O O O O O O O O O O

O O O O O O O O O O O O O O O O O O O O O

~ ~ ~ ~ x x x x x x x. x x x x x x x x x M ~;~ ~o N <'~~O _ M~

i~ O ~ ~ ~, ~ ~ 7, j~
~ b~ ', x~ .~ ~'H~ ~~ a ~ ~HC~HC~ a ~

0 0 ~ 0 0 0 0 0 0 0 0 ~' ~ ' +~ 0 0 ~ +.~,''' a\ oo " N O ~O v~

~' ~ N '~ ~ ~' , i .-.~,~ N yp M l~ ~t 00 ~ ~ ~ ~ .~~ ~ ~; ~.
N N W o H a ~ C7 a.~ H ~ w H C7 c~ H
x v~ v~ v~ ,~
a Q\ O ~ N M ~ ~ ~Ol~0001O ~ N M ~ ~ ~Ol~ 0001 M ~ ~ ~' d'~ ~'~ ~ ~ ~ V7 V7V7 V7V~V7~ ~ ~ V'1 ~ O ~ ~ ~ ~

M .--r~OI~O l~V~ M ~ Q\ M

N N ~ N N ~ N ~ M N M M M ~ N M O , M N O 0~1W O ' ~ ~ ~ .~~ d' t M M N N ~ . N N
.~~D N

N N ,~.-~ N

O ~ N M ~ ~ ~Ol~00O~O - N M ~t~WO l~00 a\O

N ~ ~ l~ ~ N ~ N ~nM N M O oo N M v~.~O O l~

M 00~OO ~ ~--~~O~ M l~~ ~ N O O O O l~O ~OO~

l~(~lp O ~ 0141~ ~ 0000 0000 000000~O00 ~ I~

N O W O -at~ oo~'~ O o000 0000 0000000000 0000 v~ ~n~ ~n O~N O o0~O~OO O O O O O O ~OO O O

~ a1~ ~ ~ ~ ~ ~ ~ a1~

I~ ~ N M I~~p N Q1~DooM ~ N ~ M .~01~'l~ ~Do0 ~OO ~' ~ .~ M M N O N ~ O~'"' N M 00 ' ~ d ~ ~ ~ d a H H H H H H H H H H H H H H H H

O O O O O ~ r~r~v~v~v~m r~v~ v~v~vovav~ rr~rr~

x x x x x x x x x x x x x x x x x x x x x vs o ,W O ~ '.-: ri o t~ oo vo vo t~

N O l~ M 00 ~ V7 l~ ~
M ~ ~ l~

I~ l~ -~ 00 01 ~ Qv O
01 N o0 N

00 O M lCr V1 (~ ~ 00 O ~ .~ N N

O N M M '~ ~ ~ ~ O O O
O O

M

M

~ O

a x x x .b N N N N N N N N

0000 00O O O O O O O ,~

O O O

O O O O O O O O O O

x x x x x x x x x x a . . .

N

pp" N ~ 00 ~ pp O ,~

M ,~ N N v-i ~ ~ ~t I~

_~~ ~r ~i ;' _j, _~, _c~

H c7 v~ ~" ~ c7 w c7 d O O O O O
O O

O O

N ~n I
,~ M

Q1.~ ~' r' ~' l~
V7 M ~
~O

cad ~ ~ ~ ~ ~
~ ~ cd a . . , ~ ~d>
, c~c~ U
x>

O ~ N M ~ ~ ~O l~ 00 Ov O

~D~

Q1 00~O M N

~

M 00 N M ~ M N ~ M

M N ~ i ~ ~ i ~ ~ ~ i ~

,~N .~.--~ ,-.wp O N o0 N N

N N ~ ~

~ N M ~ ~n~ I~ o0 01 O -r 00~ 00.--WO N ~ ~ O 01 O

~Dl0 O I~ l~M I~ M N

M 01 ~ ~ l~.-r~O N ~n M

00~O ~ l~ N ~ ~O l~ 01 M M

O O N N ~ ~ t~ N l~ N O

v'1vo V101 l~I~~ 01 ~O 01 I~

U U ~ O ~ i ~ d d C7~ O d ~ d d d pq ~

x x ~ ~ ~ ~ ~ ~ x b t~

0o Q\ ~p o0 0o N , ~

i O ~ O O

p p ox x a x x ~o Mg a a ~

N N N N N N N ~ -r N N N , N
-V

O O ~ ~O ~O ~O vDN v0 \O~O ~O I~
..

'-' O O o0 00 00 0001 00 0000 00 00 O .~ I~
~

WD vDvO v~ ~n ow n vm n ~n~n ~ oo ~n oo ~

O O O O O O O O O O O O ~1 O
O O ~

x~; x x x x~ x~ x x~ x~ x x~ x~a x O

00 00 ~ a; ,~ N oo p~ -~
~ M M ~-~' .d: M M
41 M d1 ~ V1 00 ~ N N . r1 ~ 7 ~
~

i~ bA ~ W , ?~ ?, ~ v i ~ ,~
a~ ~ ~ ~ o ,~ ~, ?, .~ U

d~~x ~ a.,HaH H
o C7 C7 ~ d ~ 0 0 0 0 0 0 o 0 0 0 0 0 o 0 0 ~ ,~ 0 ~ N ~' M O O

i' ~ l ~ ~ l ~ ,-. ~ ~ ~ N
d ~ ~ ~ N ~
' ~ ~

~' ~ ~ ~ N ~ ~ N ~ N ~ ~
~ ~ ~ ~ N N
~ ~

c7d da,HH , - v~dv~ v~ c7av~ d a a~aH~c~

N M ~ ~ ~O I~00 ~ O ~ N M

l~ l~l~ I~ I~ I~ I~l~ I~ 0000 00 00 ~O ~O~O ~ ~ ~O ~O~O ~D ~D~D ~ ~O

a1 M O O ~1 M .~01 , ~O v~\O M O

N ~ ~ Q1 ~ ~ M M ~ ~ M M

.~M , .~ , M ~ , N

i ~ ~ i ~ ~ ~ i ~ i 00 .~N vD ,~ ~ p~,~ N l~-~ M N

N .-~ N

N M '~ ~ ~ l~ 00a1 O ~ N M

00 0000 00 00 00 000o a1 OvO~ O~

Ov 01O ~n N ~ ~ l~ N oo~- ~t N

v7 O M ~n O O~ ~ON N oo~n ~ O

o0O ~ O oo M N ~O t~N O ~n 01 l~~n 01 ~ 00 ~Oo0 O ~n~ Ov Ov N O ~ ~O ~n a1 ~ O (~ ~ON ~D O

l~ l~O~ ~ O~ ~D l~Q\ ~D ~OO~ 41 t~

<t M ~ O -~ N ~ N
~

M O ~ M
~ ~ ~ ~ d d ~ ~

~ ~ c c c 3 3 3 ~ ~ ~ ~ ~ 3 ~ 3 x x x x x x x x x x x x x b b so 0 0 ~ o x x ~ ~

o x ' x ,~ ~ ~ ~

O1~j N -~ N ~ N N N N N N N N N M N
., l~
, t~pp l~l~ I~l~ l~~ l~ l~l~l~ l~I~ l~ I~l~
, I~ , O ~ 0000 0000 0000 o0 000000 0000 00 0000 I~ O
,~

~ W v~ V7V7 v~~ ~ ~ V7V7 V7V7 V1 ~ V7 00 ,~

~ x x x x x x x x x x x x x x x ~ a ~ a a ~ ~ ~

~ O
~

N ~ M M ~ ~ ~ vp ~ ~ ~ ~ N
~ ~ Q\ ~ N

'O ~ bD ' dp _~" ~., ' ~' c'~''nt ' >, b0 ~' O ~ ~ ~. ~
~ ~A

~" ~ ~ ~ ~ C7 F-~ ~' ~ d v~ ' _ ~
H ~ ~ a ~ d C7 a ~
., O (~O. p p O O ~ ~ ~ 0 ~ ~ O
O O ~ O ~

~ ~

O ,~-~ ~' a\ N .--. M
0 ' .~ N d1 ~ ~"'.~ ~ N N ~ ~ M ~D ,~,~ r, M M v7 oo ~

~. >:. O O ~ L: ~ ', d0 y~, ci> ~ ~ ~i p a~

v ~ C7 v rio i w 1 C7 a c7 a x d d ci ~ v~d ~' ~ ~ r -, V7~D I~00 01O r N M ~ ~ ~O l~ 0001 ~ Q1 ~ ~ ~ D1 d,Q\ ~ ~ Q1 Q1 ~ ~ M \O O M 00 ~ ~ ~ ~D,~ M ~ 00 ~

'~N , N , M N N N v'1 N M , ~

i i ~ , i ~ i N ~ ~ ~

~ ~ o ~ ~ M M N ~ ~ ~ N N
o M N M N

~W O I~ 0001 O .-. N M ~ vW D l~ 00 a1O

a\ Q\Q1 O~a1 O O O O O O O O O O

.-. .~.~ .~.~ N N N N N N N N N N N

M O 00 ~ O O O ~ ~ ~ V1 N V7 00 t~

O N M 00M ~OO~ V7 O ~ ~1 00~O Q1 0000 t~ I~,~ N N ~O~ ~ l~~ ~ l~oo ~ O~M

~n eto0 v~Ov N O d' oov0~ M ~ ~O o000 t~ ~ ~ 00M V)l~ t~ V~V~M O l~ ~D N O

~O t~~O l~l~ O1~O ~O 01~Dl~ t~t~ 01 ~OI~

O M W O N I~M ~) 00M (~ d'I~ .-r 00lc, p ~ ~ 00~ O M N O ~ N M l~ ~ ~--~M

w ~ H d ~ O O a., > ' > ~C~C

a a s a a a ~ ~ ~ ~ ~ ~

x x x x x x x x x x x x O O O O O O O ~ ~n O O O v~ O O o0 N ~' ~t 00 00 00 00 00 N o0 O\ V1 O ~ M N
~n ~' ~' 00 l~ t~

~O ~O ~O ~ t~ l~ l~ N O l~ 00 O I~
~ O M v7 v7 t~ t~ I~ t~ ~ I~ M l~ ~ 00 ~ ~ 00 M --~ O ~ N N

O O O O O O M ~ ~n ~ o0 00 0o O ~O
O O O O

.-r .~-n .~ ~ ~--r .~ ..-~ ..r ~
~ .~ .-~ .~ N ~D ~ ~O ~D

M

N

a' 'O

\p . .--i V~
.-r ,~ o ~i~i x o;

x a a , N N N N N , ~ ,--~ .
I~ l~ N N ,--~
~

v1 v7 W O v0 O x x x x ,..~
, O O
O

O ~ ~
t~

~nN ~

~
>, ." '-',_,~ N
O

r' N r,M

~ O ~
O

O ~ N M ~t v~

O O O O O O

M

O M ~ N M M

N ' ~ i ' i i M O

~ M ~ \O

.-~ N M d'VW O

N N N N N N

et ~ O ~ O t~

M N N 00N O~

M V'1 ~O'OV7 01 ~O a1 .~ 00 ~Ol0~ O

Ov a x x x x x x ~

M 00 ~' ~' M ~' ~ N 00 00 00 "' ~p a ~ ~ ~ a ~o ~ _ x x x ~o b o x a~ a a M N ~' '-' M ~' M ~ N M N N
~ ~ ~ ~ . '-' ~O v O

vo .. , o ,~ , ,-, .. ,_, ~n ~ ~
~1 O O O O ,.O O O O ~n O O O O
.b oo .b t~ ~d x a ~' x ~ x ~~ x ~~ a a ~ ~

~ ~ ~ ~ ~

0o ~ M ~j ~ O N
p p ~ Vj 00 .
l~ ~O ~

,~ ' ~O N M , r ' ' ~ ~ ' ~

~ ~ ~ ~ i ~
O ~ ~ ~ N ~, ~ cd _ ~ ~ ~, by C7 ~' x '-' ~ ~ C7 ~ '-' C7 d '~ x C7 ~ a ~
O p O O
O

_O _ ~ O O _ ~
_O ~ O ~ ~ _ O
~ ~

M ~ ~' ~ O N ~ ~
~ O~ ~ N .~ M
~ ~ N
l ~

', n ~ cn t' v bA ~ ~
'' p ci~

t7~- a~ iad v~a,x C7aa Hd lp t~ 00 01 O ~ N M ~ ~ ~ (~

O O O O .~ .~ .-~ ,~ --r .-r ~D ~D ~D O M \O ~ N 01 l~ ~ , N ~ ~ ~ M VW O
N

, N N N M , ~ , , N

N M .~ N N O ~ O

M -~ N M

l~ oo O~ O ~ N M ~ ~ ~D t~ o0 N N N N N N N N N

N N N N N N N N N N N N

O 01 N t~ ~ a\ M ~O ~n Ow e N

00 .~ ~n 01 N O~ v~ N o0 0o M o0 l~ I~ ~ o0 O N ~ o0 v0 ~~ ~n l~

.-r ~O v~ ~ t~ .-W ~ d' -~ 00 N

vD ~D o0 O O~ O~ ~D O I~ 00 01 v~

00 M 01 N \O .-, ~p N ~ 00 41 00 a o ~ a ~ ~ x a ~ ~ o 3 ~ ~ ~ 3 x x x x x x x x x x x x b ~

t~ ~ t~ o ~ os o ov 00 d' 00 ~t oo N V ct .-r O

o ~.-~x ~) x ~ y .-~ ~) ~ ~n x x~ ~ ~ ~ a ~ ~ a ~

cd cd a M N r'' ~ '-' N M '-' '-' '-' '-' v~
~ . .

~ ~ ~ ~ ~

O ~ 00 ~ ~ 0 o 0 0 N
O N ~ I~ 0 0 0 0 ~
~ m n ~n 01 oo v~
~

p t I ~
n a a x a x a a~ x x x xoo a ~

. . . . . .

N oo ~O oo ~ O
N ~ ' ~ 'O ~ ~
' N ~i v o ~ ~ N I ~ M
O ~ 0 ;

E-' ~ b-0 bA b4 _ >, ~ 0 _>, ~ ~" >, p p s.;
~ O , H ~ ~, C7 C7 a C7 x H C7 a~ U v~
,~

o 0 0 0 0 0 1 0 0 0 0 0 0 o ~-~ 0 d' d1 M 0 ~ 0 M ~ _ M
~ ~ ~ l~ 01 ~' I~ O
N
~D

l v; _ _ i ~ N
~

~ ~ ~ ~ ~ ~ ~ ~, ~, -' pp ~
b1J

w C7 ~ ~ ~ H C7 a ~-7 v a v~ C7 ~n ~ ~ v~ H

0o O~ O ~ N M ~ ~n ~O l~ 00 01 N N N N N N N N N N

N O ~_ ~ l~

M ~ O M M ~O ~ M N ~ ~ ~i' i i ~ ~ i ~ ~ N ~ ~ ~O
.. D1 N ~ .~ .~ N M N O

N N .-, .~ ~ M M ~, ~ N

O - N M d' ~n ~D t~ oo a1 O

N M M M M M M M M M M

N N N N N N N N N N N N

01 O r..~~ l~ I~ O O N ~--~~n M
~

Q1 V7 ~ O \O ~O ~ O M 00 t~ ~O I~ ~ N ~ ~ 01 ~O N l~

N v1 Ow e .-r M ~ d' ~ ~n ~ V1 00 .-m D N o0 l~ ~O ~ ~ ~O l~ ~n Q\ Q\ a1 00 ~O <t ~--i,~, M 00 '-' ~

a ~ x ~ w A v ~

a~ ~, x x x x x x x x x x x x M

M

M

N

t~ t~ ~ t~ oo vD oo v~

~ I~ ~' N oo I~ ~' d' oo ~ O t~
vD vp p ~ O

. O a a x I-O ~ a x -1 ~

~ ~

~-~ x~' ~ a~ ~ ~ ~ a~ ~ '-' . .

j .~ .~ ~ N ,~ ,--~ ,~ N N
o0 ~ O
~

O V'1 00 00 00 , 00 00 0 O M ~
O 0 ~ O O 0 M ~ I~ Vl V~ ~ [w ~ V~ ~ V'1 ~ ~ V'1 ~ I-~ I-~ O

V~ O O O O O O O O O O
~

ppO a x x x a x x ~ x a ~ ~x .
~ . ~ . . . . .

O v~ ~ o0 ~ M N ~ r"' N ~
oo I

c~ ' j, W o b0 y; ~
~ ~

~d r t7a a ~
, v~~e7 ~ N I~ ~ N M ~
~t -, M ,~

' N ~." b-0 v~ _~ ~ _~

H

O ~ N M ~ ~ ~O l~ 00 O~

M M M M M M M M M M

N ~ ~ O O

00 O M M ~ _ ~ 00 ~' ~ 01 ,"'~ i ~ i i i ~ ~

00 M 00 ~ I~ M V1 M M O

M ~ N N ~ N -N M d' ~n ~O l~ 00 O~ O

N N N N N N N N N N

a1 00 00 O~ oo N N O ~ N

00 l~ ~n ~n ~ O ~ N N 01 O d' Ov o0 N ~O O ~ ~D

V7 V ~ M 01 01 O ~ M

~D N M ~O ~O -r l~ l~ M d' Q~ l~ l~ D\ l~ Q1 01 I~ l~ 01 p ~ O ~ N ~O N ~D N

'-' ~ ~ ~ l~ O o0 00 U1 ~ ~

x x ~x x x x x x x x x x x x o v~i 0 0 0 0 0 00 ~n o N .~ .~ ,~ .~
l~ O I~ O

N ~D -V --~ ~
M ~ M .~

O O M M M M ~
V1 ~D

M

N

M l~ ~ I~

O O ,~ ~ ,~ O
x 01 ~. M N O r., ,~
N

M
O ~ 0 ~

V~ V~ V7 ~
~

V~ V~ N N
0 0 0 I-~

~ ~ ~ x x ~ a ~

~ ~ ~

MM

N d' I N M o0 .--y ~ N N
~ Q1 ~ >' p j ~.., ~ ~ ~ Q, p..

--~C~v~~w~ E--~C7d v~d~ d ~

o o 0 0 ~, 0 0 0 0 0 0 _~'MM~N N ~W p~ py O
~ ~'' c~ ~, ~ wi~ b~
c~ ~ p ~
~, v _ _ _ _ _ C~ C7 ~ P-~ x ~-1 _ C7 d d d Pr N O ~ N M ~ ~ ~O

00 ~ ~ ~ ~ ~ d' Wit' N

' N

M M

~ O N o0 N Q1 M .~ M

N M ~t v'1 ~O l~

Q1 V~ V~ ~ V7 V'1 N N N N N N N

00 Q1 M .-~ 00 M ~ 00 V7 M ~ ~' O 01 M M 01 l~ ~n vW ~ ~n ~n N Ov N Q1 ~ .~ .~ ~ o0 Q\ ~ ~ ~ ~ D1 Q\

tn V7 ~ M l~ N

~1 ~ Q1 O Q1 V7 00 x x x x ~ x x as ,-~ .~ .-: ,~ o .~ ~i .~
0 0 o wi as oo ~

O ~D ~p ~p N ~ 00 M O O O I~
~ ~ N o0 O ~ et ~ t~ 00 ~ N a\ ~O t~
t~ O M ~n o0 d' 00 00 0o O O ~ M a1 N Ov Q\ O O O

~ ~D lp ~O 00 00 01 Q\ O M
V'1 v7 O O O O

N ~O ~O ~O ~O

. ,...;
~p.. pp. _~
.--~..

O O O O O
~

,~~ ~ ,~a ..d~ 'db --~--~--i~ ,-i cd . .

~

O ~ O O O O O O O
O N

O O O O O O O O O O
p O

x H ~ H H H H H x x0 OH .

-, ,-, M
~

~ i i O O
~

O O p O

M ~
M

N
~

_j, O
DO

~
E-I~0001 O ~ N M d ~ ~O

~n~nv m ~n~n ~n O

~ O

i N M N M M M ~
~ i i O ~ ~ ~ N M O ~ ~ O
~

N t~,~ ~OQ\N ~D ~O
~ ~

M ~ .~ .~~ ~ON N

0oO~O ~ N M ~ ~ ~ l~

N N N N N N N N N N

N ~ 00 O M I~~ M ~ 00 ~ O t~M ~ 00d'N O

I~~nN O v~~t~ a1O N

~ON ~ ~ v~~ ~ M d' l~

~nM ~ 01h O O V~~ N

~Ol~v1 t~~n~nV'1l~l~ 01 M ~ O ~DQ1~ ~fV~~ O

' ~ ~ W

a~x ~ A ~

3 3 3 3 3 ~ ~ ~ 3 x x x x x x x x x x -b .. o l~ O 00 O ~ ~ Q1 00 M 01 ~ ' ' -r ' ~' ~ O ~ O ~' fir ~ 00 M .
.

l~ ~O l~ ~O .-~ .-~ W ~ l~ l~
.--O p p O O O " O D D O ~
x i.-a,.~,~ M d' ~ ~ t-.~~l O
O V~

~ ~ ~ ~ x O O O

a ~ ~I

ri I~ ~ N ,-, N N r, ~ ~ .~ .--rO
.--r O ~ O ~ O pp M ~ ~ ~ O O O O O ~
o0 a ~ ~ ~ M ~ ~ p ~

O O O O O O O O O
o x~ x a x aaa x x x x x x a ~ ~ ~; ~ ~ ~ ~ ~

N l~ v'i O -~
O

~n N ~ N W O
~ ~O

~ v ~ >

vW , .~ ' >
C7 ~ ~ ~ ~ , 0 o ~' x ~ w '-H

~ o o o 0 ~ ~. ~, 0 0 ~ o N 0 0 ~ ~ N ~' N w ~ ~ ~ v> 00 p '-' ~ r-' ~O ~--,M -~ 00 N M M

~ ~ ~ i , ~ ' i ~

N ~~
y., H > H ax w ~~ ~H
~~c7a ~

l~ 00 Ov O ~ N M ~ V1 ~D l~ o0 v~ vW o W D vD W O v0 ~D vD

~ M O

D1 00 \O M N d' 00 N I~

O~ N O V' M M M N 'cY d' N N M , , , , , , M

lp ~ ~D ~ M M N

N -~ - M ~ ~ oo l~ ~n .~ N .~ N ,~ N

00 a\ O ~ N M ~t ~n ~D I~ oo Ov N N N N N N N N N N N N

M M ~ ~ 00 OW E N M ~ M

M ~ ~ 01 N 01 I~ ~D M .~ 01 N l~ v~ ~n ~D O O N N ~ a1 N O v~ oo ~ oo M ~O Ov .~ .~ 00 ~n l~ Ov oo ~t OW ~ ~O N o0 M O

Q\

l~ .~ p~ O M N v~ N d' O m O N o0 01 ~ M l~ l~ v~ W n d x x x x x x x x x x x x v, ~0 0 0 ~ ~ ~ ~ 0 0 0 ~

QW 1 v~ N ~ M l~ l~ ~ ~ M

O O O M 00 00 .-r .-~ ~p ~D 00 l~ O N

N N M M M ~ I~ (~ l~ l~ l~

M

N

M

~, N 00V'1 ~ O l~

~i' M I~l~ ~' O l~l~

O O O O O O O

v~a a a ~ a a b b b ~ b i b N .~,~,~,~ .~N

~ O O O O O 01O O

a ~ ~ ~ ~-~r o o ~ 0 0 0 0 0 ~no 0 x x a v~v~vav~ v~a v~v~

(~j ~p p .~

i M
i ~~ ~~' ~
~

a x ~H

0 o d O O ~
O

N ~ ~ ~..~ 0 ~ O
+.

~n O ~n M r-..~
o0 ' N
.~

0o ~ M ~ ~ N
Ov ~ ~
.~

v c~f-~.b4 ~',y~
~ ~ ' N
~
~, _ _ ~ ~ ciW
x W U H
H
v 01O ~ N M ~1'~ ~ l~00~

t~ ~ ~O~ ~ ~ 00 ~ N O

O ~ l~Q\ N M N O
N

M i r',~~ i ~ ~ M ~

M V7~ M N ~ M

~ M

O ~ N M ~ ~ ~ I~o0O~O

N N N N N N N N N N N

l~l~ Q\O M ~ 00 ~ ~ N 00 N o0 N ~ l~N M ~ 00~ M

N ~ Ov~ -rM O o0V~N ~

M 00 ~ O ~ ~ ~ 00.~V7\p 00I~ v~\O\O~nN OvOvN N

p ,_,.~M l~O N M 01N

~ La~ U

O O O O O O O

O

x x x x x x x x x x x N O

O v~ ~n l~ l~ O O N O
N N M O

--i ~D O O ~ O N ~ 00 ~O t~ 00 I~

O O O t~ l~ N oo I~
O O M O

O O O t~ l~ oo .-~
O O N '~t' O O

C

.--.. .-..-r .-. , ,--r ~

n1 ~ vD~' l~ ~ ~ M ~O~

l~~ ~ l~~ O O C~C~M O

~ ~ ~ o ~ o n-1a n-1~ ~

~ b ~'~ ~

~

M ~ .~N N N .~.~ .~.~.--~.~N
~

O D1 O O o0 . O l~ O O O O O O ~D
..

~ ~ ~'~ N ~tI~
O

O l~ O O I~ I~ t~l~ O O O O O O t~
~

O O O O O I~ O O O O O O O O O
O

C/~~--~C/~C/W.-~ O ~l~1 C/~C/~ C/~V~Cf~V7 O

~ pp O
O

V'~1M ~O ~ M
.~ ~

~. , ~ v~
bn ~~

d _ ~ a N
~ C7 O O O p O

M N '~ Q W ' ~

M '~h M ~' M
~

',b0 ~

~ s..
c7 C7~ ., d H a w O ~ N M ~f ~ ~O l~00~O01O .~N M

~ ue N ~ O ~

O N ~nd oo ~ '~t ~ ~ o 00 ' 0 V7M N ~OM M o0 ~,,N -~M N ~,,M \O

N i ~'i ~ M i i ~ i M

O ~ ~ ~O O ~ ~ ~ a1.~~ ~ O

N 01 ~ .~~ ~ N ~ ~ON ~ ~ ~ O M

N M ~ ~n ~Ol~ 0001t~O ~ N M

O~01 0101~ 01p1 0101Q1O O O O O

N N N N N N N N N ~ M M M M M

N t~lp~ O M M 00V'1O M 00 Q1.~ ~ .-~d~ N M l~~O01M 01O

0041 <f0000414100OvO

N t~ N t~~n ~ M v0O o0M D1l~00v~

a1I~ t~l~~O N o0 l~-r~nI~~n~nvWD

Q\ ~ ~ ~ ~ ~ N ~ ~ ~ Q\

M 00 I~d'~ .~V~ 01O O V7Q100~

~ N O ~ oo M ~D

x x x x x x x x x x x x x x N - ,-;

Qp ~ 00 00 I~ 01 00 N ~ .--r ~ ~ M N M

_ ~.

~ O O O O O O
~O o0 p~ ' M

~ a a a a a a O O ~

b ~~oa b b b ~u -d -d ' a ~
~

--i,~M N ~j ,~ ,--m-n ~.-i ~

N ~
~ ~ M

~ ~ ~ 0O
O
O O ~

O O O O p O O O O O O
p v7 p ~

~ ~ x x x x a x x x x x x o . . . . .

MM

_ N O ~ N N M ~j Ov N
'-' N ~ , ~ ~ i i ~ , , bA
01 bf7 M ~
N ' ~
~
~
~' , ~ F. _J-, N ~ ~ ;
r " _cd _.

H

O~~OO~ ~ O O OO D OO O

~

00 ~O ~ ~ N V1 ''-'N .'.' O ~ p '-' N N l~ -, N '-' M v~ ~ oo ~O N
O~ .-~ v~ N

by c~'n ~ ~ j O ~ p 'n ~ s'' O s~.

H~~ac~~ ~ , , ww >
~ v dwn ~Ol~00O~O ~ N M ~ v~ ~O

0101a10101d1O O O O O O O

N '-'M M N ~' I~ M M ~' vp N ~n ~ r.M M ~ ~ N

N M ~ .~ N M

,~ N ,~

v7~ l~0001O ~ N M ~ ~n ~O l~

O O O O O .-..-r M M M M M M M M M M M M M

00O ~ I~N ~ l~ M O O l~ O ~D

N ~ 00O wtM ~ ~O N ~ ~ ~ O

~pvWD ~DN ~ ~' I~ O M O 00 M

M ~O~ D\~nN l~ ~ ~ t~ M 01 01 ~nN ~Oo0~ N oo ~O M ~n N O t~

~ d'~ ~ a1 ~ ~ a1 00~ M .-~M ,., M O .~ M O1 \p~OI~~ ~ M

~ O O 0 a a a a o ~

0 x a x x x x x x x x x x x x 0 o vi o o ~ o o ~n ~n ~ t~ o o ~ .-, o t~

v7 ~ v7 V~ N ~
~ n 01 O O O O M M C~
~O O

N N N M M N \O
~ O

N

M

01 ~ ~ N W O

D\ 01 ,~ .-~ ~ ~ \O M ", O ~ O O ~ ~ ~ O Q O M
~

x a ~ ~ ~ a a 'O ~ b O

x ~", M

Q\ d1 M 01 Q\ ~p 01 pW 1 01 Q1 p O O O O

r~

O O O O O p O O O O O
~ .b .b ,~

x xa a x x a x x x x x . ~ ~

~ ~ ~ ~ ~ ~

M ~ 00 ~ M M ~ V7 N I~
~ t~ N M
~

N M N V) , ' >, >, ~ >, ~ O
tiA ~

_ _ _ r-~~

O O O
O ~ ~ p ,~ 0 O ~, p O O
O .,~

00 00 M 0O M I~ O
O

,~ ~ N ~~ N N ~ N
~N d.
~
.

p ~ t1, y, ~ ~ p ~ r=, ~ _>, p ~

h, ~ d v~ ~ ~ P-~ P-r V~ ~ C7 P1 d I~ 00 Q\ O ~ N M ~ ~ ~ I~

O O O -r .~ r.r I~ ~ O N
M ~ ~ M O N ~ ~ ~ N

~O N a1 ~ d' ~O ~,,~M I~ , N ~ i i ~ , ' i i W n M ~ M ~O O ~ d' ~ d' M N N N N ~ ~ ,-, 00 ~ O ~ N M ~ ~ ~O ~ 00 N N N N N N N N N

M M M M M M M M M M M

V7 00 O OW E O ~O ~ 00 00 ~

N ~O M I~ O o0 ~D ~ O v~

V~ 00 M ~ ~ (~ M O ~ Q1 M ~D .-~ l~ N v0 .w 0 ~ t~ v7 00 ~ ~ v~ ~ ~ ~ 01 N ~ N

01 I~ 00 01 O~ l~ l~ 01 01 Q1 a v ~ ~

. w ~ ~ oa ~ c~

~

x x x x x x x b N .. ..

M ~ ~ ~ ~ .~ O~ v0 ~

p 0 0 x ~ O~ ~l ~1 r~ 0 ~ ' ~ '~ 'b 'b O "C3 'b ~ O ~ ~ .
x 0 ~ .--~ N ~ a c .. C

~O ~ ~ ~D ~' N M ., ~-'' ,~
r .
~' , ~ ~ ~ M 01 O
'~' O O M ~ 01 ~ O~ 01 01 V N 00 .~ O

) ~. 00 00 ~ ~ d' d' ~f ~. O ~D
O O O

O ~ O O M ~ O O O O O
~x O a ~O x x x x ,~ x x ape x ~ . . ~

01 00 ~ l~ ~ ~ N o0 l~ ~O ,-; ~j N .~ ~.,j ~ ~ ~ ~' ~ ~O ~ M

N N ~ ~ ~ N ~ N ~ ri M_ M M C~1 N N
~ ~ ~ v~ M ~ ~ N
~

~. cd ~," O ', d0 >, ~ ~. ~ ~ ~ O
c~ ~ a ~ ,~ ~ ~ ~ ~" 'n (7 j, Hd ~Hwc7 ~c7 ~t7 y H~ >da ~ c7 ~x o ~c7 O 0 0 o O O ~ p O O O ~ O p O O ~ O O ~ ~-' ~ O O O
N ~

~ ~ ~ p p ~ ~
~ ~

M ~ ~ ~ ~ N ~ N -~ ~ ~
~ ~ v~ d' .

p dp ci~ p p c~ i~ pp ~ ~ Q. N y', ~, ~. ~ ~ ~ ~ ~ 'O
b-0 wH a~~xwc7v~w wda ~v~ dtS a,H Hai~ d x ~H

00 ~ O ~ N I~ M ~ ~ ~O l~

N N N o0 N N N N N

N ~O
M ~O ~ ~ Q1 M l~ ~ I~

M ~ ~D ~ i ~O M
00 ~ N N , , N , i ~

i i '~ ~
M M

~ ~ N t~ ~ M

Ov O -~ N M oo ~ ~n ~O l~ o0 N M M M M ~ M M M M M

M M M M M '~ M M M M M

~p ~p ~--W ~ ~ C~ M M V~ O

O V~ ~ ~O 41 00 01 I~ ~ V7 V'7 M a\ ~n ~ 00 t~ N O oo N

O w 0 ~ 00 0o O ~ ~ Cn N

O l~ ~ ~ M ~O l~ ~ l~ ~ N

00 41 Q\ Q\ ~ Q\ D\ l~ ~D O~

~ ~ N ~ ~ N O

0 ~ ~ 0 ,. o 0 0 ~ o U U U U U

x x x x x x x x x M

O ~' ~' ~ Ov 00 ~ a M ~ a x a O '~ ~ O O ~ ~d 'b ~ ~ ~

.~ ..-~ .~ ,~ ,~ .-. ,-1 .-r~ ~ N

M . . . .

~h d' ~ ~ ~ ~ ~ ~ p d' ~ ~

O~ 01 O~ 01 Q1 Q\ 01 01O~ O~ tn Q1 O O O

~r ~a ~ ~ ~ra ~ ~a ~r x x x~ x x~ x~ x x x x a~ x ~ ~ ~ ;

~ ~

N M
~

~ ~ ~ ~ ~ N
~ ~
~
~

N . . ff ~ aHC
.
~
~

o O O O O O O ~ O ~, O O O
O O O

~ ~ ~ ~
~

'-'' N ~O ~ '~ N ~ '."" M ~
N ~ ~ ~h ~

7, ~ ~ ~ x; ~ s~, ~ c~ b-0 r~ s~, ~ ~ ~
~. ~

C7 d ~ ~ _ _ d _ ~ CJ

~, v~ 0.m-, v~ v~ p...
~n cl~ P~

00 01 O ~ N M ~ v7~D I~ 00 d1 N N M M M M M M M M M M

\O V7 M M M

_ _ .-a ~p I~ M ~ ~ ~ N M v7 V~

M , ~ , M , ~ , N
~ ~

, , ~ , M , ~ , ~ , N ~ 01 ~ \O N M M M M ~D M

~ ,~ N

Q\ O ~ N M ~ ~n ~ ~ ~ ~ O

M ~ ~t ~ ~ ~ d' d'~ ~ ~ ~n M M M M M M M M M M M M

00 v0 d' ~ O ~n N --~00 O ~ N

.~ V7 Q1 ~ M ~Y V7 M 00 O 00 V7 I~ ~ ~ (~ ~?' O 00 M 00 ~ M

.-i ~' ~t ~ 01 ~t O M 0O Q\ I~ M

00 .~ l~ 00 00 N V7 t~00 l~ ~n O

00 ~D ~O 00 l~ Q1 ~ 01l~ 00 00 l~

N o o a ~ ~ ~ ~ d P ' U ~
-~ ,~, ~

x x x x x x x x x x N

O~ ~ ~ N ~ ~O N

r"' M ~ ~' 00 M 00 ~' \O

_ O O ~"' O O O ~ O ~ O

a a ~ x ~ x ~ ~ o a -r' ~

~ a .~ .~ ,~ ,~ ,-, ,~N ,-~ ,-, ,~ .~ ~ N ,-,--~ 00 ~n ~

. .

O1 01 00 00 00 o0 00 o0 o0 ~ O 00 O O O o0 <t d' O O O O ~ O M O O ~ ~ O
r-~ ~1 ~ y~ ,~ O

x x x x x x x ~ x x o o x ~

. ~ . . ~ a . -1 ~ .
. .
~

ri N ., M M ~np~ ~ y p N
,-; ~ ~ N ~ ~' O M
N

M ~ ~ N N .~ N v~ N ~p ~ ~ M ~
,~

~ ; ~ b~ ~, ~ ~ O ~ ~ ,j, N
~1, 1 ~ v~ ~
J, HRH ~~ ~~~ H xa ~ ~ H a ~c~

~

~

M

~" by bD ~ ~ ~ >, ~.
N p >, ~ ~ H ~ ' ~ G7 ~ H ~
~

O ~ N M dw v0 l~ oo O~ O ~ N

~t ~ ~ ~ ~ ~ ~ ~ ~ v~ ~n ~n O

M M 01 M l~N ~ ~--~00 M r' N M

M ~ N M .~~ ~ i ~

r, ~ i i ~ ~ ~ M
l~ N ~ ~'O ~ N ~ M M ~n -r N

N M ~ V'1 \Ot~ 00 ~ O ~ N M

N

M M M M M M M M M M M M M

00 V7 N N 00 N M ~ ~ 01 00 01 t~ ~ O~ ~ N Q1N N o0 O O ~ M

O v~ N M ~ O ~n t~ ~ ~O ~ M

N l~ M Q\ ~D ~ M ~O ~n O v7 V~ O

01 00 l~ M ~ t~v7 ~ M ~O N V7 O

00 n 01 I~ O~ ~na1 01 O~ 01 D1 00 I~

M ~ M 01 ..~ ,~l~ --, ~O ~D M
N

w ~ w w w w w w ~ ~ x x x x x x x x x x ~, a' N

N

C

M

01 ~ V7 ~ 00 V~ O

~' O O O O O O ~ O O

o a x x x a x a x O ~ b b b 'b -d ~o '~ b . .~

x~

I~ .-r M ,~,-r.-..-~M N .-i ,~ ,_, N ~ ~ ,~,-, ,~

~ pp pp N V7 Vj .. V7 ~ p O

'~

O o0 ~ 0000~ ~ ~ ~ 'ct ~ ~p ~' ~t ~ N

O ~ O O M M ~ ~ M M M ~ M M M

x x x x x x x x x x a . a a a . a ~ . a ~ . .

N oo ~ ~ ~ N ~
yD ~ ~ N

~ ~ 0i ~ y~ .~f., by ~ J~~, H _ p 0 ~O a ~0 0 O
O ~

O 0 O 0 O O ~
~-' -' ~ O
M ~O ~' M ''-'d. I~ r.~
O ~ N M
~

i ~ .~ M .-r ~ r, V~
M ~ M

i i i i i i ~
i C7 C7 ~ wC7 ~ v p nU a i U

~ . r a , ~

M ~ V1 ~Ol~0oQ\O ~ N M ~t N N ~ N ~ N N ~

M 01 ~D .--~.-r 00 ~ N ~ 01I~N ~ l~ M N

(~ d' ~ ~,M M ~--~~" M l~ M N M N M
.-r ~ ~ ~ .-,~ ,~ ~ .-i ~ i ,~

i i ~ ~ ~ ~ i ~ i N N V7 ~ ~ W O l~ N
0 v~ N d1 O M

N ~, ~ ~ N

V ~w 0 I~0001O ~ N M dw v0 t~ 00a1 M M M M M M M M M M M M M M M M

Ov o0 ~O O M .~N N ~O t~ ~t ~ ~ .w p N ~ O~ M ~nt~M ~O M M ~ I~ ~ 01 ~ M

O V1 I~~ ~ ~ ~ N V7 I~ \O .~ ~1 V7~p N ~O ~ 00l~N ~ 01 ~n ~n M ~ 01 ~O ~n~-N .-~ O .~M l~l~M v n t~ ~n N a1 ~nN

t~ a1 ~n ~n~nt~a,t~ 00 00 a, a1 ~n t~ 00a\

v~v1I~Q\~ N W O ~O N ~O O

~ ~ ~ ~n N ~ z l~ 01 O

d d C/~C/~

x x x x x x x x x x x x x x

48 ~i oo ~i cYi ~i .. oo cn o0 M ~ N ~ ~ d' O ~n ~ ~' d' M .. C~ I~ ~ l~ V~ l~ l~ l~
"'' O
~

,.-ax O ~ ~ ~ ~l ~l ~l ~l ~l O

d b "C~ p M 'b '~ 'C1 'G ~ '~ 'd 'L7 M "'~

, c~3 --~ -r ~ ~ , .
N

~ d, ~ ~' ~' ~' O ~' ~ ~ Q\
~

O O ~ ~ ~ O O O O O O O O
ox x a ~ x x x x ~x x x x . . aao ~ ~ ~ ~ ~ ~ ~

M ~ M j M
~ ~

~ D
t ~ i..
~ a a U d dd v~C7 ~

M ~

M .--r .~ M
~ ~
N

_~ ~ ~ ~ O _~ _cd ~ ~ _cd C'7 ~ a Hwa C7a dd 01 O ~ N M dw ~ ~D t~ 00 Q1 O

pp d' N ~ O ~ 00 00 O_ O

M ~ ~ N M ~ ~ ~' M ~ ~ ~ i ~ i ~ ~ ~ i O O ~ ~ ~ ~

N M ~ l~ ~ ~ M

M ~ -~ N d' N

O ~ N M ~ ~ ~ I~ 00 Q1 O -~ D\

M M M M M M M M M M M M

~n M ~n N ~ O~ 00 O O M M N

.-W ~ l~ l~ ~ ~ ,- N l~ N l~ l~

V7 ~ ~ ~ ~ ~ O M -~ M d1 00 ~ O ~' 00 ~ 00 M O t~ l~

N O o0 ~n N M l~ O~ N ~ N ~O

Q\ I~ 00 01 l~ l~ ~D l~ N ~ I~ 01 ~n t~ ~O ~O M O~ ~T' ~ ~O O~ M O

00 M -~ O ~ ~ N O\ N o0 V7 ~., x x x x x x x x x x x

49 ~t~~~'~tONN

M M M M M O
t~ h 01 a\ 01 01 01 .-~ O O

~ M O O

M M M M M ~

M

~D

~L

'b .. 0 M M
r., .. ,-, N cd ~

h ~!1 ~ ~ ~ ~' O ~

01 x 01 M 01 'C O '~ O p O

M x ~ ~ O x x a o x ,_, 00 O N N ~' "" ~ N . .-i Ov .. ~ N ~
.~

M ~ M ,~ 00 01 M 01 00 O h h h ~

O h v7 ~ h ~ M h h O
~ ,-~ ,-~ ~n O ~ O O O O O O h O =J
x O a -b a x ~ h x a ~ ~ ~ ~ ~ ~

~; O
M

~ 01 ~ ~ M ~ N ~D
~O d1 ' i ;

~ on ~ GO , }
~ ' ~

" C7 ~ . ~ ; ~ o ~
U ~ ~ , ~ H

0 0 0 0 0 ~ ~ ~
0 " 0 0 ~ o O O ~' ~' t' Y ~' ~ 00 N 00 ~ N 00 ~O V~ ~ N N r' ,., ~ p~ N r", .~ M b0 . b4 ~
d bA ' a -~ _c ~ _>, ~., ~. O
~ O O
y ., r~ ~ ~ ~ ~ H
~ ~

N M dw n ~D h oo O~

O O o0 00 M h 01 ~ N O ~ h N O h 'O h ~ ~ ~O

N ~n M , , , N , , ' , ' 00 Q\ O ' N ~O

M M N

W

N M 'ch ~ ~O h o0 Ov O

O~ a\ a\ O~ ~ ~ O~ O~ O

M M M M M M M M

..-, .~ h h N <h ~ Ov 00 M ~ V7 M M V7 h 00 O~ ~ ~n ~O ,- ,-.~ h t~ O

M ~ ~ M M O O

h ~p ~ h h M M ~O

O~ a1 Q1 01 Q\ O~ Q\ D1 Q\

w x ~ x ~ x x x x 0 0 0 .~ ~ .~ t~ 0 0 0 0 ~n 0 0 0 01 O O ~ - ~ N ~ O O O
O ~O O O

00 00 O O O ~ ~ I~ I~
00 O~ I~ t~ l~

Wit' N O O O O ~n M M M
N N O M M

01 V~ O O O O O ~
V~ V~
O

.---i M M M M M M M M
~p M M

N

cd ~ p ~ ~O '~ ~O~' .~ ~pOW '~ ~' ~ 00 O v~ I

o H a a ~ ,tea ~

a~ a a ~ ~ a .--iN .~ ,~ ,~ .-i-~
. , ~ ~ ~ O

O ~
O O O 00O O O \O
~O ~O ~t~O ~D ~O
,~ ~t 00 a O O O t~O O O O M
M 01 ~' O O O O O O O O O O
I~ v7 I~

x F-'H H a H H H H x ~ o o O

N ~ ~ vp M
O N

1 Ov ~ O ~ ~.
N bA

N a ~ d ~

~ d U

~ .~ N M
~ N

v~ ~, v~ ' ~ ~ by ' O
O ~

H

O ~ N M ~ ~ ~O I~ 00Q1 Q\ 0141 41 O~01 41 01 Q1Q1 ~

O ~ 1 l~Q1 N ~ M .~ N N
' ' ' ' ~ i ~ ~ ~

N M d' ~ ~O I~ 00 01O

O O O O O O O O O

~t ~ '~ ~ ~ d' ~ V

O vW O V1t~ l~ N t~

01 ~O~ l~ ~ O~ M \O .~

01 ~nO ~ N ~ ~ t~ ~ O

t~ Wit'v~ N Ov.- ~O O ~ ~n O o0 O N I~ .~ ~O ~Ol~

N

0o M N M ~ ~ N O N N

, _, d 3 ~

a ~ ~ ~ d ~

x x x x x x x x x b M cd ~' ~' ~' vp ~ ~ d' ~h f~j "

M ~ l~ I~ M I~ ~ l~ l~ N ~

O O ~ O O O O O O p~
a a ~ a a a x a a oo;~~a:

~ ~ '~ -d o ~ ~

...: ~ ~ ~
a ~ ~ ~ ~ asaaa ,n ,~ ,~ ~ .~ .~ ,~ N ~ .-. N
l~ .. . N N '"
I~

01 01 41 Q1 O~ 01 M 01 ~ p 00 .~
O C~ p~ M

tn I!7 V~ N N Vl ~ ~ d' 00 ~ M N
a V7 \O

p O O O O O O O O
.~

a x x x x x x a . x x aaaa . . . ~ . . . . .

M ~O ~ M D1 [w N
N 00 O~

j~ OW , 7, ~, bA bn D
by j~ ' ~" Q ~
, G

_ N ~ _ ~ s..
~ ~ ~ ~ ~ [7 N d > d ~

O o O ~ O O O ~ O O O O O
p O p 00 01 v~ ~' O a\
~ ,-, O

M ~ ~ ~ ~ I~ 00 ~-, 00 N M N
~ ~ M , , , , ~ , , , .--, , , , _, , N ~ ~
~

N ~
c~a~a H ~ ~ N t7~da ~c7a>ac7 dv~

O ~ N M d' ~ ~O I~ 00 O1 O

O O O O O O O O O O

p~ Q1 01 01 01 ~ ~ ~ Q~ 01 N N ~O N
N ~n ~ O o0 v7 v~

O O ~ .~ ~ ~' v1 01 M N

N ~' , , , N , , M M ~!1 ~ 00 00 ~ ~' -~ 00 V7 N ~ 00 01 M M M N ~ ~ N

N M ~ ~ ~O l~ 00 O~ O

.-, .-.,.~ ,~ ~ .~ .~ N N

W t' -, I~ N ~n ~ N o0 a\ l~ I~ U1 D1 ~ M I~ ~ ~ ~ ~O M 00 ' N ~ N ~O ~O ~O Q\ M ~ N l~

N N M v~ M ~O 01 ~n N ~n 01 l~ l~ l~ I~ 00 l~ 0o l~ I~ l~

M
~ OM

v~

>

d x x x x x x x x x x x 00 I~ 00 Q1 l~ d' ~ o0 I~ N

N ~ .. d' ~ ~ ~' ~ N O v~ M N ~O
~

~ I~ I~ I~ t~ 'd' ~O I~ d' ~ M
O O O ~ O O O O O O O

. . a a a ~ a x a a ~ x a a~~ ~ T' ~ ~ ~ ~

x a a ~ N ~ ~, ~ ,~,~ .~ N ,-.

00 . . ~ l~ t~ t~ l~ ~j I~ l~l~ t~ pp l~
~n j N N N N N N ~p N N N N ~ N
O

~ M 00 ~ ~ ~ ~ ~ y~ ~' d'd' ~ [w d1 ~

x x x x x ~ ''~x x a x 0 0 0 ~
o , . . . . -1 ,.. . .
~ .
r M

cri O ,n ~ ~ C7~ UUrs'.' 'i' d U' d o x ~ ~ d Y

0 o ~ o o ~

0 0 o ~n o Y i.~ y., Y Y i' Y Y

N N M M ;' M ~ p vp ~ ~ ~ ~ N d y cn i ~ ~ 7, ~ ~ ~.. ~ d4 ~ v~
p v~ p a.', ~ ~
c~

a ~ c7 c7 c7 x c7 d a v~ ~ a a a~ ~ a~ d ~ a N M <t ~ ~O l~ o0 O~O ~ N M

.-. ,--.~.-~ .~ ~ .~N N N N

D1 ~ ~ d\ D1 ~ Q1 ~ ~ Q1 ~ d1 o~0 ~ N 41 ~ N ~O ~ Q~O ~ ~ N

M M N V7 ~ ~.,~ Vii'M ~.,~d' M

, i ~ i i ~ N , ~ i O ~ ~ M ~ o0 O ~ ~D
O ~ a\ N ~ I~ a1 M ,--~v~ M N .-., N ~ .~ N ~ .~ N M .~ ~ N

N M Wit'~n ~O t~ o0 O~ O ~ N M ~t N N N N N N N N M M M M M

d' ~ ~ ~ ~t .--W ~ l~ ~ l~ O M O I~00 l~ N I~

N ~ ~O ~ M 00 00 d' M V7 01 lp l~

N M M ~1 Ov Ov M ~ -r N o0 N

V' vW ~ \O O o0 00 v7 N l~ a1 00 lw 0 ~ ~ N ~ ~ M l~v~ ~n o0 v~

~p o0 ~D O~ ~ 00 00 Q\ 01I~ Q1 l~ I~

N ~O N ~ .~ ~ ~ 0 ~ ~ ~

~ ~O
~ ~ ~ ~ 3 O p ~ ~

c C .7 c w w w r w w w r. w 7 7 t 7 ~, ~

.

x x x x x x x x x x x x ~.

..

v0 cri ~ oo .. .-. vp M N a r-a a a N a a D

'b 'b 'd p ~ "C7 a a ~ ~ ~ ~ a b a ~

.-, r-. .~.~ .~ ,-- ,~ ~-r.~

.
r, p l~ N N N N N N N N N N N
p~ p~ ~

M ~n O O O O O O O O O O O
op ,a~OO x x x x x xa x~ x x x x~
. . . .

.. N r., M

.-~ ~' v~
,-, ~ N
,--~

M M O ~ N V'1 lp [w -S.~" ~ ~ ~ ~; ~ ~
~ ~ v~ ~., ~ ~ N
'-' s~.
ac7 ~~ E..,a aE-, ~,c7v~.~~

0 0 0 0 0 a o C7 a 0 0 0 ' o o o o ' ' ~ M ~ ~ Q
r.-a p ,~,, ~ '-' M l~ V1 r.. M

N ~ ,-, N N ~ M
,~ v~ ~n ~O
,~

_c~ p bA by _~, s~-~
f~. v~ bA _.~~'.,~
by b11 bA

a ~ C7 C/~ H
~ C7 ~ ~
~

C

FW D I~oo Ov O ~ N M ~t ~n N N N N N N M M M M M M

U ~ Q1 D\Q1 ~ Q\ ~ Q101 01 ~

N ~, l~ OvO v~ .~ Q\ ~ ~ ,-, 00 M N ~ N

~ ~ N N ~ N N N

l~ d'~O
~

N N

~n ~ I~ 00a\ O ~ N M ~ ~n ~D

M M M M M ~ ~ ~ ~T'd' ~ d' d' ~ ~ ~ ~ d' 0o vp M v~ O ~ a\ ~ON O Ov o0 I~O o0 O~ o0 ~ O O O

M ~O N ~ ~O .-~ M 0000 O ~D

D1 O ~ 01v~ l~ Q1 ~O O O O~ O~

V7 ~ M ~ N ~ 01 O ~ l~ d' ~O

l~ I~ OWO t~ ~ O~ ~DvD o0 ~D

m O ~n o0O~ v~ N o0 ~n.~ l~

~D ~ O O ~O M ~p l~N O

U G~1 ~1 ~4~ ~ ~ H r~v~ vo w w w w w w w w ~1~1 ~1 ~1 x x x x x x x x x x x x --.

N v~ ~' ~ ~ N
.-i ~ ~ a ~ ~ ~
o o ,- l 7 ~-o -d b -d o a '-~

a op I~ I~I~I~I~ t~ l~ r t~l~ l~I~I~l~l~ ~p ,.., N N N N N N N N N N N N N N N
pp d' ~t~ d'~ ~ Wit'~ d'~ ~ ~ Wit'~
~

a x x x x x x x x x x x x x x x a aO
. . . . .

M ~" M
, ~ a~
'~ ~
bD
~
, p ' O ~ , >, cn cn cn ~

p p ~ O ~ p O
O O

O
O a..~ .'.~ O ~n N ~ O
01 .~

00 ~ ~ '~ M M
N M
~

N ~, ~ ~ ~ S bn ~~~, t=. ~
~.

_ _ I~ ooO~O ~ N M ~ ~WO I~o0a1O -~ N M

M M M M ~ ~ ~ ~ ~ ~ ~ d'~f~ N N V7 V7 d> ~ ~ ~ ~ ~ 01 ~t ~ ~ ~ N v~ O ~ ~ 00.~ 00N ONM ~ ~ ~D

M ~ N N , ~ 61 V7 N l~M ~ N M 00~, ~.,~M

~ , ~ ~ N ~ , , ~'N , , , '-', , ' N O -~' ' M o0 ' ' O ~D~O' 00 Q\ ~ .~~ .~M N vD O N l~ l~O~~ ~ O ~ 00 M .~~ N -~ N N N ~ M

t~ 00 01O ~ N M ~ ~ ~ t~ 00O~O ~ N M

~ ~n~nv~ ~n ~n ~n ~nv~ ~ vwo ~ w o ~t ~ Wit'~ ~t~i' d' ~ ~ d'~ ~ ~ ~ ~ d' O I~ o0O O N N ~ M ool~ ~O~OO t~~ O 01 ~ M I~~DO o0 d' ~ ~ N ~n ~O(~N N O M a1 M ~ N l~I~M v7 N ~ O~~O O d'-r~OM O M

M O V7~ M M N V' d1 N M N M I~V~01 M I~ ~ON ~ oo ~n Ov ~ '-..~ M V'v~N v~ O

l~ 01 00l~I~~O OW E ~t l~l~ OWE ~OQ\01 l~ 01 ~O N ~ ~ N l~ N ~ <t ~ N V7N M ~ 00 O

~n .~ ppv~~ N r. Ow 0 ~ ~ O ~ ~ O O M
d ~, ~ ~ ~-W W U U U U U G~G~ld N ~C

U

x x x ~ ~

x x x x x x x x x x x x x t~ oo ~ oo t~ o;

dw ' t~ ~Y N ~ N D\ N

O ~ O O O O O O O O

a ~ a a x a x a x a r, ,~ ,~ ~ .~ ,~ N ~ M
.--i M

l~I~ l~ I~ I~ l~ .~ ~ l~ p l~ .~ I~
l~

~ ~' ~ ~ ~ ~ ~ d' ~

O O O ,- O O p O O p O p O
O

x x x~; x x~ x~ a~ x~ x a~ x~ a~ x ~

' M ~ N ~ .~ N ~ ~ 00 ' ~ b ~. O~ v' C
b 0 , v~ N ; ~ by ~ , ~, N ~ A ~, ~, ~ cn ~, a ~ ~ ~, ~ O y cad ~, U 7 x ~ d ~

~ ~ w rr~
~7 ~l C ~
>

0 0 ~ 0 0 0 0 0 0 0 ~ M ~' O O
~ M ~t CV' M
.

~ ,~ ~ r, .--,M ~ ~ ,~ ,~
O ~' N (~ ~ l~ .~'~,, ~ O s; W, cd ~' O s~. v~ p n j, , H
cd O
c d>w C7 U C7v~wa~ v~v~C7dxr~ C7d d'N \O I~ 00 ~ O -~ N o0 M ~ V7 N ~n ~n ~m n v~ ~p ~D ~ oo vO vo vo Q\ Q\ Q1 Q1 ~ 41 ~ Q\ D\ Q1 ~O~ M O O ~ ~ ~ M 01 Q\ 01 ~ I~

N V1 ~!1 M M ~ N , ~t M M d' , i N , , , , ~ ~ , , , ,--', 00O\ ' \O V~ ~ ~ I~ M ~ 00 ' V~

N O N O O .--~oo ~ N M ~n o0 N O~

M ~ M ~ N N N --~ N

~n~O l~ 00 01 O ~ N M a1 ~ vW p l~

a1 ~ d' ~ ~t ~ ~ ~ ~ d~ ~ ~ ~ ~t 00 M O l~ N N V1 00 '~ ~ O~ M

pvo0 00 ~t O M O O v~ ~ ~ N ~O N

M l0 \O O ~O ~ O ~D ~ 00 00 M ~ 00 M ~' ~t O v0 Ov O O ~ ~ O ~D N

~ON ~ N 01 N ~ a1 01 ~O t~ d' 01 01 a1 a1 M O O M 01 l~ ~O O~ M M O ~O N
' ~

I~v~ M O N I~ ~h N pp OW 1 a..a~ o o z x A ~ v~ ~ ~ w w U U

~ ~ ~ ~ ~ ~ x x ~ x x x x x x x x 00 0 "~ O t~ oovD
N "' -' N ~' ~ ~ ~ ~' 00 l~ ~ ~O ~D
01 v'~. I~ O l~ N l~ l~l~.
~n O O O O ~ O M O O O O ~ O M
I~ oo _ O i..~~ C/~ ~ n-.~a ~ ''~0 0 ~
N

b ~ -d b -~ -~ -~ -~~ ~
o ~ ~ ~. , , ,.
,~ ~ ~ ~ ~
o ~~
a (~j.-~.~ .~ N .-r.~ M
~ . ~

.- . . . . . r, . . . N
.-~ .-. ~--r ~

~p p ~ O t~ O O . Q\ O O N N N ~
.. .. . ..

~n ooO oo ~ o000 ~O M o0 0oN N N ~
O ~ O ~O

I~ N O N ~ N N '~ ~t N N O O O I~
d' ~ 00 ~

O O O O O O O ~ O O O ~O~O ~OO I~
I~ I~ N l~ l~

~ C/~,~ C/~r-~ C/~C/~ a ,-~ C/~ C/~C/~C/l C/~H..~
a O ~ O a O

O p1 M 01 ~ v~ 01 00 01 O ~
O

M ~ N ~ ~ N M v~ ~ l~ d' ~ ~ ~' ~n ~1,~ ~ ~ ;' c~ O ~ ~ p ~ 'O~ 4j ~ ~, O ~ ~ ~
~

H ~ ~ a i _ ~. ~ C7 ", a H d as H a > P~ C7 C7 O ~ O O ~ O O O tj .~
O O O O O O
O O ~

~D 01 ~ l~ M O V~ M N " '''I~
vp I~ 01 00 p~ ,-..~
M

M ~ ~~' ~ pp r-.. ~ M M v1 M M M '~Y ~p a1 ~ 01 b ~ ~ s' p ~, ~ 7, ~ p ~iv~bA
A ~ ~ v ~
~ r' ~, ~
~

x ~ C7 ~ v~ ~ _ H ~ U
d H ~l ~; C7 ~ C7 ~ ~
H C~ C7 x ~- , l~ 0001 O .-. N M ~t ~ ~Ot~o0 01O

Q\ D\Q\ ~ 01 ~ ~ Q\ O1 Q1Q1~ Q\~

V7 M N ~ O ~ M -~ ~ ~ 01~D ~ 01 M N ~ \O ~ l~ d' d. N M ~,,~O~

i i ~ ~ ~ M ~ N , ~ 00~ '~ M ' ~ i ' N ~ ~ ' i ~ 0 N ~ M

M lp M O M M N M N
M M I~ M M

~ ~ ~, 00 a1O ~ N M ~ ~n ~O l~00O~ O --~

I~ l~00 00 00 0000 00 00 000000 ~ 01 d~ d'~ '~t~ ~ ~t ~ ~h 00 \O~ ~ 01 ~ M ~ ~ O N l~ M l~

01 00~n O ~n .~~O I~ v~ O o000 ~ N

00 00O O ~ O N M O~ 00M 00 ~ 00 dW 0 ~ M d N N M M 00N M

l~ ~O~t 00 \O l~I~ \O M 01a1M N I~

I~ O\00 ~O ~ 0000 ~O 01 ~Ol~l~ Q\~O

~O O - M ~ N M M I~ M

l~ ,_,l~ N .-~ t~pp ~ ,~01~n O

U U p ~ ~ ~ ~ ~ ~ ~ ~ ~ ~

q C7 U U U U U U U U U U U U U
d d d d d d d d d d d x x ~

x x x x x x x x x x x x s~

b t~ ~n as Mo a [048] The first column in Table 1A provides a unique "Clone m NO:Z" for a cDNA
clone related to each contig sequence disclosed in Table 1 A. This clone >D
references the cDNA clone which contains at least the 5' most sequence of the assembled contig, and at least a portion of SEQ >I7 NO:X was determined by directly sequencing the referenced clone. The reference clone may have more sequence than described in the sequence listing or the clone may have less. In the vast majority of cases, however, the clone is believed to encode a full-length polypeptide. In the case where a clone is not full-length, a full-length cDNA can be obtained by methods known in the art and/or as described elsewhere herein.
[049] The second column in Table 1A provides a unique "Contig m"
identification for each contig sequence. The third column provides the "SEQ m NO:X" identifier for each of the connective tissue associated contig polynucleotide sequences disclosed in Table 1A. The fourth column, "ORF (From-To)", provides the location (i.e., nucleotide position numbers) within the polynucleotide sequence "SEQ m NO:X" that delineate the preferred open reading frame (ORF) shown in the sequence listing and referenced in Table 1A, column S, as SEQ >D NO:Y. Where the nucleotide position number "To"
is lower than the nucleotide position number "From", the preferred ORF is the reverse complement of the referenced polynucleotide sequence.

[050] The fifth column in Table 1A provides the corresponding SEQ >D NO:Y for the polypeptide sequence encoded by the preferred ORF delineated in column 4. In one embodiment, the invention provides an amino acid sequence comprising, or alternatively consisting of, a polypeptide encoded by the portion of SEQ ID NO:X delineated by "ORF
(From-To)". Also provided are polynucleotides encoding such amino acid sequences and the complementary strand thereto.

[051] Column 6 in Table 1A lists residues comprising epitopes contained in the polypeptides encoded by the preferred ORF (SEQ >D NO:Y), as predicted using the algorithm of Jameson and Wolf, (1988) Comp. Appl. Biosci. 4:181-186. The Jameson-Wolf antigenic analysis was performed using the computer program PROTEAN
(Version 3.11 for the Power Macintosh, DNASTAR, Inc., 1228 South Park Street Madison, Wn.
In specific embodiments, polypeptides of the invention comprise, or alternatively consist of, at least one, two, three, four, five or more of the predicted epitopes as described in Table 1 A. It will be appreciated that depending on the analytical criteria used to predict antigenic determinants, the exact address of the determinant may vary slightly.

[052] Column 7 in Table 1A provides an expression profile and library code:
count for each of the contig sequences (SEQ ID NO:X) disclosed in Table 1A, which can routinely be combined with the information provided in Table 4 and used to determine the normal or diseased tissues, cells, and/or cell line libraries which predominantly express the polynucleotides of the invention. The first number in column 7 (preceding the colon), represents the tissue/cell source identifier code corresponding to the code and description provided in Table 4. For those identifier codes in which the first two letters are not "AR", the second number in column 7 (following the colon) represents the number of times a sequence corresponding to the reference polynucleotide sequence was identified in the tissue/cell source. Those tissue/cell source identifier codes in which the first two letters are "AR" designate information generated using DNA array technology. Utilizing this technology, cDNAs were amplified by PCR and then transferred, in duplicate, onto the array. Gene expression was assayed through hybridization of first strand cDNA
probes to the DNA array. cDNA probes were generated from total RNA extracted from a variety of different tissues and cell lines. Probe synthesis was performed in the presence of 33P
dCTP, using oligo(dT) to prime reverse transcription. After hybridization, high stringency washing conditions were employed to remove non-specific hybrids from the array. The remaining signal, emanating from each gene target, was measured using a Phosphorimager. Gene expression was reported as Phosphor Stimulating Luminescence (PSL) which reflects the level of phosphor signal generated from the probe hybridized to each of the gene targets represented on the array. A local background signal subtraction was performed before the total signal generated from each array was used to normalize gene expression between the different hybridizations. The value presented after "[array code]:" represents the mean of the duplicate values, following background subtraction and probe normalization. One of skill in the art could routinely use this information to identify normal and/or diseased tissues) which show a predominant expression pattern of the corresponding polynucleotide of the invention or to identify polynucleotides which show predominant and/or specific tissue and/or cell expression. The sequences disclosed herein have been determined to be predominantly expressed in connective tissues, including normal and diseased connective tissues (See Table 1A, column 7 and Table 4).

[053] Column 8 in Table 1A provides a chromosomal map location for certain polynucleotides of the invention. Chromosomal location was determined by finding exact matches to EST and cDNA sequences contained in the NCBI (National Center for Biotechnology Information) UniGene database. Each sequence in the UniGene database is assigned to a "cluster"; all of the ESTs, cDNAs, and STSs in a cluster are believed to be derived from a single gene. Chromosomal mapping data is often available for one or more sequences) in a UniGene cluster; this data (if consistent) is then applied to the cluster as a whole. Thus, it is possible to infer the chromosomal location of a new polynucleotide sequence by determining its identity with a mapped UniGene cluster.

[054] A modified version of the computer program BLASTN (Altshul et al., J.
Mol.
Biol. 215:403-410 (1990), and Gish et al., Nat. Genet. 3:266-272 (1993)) was used to search the UniGene database for EST or cDNA sequences that contain exact or near-exact matches to a polynucleotide sequence of the invention (the 'Query'). A
sequence from the UniGene database (the 'Subject') was said to be an exact match if it contained a segment of 50 nucleotides in length such that 48 of those nucleotides were in the same order as found in the Query sequence. If all of the matches that met this criteria were in the same UniGene cluster, and mapping data was available for this cluster, it is indicated in Table 1A under the heading "Cytologic Band". Where a cluster had been further localized to a distinct cytologic band, that band is disclosed; where no banding information was available, but the gene had been localized to a single chromosome, the chromosome is disclosed.

[055] Once a presumptive chromosomal location was determined for a polynucleotide of the invention, an associated disease locus was identified by comparison with a database of diseases which have been experimentally associated with genetic loci. The database used was the Morbid Map, derived from OMIMTM (supra). If the putative chromosomal location of a polynucleotide of the invention (Query sequence) was associated with a disease in the Morbid Map database, an OMIM reference identification number was noted in column 9, Table 1A, labeled "OMIM Disease References)". Table 5 is a key to the OMIM reference identification numbers (column 1), and provides a description of the associated disease in Column 2.

Clone ID SEQ ID CONTIG BAC ID: SEQ ID EXON
A

NO:Z NO:X ID: NO:B From-To ~HADDB75 31 757028 AC006405 1023 1-96 ~

HARMG60 46 933284 AC025821 1055. 1-552 HARMM43 47 714763 AL138499 10'58 1-386 HARND80 56 864604 AP001459 1077 1-5?5 HER.AC92 69 973454 AC023498 1112 1-300 HER.AJ78 74 973676 AC009949 1120 1-848 HKAF042 113 713722 AC022059 11?1 1-1250 HK.AIL12 119 893937 AL157697 1178 1-286 HKAll~l2 119 893937 AL035588 1179 1-292 HKA~,12 119 37 AL035588 1180 1-313 HKAJG02 121 ~ _ AC073984 1181 1483 ~

~HOUHU87 154 791044 AC003692 1246 ~ 1-44 . 695-1033 HWDAP03 180 9233'19 AC073186 1297 1-371 .1210-2684 HWHGB20 185 ~ 669455 AF037222 1314 1-1736 _ X622-7896 .

HWHJJ11 241 965189 AP000217 413 ~ 1-125 HWFAD94 263 504477 AL355305 1448 ~ 1-439 HWDAP89 270 ~ 795713 AL138742 1464 1-350 ~

. 2375-2496 HSTAG60 281. 578487 AF139813 1482 1-1066 HOUB069 302 757808 AL035702 1530 ~ 1-1829 385_57-39353 HK.AFQ61 324 741786 AL355800 1583 1-1517 HK.ABR92 348 879400 AC005237 1626 1-506 HFEBY03 353 973292 AC007944 1638 ' 1-281 HFEBJ61 35fi~'' 576092 AC022488 1651 1-676 HFEBDOl 358 916725 AC010311 1654 1-774 HER.AN59 371 739562 AC016953 1680 1-215 HER.AL72 376 529196 AP000183 1689 1-348 HER.AK96 377 796591 AC027306 1700 1-104 HER.AH16 382 880475 AC023592 1704 1-568 HER.AH06 383 954672 AC013633 1713 1-574 HERAH06 383 954672 AC011138 1716 ~ 1-575 HADFW06 429 935340 AL359878 __ 1-138 1794 ~

HADDE27 452 683382 AL356365 1899 i-531 HADCWOl 464 916399 AC012431 1924 1-89 HACAA57 489 733887 AC027384 1989 ~ 1-403

[056] Table 1B summarizes additional polynucleotides encompassed by the invention (including cDNA clones related to the sequences (Clone 1D NO:Z), contig sequences (contig identifier (Contig >D:) contig nucleotide sequence identifiers (SEQ >D NO:X)), and genomic sequences (SEQ m NO:B). The first column provides a unique clone identifier, "Clone m NO:Z", for a cDNA clone related to each contig sequence. The second column provides the sequence identifier, "SEQ m NO:X", for each contig sequence. The third column provides a unique contig identifier, "Contig >D:" for each contig sequence. The fourth column, provides a BAC identifier "BAC m NO:A" for the BAC clone referenced in the corresponding row of the table. The fifth column provides the nucleotide sequence identifier, "SEQ
>D NO:B" for a fragment of the BAC clone identified in column four of the corresponding row of the table.
The sixth column, "Exon From-To", provides the location (i.e., nucleotide position numbers) within the polynucleotide sequence of SEQ >D NO:B which delineate certain polynucleotides of the invention that are also exemplary members of polynucleotide sequences that encode polypeptides of the invention (e.g., polypeptides containing amino acid sequences encoded by the polynucleotide sequences delineated in column six, and fragments and variants thereof).

M ..-,~t ~ ~ ~O I~ l~ I~ I~ l~ ~O I~ ~O
M N l~ l~ l0 l~ l~ l~
.-, t~ ~ N M ~ ~ ~ ~ d' 00 ~ oo ~t ~
M O oo ~
O

M N N d' v~ v> ~n ~n ~n ~n ~n ~n ~' ~n ~ v1 v~ v~ ~n ~

H

z ~n t~ ~p I~ ~O ~O ~ ~D ~O ~O ~O ~ ~
M 00 ~n ~O ~O ~ M ~
V~ ~

l~ 01 N O~ ~n ~n ~n ~n ~n v~ ~ ~ v~
t~ 01 ' oo ~ v1 ~n ~n ~n I~ d' M N ~ d ~n ~n w H

z ss sss s sss sssssssswssss 00 000 0 000 oooooaooooo000 O N ~O O ~ ~O ~ ~1 d' M ~ ~ M O ~ M O
00 ~O M ~ N
~O

d' ~ V7 ~D l~ V~ N ~ V7 ~ ~ In V7 V7 ~1 V7 ~ I~ In ~ N N

~. v v '_'. o ., a~ .~ ,~ ,-, ,--.

U

O M

.b N I~ ~O

~I M ~ V7 00 M

N'-' U U U

a z ~N wo y ., y O ~

N
v~

b0 O v~ O O
s4, O

44; ~ .. .fir C/~
N ' O p.. -~' .~., V ~

_ cd cCi ~ N
' N

O M cd ~ ~ 00 U C7 U ~ ~

~ a~ 0.~.~ ~ o O ~

M ~. ~ O , O ~
~, ~ O S

O ~ O ~ O
V ~

CG O ~ (1~ ~ N O
~ ~ O
v~

G.~ N

N ~ N N N

k k p ~ c~ c~3 cd c~ c~

_ _ _ _ _ a O

V~ ~ ~ ~ ~ O N

C~

O
A N ~

U , ~ ~ ~ , o o M M p~ M I~

N N ~ 00 M

x a p U U
q U

r U z x x x x x M ~ ~ ~ t~ ~ t~ ~ wo t~ oo ~ oo r~ ,~ ,~ .-. .-. ~ ,-. vo vo ~ wo ~ .-wo ~
N 00 00 00 ~' 00 ~ 00 00 N ~Y ~ ~~ ,--~ .~ .~ .~ ~ .~ p\ .~ p~ 01 01 ~ 01 01 .-~ Q~ 01 ~n v~ ~ v~ ~n ~n ~n ~n v» V~ '~ d' ~t t~ t~ I~ ~ Iwo t~ W o vO v0 vD v0 n ~D
v0 ~D ~ ~ M ~O ~O ~O ~D ~D ~ M ~D \O ~O 01 t~ l~ M M I~ l~ I~ l~ I~ I~ l~ l~ l~
I~ l~
V1 ~ V~ V~ V'1 V1 V~ V~ l~ V~ ~ V~ V7 V~ ~ ~ ~ O~ 41 ~ r-.~ ~ ~ .~ ~ .~ .~ .~r .-1 .-r V~ N ~ ~ N V~ N N ~ V~ t~ V'1 V~ V~ ~
0 0 0 0 0 0 ~ 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 ~ 0 0 0 0 0 M 00 00 t~ 41 lp l~ ~ ~D O~ O 00 ~ 00 O ~ ~~ 01 01 V7 M ~ M ~1 M M M O ~ V1 V7 V ~ ~ ~ ~ ~ ~ ~' ~ ~ d' ~t M ~h '~t ~ M M ~ Wit' ~f ~ ~ ~ '~t ~O N ~O ~O ~D ~ ~Q O N O~ ~p ~p ~p O ~ ~O 0o ~ M N N O
.~ 01 O ~ 01 01 01 ~ O l~ D1 00 00 l~ l~ 00 ~O N ~-~ 00 l~ M
I~ ~O h ~D ~O ~O ~O ~ <t N N ~~ ~ ~~ M M .~ N U1 N
I~ l~ l~ l~ I~ t~ t~ I~ l~ ~ ~ O ~D ~O N M 00 O N O~ t~ O o0 W V'1 V7 V7 V'1 t/7 V7 ~!1 ~ M N M ~t N ~-i \° \ \° \° \° \° \° \°
\° \° \° \° \° \° \°
\° \° \° \° \° \° \°

~D M M M O ~O M 00 ~ N N N ~ ~ ~O ~ l~ ~ 00 00 ~t ~t ~' d' ~ d' ~ M N l~ ~ ~O M M Wit' ~ ~O 00 Q1 ~ ~ I~ O
O O
p V1 ~ ~ ~ O
O ~ ~ ~ Do N ~ N ,-, N N ,-. vp U ~ ~ W
~G ~ ~ ~ Gq U ~ o'oo op o, ,~ ~ N o b b0 ~ O .b0 _Uv ,~ N
a~ ~ ~ x ~ ~ i ~ ~ U

a ~ ~ a ~ ~ U r-~
Cn ~ Ai ~ ~ ~ O
U
0~0 ~ ~ ~ N ~~ ~ ~ ~ ~ O \O V
a ~ p.
o ~
O ~ O ~~ ,N O ~ V ,~ O ~ .N N ~ O
w ~ o ~ w o ~ ~ o ~ ~ o a.~dx'N ~ ~ a,~ d o. ~~ a-, N N N N N N N
_~ ~ ~ _~ _~ ~ _~
00 ~ V) l~ ~ M \O lp N M M M M d' ~ ~D
O ~ N ~D O ~~ ~' ~O
~O I~ O M M t~ O f~
M 00 O V7 00 Ov \O t~
~O ~O Q1 l~ ~n 00 ~ O
~O o0 ~ N l~ <h ~O M
vp ~ oo W O o0 0o O~
O_O p~ ~ pp y1 ~~ O
r-'7 w a ~ M P~ O
U ~1 ~1 w w t7 0 t~ ~n ~ ~n ~ ~ ~ t~ ,..., vo ~ o Wo ~n oo a1 ~n 00 O Ov o0 N ~ O Q\ N M O M l~ N oo dW ~ M
M M ~ N r' M M M 00 ~ .~ V7 . N .-~ V7 M l~ ~ O ~-~ N 00 01 .--i N M 00 ~~ 00 M O N 01 O ~O o0 O l~ ~~ ~ ~ N ~D M N o0 d1 t~
~M ~ ~ M
\o~~~

~ O ~ ~~ W Y ~ 00 N v7 \D l~ 00 00 O l~ 00 V1 N
~ 01 ~ WO 00 ~ ~O M N V1 ~ M M ~ ~D ~O
.-r ~ ~ ~
pNp O ~O p ~ I~ M vp O oo N '""' N
N Q\ ~ N ,~ N w ~ O
01 N 00 ,~ ~ N ~ O ~ N N
O N .~ ~ M .~ ~ r' ,~ O N N
~I U ~I ~I ~I
M ~ ~ ~ due' ~ ~ ~ ~ p ~--i ~ .~, ~ 00 .fl .fl ~ ~ .~ ~ 00 by bW O '~ N d4 Ov 'b bl~ d0 U1 bQ b~ .--~
H
O M V ~ O ~ N ~ W N
~O ~ O V . ~ ,~ . G,~ M ~O 4-~ ''b~''D . o W a1 U 7, N ~' p, ~ '~ ~' o ° ~~~ ~ ~x ~ ~ ~ ~ o~ ~°
' a~~
o°o~oø'~° ~~'~' ~~o0~"oNO
Mø,,-,YCnMcn O~V ~x~~NO.,-,N~U V~~M~,~Oc~n o°~~ ~o~o ~oa~~~.~o°°,oo.~~ °.~~n ,~~~'oo°~°o 'a~a~ s~~~~ ~~~vOa~'-,~oa~a~.~a~~ .~~n0 '_' ... a~ o w o ;~ ~ a ~' o ~' . ~' ~' ~ ~ ~ 0 0 ~ t1, Vl s-a a a a !n C/~ Qr '~ a S~. ~ a a (1, V1 a f3~ Z," a a a 4. a a a N N N N N N N N N N N
cd c~ _cd cd cd _cd cd _c~i c~3 cd _cd .fl .fl .fl ~ .fl Q ..Q ~ .~ .O
~D l~ M
O~ ~t v7 ~O D l~ d1 O O D
Q\ d1 U
~D ~n N ~n M N N ~t ~n N
~n I~ v~ O M ~ t~ N d1 M N
d' ~O M ~ ~ 41 M O 01 N I~
M M ~ ~ d' ~D N oo ~h v~ M
I~ I~ N V~ twp O O ~t ~D ~
'_"'' op M N
U
U O
W W W G.W~, GWI, x x ~x x x x x x x o, .-. V, ~ wo .~ ,-. ~ 0 00 ~ av o N n N ~ ~ ~ ~ ~ M O~ M 00 O
M .-.~ ..-i ,~ M N .~ ~ M N
M ~D 01 N N ~ O ~ r-~ O M N 00 M
.-~ O O O I~ ~ ~i' ~O Ov M N
N N ~ ~D N

~D D v'1 V~ M l~ 01 O a\ C~ M d' O V1 O~ O ~O v7 ~ ~O I~ O 00 \O 01 Q\ O M
O .~ O O
.--i .~ r' ~O Q~
.-a ,~ ,-~ N ~ ~ ~ oo t~ ~n M
O M_ ~ ~ M ~n p p 00 ~~ O~ M
N M O ~ ~ O N
I I I~ ~ IU
~0~~~~1~~~ G~1~1~1~,.fl cp vv on ~n cn a~ a~ -o an o, op -d ~d b a~ a~
o ~ o b x ~x ~x ~ v ~ ~ ~ v ~ ~ .~ ~ ~ ~ ~ ~,-; o 'd G' P-i 'S-." ~ w ° ~ U Pr ~ o p.. ~ U ~ U ~ U
U o~0 O A"' ~ '~ '~ c~i ~~ '~ 'd °
.-~ N ~ ~ O Q. M ~ ~ Q1 ~.~ "C~ 00 ° p . ~' ~ U ~ 01 ° \D
° O1 ° ~ ~ .~Ur can ~ U O\ ~ 01 ~ y ~ p., ,-, ~ ~ N ~ ~ U ~ Q., ~ Q., ,-, ~ gyp., ,-, cd ~ o ~ d ° ° ° ~ > o ~ ~ a°, ° °o .~
~~ o ,~ ~ o ,~ ~ o ~ ~ M .~
o ° ~,~Zo ~o ~.~,o~a;.~o ~o 0 ~ ~x~~~x~~d~ ° ~~x~ ~w~ ° ~~ ° ~~ ~,~
N N N N N N N N N ' N N N
k k ~ k w cd c~3 c~ _cd cd cd c~ c~ _cd c~ cd c~
.fl .fl ~n M t~ Q\ ~~ 00 N ~ .~ ,~ N Q1 N N N M M d' ~ ~n t~ l~ l~
~D l~ M l~ O l~ o0 .~ ~ o0 .~ O
00 00 ~ ~' N ~~ ~ v~ 00 'O ~D N
M ~ ~ N N l~ N N I~ M Q\ ~
I~ ~O M O~ M ~O Wit' ~ O o0 ~O 01 o0 M ~O t~ I~ N N M ~n O O (~
N
3 O O Z U x a7 N ~' U d O ~ O O
x x x ~ x x x x x x xx v1 ~O ~O O ~ ~ ~O ~O ~t l~ o0 N OWE v7 ~O oo O_ N
N t~ l~ O M I~ ~ ~ N ~ N .--.
M M M d' ~h M ~' N ~ N 00 00 .-a N N M N ~
O M I~ \O ~ 00 .-r ~ ~ l~ ~O I~ ~ I~ N ~ ~ 01 00 v7 ~ O~ O O~ V~ ~O 00 ~--~ 01 v7 ~ I~ M ~n O
.. .~ N M N N M ~-~ v~ ~ r. .-~
0 0 0 ~ ~ 0 0 0 0 ~ 0 0 0 0 0 0 0 00 v~ t~ v~ ~p ~ o0 l~ O N ~' M O N ~ ~ O ~ ~O
N M M M ~ N N 01 ~O o0 l~ ~ O ~ ~O t~ 00 ~ ~D
O
-t~ 00 N ~ O ~ M ono N M
O ~_ .--. O
N
O ~ ~ N ~~ N ~
O ~1 ~ ~ P~.1 ~ ~ O ~ ~1 ~L, b 'b by '~ d0 by ~~ GD 'b bQ 01 r-~ "C7 ~ N 'd ~ U ~ N i-~.~ ~d 0 M
U cad v ~ ~ ~ cd ~ ~ at ~ ~ ~ y,y ~ ~ O
x ~ ~x ~ ~ o ~ ~x o ~~x ~ ~ ~ o ~~ x~u~w ~ ~ ~ ~ a., ~
.din ~'Q,M~~o x. a~,o ° oy~.~v.'~on~ oo ~v, °' '~ '~ ~' N° ~ o ø. ,~ ~ o ~~ ,-~ °~ an o ~ o ~ ;~ . ~
o u. " ono ° ~,~~o~~.~ o~~o°u,o°,~°o~~o.~~°o o ~, o ~ o ' a~ a~ ~ o ' a~ a~ o o ,~ ~ .-~ ~ ~' o a~ a~ o o y,,U ~ N O o ~ ~> ~ o yw o w~:~ ~~ ° ~.~ ~ ~~~y3x~x~od ~,~~x x N N N N N N N N N
w x N ~ ~ ~ .fl ~ .fl N d' O v~ 01 O .~ l~
00 00 01 Qv 01 O O O
.-r .~ ~ N N N N
O~ O N M O O O ~n V7 M N O M ~O 01 \O M
.-. O ~ l~ N ~ ~~ o0 M
01 V1 O ~n 01 N O ~ ~O
N ~O l~ I~ M v1 I~ I~

c~ a~ a a a a x x x x x x x x x ~O '.D N l~ ~O V7 O1 M 0o M ~ O~ ~ N O M 01 O d' N a1 N ~ oo v~ Q\ N v~ M N ~ oo v~ a1 N M ~t ~-~ N N N M o0 01 l~ l~ N N
I~ N l~ ~O ~ ~ ~-~ I~ v> M l~ O ~ V1 N a\
~t V'1 N v~ v7 l~ o0 ~ 01 v'1 M v1 I~
~-~ N '--~ ~ N N N
-, 0 0 0 0 0 0 0 0 0 0 0 0 0 ~ \ ~ \ \ ~
O V~ O O ~ O 01 ~ O ~ ~~ M O 01 O ~n ~ O~ O Ov O l~ ~ O M 01 ~ O~ O O M
-r <h O
M
O 01 ~ ~ O
O M
00 00 ~ ~ N l~ Q\
00 I~ .~ d' r.-~ ~ ,~ O
NI ~ ~I
o~0 ~ O N ~ O
o ~ P~1 W ~ o° oc~o o No G~1 0 0.1 w ~ ~'-' ~'-' ~ w ~ o w y.', :fl~' w P-i b~ 'd 'G N ~ b4 01 P-~ bO M 'C
Y, O
n cd N ~ N
Ox~~', .S-'.~0~ O~ N~ ~ ,~ ~~M
~~~~~,ox ~x O ~ x ~ ~ ~ ~x N
M .~ ~ V . ~ ~ ~ O (1, ~, ~' . cr'n x O ~ N
M O O ~ 01 O E..~ N x ~, ~D ~ "" O "O
O M s., ~O ..~ cd S.r ~n O ~ ~ ~ ~ N a N p ~ ~ O (, O
N dl] ~ ø" ~ ~ ~1, ,-, ~~ a 00 ~ ~ O ~ (~ ~1, r-, r-,~10~~,'O~"' .~~r.,pOp~'S"'~' ~~ M~OO.S~~ O
cd0 NO.NNOa7 ~ ,-~ø, ~Q..~Op~~ O
a~1 x ~ N a Q~ a U V7 '/ r3, tW ./ ~ ~1 a Y~ A-1 ~/ ..-r a '/ ~2, !!J A-I
N N N N W N G,7~ N N (~ N
x ~ ~ ~ ~ ,~ ~ N ~ ~ ~~ ~ ~ x M ~ 00 N 01 O M ~ ~O
N N M ~' V~ v~ ~n v~
N N N N N N N N N
O ~ N ~O ~ N O~ M o0 N N o0 v~ N Ov ~ M o0 ~O O l~ ~ ~~ ~O O 01 M
N ~n M ~n l~ ~n N
~O 01 v7 N M ~ ~' ~~ <t' l~ ~O I~ l~ l~ Q1 Q\ 01 D1 N
U d o ~ ~ c~,7 Cw7 ~ CU7 x x x x x x Ov M O o0 N o0 0o N v~ v~ ,-. 00 00 01 v~ O~ oo d' oo ~n ,-.-W ~ v~ 00 0o M
O O O 00 ~ ~ V~ ~ O~ 00 00 00 \O l~ ~ 00 Q1 00 l~ V7 00 l~ 00 00 01 ~D vW D v0 ~ N 01 N ,-.~ .~ ~ .--~ ,~ ,-. .~ N
N M O O O ~n M N ~n ~--~ ~O ~O ~D ~ ~O ~O ~O ~O ~ ~O ~O ~O ~D O 00 ~1 N 01 ~1 Q~ O 00 V7 00 00 ,-r .---n ,~ ~ .~ ~ .~ ,~ .~ ~ .~ ~ ,~ V7 01 ~O M ~ N ~~ V1 v~ v~ v~ v~ v~ ~n ~ ~ ~n V1 Vmn ~ ~t ~t 0 0 ~ ~ 0 0 0 ~ 0 0 0 0 0 0 0 0 0 ~ o o ~ 0 0 0 0 0 ~O ~ M ~ N 00 M O ~ I~ DO M O 00 p~ \O ~~ 00 ~ ~O O ~O ~ ~~ 01 00 M M ~ M ~ QWD l~ 41 41 ~ V1 V1 t!7 V7 V7 V7 ~ N ~ N ~ ~ d' N 01 N N p N ~ O
O N v~ '-' 01 N r., p~ ~ ~ Q\
--~ N N I ~ O .-. I ~
0 ~
b .'.
_N .-, O
O U ~ ~" O ~. . a; .~
p.., ~ c~ x O c~~ . ~'~ . ø, cd y_, C/~ ~ ~ ~ ~, ~ ~, Vi U
x ~ Q., ~ ~ ~ f~.~ O ~ p ~.~~yo ,.d ~~ '~ f O ~ r~", ~ ~ ~ N 4 ~p O .~ p o0 ø, ,-, N cn O v~ '~ tan ,-, ~O x ~ x 00 N ~ ~ N a N~ ~~,r'~ONNr"'N Np O
~ ~ c~ ~ ,~ ~1..~ U
Q,i V1 a a a a a Q., V1 a w S~ 4-1 N N N N N N
k ~ k k ~ X
cd _cd c~ cd _cd cd ~ .fl l~ N ~ o0 ~ l~
N N N N N N
V7M01 .-~M M
N M 00 '~h M M
O1 ~n ~ ~ N N
a1 v7 ~ N O
00 l~ O v~ l~ 00 Q1 ~
0~0 O o~0 O N
x x x x x x d1 N Q~ N M M a1 N M ~ vD N M O ~ d' ~t v0 ~n ~n M d' M ~ M M M ~ M N N M O Q\ ~D O ~p M M M M M M M ~ <t ~ N M N N ~ N
~ ~--~ M

00 'd ~ ~ Ov \O M M .~ O ~ V1 \O ~ 00 M .--~
~p I~ .-mn ~O N v~ v~ v1 ~ M N ~O t~ ~ V~
O O~ ~

N M M M M .--~ N N

N \ \ \ \ \ \ \ \ \ \ \ \ \ \ \ \ \ \
\ \ 0 0 0 0 0 0 ~n oo O N O ~D ~n ~ oo O O ~ ~ ~t ~O o0~O
~ 01 O O

l~ ~Y ~ ~' M M ~ M ~ ~1 00 ~O 00 ~fi N M l~ V7 O

-- -.~ --~ N ,-, ~ '-' M a1 O ~

U ~ ~ U

po ~ U ~ U

a, ~~ ~ b N ,-, x N
O ~ M , ,b ~ a~O

' N 'p ~'O

O ~ > 9 . c O
... -~ O

, cn N O a ' ~ O
' cd .-~ O ~ -~-m' ~ G ."'~ ~
~ 'Zj ~~
~r ...
~

c W.. G~ p ~

~ . x .

O N '~ . ~ .--i ~ ~ ''-'~
n Q, ~ O
.~"' ~
Q., ~ ~ ~ I~ 41 yes., ~' a In ~ U n O _ . ~

~ ~ W p ~ ~

o'~' o ~ y ~ '~ y o ~ o ~
'' x N N N N N N N

xN ~

01 O .~ v~ o0 .~~O

l~ o0 01 01 01 O O

N N N N N M M

M I~ V7 O~ M M O

O\ N ~ ~ l~ O~

Ov N ~ ~ Wit'O w1 00 M N ~n 'O

O oo ~ ~ I~ ~nN

a1 D\

x x x x x x O~ 01 01 ~ (~ O '~ v~ N V7 O O~ o0 o0 N Qv N 00 Q1 M M M N I~ O~ v~ M 01 I~ N l~ .~ I~ ~-r ~ l~ M ~ 00 ~ M .-w0 ~ ~ ~ ~O ~-i N M ~~ M N ~ M N ~ ~O O
o0 vD I~ M ~n O~ O N d1 N N O~ O~ M o0 O~ N ~ o0 'O
~~ M l~ V7 ~ O l~ ~ 01'~ ~O ~ ~ ~ ~ l~ Q~ 00 M
M r~ M V7 v1 N ~ N N M M ~ ~ N N l~

~ \ \ \ ~
d' ~n N ~ ~ 01 00 M 01 Q1 N t~ Ov ~ ~O l~ ~ ~O
O M ~ 00 \O ~O V7 I~ V7 V7 'Ch ~ ~' O ~' M ~ O 00 00 .-~ - ,~
.-~ . ,-, ~ . M .-~ '-' r.r 00 ~ ~ ~' ~n ~ ono O O
--~ 00 01 00 ~ N V1 M N
N v_0 U ~nlo ~ ~ ~ U
-. ~ ~ d ~ o ~ o :fl .n .~ ~ M ~ ~ ~ M w .~ ~ w en -d cwo on oo 'd a~ cn r~ a, tin -o a. a ,b O M ~ .b O ,-, ~ .~ O
N ~ ,~ N ~ ~ ~ v~ N ~ v~
° ~ ~ o ~ °' ~ o N
.~ ~x ~g a~ ~ ~~x x x ~ W .~', _ V ~ ~ ~ ~ ~ ~ U ~ M
O ~ O ~ ~O "., O . ~ b ~ ~! . ~ 0~0 b ~ %~
a t/7 ~ ~ ,~ ~ N ~ '~ ~ ~ ~ 0 1-~ ~ n -, ~O by ~ P, ,-, M ~ ~ ~ ~ ø, ,-, ~ ~O ~ a ~ ~ '~ ø, ~ p.i yn ~Oi,"~~O ~OO,G'~u~0 'cad ON~',~ ~'' a~0 o O~.G~~ ~' O ~Oy.a~~.~ ~ ~00 O~~~ ~ iG
w ~ ~ ~, w o .~d~ ~ ° ~~x ~x~~~ ~~xw~1 W '.ss.~a~b:~
N N N N N N N (~ N N W N
~C >C X k ~ k ~G ~ ~G _ X
x~.~ ~ xN~
.~ M t~ O~ ~--~ N ~O o0 O~ O
.~ ,~ .~ .-. N N N N N M
M M M M M M M M M M
l~ O ~O 00 01 I~ 00 .~ ~O ~O
N O N M I~ ~ ~ O
O M ~ M ~O O O~ ~ M
l~ ~ O~ ~ I~ N ~t v7 O
00 M t~ ~ ~ ~~ N N O l~
00 O~ t~ O~ 00 01 41 01 00 O
O ~ N O~ ~ N p (~ ~ P

~a x x x x xx x x 00 N ~ M V7 o0 .-. Wit'N ~n OWE l~ Q\
Q\ M v1 Ov N d1 l~

~ v~ M O ~O ~ N v~ v~ O N ~ M
M t~ v7 O N oo ~

.-r N ~ M M M ~ M M -~ N V M d' V7 N ~ ~

00 ~ M ~O O~ ~ ~ ~ ~O 00 .-r ~ N V7 ~
M ~ M 41 ~ M

t~ N ~ ~O o0 o0 N
~ ~ Q1 M

~ d' N M ~ d' M d' ~ M
00 ~ N
O

\ \ \ \ \ \ \ \ \ \ \
\ 0 0 0 0 ~ ~ 0 0 0 0 0 0 \ 0 0 0 0 0 0 ~ o0 I~ ~ o0 I~ ~O ~ D1 O N ~ o0 ~t ,~ o0 O ~O ~O
O

00 t?' C~ I~ l~ t!1 ~!1 V1 ~ O ~ ~O t~ 00 M V7 V7 M N ~' ~

f'rj'~ ~--i ~--i~ ~--y Q .-n ~O ~ I~

~

O ~D vp O o0 U U U

~

d b ~ ~ ~ ~ ~ ~ b H o o ~ z ~ -d o . . ' a. ~ ~ o x ~ o ~

O ~ x ~ O _ ~ OU ~ O ~ x O

cd 0 M 'b cd N ~ N . N s~ N 'd O .~

M ~ ~ ~O ~ cd ~ U ~ ~ ~. pp O
~. ~
~

~. N . ~-~'~ ~ j, '-' ~ ~ r-~ M
r-, ~ r-, ~

. p ~ ~ 0 0 , O ., =
.~

~ ~ ~ -i~ cn '~ '~ O N O N
, . ~ ..,, ~ ~ O ,b N N
-, O N U ~ ~ O cd , ~ O y N ~
N U

0 0 ~ ~ o ' W > ~ w ~' ~ .~ ' o ' . a.. ~ o ' a. ~.
~ ~ ' ' ~., ~

~. s.. o cd U aW Q , ~ c~ s..
cd ~ U ~ x cd ~ cd ~ ~ ~1 a ~ '/ U a y~ a a .N , V7 , V1 V1 V7 C a , a !n N N N N N N N N N

y ~

C~ CC3 C~ _(d _C~ C~ C~ _C~ C~

_ _ N O~ O N ~O N ~ O O

M M d' ~' V7 ~O \O h 00 M M M M M M M M M

I~ 00 'O ~ N O~ 01 N v~

l0 .~ V7 ~ Q\ .-~ N M

I~ .~ y ~ O M

r-. d. .~ ~p ~n N ~t o0 00 .~ oo l~ ~n N I~ N

41 00 ~O 00 V~ 00 l' 41 M ~ M

o o ~

w a W ~ U, U U U ~'' ~' w w w w ~

x x x x ~ x x x 00 N N ~ ~ l~ N N t~ M M O I~ M V7 ~ N 01 ~ M
00 ~ ~O O I~ ~n o0 ~ ~O ~O ~~ 01 N N ~-~ l~ v~ ,--~ M
.~ M M M V7 l~ ~' Q1 ti' M .-~ ~ .~ V7 M l0 M M ~O ~ ~ ~p V7 O 00 O ~O .--~ lp N 00 ~O ~--W ~ I~ 01 00 0o M l~ t~ o0 N l~ N a\ 01 M ~' ~ N l~ N ~ ~ ~ ~-~ N ~-~ M
0 0 ~, 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 ~.
~D ~ ~ O ~ M ~ V'1 01 00 ~O O ~ N O ~ M 00 Q\ ~' [~ \p N ~p M M M N N O~ ~D O 01 00 00 ~ l~ 00 ~n ~O
O ~ '"_' "'' O ~ .-r M 0O ~ N N 00 ~
O .~ O~ ~, ~ O 00 ~O ~ N ~, N ~n N ~I ~ N N U N NI ~t ~ a1 d ~ U
ono o d o d ~ .a o ~ o w ~ w ~ ~'-' ~ ,.fl oo ~ v~ .~
an o, p.~ on p, on ~d a~ on ~ an ~n c0 r._, oo d O cn N ~ ~ V7 ,-, o >, ~ .x ~ .b o ~ N
x o ~ an ~ ~ , a; ~ ~ .~ ~ ~ o .~ ~ s?, . ~ ~ ,~ 4, p w ~ ~ ~ a~ ~ '~ o -~, ~ ~ x .~ p . ~ ~ ,~ b d o ~ ~ ~ o r~ ~ U o z Q., ~ ~ Y x ~~ .fl ~ o ~ ~ > ~ o ~. ~ N ' p., '~ o ~n .~ ., cd ~ M ~ o ~ ~ ' ~ ~ s~ a> o ~ cd ~ x p '° ~n 'i' ~ ~ ~_ ~''''~ N ~ O '5~ ~ ~ -~~, N O ~ ~ ~ CD ~ O ~ ~ M N ~"~ 4~'' bA
Ow~w~~~~(11 ~''~l,x,~' OQ.~ O '~w~'~MO
x ~ x d -~ U ~ U a, ° ~: ~ U ~ c~, ~ 'N ~ ~ x ~ ~ d ~ o .x~ U
N W N W N N N N N N
x N ~ ~ .~ ~ .fl 00 M ~D V1 00 01 ~' ~O
00 ~ Q1 O ~-~ N M M
M M M
O ~~ l~ ~ 01 O I~ 00 I~ M M ~ ~D
M .~ .~ N M M N M
O M M 01 ~f ~ ~h O
N t~ t~ N N M ~n ~t Q\
V) M
N ~ ~ ~, N~-' O
O O ~ w w~
x ~ d x x x x x N V~ Iw0 ~ ~1 O I~ V~ ~mn ~ l~ Q~ N O I~
O V> M N M O o0 ~O N ~ ~O N d1 O ~n ~ N
N ~O ~~ N N M N I~ M N M ~ ~O M
o0 o0 01 ~O Ov d' ~ ~ ~' l~ N ~ 00 ~ ~ ~ l~
vW O ~O oo ~ M N ~ o0 .-~ ~n O a1 O~ ~t N
N ,~ N
M
pp o 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 N ~t ~n d' O l~ l~ oo ~O M v1 ~ O o0 I~
N ~ v~ 01 v~ I~ v7 l~ l~ ~ l~ ~h ~ O o0 I~ ~n -, ~ ' p pip ~ M ~ ~ 00 00 d' M oo ~ O ,~ N p~ O N N
O ~ N ~ I~ ~ p~ oo ~t M o0 00 O O N .~ N N N N ~ N ,~ U M .~ N
O 00I ~0ll~ctl ~ ~vll~ ~~_OI
O _ _ _ _ _ _ _ _ _ .D M ~ 000 .D o~0 ~~' .~ ,~ ~ .D ~
P-~ bD by bA ~~ b-0 --~ b1~ ~ 'b d4 ~O by by N b0 :b ,.-~ ~ N Z ~ ,-, ~d O M ~ ~ O
0 .O ~ ~ O ~ > vy ~ ° O C
'G O ~ ' .~ ~ ~ ~~ ~~~= ~x pU.., ~~ ~x N
o ~ ~ ~ ~ ~ ate. ~ a~. ~ ø~~ ~ ~ ~ x ~ ~ o y '~ ~. ~ ~. . ~ ~ ~ . ~b .~ .-. r..~ _ .-. - .-.
'fl~(7oc~o~o~c~ o~oc~~c~d~.N° ~.~o~oc~,c~"~Q., n M ~ ~ ~ i ~ ~, ~O v~ O w ~ .~ O '~ ø, ~ M v~ M cn ~~ ~ N
-~a ~ ~ ~ ~ ~ ~ ~ ~ ° ~ ° ~ ~ '~ ° ~ ~ ~ O oMO ~ O N ~ ~
N
'~wo ~oU- ~o~aia~,~ooyo.-.,~°
'x',aw.,-oW ~~Nu~ ~dx ~xd ~~~ ~~~~x~
(,1,~ N N N N N N N N N N
0o N ~ ~n O~ M N O ~ ~ ~D
M ~ ~' ~ ~ V7 ~O 00 00 00 00 d' ~ ~ ~i' ~ V' ~ ~Y
M a1 N O o0 N ~ ~ ~O M
l~ 00 O O l~ ~ O ~ O ~O V1 N M 00 O N ~1 M O O N °~
Q\ ~O O 01 ~n N a\ O~ M ~ M
V7 O l~ ~ ~O V'1 V7 ~ 00 l~ M
00 00 ~ l~ ~ ~ ~0 ~ ~ M N
~_ w W A A ~ ~ U 0.31 ~ act U U U U
x x x x w D v~

N

N .-a O o0 00 O l~ Q1 O
M

N
I

_ U
'n pq N b~ N

N
~ .

xi Q.

N c~d ~ O
~

O ~ O
F~
~
Q

.. N
b O

x V1 .~ n~
Q., a O O
~ NN
O
N

.N
O

o C~~

U a Ina a Q.r Qr N N

k _c~ cd O l~

O N

N

x x

[057] Table 2 further characterizes certain encoded polypeptides of the invention, by providing the results of comparisons to protein and protein family databases.
The first column provides a unique clone identifier, "Clone ID NO:", corresponding to a cDNA
clone disclosed in Table 1A. The second column provides the unique contig indentifier, "Contig >D:" which allows correlation with the information in Table 1A. The third column provides the sequence identifier, "SEQ II? NO:X", for the contig polynucleotide sequences. The fourth column provides the analysis method by which the homology/identity disclosed in the row was determined. The fifth column provides a description of PFam/NR hits having significant matches identified by each analysis.
Column six provides the accession number of the PFam/NR hit disclosed in the fifth column. Column seven, "Score/Percent Identity", provides a quality score or the percent identity, of the hit disclosed in column five. Comparisons were made between polypeptides encoded by polynucleotides of the invention and a non-redundant protein database (herein referred to as "NR"), or a database of protein families (herein referred to as "PFam"), as described below.

[058] The NR database, which comprises the NBRF PIR database, the NCBI GenPept database, and the SIB SwissProt and TrEMBL databases, was made non-redundant using the computer program nrdb2 (Warren Gish, Washington University in Saint Louis). Each of the polynucleotides shown in Table 1A, column 3 (e.g., SEQ ID NO:X or the 'Query' sequence) was used to search against the NR database. The computer program BLASTX
was used to compare a 6-frame translation of the Query sequence to the NR
database (for information about the BLASTX algorithm please see Altshul et al., J. Mol.
Biol. 215:403-410 (1990), and Gish et al., Nat. Genet. 3:266-272 (1993)). A description of the sequence that is most similar to the Query sequence (the highest scoring 'Subject') is shown in column five of Table 2 and the database accession number for that sequence is provided in column six. The highest scoring 'Subject' is reported in Table 2 if (a) the estimated probability that the match occurred by chance alone is less than 1.0e-07, and (b) the match was not to a known repetitive element. BLASTX returns alignments of short polypeptide segments of the Query and Subject sequences which share a high degree of similarity; these segments are known as High-Scoring Segment Pairs or HSPs.
Table 2 reports the degree of similarity between the Query and the Subject for each HSP as a percent identity in Column 7. The percent identity is determined by dividing the number of exact matches between the two aligned sequences in the HSP, dividing by the number of Query amino acids in the HSP and multiplying by 100. The polynucleotides of SEQ
>D NO:X which encode the polypeptide sequence that generates an HSP are delineated by columns 8 and 9 of Table 2.

[059] The PFam database, PFam version 5.2, (Sonnhammer et al., Nucl. Acids Res., 26:320-322, (1998)) consists of a series of multiple sequence alignments; one alignment for each protein family. Each multiple sequence alignment is converted into a probability model called a Hidden Markov Model, or HMM, that represents the position-specific variation among the sequences that make up the multiple sequence alignment (see, e.g., R. Durbin et al., Biological sequence analysis: probabilistic models of proteins and nucleic acids, Cambridge University Press, 1998 for the theory of HMMs). The program HMMER version 1.8 (Sean Eddy, Washington University in Saint Louis) was used to compare the predicted protein sequence for each Query sequence (SEQ >D NO:Y in Table 1A) to each of the HMMs derived from PFam version 5.2. A HMM derived from PFam version 5.2 was said to be a significant match to a polypeptide of the invention if the score returned by HMMER 1.8 was greater than 0.8 times the HMMER 1.8 score obtained with the most distantly related known member of that protein family.
The description of the PFam family which shares a significant match with a polypeptide of the invention is listed in column 5 of Table 2, and the database accession number of the PFam hit is provided in column 6. Column 7 provides the score returned by HMMER
version 1.8 for the alignment. Columns 8 and 9 delineate the polynucleotides of SEQ >D
NO:X which encode the polypeptide sequence which shows a significant match to a PFam protein family.

[060] As mentioned, columns 8 and 9 in Table 2, "NT From" and "NT To", delineate the polynucleotides of "SEQ ID NO:X" that encode a polypeptide having a significant match to the PFarn/NR database as disclosed in the fifth column of Table 2. In one embodiment, the invention provides a protein comprising, or alternatively consisting of, a polypeptide encoded by the polynucleotides of SEQ ID NO:X delineated in columns 8 and 9 of Table 2. Also provided are polynucleotides encoding such proteins, and the complementary strand thereto.

[061] The nucleotide sequence SEQ ID NO:X and the translated SEQ ID NO:Y are sufficiently accurate and otherwise suitable for a variety of uses well known in the art and described further below. For instance, the nucleotide sequences of SEQ >D NO:X
are useful for designing nucleic acid hybridization probes that will detect nucleic acid sequences contained in SEQ m NO:X or the cDNA contained in Clone m NO:Z. These probes will also hybridize to nucleic acid molecules in biological samples, thereby enabling immediate applications in chromosome mapping, linkage analysis, tissue identification and/or typing, and a variety of forensic and diagnostic methods of the invention. Similarly, polypeptides identified from SEQ m NO:Y may be used to generate antibodies which bind specifically to these polypeptides, or fragments thereof, and/or to the polypeptides encoded by the cDNA clones identified in, for example, Table 1 A.

[062] Nevertheless, DNA sequences generated by sequencing reactions can contain sequencing errors. The errors exist as misidentified nucleotides, or as insertions or deletions of nucleotides in the generated DNA sequence. The erroneously inserted or deleted nucleotides cause frame shifts in the reading frames of the predicted amino acid sequence. In these cases, the predicted amino acid sequence diverges from the actual amino acid sequence, even though the generated DNA sequence may be greater than 99.9% identical to the actual DNA sequence (for example, one base insertion or deletion in an open reading frame of over 1000 bases).

[063] Accordingly, for those applications requiring precision in the nucleotide sequence or the amino acid sequence, the present invention provides not only the generated nucleotide sequence identified as SEQ m NO:X, and a predicted translated amino acid sequence identified as SEQ >D NO:Y, but also a sample of plasmid DNA
containing cDNA Clone >D NO:Z (deposited with the ATCC on October 5, 2000, and receiving ATCC designation numbers PTA 2574 and PTA 2575; deposited with the ATCC on January 5, 2001, having the depositor reference numbers TS-1, TS-2, AC-1, and AC-2;
and/or as set forth, for example, in Table 1A, 6 and 7). The nucleotide sequence of each deposited clone can readily be determined by sequencing the deposited clone in accordance with known methods. Further, techniques known in the art can be used to verify the nucleotide sequences of SEQ >D NO:X.

[064] The predicted amino acid sequence can then be verified from such deposits.
Moreover, the amino acid sequence of the protein encoded by a particular clone can also be directly determined by peptide sequencing or by expressing the protein in a suitable host cell containing the deposited human cDNA, collecting the protein, and determining its sequence.
RACE Protocol For Recovery of Full-Length Genes [065J Partial cDNA clones can be made full-length by utilizing the rapid amplification of cDNA ends (RACE) procedure described in Frohman, M.A., et al., Proc. Nat'1.
Acid.
Sci. USA, 85:8998-9002 (1988). A cDNA clone missing either the 5' or 3' end can be reconstructed to include the absent base pairs extending to the translational start or stop codon, respectively. In some cases, cDNAs are missing the start codon of translation.
The following briefly describes a modification of this original 5' RACE
procedure. Poly A+ or total RNA is reverse transcribed with Superscript II (GibcoBRL) and an antisense or complementary primer specific to the cDNA sequence. The primer is removed from the reaction with a Microcon ,Concentrator (Amicon). The first-strand cDNA is then tailed with dATP and terminal deoxynucleotide transferase (GibcoBRL). Thus, an anchor sequence is produced which is needed for PCR amplification. The second strand is synthesized from the dA-tail in PCR buffer, Taq DNA polymerise (Perkin-Elmer Cetus), an oligo-dT primer containing three adjacent restriction sites (XhoI, SaII and CIaI) at the 5' end and a primer containing just these restriction sites. This double-stranded cDNA is PCR amplified for 40 cycles with the same primers as well as a nested cDNA-specific antisense primer. The PCR products are size-separated on an ethidium bromide-agarose gel and the region of gel containing cDNA products the predicted size of missing protein-coding DNA is removed. cDNA is purified from the agarose with the Magic PCR
Prep kit (Promega), restriction digested with XhoI or SaII, and ligated to a plasmid such as pBluescript SKII (Stratagene) at XhoI and EcoRV sites. This DNA is transformed into bacteria and the plasmid clones sequenced to identify the correct protein-coding inserts.
Correct 5' ends are confirmed by comparing this sequence with the putatively identified homologue and overlap with the partial cDNA clone. Similar methods known in the art and/or commercial kits are used to amplify and recover 3' ends.
[066] Several quality-controlled kits are commercially available for purchase.
Similar reagents and methods to those above are supplied in kit form from GibcoBRL for both 5' and 3' RACE for recovery of full length genes. A second kit is available from Clontech which is a modification of a related technique, SLIC (single-stranded ligation to single-stranded cDNA), developed by Dumas et al., Nucleic Acids' Res., 19:5227-32 (1991).
The major differences in procedure are that the RNA is alkaline hydrolyzed after reverse transcription and RNA ligase is used to join a restriction site-containing anchor primer to the first-strand cDNA. This obviates the necessity for the dA-tailing reaction which results in a polyT stretch that is difficult to sequence past.
[067] An alternative to generating 5' or 3' cDNA from RNA is to use cDNA
library double-stranded DNA. An asymmetric PCR-amplified antisense cDNA strand is synthesized with an antisense cDNA-specific primer and a' plasmid-anchored primer.
These primers are removed and a symmetric PCR reaction is performed with a nested cDNA-specific antisense primer and the plasmid-anchored primer.
RNA Ligase Protocol For Generating The 5' or 3' End Sequences To Obtain Full Length Genes [068] Once a gene of interest is identified, several methods are available for the identification of the 5' or 3' portions of the gene which may not be present in the original cDNA plasmid. These methods include, but are not limited to, filter probing, clone enrichment using specific probes and protocols similar and identical to 5' and 3' RACE.
While the full length gene may be present in the library and can be identified by probing, a useful method for generating the 5' or 3' end is to use the existing sequence information from the original cDNA to generate the missing information. A method similar to 5' RACE is available for generating the missing 5' end of a desired full-length gene. (This method was published by Fromont-Racine et al., Nucleic Acids Res., 21(7):1683-(1993)). Briefly, a specific RNA oligonucleotide is ligated to the 5' ends of a population of RNA presumably containing full-length gene RNA transcript. A primer set containing a primer specific to the ligated RNA oligonucleotide and a primer specific to a known sequence of the gene of interest, is used to PCR amplify the 5' portion of the desired full length gene which may then be sequenced and used to generate the full length gene. This method starts with total RNA isolated from the desired source, poly A RNA may be used but is not a prerequisite for this procedure. The RNA preparation may then be treated with phosphatase if necessary to eliminate 5' phosphate groups on degraded or damaged RNA, which may interfere with the later RNA ligase step. The phosphatase, if used, is then inactivated and the RNA is treated with tobacco acid pyrophosphatase in order to remove the cap structure present at the 5' ends of messenger RNAs. This reaction leaves a 5' phosphate group at the 5' end of the cap cleaved RNA which can then be ligated to an RNA oligonucleotide using T4 RNA ligase. This modified RNA preparation can then be used as a template for first strand cDNA synthesis using a gene specific oligonucleotide.
The first strand synthesis reaction can then be used as a template for PCR
amplification of the desired 5' end using a primer specific to the ligated RNA oligonucleotide and a primer specific to the known sequence of the connective tissue antigen of interest.
The resultant product is then sequenced and analyzed to confirm that the 5' end sequence belongs to the relevant connective tissue antigen.
[069] The present invention also relates to vectors or plasmids, which include such DNA sequences, as well as the use of the DNA sequences. The material deposited with the ATCC (deposited with the ATCC on October 5, 2000, and receiving ATCC
designation numbers PTA 2574 and PTA 2575; deposited with the ATCC on January 5, 2001, having the depositor reference numbers TS-l, TS-2, AC-l, and AC-2;
and/or as set forth, for example, in Table 1A, 6 and 7).is a mixture of cDNA clones derived from a variety of human tissue and cloned in either a plasmid vector or a phage vector, as shown, for example, in Table 7. These deposits are referred to as "the deposits"
herein. The tissues from which some of the clones were derived are listed in Table 7, and the vector in which the corresponding cDNA is contained is also indicated in Table 7. The deposited material includes cDNA clones corresponding to SEQ >D NO:X described, for example, in Table 1A (Clone >D NO:Z). A clone which is isolatable from the ATCC
Deposits by use of a sequence listed as SEQ m NO:X, may include the entire coding region of a human gene or in other cases such clone may include a substantial portion of the coding region of a human gene. Furthermore, although the sequence listing may in some instances list only a portion of the DNA sequence in a clone included in the ATCC
Deposits, it is well within the ability of one skilled in the art to sequence the DNA
included in a clone contained in the ATCC Deposits by use of a sequence (or portion thereof) described in, for example Tables 1A or 2 by procedures hereinafter further described, and others apparent to those skilled in the art.

[070] Also provided in Table 7 is the name of the vector which contains the cDNA
clone. Each vector is routinely used in the art. The following additional information is provided for convenience.
[071] Vectors Lambda Zap (U.5. Patent Nos. 5,128,256 and 5,286,636), Uni-Zap XR
(U.5. Patent Nos. 5,128,256 and 5,286,636), Zap Express (U.S. Patent Nos.
5,128,256 and 5,286,636), pBluescript (pBS) (Short, J. M. et al., Nucleic Acids Res. 16:

(1988); Alting-Mees, M. A. and Short, J. M., Nucleic Acids Res. 17.'9494 (1989)) and pBK (Aping-Mees, M. A. et al., Strategies 5:58-61 (1992)) are commercially available from Stratagene Cloning Systems, Inc., 11011 N. Torrey Pines Road, La Jolla, CA, 92037. pBS contains an ampicillin resistance gene and pBK contains a neomycin resistance gene. Phagemid pBS may be excised from the Lambda Zap and Uni-Zap XR
vectors, and phagemid pBK may be excised from the Zap Express vector. Both phagemids may be transformed into E. coli strain XL-1 Blue, also available from Stratagene.
[072] Vectors pSportl, pCMVSport 1.0, pCMVSport 2.0 and pCMVSport 3.0, were obtained from Life Technologies, Inc., P. O. Box 6009, Gaithersburg, MD 20897.
All Sport vectors contain an ampicillin resistance gene and may be transformed into E. coli strain DH10B, also available from Life Technologies. See, for instance, Gruber, C. E., et al., Focus 15:59- (1993). Vector lafmid BA (Bento Soares, Columbia University, New York, NY) contains an ampicillin resistance gene and can be transformed into E. coli strain XL-1 Blue. Vector pCR~2.1, which is available from Invitrogen, 1600 Faraday Avenue, Carlsbad, CA 92008, contains an ampicillin resistance gene and may be transformed into E. coli strain DHlOB, available from Life Technologies. See, for instance, Clark, J. M., Nuc. Acids Res. 16:9677-9686 (1988) and Mead, D. et al., Bio/Technology 9: (1991).
[073] The present invention also relates to the genes corresponding to SEQ >D
NO:X, SEQ )D NO:Y, and/or the deposited clone (Clone )D NO:Z). The corresponding gene can be isolated in accordance with known methods using the sequence information disclosed herein. Such methods include preparing probes or primers from the disclosed sequence and identifying or amplifying the corresponding gene from appropriate sources of genomic material.

[074] Also provided in the present invention are allelic variants, orthologs, and/or species homologs. Procedures known in the art can be used to obtain full-length genes, allelic variants, splice variants, full-length coding portions, orthologs, and/or species homologs of connective tissue associated genes corresponding to SEQ )D NO:X or the complement thereof, polypeptides encoded by SEQ 1D NO:X or the complement thereof, and/or the cDNA contained in Clone ID NO:Z, using information from the sequences disclosed herein or the clones deposited with the ATCC. For example, allelic variants and/or species homologs may be isolated and identified by making suitable probes or primers from the sequences provided herein and screening a suitable nucleic acid source for allelic variants and/or the desired homologue.
[075] The polypeptides of the invention can be prepared in any suitable manner. Such polypeptides include isolated naturally occurnng polypeptides, recombinantly produced polypeptides, synthetically produced polypeptides, or polypeptides produced by a combination of these methods. Means for preparing such polypeptides are well understood in the art.
[076] The polypeptides may be in the form of the secreted protein, including the mature form, or may be a part of a larger protein, such as a fusion protein (see below). It is often advantageous to include an additional amino acid sequence which contains secretory or leader sequences, pro-sequences, sequences which aid in purification, such as multiple histidine residues, or an additional sequence for stability during recombinant production.
[077] The polypeptides of the present invention are preferably provided in an isolated form, and preferably are substantially purified. A recombinantly produced version of a polypeptide, including the secreted polypeptide, can be substantially purified using techniques described herein or otherwise known in the art, such as, for example, by the one-step method described in Smith and Johnson, Gene 67:31-40 (1988).
Polypeptides of the invention also can be purified from natural, synthetic or recombinant sources using techniques described herein or otherwise known in the art, such as, for example, antibodies of the invention raised against the connective tissue polypeptides of the present invention in methods which are well known in the art.
[078] The present invention provides a polynucleotide comprising, or alternatively consisting of, the nucleic acid sequence of SEQ 117 NO:X, and/or the cDNA
sequence contained in Clone >D NO:Z. The present invention also provides a polypeptide comprising, or alternatively, consisting of, the polypeptide sequence of SEQ
>D NO:Y, a polypeptide encoded by SEQ >D NO:X or a complement thereof, a polypeptide encoded by the cDNA contained in Clone >D NO:Z, and/or the polypeptide sequence encoded by a nucleotide sequence in SEQ >I7 NO:B as defined in column 6 of Table 1B.
Polynucleotides encoding a polypeptide comprising, or alternatively consisting of the polypeptide sequence of SEQ m NO:Y, a polypeptide encoded by SEQ m NO:X, a polypeptide encoded by the cDNA contained in Clone m NO:Z and/or a polypeptide sequence encoded by a nucleotide sequence in SEQ m NO:B as defined in column 6 of Table 1B are also encompassed by the invention. The present invention further encompasses a polynucleotide comprising, or alternatively consisting of, the complement of the nucleic acid sequence of SEQ >D NO:X, a nucleic acid sequence encoding a polypeptide encoded by the complement of the nucleic acid sequence of SEQ
m NO:X, and/or the cDNA contained in Clone >D NO:Z.
[079] Moreover, representative examples of polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more of the sequences delineated in Table 1B column 6, or any combination thereof.
Additional, representative examples of polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more of the complementary strands) of the sequences delineated in Table 1B column 6, or any combination thereof. In further embodiments, the above-described polynucleotides of the invention comprise, or alternatively consist of, sequences delineated in Table 1B, column 6, and have a nucleic acid sequence which is different from that of the BAC
fragment having the sequence disclosed in SEQ 1D NO:B (see Table 1B, column 5). In additional embodiments, the above-described polynucleotides of the invention comprise, or alternatively consist of, sequences delineated in Table 1B, column 6, and have a nucleic acid sequence which is different from that published for the BAC clone identified as BAC
>D NO:A (see Table 1B, column 4). In additional embodiments, the above-described polynucleotides of the invention comprise, or alternatively consist of, sequences delineated in Table 1B, column 6, and have a nucleic acid sequence which is different from that contained in the BAC clone identified as BAC m NO:A (see Table 1B, column 4). Polypeptides encoded by these polynucleotides, other polynucleotides that encode these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides and polypeptides are also encompassed by the invention.
[080] ' Further, representative examples of polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more of the sequences delineated in column 6 of Table 1B which correspond to the same Clone >D
NO:Z (see Table 1 B, column 1 ), or any combination thereof. Additional, representative examples of polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more of the complementary strands) of the sequences delineated in column 6 of Table 1 B which correspond to the same Clone >D NO:Z (see Table 1B, column 1), or any combination thereof. In further embodiments, the above-described polynucleotides of the invention comprise, or alternatively consist of, sequences delineated in column 6 of Table 1B which correspond to the same Clone >D NO:Z (see Table 1B, column 1) and have a nucleic acid sequence which is different from that of the BAC fragment having the sequence disclosed in SEQ
>D NO:B (see Table 1B, column 5). In additional embodiments, the above-described polynucleotides of the invention comprise, or alternatively consist of, sequences delineated in column 6 of Table 1B which correspond to the same Clone >D NO:Z
(see Table 1B, column 1) and have a nucleic acid sequence which is different from that published for the BAC clone identified as BAC ID NO:A (see Table 1B, column 4). In additional embodiments, the above-described polynucleotides of the invention comprise, or alternatively consist of, sequences delineated in column 6 of Table 1B
which correspond to the same Clone ID NO:Z (see Table 1B, column 1) and have a nucleic acid sequence which is different from that contained in the BAC clone identified as BAC >D
NO:A (see Table 1B, column 4). Polypeptides encoded by these polynucleotides, other polynucleotides that encode these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides and polypeptides are also encompassed by the invention.
[081] Further, representative examples of polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more of the sequences delineated in column 6 of Table 1B which correspond to the same contig sequence identifer SEQ >D NO:X (see Table 1B, column 2), or any combination thereof.

Additional, representative examples of polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more of the complementary strands) of the sequences delineated in column 6 of Table 1B
which correspond to the same contig sequence identifer SEQ ID NO:X (see Table 1B, column 2), or any combination thereof. In further embodiments, the above-described polynucleotides of the invention comprise, or alternatively consist of, sequences delineated in column 6 of Table 1B which correspond to the same contig sequence identifer SEQ >D NO:X (see Table 1B, column 2) and have a nucleic acid sequence which is different from that of the BAC fragment having the sequence disclosed in SEQ >D
NO:B (see Table 1B, column 5). In additional embodiments, the above-described polynucleotides of the invention comprise, or alternatively consist of, sequences delineated in column 6 of Table 1B which correspond to the same contig sequence identifer SEQ >D NO:X (see Table 1B, column 2) and have a nucleic acid sequence which is different from that published for the BAC clone identified as BAC ID NO:A
(see Table 1B, column 4). In additional embodiments, the above-described polynucleotides of the invention comprise, or alternatively consist of, sequences delineated in column 6 of Table 1B which correspond to the same contig sequence identifer SEQ >D NO:X (see Table 1B, column 2) and have a nucleic acid sequence which is different from that contained in the BAC clone identified as BAC >D NO:A (See Table 1B, column 4). Polypeptides encoded by these polynucleotides, other polynucleotides that encode these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention.
Additionally, fragments and variants of the above-described polynucleotides and polypeptides are also encompassed by the invention.
[082] Moreover, representative examples of polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more of the sequences delineated in the same row of Table 1B column 6, or any combination thereof. Additional, representative examples of polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more of the complementary strands) of the sequences delineated in the same row of Table 1 B column 6, or any combination thereof. In preferred embodiments, the polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more of the complementary strands) of the sequences delineated in the same row of Table 1B column 6, wherein sequentially delineated sequences in the table (i.e. corresponding to those exons located closest to each other) are directly contiguous in a S' to 3' orientation. In further embodiments, above-described polynucleotides of the invention comprise, or alternatively consist of, sequences delineated in the same row of Table 1B, column 6, and have a nucleic acid sequence which is different from that of the BAC fragment having the sequence disclosed in SEQ >D NO:B (see Table 1B, column S). In additional embodiments, the above-described polynucleotides of the invention comprise, or alternatively consist of, sequences delineated in the same row of Table 1B, column 6, and have a nucleic acid sequence which is different from that published for the BAC clone identified as BAC m NO:A (see Table 1B, column 4). In additional embodiments, the above-described polynucleotides of the invention comprise, or alternatively consist of, sequences delineated in the same row of Table 1B, column 6, and have a nucleic acid sequence which is different from that contained in the BAC clone identified as BAC m NO:A (see Table 1B, column 4). Polypeptides encoded by these polynucleotides, other polynucleotides that encode these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention.
[083] In additional specific embodiments, polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more of the sequences delineated in column 6 of Table 1 B, and the polynucleotide sequence of SEQ >D NO:X (e.g., as defined in Table 1B, column 2) or fragments or variants thereof.
Polypeptides encoded by these polynucleotides, other polynucleotides that encode these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention.
[084] In additional specific embodiments, polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more of the sequences delineated in column 6 of Table 1B which correspond to the same Clone >D
NO:Z (see Table 1B, column 1), and the polynucleotide sequence of SEQ 1D NO:X
(e.g., as defined in Table 1A or 1B) or fragments or variants thereof. In preferred embodiments, the delineated sequences) and polynucleotide sequence of SEQ m NO:X
correspond to the same Clone )I7 NO:Z. Polypeptides encoded by these polynucleotides, other polynucleotides that encode these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention.
[085] In further specific embodiments, polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more of the sequences delineated in the same row of column 6 of Table 1B, and the polynucleotide sequence of SEQ )D NO:X (e.g., as defined in Table 1A or 1B) or fragments or variants thereof. In preferred embodiments, the delineated sequences) and polynucleotide sequence of SEQ m NO:X correspond to the same row of column 6 of Table 1B. Polypeptides encoded by these polynucleotides, other polynucleotides that encode these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention.
[086] In additional specific embodiments, polynucleotides of the invention comprise, or alternatively consist of a polynucleotide sequence in which the 3' 10 polynucleotides of one of the sequences delineated in column 6 of Table 1B and the 5' 10 polynucleotides of the sequence of SEQ >D NO:X are directly contiguous. Nucleic acids which hybridize to the complement of these 20 contiguous polynucleotides under stringent hybridization conditions or alternatively, under lower stringency conditions, are also encompassed by the invention. Polypeptides encoded by these polynucleotides and/or nucleic acids, other polynucleotides and/or nucleic acids that encode these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides, nucleic acids, and polypeptides are also encompassed by the invention.
[087] In additional specific embodiments, polynucleotides of the invention comprise, or alternatively consist of, a polynucleotide sequence in which the 3' 10 polynucleotides of one of the sequences delineated in column 6 of Table 1B and the 5' 10 polynucleotides of a fragment or variant of the sequence of SEQ >D NO:X are directly contiguous Nucleic acids which hybridize to the complement of these 20 contiguous polynucleotides under stringent hybridization conditions or alternatively, under lower stringency conditions, are also encompassed by the invention. Polypeptides encoded by these polynucleotides and/or nucleic acids, other polynucleotides and/or nucleic acids encoding these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides, nucleic acids, and polypeptides are also encompassed by the invention.
[088] In specific embodiments, polynucleotides of the invention comprise, or alternatively consist of, a polynucleotide sequence in which the 3' 10 polynucleotides of the sequence of SEQ 1D NO:X and the 5' 10 polynucleotides of the sequence of one of the sequences delineated in column 6 of Table 1B are directly contiguous.
Nucleic acids which hybridize to the complement of these 20 contiguous polynucleotides under stringent hybridization conditions or alternatively, under lower stringency conditions, are also encompassed by the invention. Polypeptides encoded by these polynucleotides and/or nucleic acids, other polynucleotides and/or nucleic acids encoding these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides, nucleic acids, and polypeptides are also encompassed by the invention.
[089] In specific embodiments, polynucleotides of the invention comprise, or alternatively consist of, a polynucleotide sequence in which the 3' 10 polynucleotides of a fragment or variant of the sequence of SEQ >D NO:X and the 5' 10 polynucleotides of the sequence of one of the sequences delineated in column 6 of Table 1B are directly contiguous. Nucleic acids which hybridize to the complement of these 20 contiguous polynucleotides under stringent hybridization conditions or alternatively, under lower stringency conditions, are also encompassed by the invention. Polypeptides encoded by these polynucleotides and/or nucleic acids, other polynucleotides and/or nucleic acids encoding these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides, nucleic acids, and polypeptides, are also encompassed by the invention.
[090] In further specific embodiments, polynucleotides of the invention comprise, or alternatively consist of, a polynucleotide.sequence in which the 3' 10 polynucleotides of one of the sequences delineated in column 6 of Table 1B and the 5' 10 polynucleotides of another sequence in column 6 are directly contiguous. Nucleic acids which hybridize to the complement of these 20 contiguous polynucleotides under stringent hybridization conditions or alternatively, under lower stringency conditions, are also encompassed by the invention. Polypeptides encoded by these polynucleotides and/or nucleic acids, other polynucleotides and/or nucleic acids encoding these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides, nucleic acids, and polypeptides are also encompassed by the invention.
[091] In specific embodiments, polynucleotides of the invention comprise, or alternatively consist of, a polynucleotide sequence in which the 3' 10 polynucleotides of one of the sequences delineated in column 6 of Table 1B and the 5' 10 polynucleotides of another sequence in column 6 corresponding to the same Clone ID NO:Z (see Table 1B, column 1) are directly contiguous. Nucleic acids which hybridize to the complement of these 20 lower stringency conditions, are also encompassed by the invention.
Polypeptides encoded by these polynucleotides and/or nucleic acids, other polynucleotides and/or nucleic acids encoding these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides, nucleic acids, and polypeptides are also encompassed by the invention.
[092] In specific embodiments, polynucleotides of the invention comprise, or alternatively consist of, a polynucleotide sequence in which the 3' 10 polynucleotides of one sequence in column 6 corresponding to the same contig sequence identifer SEQ ID
NO:X (see Table 1B, column 2) are directly contiguous. Nucleic acids which hybridize to the complement of these 20 contiguous polynucleotides under stringent hybridization conditions or alternatively, under lower stringency conditions, are also encompassed by the invention. Polypeptides encoded by these polynucleotides and/or nucleic acids, other polynucleotides and/or nucleic acids encoding these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides, nucleic acids, and polypeptides are also encompassed by the invention.
[093] In specific embodiments, polynucleotides of the invention comprise, or alternatively consist of a polynucleotide sequence in which the 3' 10 polynucleotides of one of the sequences delineated in column 6 of Table 1B and the 5' 10 polynucleotides of another sequence in column 6 corresponding to the same row are directly contiguous. In preferred embodiments, the 3' 10 polynucleotides of one of the sequences delineated in column 6 of Table 1B is directly contiguous with the S' 10 polynucleotides of the next sequential exon delineated in Table 1 B, column 6. Nucleic acids which hybridize to the complement of these 20 contiguous polynucleotides under stringent hybridization conditions or alternatively, under lower stringency conditions, are also encompassed by the invention. Polypeptides encoded by these polynucleotides andlor nucleic acids, other polynucleotides and/or nucleic acids encoding these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides, nucleic acids, and polypeptides are also encompassed by the invention.
[094] Many polynucleotide sequences, such as EST sequences, are publicly available and accessible through sequence databases and may have been publicly available prior to conception of the present invention. Preferably, such related polynucleotides are specifically excluded from the scope of the present invention. Accordingly, for each contig sequence (SEQ m NO:X) listed in the third column of Table 1A, preferably excluded are one or more polynucleotides comprising a nucleotide sequence described by the general formula of a-b, where a is any integer between 1 and the final nucleotide minus 15 of SEQ >D NO:X, b is an integer of 15 to the final nucleotide of SEQ
~ NO:X, where both a and b correspond to the positions of nucleotide residues shown in SEQ m NO:X, and where b is greater than or equal to a + 14. More specifically, preferably excluded are one or more polynucleotides comprising a nucleotide sequence described by the general formula of a-b, where a and b are integers as defined in columns 4 and 5, respectively, of Table 3. In specific embodiments, the polynucleotides of the invention do not consist of at least one, two, three, four, five, ten, or more of the specific polynucleotide sequences referenced by the Genbank Accession No. as disclosed in column 6 of Table 3 (including for example, published sequence in connection with a particular BAC clone). In further embodiments, preferably excluded from the invention are the specific polynucleotide sequences) contained in the clones corresponding to at least one, two, three, four, five, ten, or more of the available material having the accession numbers identified in the sixth column of this Table (including for example, the actual sequence contained in an identified BAC clone). In no way is this listing meant to encompass all of the sequences which may be excluded by the general formula, it is just a representative example. All references available through these accessions are hereby incorporated by reference in their entirety.

~i o0 .~ ~i cri ,~ v~ '~ ~j ~ ~ ~ ~ M N
t~ ~ ~~ ~ oo a\ '° ~n ° ~t ~ °' 0 0 ,~ l~ ~ .~ M N M 00 V7 '-' M ~ ~_ r' -.yes O~ Q1 ~ ~ 0 ~O ~ W M O N O N p ~oMO ~~oNO3M3ooo~ooUao ~~~d~d~~~Ud~d w ,n N oo t~ N o ~ N °~ t\ ° ~ d °° ~ o N .-m ~ p~p~M~~01\pM~N~p~~
ca ~° r' ~ ~ ~ ° ~ ~ p ~ ° ~°.1 U o~n ,.°-~
U o ° ~ d ~ ~ d d d d d ~_ U d d V ~ M l~ ~ ~ 01 ~ O 00 D1 ~ ~ [~ d M
M~ ~M~~N~o~oN~~O~~N
v0 d N M O o~o O ~ ~ O ~ N ~ O ~ p O
d U a U
"' dNdd d v0 M .~ ~ t~ f~j ~ ~ ~ M N ~_ ~ M ~V o0 p N ~ ~ o0 N O ~ O ~ ~ ~n l~
M ~ d' v~ ~ ~ .~ ~p ~O O O O O O N
H~ d~d~~~~~~~d~dd~d W O M .~ 00 ~ ~ ~O ~n O N
O N v~ ~n O ~ ~O o0 C .~ ~ N ~' M M ~ ~ ~ ~O
R ~ ~ i i ~ ~ i ~ i i L~.I !~ V~ In V'1 V~ V~ ~ In ~ l!7 ..r V
.,.., A
H
W
R
W
N ~ l~ ~ N N N W O o0 ~ N o0 .-r ~ ~ 01 V1 ~
~ M M M ~ ~ \O
R ~ ~ ~ ~ i i ~ ~ i .-r rr ~ ~ .--i ~ ~ .--r A
H
~ V1 ~ ~1 O ~ ~ O M ~D
I~ N ~ 01 Q1 ~
~,' V7 M 00 00 l~ ~~ ~O ~-~ I~ 00 O N ~O ~n N v~ ~n r~ O~ ~n Ov (, ~n N l~ N V1 d' M 01 O O
d1 01 ~n ~ 00 0o I~ ~O IWO
ao Wz .-~ N M ~ ~n ~O l~ 00 01 O
~ N M
W N ~ ~ H ~ M ~ M
C7 ~ ~ f~ G~l U ~ ~ ~ U
d dddd~~~~
~x xxxxxxxx ~ ~ M ~' N m 00 ~ ~n ~ ~h O ~ N ~ ~ ~p ~ M M o0 N N M ~ ~ ONO .-~ N_ ~ p '_-' M O ~ ~O l~
O~NO~OO~OOOOM~..~~~M OpN~'pO~O~po~O
M O .~ M
O O ~ O I~ O O '~ O O p O ,~ p~ .~ ~O O,~ .~ O o0 O ~ O O ~ ~ O ~' ~~d~°o~°odd~d~d~~~~U~od~~NddNUdU
o ,-~ c.i oo d v; U vo 00 ~ o o ~ .~ ~ d .~ d c~i o ~ ~ o v; ~ o; o d M d ~n d N p ~' ~.,~ M Q1 ~ [W ~ l~ O O V_~ ~ N ~n '~ O M
~ N l~ N ~ ~ ~ ~ ~ p ~ ~ N ~ ~ ~ N ~
~n O O p ~ p ~ O O ~O p O O ° O ~ ° p O M ° O ° O
O O p M N
~ U U ,~ .~ U U U ~ ~ U ~ U U o U N U ~ U U U U U U '° 0 0 ~dddUdUdd~dddddUdUd~dddd~dd~~U
~~°~n°~d~do~N~~~NNd~d°°~°M°o'°
o°~°o°o~~~~d O ~ l~ gyp" N ,~ pp ~ O ~_ ~ ~O [w ~ M ~ V~ a O M M ~O ~ ~ v~ d' I~
O O O ~ O ~ O O ~' O O N O N '~ O vp O d O p p '-' ~O M v~ ~ Ov N
O O O ~ O ~ O O ~ O O O ,~ p ~ O N O M O O O p O O O O O
,~ U W U o~ U o U U o U ~ a ~ a~ U o U ~ U U ,~ U U U U o .~dddNdoddUd~dddNdUdU,d~dddddd~o c.i vi ~ ~ c~i ~ oo d Vi ~ o ~ ~ ri M d -,~ N '~' °' '° o 00 ,~
~ o0 00 ~
N ~ 00 ~O p~ p~ t~ N ~n O ~ M p~ ~ M v~ 00 ~ 00 O ,~ 00 ~O
O p ~ ~ M ~ d1 ~ ~ N N v~ ~ ~ ~ ~ p ~ ~ pN ~ O v~ v0 O ~
O O O O O ~ O ~ ~ O O O O p O p ~ O ~ O O O p ~ p p O O
Ndd~ddNddNdddd~ddNdNddd~ d~ddN

-rs 00 ~ M O ~ N Op M M M ~ .-' Vj N ~ ~ V7 M O ~ O ~ ~ O O O v~ ~ p O ~' ~' ~ v1 p ~ 0~1 o ~ ° ° ~ ~ U U ° ~'' ~ U U ° ~ N o~°o ~ ~
av a U ~ ~ U o U N o, dd~ddU~ddda~dQ~ ~ do~~d p~pl~ap~p~~v~MMOdO~N~ d ~ dN,-, \O N_ M ~ ,~ Oy~ ~ N ~ O M M
O O M M N ~ O O ~ O O M ~ p N o0 ~ O ~ M N
.--~ O rr p O ~ O O O O V~ O O O O
U Q. ,~ ,....a U U U ,~ ~ U U U ,-.a ~dxdd~ddd p~ddd _ ~p ~~ l~ ,-i ~ ~ I~ N \O ~p ~ ,-, ~p o0 N ~' 00 O~ ~-.~ M Q1 I~ M 01 N N o0 ~O ~ ""' ~ d N ~h v'7 v~ O ~ 00 ~ 00 O O ~.,~ ~ ~_D O N O p N ~ 00 O O O O O ~ O O O M N M O O O ~ M p ~?' OOOpO~OOON'"OOOO ~ ~M"'' U U U U U ~ U U or~, a°~, a U U U ~ ~ rM,~ ~ 00 dddddNd~d~Ndddd ~ ~d N v-i l~ p p~ wi Ov ~ ~O l~ d' M ~j O ~n ~ M rj ~ ~ d 01 I~ ~ O M V7 N d1 ~O N o0 ~ 00 ~ t~ ~ O~ O M ,-, ~ N 0o ..-~ 00 O O N N ~ ~ M o0 ~-~ v~ Q1 00 p O o0 pp 'n ' Q\ M ~ ~O v~
~n v~ N ,~ 01 N ~n O ~ ~ ~ ~ o O l~ ~ ~ '-' d' O ~ ~ M O~ ~ ~h O O O M ~ O O N O M O O o O O N O ~ O o0 o N ~ ~ M
O O O O p O O O O p O O O O O O '-' O p O ~' M l~ N '~
U U U ~..a ,~ U U pq U a U U a., U U '~ ~ ~ '~ ~ N
dddd~dd~dddd~dd~dNd d~ ~d~~~
o, ~ .~ a, ,-. ~ o N
O M O l0 M ~ ~n O
M M ~ M M 00 ~ M
i i ~ ~ ~ i i V7 ~ ~ V~ V7 ~ ~ V~ V~
N ~ O I~ ~n l~ M ~O o0 O Q1 N 00 V1 ~ M M 00 N M M M M 00 ~ N
i i i ~ i i i i i l~ N Q1 ~D l~ v~ ~ O O
N ~ v~ O ~ O ~O
N M .-r I~ t!1 M 00 ~l' l~ ~O v7 I~ ~ O ~D a1 t~ V~ .~ ~p ~O M I~ ~O ..~
N M d' vW D (~ 00 ~
N N N N N N N N N
~ o0 V' n I~ 00 .~ <t '~ N V1 M .--i a 000000'~~
U U U U U U U U U
xxx xxx 00 gyp" ~ Ov N M N
~ 00 ~ ~ ~ ~ O
~ 00 M M 0O ~ ~ ~ ~ ~ ~ ~ ~_ ~ ~ '~ M
O ~ ~ N N 01 ~ O ~n v~ O 0~1 r.. M
~ N Oy N ~ ,~ M ~ ~~ ~, 00 a ~ ~ ~ N O
N y--.i N ~ ~ o d ~ ~ err' 3 ~ ~ o ~ d M ~ d ~i N d d d v'i .~ ~O N O O '-' M v~ '~ ~ ~t v~ N N ~ ~ d v~ ~ ~ M ~ ~ ~ N .-. O d' ~ N (_~
Wit' I~ ~ l~ N ~ d1 l~ ~ M d' M O N ~ D1 v~ ~ ~D '~ ' O ~ ~ ~ ~ ~ ~ 0N0 ~ pOp d r-m~ ~ ~ ~ ~ V, ~O H a d ° .-., d '_°
d d a, ~ d ~ ~ ~,,~ d ~ "~ oo ~ d ~ ,r; f~ d fV d ~ M ~ O O M (\ M ~ a O~ ~ ~ O ~O ~ N ~ M
O ~ ~n ~ ~ ~ N O~ ~ ~ ~ ~ ~ N M j d M y0 ~ ~ d' ~ 000 0o M_ oo N O ~ ~' M ~ ~ V7 ~ M ~ M ~ ~ ~d ,.M~ ~ '"'' ~ ~ O O pp ~ ~O r-~d~°°~'~° ~~~Nx ~ d~~d~~d~~~dd~d o°° ~ ~ ~zM°°
00 M O M (w ,--i 01 '~ M ~ ~ M O o0 ~ ~' N O ~' N
O r" oo V' pMp ~ O N oo ~ d' N I~ ~ ~ I~ 00 00 ~ O_1 ~ ~ ~n N
N~~~N MO~~d~'N ~ ~MMNN~opoM~~~O~oMo O O I~ ~ d' ~ d N ~p ~ O ~ l~ ~n ~D Q OM N M
ddar~~~~~~3~~ ~ ~~ddH~~~~d~~~~
o t~ t~ .~ 00 d' I~ 0o N O
M
i i ~ i ~
V'1 V~ V7 ~ V~
~O M M l~ ~T
N ~D l~ O ~
'Wf ~' M M
i ~ i i i 00 .-, N ~
N O ~ l~
I~ O M ~ 00 V~ I~ I~ M ~O
O V1 00 l~ 00 O ~ N M ~
M M M M M
N~
M I~ ~D I~ 00 ~C al U W w U ~ ~ A f~
xxxxx ls~

~ v~ M ~ ~ ~ N ~t I~ ~n M a1 ~ N
wj ~1 ~ ~ .-r M ~.,~ ..~ ~ ~p ~ 01 O ~!1 tn ~ O Q1 Q1 ~ 'O M
M 00 ~O N lp O 00 00 O 00 ~h 00 M O ~ .~ N M 00 00 ~ ~ ~ O O\ Vl N M ~ p ~ ~ ~ ~ ~ N ~ ~_ O oho ~ N ~ ~ ~ ~ '"N"~ ~ ~ ~ O ~ pv Ndd~ ~d~~~ ~~~~d~~H~~~~~oo~~~~~F°
d a, o o ~' ~ ~.,; d c~i d d ~ o d ~ d ~ N d ~ i ~ ~ ~ a; vi ~ ~. ~ ~ ~ ~ ° d a. ~ ~ ~ N ~ ~. M ~ ~ "~ ~ ~ o ~n oo t~ o 00 ~ o ~
~p O N
M ~ ~ ~ ~ M ~ ~ ~ ~ ~ ~ Q ~ ~ ~ O N d~ 3 ~ p N v~
dddd~~~~~~~~~~~d~~~~~~~d~~d~~~
0o a1 O ~ ., M ~ p~ ~ ~ O N N M ,-. ~ O r.i a;uivN~,o'_°oooMod~~a;o ~~~~o~oo_oo~oc~o~°_°~'~''°d~ o N ~-M-~ 0~0 M ~ ..-y ~ ~ ~' pp 0M0 ~ N ~t N O N ~ 00 ~ ~ N ~ ~--r O~O M
~ 00 ""' ~p ~ ~ O~ .~ l~ O ~ O ~ O ~' M M ~ M ~ l~ ~"' M ~' 00 00 ,~ 00 ddddddd°~d~d~~d~dd~~~~d ~d~d°_~~
M ~ t~ ~ N ~O ~ vp vp ~ M ~ <t p y0 0~0 d d1 ~ ~ ~ O O O d N M
00 M ~ ~ M ~ M O 00 ~ ~ 01 O N V' ~ ~ 00 ~ N ~ 01 M
O O O_O 0o I~ M ~ ~ ~ M V7 M N M v7 ~ ~ \O ~, ~O l~ ~ 00 ~ ~p ~O ~ N N ~O
\O O l~ ~ ~ O ~ ~ ~ ~ M O ~ ~ N ~ O M ~ ~ 00 ~ vp N O O ~ M
~dd~~ddxd~ddz~~d~dz~dd~ddddH~~

ri o "' 'D ~ ~ o, ~ ~n ~ M ° ~ ° ~ o'no -; l~ V7 M c'n ~ ~N O 00 ~ ~ ~ ~ M ~ N 00 Crj I~ ,~ N ~ M ~ l~ 01 ~ M O O ''' O O p~ 00 M
l~ N M N t_~ O ~ ° M M M 0o ~p O 'ct ~ O ~ O O O O ~ ~n ~ '-' °
~ ~n O
,.°~ o M o a o ~_ I~ oo ~ M ,~ ° '"'a O ~ d d d x °
° '~ ~ N V, ° d ~ ° ,'~ N vo 0 d U U ri t~ ~ oo a\ °o '"~ d U v~ d o t~ ~t oo r~i d o ~ ri a~
°o M~~d~dN,~~ °~UdNd,.~ ~~~~,,~ UM~.,,.-. O
O O~ 01 M N '~ .~ V~ M N o0 ~ d1 O 01 O M O l~ ~' op ~ U1 ~ ~ ~_ O O ~ O o O pip l~ O M ~ .-, O v7 ~ 01 ~ M O O ~~ ~ ~ pp N O ~ M
a~ O O ° N_ ° ° ~ ~n O o ~n O o O O O N O O
d U o U ° ~ U U
~~~°~C°dd°°d~~~°°d° dU
~°~U U
o"~oN~~pd~o~°o~d~;~U~~dNd_o~oMOO'°o,M~A~ oc'~o 0 O N O 00 ~ ~ ~ M N ~ ~ M O ~ O N ~ ~ ~ M O ~ I~ ~ ~' O~O V7 N pp O Q\ O N M O ~ 01 N O ~ O ~ O M O O M O O l~ ~ ° O O
O N O ~1 ° o pMp ~ O ° ~ p°p O ~ ° .~ O o ° O o ~ ~ ° O p ~n O
w ~ a., o a U ~ ~n U ,~ ~ a., o U o, ,.~ U ,~ U U o v-, U U ,..a ~ U
~dx~°ddN~C~d~N~°d~~ddddU~Cddd~d N N M ~-~ ~ ~ ~ 00 00 O ~ '~' N D1 d d1 N M ~O 01 fV ~-~ d N M I~ N N 01 N ~ O ~ ~ ~O 00 ~ l~ ~ M ~n v7 ~ ~ '-' ~ ~n ~O O O~ M 01 ~
M O M ~ .~ N O ~ ,~ ~ ~ ~O .~ ~ M O 01 ~O ~ D1 ~' d1 Ov O N ,~ N t~
M O O O ~ N M l~ N ~ O ~ M O ~ O O O O p p O M M O O O
O O O .~ O O O ~ O O ~ O O pip O O O O O O O pp ,~ O p p ~ O
~dd~~N~~~~~da~~N~~~~~~d~d~~~N~

O N .~ O ~ p ~' N ~ o0 ~O M
M p oo ~ ~ ~ 00 00 ~ ~ I~ ~ '~ N O N
00 O Q ~ O ~ ~ ~ N ~p 00 ~ 00 N ~M ~ vW~ ~ O O O O ~,.~ M
d ~G d ~ U d ~ ~ d ~ U o U N ~ o° a U U d a al ~ U ~ d ~ tj ~ o ~ N ~ ~ d c~.~ ~ ~ ~ ~ d U d ~ o U d d d ~ d d ~ d ~ ~ N
~~MM~N~N~o~pd~~~d~~~~M~pNNNN~~M
O M M ~n ~ p 0~0 O O ~ 01 N a1 O ~ N d' M N I~ U l~ O O M p M
Ua~oNUo'''oUU,~'to~°~oo-lU°'oo~~c c d~UU~-°-IUaN
ddd~,~dNddxo~N.,oddoooodoooNdddddo ~., U oo U ~ U U a ~l ~
v~ d N d o;
M i~°~~~ oN,~oo _"~,_,-,~dd~~dd ~_ ~ ~ 0 ~ ~ oNO ~ ~n '~' I~ ~ ~ N ~ O ~_ O
O O O O v~ O O ~' O .~ M o0 ~n O O M ~n 01 .~ O o0 00 O N O d' U U U U o U U ~..a U o a~ o ~ o o ..-.. o 0 dddd~dddd'~~N~~~~aN°~~a°d~dd~d~
~-a o; c~ c.~ Q ~ o; ~ r; oo d .~ yo 00 o d ~i ~ ~ d d N a; ~ c~; v-~ ~ M d O o0 O p o0 V~ 00 '~Y Q1 ° 00 M ,-, ~' ° a1 l~ N ~ N O
O O O M ~ O O p 00 O ~ O N M ~ O O O M O N ~ O O O M O O O M
O O O O ~ O O .~ O O ~ O N O O O O O M O N I~ pp O O O O O
ddddNdddd~NdNd~dNd~dN~Ndd~d~dN

_~
O N
N O ~ N ~ O O
t~ M ~O '-' O vW O N N o~0 ~ ~ 00 V~ O~ O ~n N Q_1 N ~~,-~,M ~MOOOO~d~'M~OOpOVpOOOO
0 o M N d "' N o U U U ~ o ~ ,~ U U o° U U U ,~ o U
UVa ~ ~°,~°~w~d~dddNU~dddoddd~Ud d d d ~ a; ~ °~ o; °~ o ~ °~ ~ ~ d N ~ ~ ~ ~ o°~, o ~ ~ d ~
O O O ~ ~ ~ p O ~ p O p p ~ O
~n N ~O l~ ' ,-.~ ~ v~ ,~ M O O O O N ~ O p O ~ O O p O ~ O
00 00 0 0, '''' o N N ~ ~ ,...a U U U o o U U M U U U U o U
~ N U N o ~ ~ ~ ~ ~ d ~ d d d d U ~ ~ d d a d d d d U d ~d~c~-i~~c~i~°~°°~d~N~~d~°~°M°do°o 0 0o M
O O d O I~ M M ~ ~ O O O O O ~ ~ ~ N O ~ O ~ ~ ~ N O
O O pip O .b M p~ ~ O O~O ~ p O O O ~ ~ ~ p O ~ O O p O ~ O
ddNd ~ ~Mddad a.,UUUNMr~, UUooaUUU~U
d ~~dddaa~dd~7dddd~d ,~ ~D N ~ ~ ~ ~ ~ 00 O N ~ d d O ~ v~ ~D o0 ,~ ,-; vwp ~
v7 O I~ ~ ~n O O t~ v7 O N ~.,~ ~ v~ l~ N v~ ~' M ~ v~ ~' v_~ ~ Vwp V7 O O O ~ ~ O ~ ~ M I~ ~ ~ ~ ~ O N ~ ~ N ~O ~ 00 ~n ~t M
O O O O O O O ~ .~~. ~ ~f ~ N ~ O O O O p ~ O p O O O O ~ p p O
d~~~~~~~z~~~~dd~~d~xa~ddd~~~~~
M ~ N
i N
M ~ N
N ~O ~D
O ~ M
O M
D1 M l~
M N
M M M
N
N v»n a ~ c~
xxx O O ~ ~ O~ ~ 00 ~ ~ ~p O
N N
~

01 ,~ N
Ov N O M 'n N O ,.~ 00 I~
v~ ~, O _.. p p1 V~ p ~ ~ ~ ~ O ~ ~ p ~ 'cY O ~ O ~ ~ ~ Q\ O
~ O 0 O
p O
O ~ O
~
~

O V~ Q1 ~ ~ U ~
~t ~ O
O O N ~
.
O O O
O .b U U a1 U U U U ~ ~''~ pq U ~ (~ U U o ~

oadN~ d~dNd~dddd~~
~d~dddU~

o ~n _ d ~ O O ~
p N ~ ~ ~ ~O 01 d "'~ M vp M N ~ N
0 ~ ~
~

j pp U ~O
~ O
0 0o ~ ~ O ~ N M l~ ~ ~D 00 ., ~ O 01 V~ M ~D ~
I~ M M
N
~!1 ~ v~
N ~n I~ ~n ~n ~n ~

v~ M .. ~
d N "",, N ~ v~ .~
N
,.~ N M M O N M O O O ~ ~, O O O N N N M O
~ O
O
O ~
O O O M
O

~O ~ O ~ v7 '-' 'C O
O a O ~' ~ O .~ O 00 O O
N O O O
~ "'~ ~l ~-1 U ~l ~l U U U N oo U U U M ,..a a ~ ,~ U
~
~

UUd~ dddddddo~dddo " d dd~dd 01 N 00 N ~j 01 M '-' V7 t~ M O ~D a N Ov O ~ ~ p ~' N [W M .-i ,-; ~rj ~ O ~ ~ I~ ~t d N O N N M
-.
~
O

O
~n yp ~
op ~ N .~
p~
v ~ ~ M ~ O N N ~ O ~n ~ ~n ,w0 00 pv M O o0 ~D

N M ~ ~y0 O v~ ~ p~ p O~ t~
~ '-'' O O~ N ~n 'WD O~ ~n N l~
N O ~ O ~ N M O O O O O O M O O O O ~ ~ N O O O
N O

~~Uoo~a UpaaUUUUUU~NyUUUU~p,....~UU 1U
~ ~
a~~

~~dN dddddddodddda ddddd I~ 01 ~ M ~ ~O ~ I~ ~~ 00 ~ I~ ~ N ~n o0 V~ I~ ~
M ~ l~ t~ ~~ oo v'i .-i N M ~ ~ p~ 01 N v~ ,.-~ N O ~ 00 ~ 00 tn N I~ N
M M Q ~ O o0 O
t!~ O ~ [~ ~ M O M o0 M Q\ ~ ~; O N lp ~ N N M O
O -r O ~
00 O V~ I~
v0 M O
~n _ p M O O ~ ~ O N O pOp O O O O M O O p O O O ~ O p O O M O d' O O O O O O O O O
O O O O
O
O
O

O O
~d O d O
O
O O O O
O
ddd~ddddddNdddddd~d~~~d ddd N

~t~t i I~l~ N
O M

i O
~OM

V700 l~
O V~ ~n I~l~ O

M M d' O V'1 w w d C

b cri j N 0~1 N
b o0 -; 00 M
N o 0 0 .N
M ,~ M N M
cd ~n N 00 ~ ~ N
N ~ O
M O O M O ~ 00 N
~b -b ~ .b ~ O
H b ot\O ~ ~ ~ O
00 .--~ O~O 00 N cd N ~ ~ ~ N ~ O ~
00 M V7 '-"' M Q1 O N ~ pp I~
N ~ .~ N O ~ 00 01 N ~ ~' ,-a Q1 M ~ O N O ~ v~ M ~ ~ ,~ M N
d~~d ~ d~ ~ ~d ~~ ~z ~O ~~ 00 ~ M N N o0 O O M N 01 ~ ~O V o0 I~ N ~ ~O ~ ~ ~ N
~O ~D ~O O ~~ M 00 Q1 O M 00 01 O 00 00 N ~O ~ a1 ~ ~ l~ ~~ v~
N ~ M ~ ~ ~ ~!1 ~ N M ~--~ 00 N ~ ~ ~ M M M ~ ~1 ~O M
i i i i i ~ i i ~ ~ ~ ~ i ~ ~ i i i ~ i i ~ i ~ i N ~ ~ N ~ ~ ~ ~ ~ N N ~ ~ ~ ~ N ~ ~ N N ~ ~ ~ N
N I~ ~ O 01 00 0o W p ~O ~ 00 ~ O N O ~ M o0 I~ N O ~ I~
v7 ~ ~ Q\ Q\ ~ 'O 00 00 ,~ ~D l~ 01 I~ t~ ~ ~n M l~ N O ~O O M 00 N ~' N M ~ V7 t!7 ~ V7 M ~ (~ N V1 \O ~ M M M ~t V7 ~D M Qv i i i i ~ i i i i i i i ~ i i ~ i ~ ~ i ~ i ~ i 00 ~ ~ ~O ~ M N I~ M O~ O~ v7 M M ~ N I~ M d' O1 l~ l~ O N
~D ~O l~ N O~ 00 ~O v'1 ~ I~ ~ I~ I~ ~O ~ O l~ I~ ~ N O~ N N N N
l~ I~ Ov Ov a1 N t~ N N N ~ ~ ~t OW O ~O O~ N O l~ .-. 00 0o O O
Q~ ~1' 00 .~ ~ M ~ V'1 M M N M V7 Q1 O ~ l~ ~ O 00 l~ O M ~ 00 N ~' ~ ~h O M ~~ O ~ M ~n N ~-r v~ .--~ ~O 00 N ~ O M M
~ N
N M ~ v~ ~D l~ 00 01 O ~ N M ~fi ~n ~D I~ 00 O~ O ~-~ N M ~ ~n ~ ~t v'~ vo vmn v~ ~n vo v~ ~n ~n ~p ~O ~O v0 ~O ~D
.~ a1 O M 01 N Q1 I~ M ~ ~ O O V~ N a1 M ~ O O ~ O
rr M N
M ~O ~O O ~ ~ M ~ M l~ O O M ~ 00 ,-, ~ N
C~ C7 CJ ~ ~ ~ ~ v~ v~ x ~C U O ~ ~ C7 ~ O
xx xxxxxxxxxxx xxxxxxxx xx~ x wi N M v> " O ~ ~,~ N oNO ~ M
M p~ ~Y M ,~ O~ p~ l~ ~ 00 O1 M ~ O
00 00 p ~ ~O ~ ~ V1 N ~ ~ ~ O ~ O1 00 ~! ~ ~ ~ ~ ~ ~ O N O ~ ~ N .~ ~ ~ ~ N N l~ O ~ ~D
lp Q r3 ~ ~ ~ ~ M ~ ~ ~ 01 ,~ r'~ N ~t N ~ ~ M M d1 d ~o d ~ ~ ~ "' M ~ d ~ ~. ~ ~ ~ oo d M "' a d ~ N ° ~ ~
~ _.. ~ oo d c~i d ~ vo d ~ d ~ ~ ~ v °~n° ~ d ~_ d ,..MN-, d t~
~ 00 ~ ~ N p ~ N d ~ ~ (O-, ~t ~ M N
O l~ ~ 01 00 '-' ~ ~ v7 M ~ ~ O ~O O N O N l~ O M v7 ~ ~ l~ M
O ~ ~ N N ~ ~ ~ 00 i-~-.~ OM ~ ~ ~ N N
O ~ ~ ~ O ~ O d M M N ~ '~ ~ ,-. M d1 0 N .~
ddd'"' M ~oo~OdNd~~ ~dNa~
M d d~~ ~d~d ONxd~ ~~M
00 ~_ ~ ~ o N 'r' d x c~i ~ d ~ ~ ° ~' '° v; oo M d N ~ O O ~ ~ N N O ~O ~ ~ O M O ~ ~ ~ N O ~ O N M
M ~ ~ M V7 Ov ~ l~ ~ ~ N N ~ ~ N ~ O pip 0 ~ ,M-,, ~ ~ ~ 00 pp M
Cn N .-.~ 01 I~ 00 O pp ~ 01 ~ ~ N ,-.yn ,.Nd ~~~~~~~w~% ~~°~,~°~°~ °~'rd~r3 war, d ddd x d N
O ~, ~ ~ ~~ N N ~ ~ ,-. ., vp ~-~ c~ c~j c,i v~ ~' ~ ~~ p~ O M N ~p M O I~ N d .-r d ~ .~ ~ ~ ,--~ O O N pp -; N ~h ~' t~ ~' .-~ ~ O ~ 01 ~t O N v~ t~ N M v~ ~D O o0 M ~
M M o0 ~ N ~ ~O ~ ~ p~ M ~O d' v~ ~ N ~ ~ ~ ~ d' N ~.~ ~O o0 p ~O
~' O M ~ N M ~p ~ .~ ~. ~ I~ 00 O ~ d1 V7 'p M M N V1 N I~
00 ~ 00 ~n O o0 O V7 ~p ~' O l~ l~ 01 ~ N N N ~ 'V o0 00 00 M ,-, W
d~ddd~~~~~~~~~w~wd~~~~~~~~w ~ ~ M
l~ .-V 00 O
Wit' d' ~O ~n ~n ~n ~mn l~ O N 01 v7 O I~ 00 i n O v~ v~
O ~ M M
M ~D l~ l~
O~
xxxx N
N O~~ON
~ Ov ~ t~ 00 ~ ~t ~ N ~ ~ n O ~ O~
I~ r., N 00 00 O ~O ~-' M ~ 00 l~ ~O O ~ ~ ~' QWD ~p M O M M ~ O pMp M ~O O
~ V'1 01 M ~ ~ ~ ~ pip ~ N M N [W ~ ~ M O .-~ [~ ~ O N
d O ~' pp o0 ,_, M ~ ~ O M 01 ~ O r-~ ~ ~ O
r, H d N M 01 ~ ~ 00 01 01 d ~ ate' ~ w yn .-. r-' ~ °o d N oMO ° d ~ o oo,r..,'"'~N a\~v-,~'-'M° ~C~~~'~ ~.-.U .,U
o Q, .~ ~t "' ao ~n yo 00 ~ oo N ,~ ~ ~ wi ~ ~ d v-; .~
00 ~ M o0 D\ O O N N N l~ ~ O Ov ~ ~ N ,-, '-' Ov 01 N ~ V7 N M O .--~ N ~ O p°p '~ .-r ~ Q~ V~ I~ p M ~ N \O 00 ~O M '~ .-i V~ ~
°o d ~ ~ d ~ d'n d ono ~ ~ ~ ~ ~ o M ~ ~ N ~ o ~ o ., r-.a N d ~d d ~ x ~ d ~ N N ~s., M '~' ~ aN, o ~,~ U
~~~NN~~~°°~NO°~o~oN~o~'~°~~a~
~00MMp~pO~~~d'n,~a~,o~~~o~~do°o°~
M 00 ~ M N .~ ~ 01 ~ M ~ N
<f' O ,~ ~ .~ ~ M ~O ~ ~ ~ 01 M ~ c'rj ~ I~ ~. O O 01 ~0~~~ NM ~MOp'nU~O.-.OHO
O
dd _~x ~ ~d~ °~~xd o°~U~~
rM., os ~ o°, ~ ~ ~'' '-' ~ o ~ ~ ~ o°°, v~ ~
°° ~ o ~ o ~ ~ ~ ~ d d N
000 °' '-' ~p ~ ~ ~ °' o, \° ~ o°°o ~ ~ ~
oho ~ ~ ~ '~ ° N ~ ,~ ~ o y_o °N I~ ~ ~ 01 ~ 00 M ~ ~ I~ d' M ~ ~ ~ p M ~ N
OM~_O ~~_01O~O~OONN'-'~_N~
dx~~~dd~ddd~~dd~d~dNd~Cdd~CNd~NN

wi c_~i wi ~p O
NMM O N ~OOMN M°~ ~t0~0~ .~"',00~0~ °
p ~n p O ~D ~ O N ~ M O ~ ~ N ~ M ° ° ~ ~ O
M N O o0 ~ t~ v~ V> 'n N M 01 01 M O o0 ° M ~h ° ° M ° O ~ O N N N
° ~ ° ° d' O
d o d ~t ~ M O M ° O ~O ° oo O d ° N ° ~ ~
° '~ ° d ° ~ d ~ d °M~C~r~dUd~Mdad~dU~~_vU~UdNd~oo_U~
~ o o, ,'~'~' ,'~ M ~ d ~ o ~ N d ono '~° ~ d ~ ~n '° d M d M ~n M ~_n t~ d ~
~ M ~ ~ O ~n ~ M ° ~ ~ ~ O O ~ ~ M N O ~ O ~ ~ O
U M U o yn o 0 0 o U ~ ~ M U o ~ o U M ~ ~ N U o U ~ ~ U
dNd°ooX°N°°d~C~~d°oo°~d~~Ca,od.degree .ddod °' '-' N d oo N d ~ d ~ ri os °o ~ ~ d N d ~ N ° ~ N d os d O os ~ M
o°o ,-.~, ~.r..U ~ ooUo c~ M
M N '~ N ~. ~ ~' oo ° ~ d ~ O l~ ~ ~ ~ ~ ~ ~ ~ ,~ ~ p ~ ~ y0 O ° N O~ oo ° ~O I~ ° N ~ ° p~ O N oo O O O ,-; O l~ p~ O~ ,~ M O ~ O
° ° ° O Ov ° r, p~ l~ p O ° ~ ~ O ~ ~ v~
° O ~ O ~ .~ ~-.~ N p ° ~. O
dU ~ ~ N ~ r~., o° o 0 o U ~' o, a., 0 0 o U U ,-, U ~ o ~., o a U o U
U Vor~,~d~_N~VUU~d,~d~V~UddUd yi ~o°d~d~°~t~~o°°o~ddd~~x~~d~da~,'~°d~
\p O M O r..; ~ ~ ~ 00 00 O 00 ~ d1 t~ ~ ~ ~ ° ~' ~ ~ ° pp ~ 00 ~ ~' N M
OO~c~~o~O~l~O~t~00p,~-,~ON~pO~OpMpN~OpMpN
° O O O ~ ° ~ ~ ~ ~. O O ~ O O [~ tn V7 O O M .~ O ~ O M O .-r ~
O
~~~~x~~ax~~~~a~~~~d~AddNdN~~N~

p ~ M ~ ~ N O ~O
v7 _ M l~ ~~ ~ ~O
\O .-, ~ ~' ~ O O M I~ ~ ~ 00 I~ '~ O ,-, O ° O ~ ° 00 O I~ ~D O
N M O ° ~O O N t~ oo O ~ ° N M ~' ° .-. O
° U U U ° ~ ,~ o~', ° ~o ° o, ~dd~M~UoMOO~U~N~UNdU~o d N o o ~ ~ d N U ~ d ~ N o d °,~, N d N
M ~ N O N ~ ~ d N ~ O ~ ~ ~ ~ ~ p ~ '~ ~ O
Op~O~~Np'"'O~~n~OO
° U U ~ U M o v"~., U ~ ,~ N ~ o U ,..~ o U d o~dd~daVodod°~~dda~
o ri oo ~ ~t d d o ~ ~ ri ~ d vs ~ d r. ~ ~~ o0 N ~n M O U .~ ° ~ l~ p ,~ M Vj o0 N
~0°O~~~d~'d~~~ ~~N ~~~ ~ N MM
O p ~ O N ~ O O ~ O ~ O ~ O p ~ p Ov d o U U .-~ U ~ o o U o U o M ~ U ,..a ° U o 'b N N 3 U~dd~d~U~d~d~.~Ndd~dU ~ b ~d N v'i N M N ~D ~ N ~O ° ~' O N O~ ~j oo ~
d oo W p oo vo v~ ~ M ~ oo ~ ,~ O~ D\ ,~ t~ ~ f~ cYi ri N
~ 00 0~ 01 M cri o '~ oo ~ o vo 0o yo c~ .-.
V~ ~ 01 M M ° .~ .--i ~ Q1 ~ ~ ° d' l~ (~l ~ .-~ ~ I~ '~Y M
~ l~
O O ~ O O p ~ ~ O p O O ~ O O ~ O O
nddddd~~~ddddNdddddN d ~ N dd o ~n ~n ,-~ ~ ~o M vo o~
M t~ N 01 \O M l0 t!7 00 M M M N ~O N N ~O M
i i ~ i i i i i ~ N ~ ~ ~ ~ N N
~ l~ N N 01 N
~-W O~I~V~N~~I~
M M M N ~O v~ N ~O M
i i i i i i i ~ i 00 01 00 00 l~ l~ M Q1 V7 00 00 O ~O l~ 01 O~ O I~
l~ ~ l~ ~O ~ d' ~ I~ N
I~ M ~ M M d' 01 M V'1 N l~ ~ l~ l~ l~ N I~ M
Q\ Q\ Q\ ~
O ~ N M ~ ~n ~O l~ 00 O ~~ l~ pp M ~' M N
° .-. N ~ ~ a o0 ,~
WWWWWWWWW
xxxxxxxxx M 00 ~O
~ N ~ '-' l~ O '-' M ~ 01 Q\ oho ~ ~ ~ ~ ~ ~ '~ O o~0 a1 O O N
O M ~_ ~ v7 ~ ~ ~ N ~ M ~ O ~ N M N '~
N N rN-i~ O~ ~, N ~ M O M ~ 01 O~O N a O ,M_, M O ,N
~MMdNdodd'-'~o ''' ~dddOd°
WD ,~ ~ M M ~--~ M Wn N ~ O ~ ~ 00 01 l~ M t~ ~ l~ ~ ~." ~ r3 ~ ~ ~ ~ N ~ '~ M M ~
vp ~ ~ ~ ~ 01 ~n ~ M V~ . N O M ~ ~ ~ ~ ~ ~ O N
p~ N -.-... pt~p N 01 l~ N ~ ~ l~ ~n b M Own ~D ~ ,N-, ~ O~
~_~r;~o_o ~~ '-'MOB
G~ o0 00 ~ M ~ M ~ d ~ ~ ~j ~ M O ~ Q M d' ~ Q1 ~ ~-r~ V'1 N ~ O O
00 d' ~ M N O V7 v7 ~ M_ ~ U ~ O M ~
~_ ~ O ~ ~ ~ M ~ 00 ~ 00 ~ b b M ~ ~ l~ ~ M N N ~ ~ M ~n ~~'dd OH~-~-d~'~~ ~ cad .~~[~~d~Ndd~M..
~~a;dN ~ ~Md~
~t ~ t~ N d ~ oo N N N ~ pip ~ c~j ~ ~ ~n ~ ~ N O ~ ~' p ~t N ~ ~ ~ ~ ~ 00 O O ~ ~ ~ M V7 Q~ 41 01 ~' 00 '-' ~ N 00 O "'' ,~ l~ O O M M M ~ ~ N M O p ~ ~O ~ ~D N
~~~~,~d~ ~°°~3d3a~ 3~3 ~ddddH~ddddHdd ~~dddddddd~ddd N ~ ~ ~n a1 .~
O ~n N M Q\ N
~ M N N N N M
~ i ~ ~ i 00 O O ~~ ~n I~ N
OO~~.NO000 M M N N N N M
Q\
~n d' ~ ~n oo N ~n ~n 00 M
I~ l~ l~ v~ ~ N
M M M O O M N
~n v~ ~n ~ ~n a1 v~
01 O ~ N M ~ ~n I~ 00 00 00 00 00 00 N N o0 I~ v~ N M
d1 N OO M M O <f' d d d C/~ C/~ C/~ W
xxxxxx~

cri M
O ~ 00 N ., .-~r N vp ~ '~ M O O O o0 00 N N
~O l~ O ~ ~ N _v~ oo O o0 ~ ~ O ~ O M ~ oMO ~ ~ ~ ~ ~ ~ O Ov ~' D1 ~ ,-M, M N O 00 ~ ~ ~ l~ V~'7 O N ~j ~ 0,-,O ~ ~ V1 v'1 ~ ~ O ~ ~ ~ ~O
o vo .~ o M
°° ~ ~ z d ~ ~,~, "~ o~, ~ o ~ N ° ~ ~ o ~_ o o ~ N
0 O M pp ~ '-. tfi ~ r-. ~ N ~ N ~C pMp ~ ~ ON N ~ M ~ p r~ ~ ~ d' M
~O O ~ M ~ ~ r; ~ M '-' 00 ~ O N ~ '~ ~ v~ O V~ V~ ~
.-r ,..., ~ Q1 ~ O N 00 ~ r"'i r~ r'~ x x p~ ~ .~ t~ ~D
O ~ N 0~1 N r-~ ., ~ .~ N .~ I~ ~n O
dH~ar,~'~~"'d~ 00o a~,~o'~°o~~,~~~ nU
M M M l~ ~ .~ l~
~ o ~ ~ ~ o d ~ ~'' ~ o°o ~ ~ ~ ~ ~ ~ ~ 00 00 ~ ~ ~ ~ ~ x ~ x o d i o ~ ° o ~ ~ o ~ ~ M '~° ~ N o o v~ ~ ~ ~ ~ ~C ~ o ~ '~°
aW ~ ~
00 ~ ~ ~ ~ l~ ~ N M ~ M ~ a1 N ~ 000 ~ O ~ N D ~
M N o0 M N d ~--y'1 r-~ ~ c,,~ Ov ,-, .~ .~ O _ O O
d ~ w ~ ~ ~ ~ ~ oo d d d "' ~ d ~ ~ ,'~ v~, ~ ~ ~ ~ N ~ U ~ ~ ~ U
~o; 2r o~o~~ca\.~°~~C~C~C~C~~~~dM~~Cd N c~ ,~ py0 M ~' 01 ~t ~ ~ ~ l~ N ~ ~ ~ ~ ~ ~ 01 ~ N N M ..~ D1 ~ O M l~ ~ ~ M ~
M 'p M N .~ ~ v7 ~ O O ~ ,~ ~O O~ O 01 M N M D1 O l~ .~ N 00 ~ N O
lp M M ~ M O ~ ~ ~ p ~ V'1 ~ l~ M ~ ~ 00 V~ V7 V7 00 ~ O ~_ l~ O O ~t d_' ~d~ddd°w~w~d~Zd~~~~~C~C~C~v ~~~dd~~

~W p" t~ cYi oo v-i o wi ri o~ ~_n _" ~t o ~ _., ~ ~ t~ ~ ~o °M ,~ O ~ ~ ~ ~ m ~ N O ~ l~ O N ~ N N ~ N
~n v~ ~_ ~-~ .--mp N O .~ O o0 M ° 00 O O
~ o ° ° 'r' '~, o o ° ~ ° ~ °~,°
° ~ U ~ o ~ o ° o o ~n ,r o ~ ° o ~
NU~~ooUo~~o~o~'.~°d~d~oN~d°~~~d~o N d "; ~ d ~ d ~ N ~ N o N ~ a ~ U ~ ~ ~ o ~ M d ~ c~i N ~ ~ ~
01 ~ ~ N ~ ~O Ov N ~ p N
~p O t~ O ~ N ~O v~ O O ,~ ° ~ O ~ ° r; O ° N O °
~ ~ M t~ ° O ~
O N O O v~ M ~ N ° O r-' O .~ O ~n O M O O ~ ~ O O ~ ° p ~
O
0 o U .~ ov M ,-r U U r, U M o U oo U U oo ° U U o ,~ ~l o, U o0 U~~doN~~dd°da~Ud~ddN~_dd~ddNd~N
d ~ ° o ~ ~ ~ ~ M o N d ~ d N ~ M ~ ~ ~ ~ ~ d ~i ~ ~ ~ ~
N ~n ~ ~ O O ~ ~ ~ ~ Mp ~ 0 ~ ~ t~ N ~ "~ p O ~ N ~ N
\p ~ O ~ O ° N O O N O ~ O o0 O O ~ ° r' N N ,~, ~. ° O O
O M O
~O O O v~ ° ° O O O ~p O ~1 O l~ O M .~ O O ~ O O ,_, ° O
O O O O
w ~ U U .-.a U U o U o U ~ U o U ~'''' a N U U ~...a U
Nxd~°ddddd~d~d~d~~d~od~°~~dddd 00 ~ ~ .~ O ~ N M d ~ d ° N N d N l~ N ~ l~ ~ ~ p ~ 01 M \O \O
tn M .~ 00 ~p 01 ~ .~ V7 O 00 01 01 M ~ I~ M N ~ ~ ~ V7 Q~ ~ M
,~ t~ M a\ ~ op oo ~ .-, .~ ~p O ,-; ~ O ~ ~ WO O O M ,-, v~ ~ oo vp N ~O
N N D1 ~ ~ M O ~ ~ ~ ~ ~ l~ ~ 00 ,~ Q\ N ~ M N M ~ l~ O l~ M d1 .~ O N O ~ ~p ° N O .-~ ~ O ~ O O O ~ M '~t ° ~ °
° N O O N .~ M ~
O O O O ~ O ° O O O ~ ° oo O O O N O O ° O O ~ N O O
~ ,-~ O
~~~~~~~~~~~~x~d~~d~d~da~~~~d~~
l~o M~

N

N o ooUU~ ,_, I~ N

dd ~

d ~ 'G o~d~o UM ~
~

'o M o d N ~ ~ W n ~ d ~ H ~

00 N ~ ~
v~ ,d' a1 d ~
mn oo O

o0 0 O ~ O O N N '-' O

M O
~ o o o .~ oo'b~n c~ ~
U ~ o o U U ~ Q M ~ ono a, ~
,n o0 ddd0 ~

~ 0 N N ~
d os ~ o o 0 N ~ o ~ ,-...
~ ~ ~

; ' ~ v~ r~~c~id ~~o~

r l~ O~ ~ p ~
'-' N ~O N p l~ O t~ 0 ~ ~
o O O O O ~ b '~N o0 ~ '-' ~ ~ ~ O
l~ M O ~

o U U U U
~ ,-, _ _ oddddtno -d ~ d ~y dddE-~

Q~ O O ~ o ~

(M ~ I\ M 00~ ~ p V7 ~
~t M

,~M ~ 00 ~
~D ~

O ~ O

O O O O O M ~ O M ~ ~ O ~ O ~

N ~ d d d d ddddd~ ~.. d ry! d a ~ON ~ 010000.~~ .-.~ ,~~ N M ,~~'N ~' N ~nQ\O~.--m0N M ,~.~N I~I~vwn 01 M N M M M M ~D~ M ~ M ~!1N 00~ N ~ M
i i i i ~ i i i i i ~ ~ i i i i i ~ i v7v~~nv~v~~nv~~n~nv~~n~n~nv~~n~n~n~nv~

N ooI~~n~ ~ I~O l~. l~O o0OW~ O o0O O

M N Ov~ ~ 00t~O ~f00.~O 01O v7~DM ~ 00 M N N M M M ~ ~'M M M V W 00~ N ~ M \O
i i ~ i i i i ~ i i i t ~ ~ i i ~ i i i N oo~ M ~nM O M ~n~ l~M N N oo~DO M N

O ~ N M M ~ l~0000N O v~a1I~~ ~n~ ~nN

~hM ~ ~mn t~N ~OO ~ ~ ~ ~ M ooN M ~ O

O O d wta1~ ~n~Ov~v7~O01N ~DO M N o0 V7M ~Ol~t~00l~M ~ON v7~ I~O ~ ~ l~v7O

~1v~01a101t~a101I~I~00O~00l~d'l~D\Q\Q\

O ~ N M

vDl~ooO1O ~ N M ~ ~WO l~00D\O O O O O

0000000oOv~ Q1D\~ Q\Ov01O~O~,~~ ,~~ ~, t~r'Q101~nN .-r~Oo0,~N ~'~ 00O O ~n~''~M
'-"'~D~tM o0 ~O~ 00~--~M ~ON O o0~n~p~ w rte!~1C7a ~ ~
O O'~ ~1W A w C7a ~ ~

d d d alW C~a~ x1r~

w w w w w w w rw~,w '-'~'~ ~ ~ ~ ~
~ ~ ~ ~ ~ ~

x x x x x x x x x x x x x x x m M N
N~ l~~ ~~ OM NO N ~~~ ~Oo~O ~N
00 ° 00 N 00 M M I~ M ~ ~ '~ O M ~ I~ ~ ~ N l~
O~ ~ ~ ~ ~ ~ O ~ M p ~ Q ~ ~ M V M M_ ~ '~ M ~ '~ 00 N ~ ~ ~' ~. ~ ~' ~ ~ ~ r-~ ~ OMO i--i p d OM '~ ~ (M 00 0o M 3 ~ d ~ d ~ ~ °
N M .., d ~ ~; z d a; ~ ~ ~r o ~ ~ ~ d ~ ° ~ r-' ~ d N m M N ~ -d ~ M oo ~ O ~ ~ ~ v'i 0_o O_ ~ oo ~ O d N ~ ~ p~ .~ N ,-. ~, oo ~ ~n M
°°°~°Ma°°~°-d~"d c'~.' '-'°o ~ i°~°~° ~~ oo~o ~~d~~~xN~~~~~~~ ~U~ ~zd~~d~
d ~,~~ d ~ ~ ~ o M ~ O d1 M ~ ° .~ OyD o0 I~ ~ M ~ ~ ~ ° ~ N o0 00 ~ N ~ I~ 01 v7 M o0 M ~ ~ V7 N ~ M 01 N N ~ ~ l~ M I~ ~ 00 l~
vp ~ ~ 0 N O~ ~n WO oo N M ,b ,~ Ov M ~ ~O O ° I~ ~ ~~ N d1 01 vWn ~ O N o0 01 ~ r3 O ~ ~ M ~ ~ 41 ° ~ O N
~a~~N~~x~3~~~ ~~~ ~d~~z~~~z ~
p ° ~ D o0 ~ a\ ~ O O Ov ~ N p O v0 N M ~' M N oo ~ .--~ Ov oo ~ d ~D
I~ N ~O ~ V ~ M oo ~n Q1 N oo ° Vj O l~ oo ~ ~ O ~ O ~~ O O ~O ~ N
° N p~ 01 O l~ ~ ~ ~ M O ~ 00 ~p I~ U 00 M ~ ~ pp 00 l~ d' ~ ~ O ~ ~ 00 M ~ p~ ~ l~ ~ \O ~ O~ M N O ~ ~ 00 N O 00 ~ ° 00 M ~ ~ 00 M Q1 pp O ~.,~ op .-r ,~ o0 ~ d' O a1 O O N ~ d N ~~ M N ° N oo l~ ~n N 01 ~O M ~
~d~~~ddxdddd~~~~~dddddd~~dddzd 00 ~-wo v0 W O N
V7 M 01 ~ ~ \O N Q~
M V7 V7 M N ~ M M
i ~ ~ ~ i i i ~ N ~ ~ N ~ ~
d' l~ N N N N o0 l~
,~ 00 ~ 00 V 1 M I~
M V7 N M N ~ M M
i i i i I~ ~ V'1 V7 ~ 00 M
00 O\ M V7 ~ O V7 .
t~ ~ V7 M ~O 00 M l~
N d' N t~ N N M O
~ ~O V1 ,~ l~ l~ M
00 Q1 I~ 00 00 ~O ~ 41 V7 ~D l~ 00 O~ O ~ N
O O O O O .--w ,.-~ .
U P~.~ U W
x xx x xx 00 ,~ ".~ ~ ~; ~i N vD ~ I~ of 00 ~ '~ ~ ~ ~ ''~ ~ M N
~p~O~~~ NNMM ~O~O,~j~~~~~°Od°'00 ~ N N ~ d ~° os o U ~ U o U o U '~' U U ° U
r,Md~~.,, ~ o~~:o", E''c.,~d dUdUdNddUd ~ a1 d ~ o d oo M ~'' '-' ~" ~ '° 00 0 ~ _'~ d ,~ t~ ~ t~ d vi x ,~ d~N~~N
fx ~ ,~-, E""i ~ M l~ ,~~ d ~ ~ N ~ M ~ M OMO ~ O ,-; ~ O
00 00 d Ov N l~ M O ~ N M M ~ p ~ ~O O O O ~ M O O O
M ~ ~ ~ oo ~ ~ ~ ~ ~ M ~ "' ~; N o o ~ U ~ U o o ~ U U °°
~0-1 ~~~~~°,,~~~~No ~ 3~~0~0~o~~~dadU~~dd~d N ~O ~--i cd 00 ~n ~
Z d ~ N o U d d ~ oo Gs! ,'~ o, ~~, ~ d ~ d ~ vM-, ~ ~ ono oMo 'a E-~ d ~ o; ~ ~ ~_ ~ ~ °~ ° ~ ~ o ~ o' oN, ~ ~ ~ '~ ~ r' ~ o ~
N ~ ~
~O vp ~ O N l~ ~ oN0 N yD ~ N O O O O O ~ ~ O O O O O O
x~~~o°,v~~,Q'°~d~~Mp..,'~~o~~ddodNd~dddd d o H _ x N d 01 ~ ~ d' ~ N ~ ~ ~n ~ E"' ~ ~ N '~ M d l~ O_W ~ M M ~j M o0 i oo V o0 N ~ twn ~ v~ M ~ ~ N ~ ~ v~ ~ ~ N oMO ~ O
N O O ~ ~ 01 O ~ 00 01 ~ '-' ~O a\ '~ N ~' l~ v> ~ M v> ~n ~ ~ ~n p~ N ~n M N ~ ~ ~ ~ O ON ~' ~D O ~ ~ N ~ ~ ~ O O O O ~ O O O N O ~ O O
~ av M oo U U U U ~ U U U o U a U U
~~~w~~a;HddddNddd~dddd N
N
N
M
M
N
d' l~ 00 ~''~ ~D ~ N M N ~j 00 M ~

M rr M 01 O ~ 00 ~ .-~ Ul D\ ~
V7 M O ~O \O ~' ~ ~ 00 p ~t ~n .~ ~ ~ ~ N O
O ~ ,-, tn ~--' pp O v~ O O ~
N Z Q
~ ~
~ ~
Q

i U ,~,~ U -1 U ~ ~ o U
~ ,~
' U ~
~ d ~ dd~
~

d d d o;
d d ~ N ~ ~
O d ~

a1 O M ~
N ~ ~

~ N ~ d ~ O ~ ~ N ~ O ~ v~
~

~

N O O ~ ~ M

o ~ U a ~ a ~ a; ~ ~ .-7 ~ U ~
U ~ U
ddd d ddd~ndd M ~l oo~N

~ ~

~~ No~~~
~
N

o , o N , n~ oo ~ o o ~ ~1 N ~ ~ ~ V7 o0 ~. O p N ~ ~ d M .
n O ~ Z ~ ~ O O O O O
M p N ~ M Q O O

~"'~ ' b .d ~UUUUUU
'''p , ~ ~ ~ Mdddddd N~~~Nd~ ~

~

~n d' ~n N V~ p~00 00 I~. v~ O l~ N .-.~
~ O ~ N '"'N ~n a1 ~

v7 O M o0 ~p ~ d1 pp O 01 N I~ O ~ N O~
M O ~ ~n N 01M ~ d' l~ ~n ; N O 01 ~t l~ O M

~ ~ N v~ ~ ~ ~ 00 ~
~ ~ v~ ~ ~p .~ N t~ M 01 N ~
v7 ~n .~ M N ~h o0 N
I~ ~ 00 O O M O N O a\O M ~ V~O ~ ~ ~ ~. O O O O
O N [~ O

O O O O O O O O O O ,~O M ~ l~ O O O O O
O ' O
'b N
n U U ..~ U ~..a U ~ U ~
U U ~ ~ ~ ~ -1 ~ o U U ~q U ~ U
~ o ~
~

dd d d d d dddd ddd wdd t~'~Yo0 I~~ ~D~ ~ N ~ O ~1~O

I~M ~ 41N O ~ I~00l~I~N t~N

M V1~!1N M ~ ~ON ~--~~ ~ M V7 i ~ ~ i i i ~ i ~ i i N ~ N ~ ~ ~ ~ N ~ ~ ~ N
.~~ ~ .-~.--~~ ~ ~ .~

M O ~ M t~N O I~00O ~D.~N

~ON O m .~ooO ~O~O~n~ O ~O

~ M v~~ N N ~n~D.N ~ ~n.~M ~n i i i ~ ~ i i ~ ~ i ~ i i i O ~OQ1~ ~ I~N O M l~~ I~N M

t~00lCtO -yM N M ~ 00M 0OM

V7M ~ ~ ~ O~M M M V7N N O ~!1 O ~ O 00~ M d1l~V7~OM N I~M

l~0000~nI~01I~~ ~ M (~00N

~ 00I~010100I~000100Q1l~01Q1 ~ ~n~DI~00Q1O ~ N M ~ v1~Ol~

.~.~~ .-~.~.~N N N N N N N N
~ ~ .~.~~ ~ ~ .~,~.~~ ~ ~ ~

N ~ M ~ ~nN N N ~ N O ~ v~O

w oo~ N ,-.~000 0 '~oo~ ~

N W
~~ f~ ~ c~~

O

x x x x x x x x x x ~

b ~ ~

ri ~'' ~ ~ ,-~ M ~ ~--~ p a; -~ ' ~ _--~

_~ O M ~ O ~ v~ d' ~ _~ ~j ~ rj ~ ~-' N
~
N

M O N ~ ~ O vW 0 '"'_' ~ Ov O ~ 'n M ~ O ~n ~ M N O O O O M N
O O O O
~ ~ O N N ~ M ~ O O O O O vp O ~
O O

,~ U o M av d N o M ~ ~ ~ a U U U c~.~ o a o Nddo ~~ ,-.. U U
~M
d d d d d p ~~ ~o N x d d U
U
d ..~ ~~ ~~ '-'d'n~~~~ o ~ o~osd~v;
,'o~ --. ~~M
.o ""' ~ wj wj p M '-' N o0 p N ~ f~j M p ~ ~ O ~ O ~ l~
N

O\O ~_~ M~f' ~M1~00~'~Q10O~O01MV7 ~ O
~
~

O O M ~ ~ ~ ~
O O
a1 N ~ ~n O ,~ O O O
~''~
~xa~~~~~d~d~~N~odd O ~ i M M O ~ M ~n ~ v'i O ~ ~ U ~ N I~ ~ I~

_ v O O
d ~
~
~
~
M

~ M ~ ~ 01 M ~
~ ~ M M ~ ~
~
N ' ~
N

V7 ~ O Q~ d' M
~ ~

~ ~ ~ 0O ~
U U ~ ~' '-' ~ 01 M
~ O ~ 0 O
~ ~ N ~ ~ .~
o a U
~.
U o ~ddN ~ U
~d U
o o ddt~d~
~~~d~d ~~~N

0 N ~~ _ I~ ~ ~~~
N ~ c~ ~"~ ~-~j a1 ~ d 01 V~ O p O M
M ~O ~ ~O ~ 00 p O N N ~ ~ N _ ~ p O ~ M ~ p yO ~ vW O l~ oM0 N M oho O ~n ~ ~ 00 m ~ ~D O M O ~
~p ~ V~ V~

p ~
O ~
O O O 0 ~n ~ ~ 01 .~ N ,~
O O N N M .~
O O ~ .--. N M ~ N O o0 0o O ~ M N M
O O O N

Wit' M O
O O O

O ~ pp O ~ ~
p O ~~~ ,_..., ~ p ~ O O O O
~
H
~~

ddd ddx dd x dd dd dddddd M I~ lD M
01Q\ ~n N

~nvm n ~n O~M N d\
I~oo ~ O

M l~ 01 O
O ~ I~ N

M N l~ N
M

N N M M

~'N M op x ms 00 ~t v_~ N vs ~p v~ ~ ~ r; ~ t~ t~ t~ ~_t O 00 ~ N W '-' ~ ~ ~-N~ M ~O ~ O M V~ °~ ~n O ~O M
v~ O oo N N ~''~ ~ O ~~ ~n ° V7 I~ v~ d' 00 in M ~ O~ I~
O '~ ~ O o~p o O r N o N o0 p o M ,~ O p O M ° ~ N O O O ~ O
O O ppp ~~ O ~ O p~ N ° O ~ ~ ~O ° O O d' O O ° O
° .-- .-- O p O O
U U o U o U ~ N ,~ a o ° '-' ° °p o U U U U M M U U
U ~ U
ddo~dod°;Nddooo~~°Uddd~doo~ddNd U ., ~ U U a U ., ..-a a ~ o d N ~° d ~ N M ~ ~ d d d M ~ d d ° ~ ~ o °~° d d ~ c°\~W
M (~ l~ ~ [W M N ~ 00 ~ M N 00 ~ ~ ~ ~ '~ ° d1 M ~ d1 ~ O1 ~
O O~ ~ V1 .~ ~ l~ O~ ~ ,-, d. M ~ ~ V~ O O ~O ~ 01 ~ 00 ~ ,~ M
~ ° O d' O ~ ~ O ° N ~ o ° O I~ N ° ° O O
° a1 0o O O O O O
,~ ~ U a ° U o ,~ U ° N °° U ~~.., o ° ,~
,...~,..~ U U ~ ~ ,~ a U a U
~daddU~Ur~~"~doaNd~UUd~~ddNa~dddd o'Y'o~d~°d°d~N~~~~~°~dMONO~~N~~N~~~°
vp ~ ~ O D ~ p ~ ~ p D '-' ~ v~ ~ O N ON1 o M O p ~ O O p O O O ~
O O ~ O O M p N N ° O ~ O N O O ~ I~ o O O O O O O O O O O O
a a ° U U oo U ~ ~ U ~ o ~~ a w o ~' U ,-a U U U U U U U U
ddUddNdN~~dN~-lddd~~ddd~ddddddd~
O w d ,-i O v~ .~ pp ~~ ~p l~ ~O N ~ 00 ~D ~ ~' I~ ~ ~O ~ O O ~j a1 V1 N N
O N ~ d1 00 ,~ ~ M M d1 ,~ ~~ ~D ~ O ~-W ~ ~ ~n ~ ~O N V1 ~p O ,~ 00 N
M O N .-r ,~ V~ M ~ 00 M Q1 ~ l~ ~' ~' N ~ O 01 ~ O ~ O O M ~p N M 00 l~
OOMOOOMO~NN°OMOMO~OOMNOOOOMONNO
O O ~ O O O p p O O O O ~ O ~ O ~ O O O O O O O O O O ° O O
U U ~ U U U a w U a a a.,,..~ U a ~ U U ~1 U U U ~.-a U a ,...a U
ddNdddddddd~~dd~Ndd~d~dddddddd "" ~ ~r o ~ ~~ t~ .-; os ~ j .~ M
--' ,-, .-., ~
r ~
p1 O ~ ~ O

O O p ~ ~ ~ N M N ~ 0~1 O N ~ ~ ~
, d' O p p ~p ~ M pp V~ M ~O 00 ~O ~' O N
O ,-, v0 V
~

dddd ~ ~~ ~'''dN
~U~ddNdN

~ ~ ~ d d ~ ~o d ~ N
,; O d O ~ v ~ ~ d d oo ~
a _ _ j O ~ M _ v7 ~ b O~ t~ M o0 ~ N '-' vD O
o ~ ~ O v'i WO

~ N p o v7 p N M ~ l~ ,~ M ~ I~ N ,-. p ~ N v~
l~ O v7 ~ a1 d' D O O
O p p N O M ~
0 0 o U .. ~ ~ ~ ~n O ~ N O D ~' O O l~
U O O
U a, N
aU

Uo U o . M .~
d o Uo,UUo d~ ~~~r'~~~~
d ~
~dd .~ d ~ ~ d d dd U d ~

O ~ ~ N ~ ~p ~ ~Y Ov ~ '~ (V p ~O ~ v'i 01 ,-; ~,,~ ,-;
~ ~p M
d o N W .~ ~.,~ ~, O
~' p~ 0 ~ ~ p~ N M v0 M ~ N ~O
n ,'~ o oo ~ ~ ~ ~ ~
d ~ a, ~ o o N '~ ~ ~ o ~ ~ o ~

M O ~ d _ ~ ~ ~
~ ~ l~ 00 V~
O 0 p O O O ~ ~ ~
O p U U U M ~
U O O O
U a.,r~., N ~ ~ ~ N
ddd~da d~ ~Z u.,~~~~d~Q~~~~d 00 ~ p~ t~ d , ~ 01 ~j o0 ~p ~ ~ 'p ~ d' ~
~ Q1 Ov D t~~ N M N V7 d Q\
' O d' t~ N ~ p ~p p~ ~ ~ 00 M ~ O t~ d 01 v~ ~ ~ ~ N v1 D N
.~ O .-, N ,~j ~ O ~ M O~ p~ ~ 01 M ~ ~O I~
p~ 00 ~n M O ~n ~

M ~ p~ p ~ O N ppoo v~ ~ .~ I~ p N I~ M ~ ~ I~
~ .-. ~ ~ .-~v> O ~ N
O O M ~ M "'' p~
~'' O N
o0 O
~ O O O O O

O N ~ M N p N
O O p p p ~, N
O ~ v~
.-~ ~ O p O O O O O O
~ ~ ~ ..-~ ~

dddN dd dd d ~ d d d~d~d~ddddddd dd M .--~~ V7 V~M M

M O l~O M v7~
M M ~ ~ ~ M M
i ~ i i i i i N ~ N N ~

O~I~ N ~ ~ 01a1 ~ 00 ~O~ N M M
M N ~ M ~ M M
i ~ ~ i i i i O v~ ~nV' l~N I~

N O N M I~

0100 'ctO Q1~ t~

I~I~ W O M N ~D

N N M M I~00N

~nv~ v~a\ Two v~

N M ~ ~ ~Ol~00 M M M M M M M

H ~ a ~ ~ v v x x x x x x x ,~ c~ wi Q\ o ri ~ "~ p r1 Qv (~ N N o0 _" 00 ~ ~ ~ ~ N N oo O a1 N ~ O O O ~ ~ ~' o O N ~ ~ ~ ~ .~ ~ v~ ~ ~ O O
p O O O ~ O ~ '-' O ~ p ~ ~ v~ ~ 01 ~ ,-~ o0 M
ooUUUUoa'~°oU o~~o~~~~~~,, °N
~NddddUd,~Ud ~ ~ od~d~~°~~d M~~~Mn~~d~~dooo~ dv°od~-!~ o~ dd OO~OONNO~~OO~o ~ ~~N~M~ON~~O
OOpO00~O~~OQ1 ~~o~OMr~- o~0~~NN~00 dd~~d~adaNdb ~M'~' °~ °d~~d~o°°dd ~~~~~od~o~°M_° ~ °~o~~~~°do;~ddd°N
N O~ M ~ oNO N ~ N o0 N ~n O O ~ ~ ~ M ~ ~ O O N ~ ~ O~ oho V7 l~ 00 pp N M .~ ~ l~ 41 M V) O
O ~ O O o0 O ~p O O ,~ vp ,n I~ N v0 t~ N ~ O ~ O
O O O O O O O O O O .~ ~ .b O N ~O ~ ~ N v~ O ~ ~ O O O
U UoU~U
d~~d~dWddd~N
,~ ~ o; d .~ o ~ r; ~ ~ ~ ~ r; oo " v~ ~ ~; o M O O N ~~ V7 l~ l~ \p 00 V1 ~ .-~ ~ 00 M (~ l~ ~ V7 N d .-r M
~ 01 V~ Q1 V~ ~ l~ ~O ~ M V7 ~ ~ '~ Q1 ~D ~O ~ ~ ,_, p O O O O O ~ O O p~1 O O ~ ~ ~ N ~ ~ ~ ~ ~' ~D N ~ ~ p O
O O '~ O O O ~ O O O O O ~ O ~ ~ ~ ~ ~ ~ ~ ~ ~ ''-' ~ O O
~lU°UUUo'"oUU~IUU "' U o 't ~~ ~oNO~UU
dddddd~ddddd ~dH~~~~dd~dN~dd l~ l~ N v~ 00 N O M N o0 N
~ I~ M N
~ i N
M
M M 00 ~ l~
.-r 01 01 O l~ N

~ i i i i M t~ 00 d' 00 I~ O ~ I~ V~
.--~ oo ~O N oo N
01 M ~ ~ M I~
I~ N N t~ M
Ov O ~ N M
M
~p O O v7 O
~ v~ N ~n o0 U~zxzr~
o~o~o~o~oo xxxxxx mg ~t t~ .~ oo _'~ M vo ,-: pp .~ N o ~ cri o of ~
001 ~ ~ O '~ ~ N O Q\ N ~ ~ M O O ~ ~ ~ ~rj _ ~ 00 00 N O ~ O O ,-, ~ o O O d' ~ O ~ M O M 0 ~ ~ ~ M O O ~n tI~ ~. O ~ N V~ O O O O l~ p ,-, ~-' O O M ~ O O O ~ ~ O O
0 0, ,~ U U M o ~' U U ,..~ o U M ~ U a U o0 0 ~..a U o o° N U U
UN~ddoU~dd~~Udo~ddd~NVd~dUoodd d o0 00 I~ p ,-. ~' ~ ~ r, ~ ~ ~ O ~ c.j ~ N p ~t cn N~O~~~oOM_~N~,M-,NO~~N~~0~01OO~~~UOV ~
O O O o0 00 00 ~ O ~ M ~ ~ o O ~ O U M ~ p~ O O o0 ~ d O O
"., 0 0 0 0 ,~ M ~ ,--~ 0 0 0 ~ o ~ o °p °p o d ,-r ~ 0 0 0 ~r oo ~ 0 0 o, U U U o 00 o U ~l ~ o ,...a oo U U U a, U U o o, ~t U U
~ddd~oNo~dddUdNd~ddM ~~ddoNNdd ~O ~n ~ ~ p ~ N v'i v0 ~ d Vj ~ O ~j ~p 01 ~ ~n y0 <.j ~ N p oo M
M ~. rr 00 00 ~ d 00 ~ ~O ~D ,-, N N ~ t/~ Q1 O 00 ,~ pp O 00 00 ~ct~~~~M~~d'pNpN~N~p~N~NN_01 O O O O O ~ O ~ O O O O ~ p O O p p O ~ O ~ O p p ~ O .--i O O
U U U U U ~ U ~ U U U U o ,,..a U U U U ~ U N U U ,~ oo U ~ U U
dddddNdNddddUdddd~d~dadddNdidd v0 .~ o0 .~ v'i t~ l~ 01 N ~~ M N d ~j O N p~ ~~ ~ ~ ~ d O O~ Q~ M ~ N N ~i ~ Q\ ~ N o0 ~' .~ ~ .~ O M .~ pp oo N ~p ,~ v1 N ~ ~ ~ ~ Q1 ~ I~
O ~-~ N N ~ N O ~-~ O M gyp" ~ N l~ p~ V1 .~ 01 nj ~ vW n N ~ 00 O
~O ~ ~D ~ ~ 01 ~ ~ 00 00 d' ~O N M ~ W D ~O l~ ~ ~ ,~ v~ ~ ,-~ t~ v~ ~ I~ ~' O p~ O O O O M M O O O O ,-, M O O ~ O O ~ O ,-, M M O O O ~ O O
O p O O O O O O O O O O pp .-~ O O ,-~ O O ~ O O O O O O ~ O O
U ~ U U U U ,...a ,..a U U U U N a U U U U ~ U ~ ~ U U ~ U U
dddddddddddd~7ddd~ddNd~~d~ddNdd ~
~
~
~

MMN'~ o ON~
0~ Nt~
01~N0 o ~ ~ ~ M N ~
~ ~
~ ~
~

~' ~h v~
~ ~ N
p N ~ t~
O O O M ,-, O O p N O O N ~ ~ O O N M O O O
O~ N

O O O O O O O ~ b O O O O O O M O p O O O .--~

-. U U U ,~ (~ U ~ U U U U U ~ o o U U a ~ U U
a, NddddddooddddddpQdddd~dd~ ~

l~ l~ l~ op ~..~j v~j a ~ ly0 M I~ I~ ~ ~ N O O
l~ Q1 ~

l~ l~ N pp N ~_ M_ p~ ~p I~ O V1 ~O M ti' ~ Q1 ~Y N M
~ ~ ' O
~
~
~ ~ ~ ~ ~ ~
~
ue ct ~n ,--, V
~n ~ d d r1 N vp O O
' O O O M N O O O M O O M M ~ ~ O O O O ~ O O O O

OOOOOOOOOOOOOOO~
N

p p N N d ~ ~ a Q ~ ~ ~ d W O I~ ~ .~ ~p ~.~j W t~ 00 ~p ,-- M ~ p1 I~ N
N O N l~ O o0 M
N r"' ~ ~ v~ ~ D1 ~ ~ 00 ~ 01 ~O o0 M o0 N l~ I~
N o0 v~ D1 M O O N M '~ o0 0o O .-~ ~ N ~ ~f ~ ~n M O ~' d' M M
N l~ ~ M M O N ~ ~--~ p ~ N ~n .~ ~ v~ O N N o0 ~ l~ O

O O O N M O O ~ M p O O O .-~ O O ~ N O l~ M O
O O

O O O O ~ O O O O O O O O r. O O O O O O O O O
O

N~~~~~~a~~~~ad~d~~aa~~~~~

~~o~~~~~o~~~~~o~M~~~~~~~~~ ~

O ~n l~ N N t~ O t~ ~'~ .-~ .~ ~ ~ ~O N v~ ~n l~
~p v0 M ~ t~ ~n t~

~ O v~ ,~ p1 N ~p o0 ,~ r, O M ~ 00 ~D I~ O ~ 01 ~n O N l~ 00 01 01 WO ~O V7 ~O V7 l~ ,~ ~ O O ~O .~ ~t Q1 Q1 D1 l~
~ 01 ~ N .~ N ~t o0 O O O O O O O M O O O O N O ~ ~ ~ O ~ O O M M O O
O O
p O O O O O O O p O O O O O O O O O O O O O p .~
O O

dddddddd d dddddddddddddd d ~ ~Ol~N

N N d'~

M I~V7I~
M ~ ~ O

N N d' i i i N 01~ N

~O00M ~
I~O 00~

O l~l~Q\

N ~
~t'~'~d' M p1 M [~N M
a H

x x x x lso ~_ M p ~ O ~ ~ ~ ~ o~o M ~ O N '~ N ~ °N
°°~o~~.oMN~~~ x~x~
U O .~ r.. ~ M N Uj 01 01 0 MO N oo ~ ~ ~ N ~ O ~ N ~ ~ Ova M
M O ° ~ M ~ ° ~' ~ ~ 00 ~ M '~ N ~ ~ d' 00 M O M ° ~ ~ ~ ~' M 00 M N
dM' O v7 ~ N O ~ U o0 ~ N o0 I~ x N R~'' .-. ° ON1 O O .~ ~ N N ~-~ N
v; ~ ~ U ~ U o M ~ ~ ~ ~ ~ ~ °N° ~ M ~
M O Q\ b ~ O
~ o d U ~ ~ z p,' ~ ~ ~° ~ o ~n ~ o ~ ~t M ~ ~ ~ ~ o ~ o ~, ~ N N ~ ~ ~ ° x x ° H
a ~~o~~~M~~~~M~~x~~z~~,~~o M ~ M M O N ° o O O M O~ O ~' ~O 01 t~ .-r ~ N ~ ~ N M ~--r O .~ O ~ O O O1 O ~ M M N O
'b U ° U M ~l o ~ ~ ~n ~ oo ~ oo N ~ o ~d~x~a~~~~~d~3zNxx~°~~~o oN,~~~N°~o~oMO°o°~°~~~~z~~xd~M
M y0 N N N N pp ~ N ° ~ ~ W_ p ~ M N N ~ ~ O ~ Q_v M
° ~ ~ OI~1 M ~ ~ ~ M ~ ~ 01 O ~ M l~ l~ ~ 01 ~ 01 O M M ,., O ° ° O ° M M I~ ~ O 00 ~ M N ° N ~
~O M
~~d d~~~~~~~~~~~~~~d~~~~3~~~~
O O N l~ M ~O ~O ~ ~O O
M 01 v7 l~ ,~ M ~ ~O l~ 01 Wit' N N ~-~ M N M N N ~t ~n ~n ~n ~n v~ v~ ~n v~ ~n ~n ~O ~O o0 M 01 N N O N
.~ l~ M ~ O~ N M V~ ~ l~
~ N N ~ N N M N N d' i ~ ~ i i i i i i l~ N ~ ~ l~ ~ N ~~ N
O~ M OO ~D O ~ ~ ~O <t M
N ~D I~ l~ ~ O ~ 01 01 O N O V~ ~O ~-~ t~ 00 ~ ~n ~n .~ ~n ~ N O 00 ~O
N
O~ O ~~ N M ~ ~ ~O l~ 00 N
N t~ ~ N M l~ 'p N O1 y~ O ~p O ~ 00 ,_, M M N
ca A
000 000~~
xxx xxxxx x x a;
~,.~ ~o W ,--.
M ~ 01 I~ O 01 01 ~ ~ ~ ~ O~
00 v1 ~t O N
M ~ ~ ~ ~ ~ ~ vp ~
d .~ M a~ a, a' M ~ ~ ~ p O ~ ~ Np ~ M M O O ~ 01 ~ v0 b O ~p ~ ~ O p~ ~ M a1 ~ O w0 M
O p~ ~ 01 ~_ ~" O oo M
00 O ~ ,~~, O ~ 'b ~ M M ~ b ~ ~ ~ M b fWp O O pip d ~ ~n ~ ~ ~ ~t N l~ ~ ~ ~ ~ ~j p d' 00 ~ oo v'i ,~ ~i ~ ~ N M d ~ ~ O
r'' ~ N M ~ ~ N O M ~ ~ M 00 M M
00 l~ O N O ~ ~ ~ M ~ l~ ~O ~ ~1 O~
O O ~ M N ~ ~ 01 ~ ~ 01 01 O~ 01 01 01 Q\ N ~ 01 d 00 I~ ~ ~ M N M I~ ~ ~ l~ l~ I~ I~ l~ l~ I~
U1 '~ ~ O ~1 ~ M M ~ M M M M M M M p ~~d~ddd~ d ~ ~ ~NQ_~O
d' v~ l~ ~ ~' M op ~ l~ N N ~ v~ 01 OWE M M v7 v1 O l~
00 00 .~ 00 00 ~ ~ M M N ~O N ~O ~--~ a\ N ~ ~t o0 O ~ v~ ~ 01 p~ O I~
O oo l~ M ~ '~' O ~ O v1 N ~ ~O o0 l~ ~n 01 M l~ l~ I~ 00 N M 01 I~ O o0 M 00 00 ~ N O O~ O ~ 00 O~ 01 01 O~ O1 Q~ ~ O~ 01 d1 Q1 O~ ~ M O
M ~ 00 ~ N 'O O I~ 00 M O t~ l~ t~ I~ I~ I~ l~ l~ l~ I~ I~ I~ C~ M o0 N M
\O ~~ 00 M M .~ O M M M M M M M M M M M M M .-, ~~ddddd~~dd~~~ ~~~d~~~~~dHwd~~
01 OW O N O oo ~ ~ ~ ~n N ~D t~ O
~n O M O \O ~ O1 N 01 ~ ~ M M 01 M M M M M M N M N ~' ~ ~ M N
i i i ~ ~ i ~ ~ ~ ~ i ~ i ~
V'1 ~ V~ V7 V7 ~ N V~ V~ V~ tn ~
v7 v7 N oo ~O ~ l~ O O ~ N M \O
O~ N o0 d' M I~ ~ 00 O o0 N N t~
M N M N M M N M N ~t Q1 ~ M N
i i ~ ~ ~ ~ i i ~ ~ i ~ i i M N M O 00 N M V7 ~ O O l~ 00 ~
l~ ~ ~~ N O O O O I~ O ~O Q\ ~O ~O
00 00 00 00 00 00 00 \O 00 01 l~ M O~
O 00 00 00 00 00 00 00 00 00 00 00 00 ' ~O
~O O O O O O O O ~ O O O O O
In 1n V1 V7 N ~ N 01 ~ V~ V~ V7 ~
C1 O ~-~ N M d' ~ ~D I~ 00 Q1 O ~ N
N
.-, ,~ ,-~ .--. ,--.
00 M ~ N ~O M ~ Ov ~ (~ ~O 00 O N ~O Cn ~-' N M o0 ~ ~ ~~ ~ ~n O
d ~ ~ ~°d~~~a H H E-~ E-~ E-~ H H E-~ H H H E-~ H ~
x x x xxxxxx xxxx x oho O ~
N N o0 00 M t~ ~ 00 ~ M ~ 01 00 V'1 M
O l~
M ~ .b N O ~ N ~ '~ ~ pp d' O
.ty ~ ~ M ° y.-~,-yn O~ ~p O~ M N
~ N ,~ M O ° ~ ~ d N o ~ ~ t~ ~ ~ ~ ~ M '~° ~ .~ ~
x -d °° N ~ ~ o ~ 01 ~
M M~ ~~ ~ ~~d~~ ~~~.~ x~Mo -, o; '-' O o c~ cMV M o 00 't~ M ~ ~ M M ~ ~ ~ d O N O ,~ ~-~ v7 M ~ 00 ~3~~~3 °° 3N
dQ d dd dd~dN d~Z
r; a;
v0 O N ~ ~ ~ N O N M ~ '~ vp ~ O ~ ~ O M a" ~
'"'' p p~ ~' M ~O (~ ~_ N o0 ~ (~ 00 N M O ~ V1 ~ N o0 00 00 ~ Op N M O ~ O M p M ~ O N ~ ~ O ~' ~ ~ N
d U d ~ d '~° a ~ °' oo d 'Wn d~dddddd dddd~ddZ~~ ~~d~
N ~D o0 0o O O ~ I~ ~ ~ M Q1 00 ~O ~n ~-~ O M ~O 00 M ~ l~ N 01 ~~ M O N
N ~ ~--~ M V1 M ~ M ~ ~ N ~ l~ U1 i i i i i ~ ~ i i i ~ i ~ i i ~ ~ N ~ N N ~ ~ N ~ N N
l~ 00 N ~ ~ ~O ~ O M O N ~ ~ ~t N
01 00 N V1 I~ .-~ M ~O ~~ 00 O ~--~ d1 ~~ O O
M ~~ M ~ M d' M ~ ~ N ~ ~ N
~ i i i ~ i i i ~ i i i i i 00 ~~ ~D N ~~ ~~ O 01 O Ov Ov O v> N ~ Ov O l~ l~ M I~ M N ~~ ~~ ~ O M ~ O 01 ~O
~D d' l~ ~ ~ N N M ~ ~ 00 O ~h O 00 M
~D l~ N ~D ~ l~ 01 M M O~ (~ ~n Q1 ~ 00 N N ~n ~ I~ N I~ N O N O ~O ~ ~n Ov Q\ ~ ~ ~ Q1 ~ a1 ~ ~ ~ a1 ~ a1 M ~ ~1 ~O l~ o0 01 O ~~ N M ~ ~n ~D l~ o0 I~ I~ I~ I~ I~ l~ l~ 00 00 00 00 00 00 00 00 00 rr .-mr rr ~ e-r ~--~ ~ .~ ~ ~ .~ w---~ ~
~f ~~ M Q\ ~ ~O M ~ ~ M ~~ O ~ N
~-' O t~ ~~ ~ O N O M vD Ov ~-~ N N M
~ U U C7 O O ~ ~ ~ G~-1 ~ ~1 Ga ~ddd~~d~d~~d dd~C7 ~C7 t7 x333333~~ 333 33 xxxxxxxx xxxx xx x 00 I~
~~_D~~"~NO~O
~N
M o0 ~ ~ O '"' p N p ~ O O M
'd M ~ ~ pMp O ~' O N O O O
o ~ o ~ o ~ ~ U a U O U U a rM,~~ ° Uo~~~ ~ dadUddd ~~°~o'r,d~~o~'o O ~ m ~ ~ ~ U ~ M ~ ~ M ~ t~ ~ N ~ ~ a\
U ~ 'b ~ o b 3 ~ ° U ° M ° ° ~
~b~°z a o ~ ~~ ~ d~~d~~~d~
d ~ ''~; ~ °° ~ M oNO 'M ~ ~n ~ N
N ~'M ~4loNp OIOOW~~~Do°O°
N ~ ~ ~ ~ ° ~ ° ° ~ ~ ~ ~ ° ° O O O O
°
'-' b ~ ~ '~ '° ~ ~ ~ ° ° ~ 00 3 3 a U U
~d °3 ~ ~ Uda~~ aadd~a~dd WO M H a O~ .--~ ~ 01 00 ~' pp O~ I~ l~ ~j N o0 vi ~ ~p M M M M ~ N ~ ~p ~ ~ ~ M ~'~ M N V7 V1 ~
N '~' O ~ ~ O ,~ O N ~ ~n N ,~ ~ N
d1 ~ I~ .~ O M 01 V7 V) ~D ~ .~ M ~ M ° pip ~ ~' ~D M d1 Q1 ~ ,~ O O ~ ~I' N ~ l~ O ~ 00 ''~ O N O ° ,-~ ~ 00 M M N ~. ° N O ,...
~O O~ O I~ O ~ O M .-. ~ ~ ~ d' 00 p O .~ O O ,..~ O
'b UaUMw '°'° aU~ UwU
z ~a333~~ aaaxd~~~ ~~aad~aaa WD o ~ ~ ~ ~ o0 .~ v~ ~ .-. t~ V, oo I~
d' M 00 O 41 O V~ l~ l~ M .~ ~ M ~ M N
V'1 ~ 00 M d' d' N M M M
i i ~ ~ ~ ~ i i i ~ ~ i i i i V7 V7 ~ ~ N N N V'1 V~ N ~ N tn V~ In t!7 N N ~ l~ O O M ~ ~ ~ M l~ M ~ ~ M
M N ~ 00 00 O~ ~ ~ ~n N O 01 N M N
V7 M 00 M ~ ~ N N M M
i ~ i i i ~ i ~ i i ~ i i i i i I~ N oo .-r ~f' N M O o0 .~ O O O ~-~ a\ t~
N N oo ~n ~ O O N M oo M ~D O~ ~n O d' N ~O l~ N O V1 t~ l~ ~ N N ~O .~ .~ l~ ~O
00 O v~ ~O Q\ a\ ~n ~ 00 v~ 01 N O ~ 00 ~O
O l~ ~D N ~O O I~ ~ ~n oo M ~n l~ I~ ~n ~n Q\ ~ ~ D1 a1 ~ ~ ~ ~ ~ ~ Q1 ~ ~ Q\
01 O ~~ N M d' ~n ~O I~ o0 01 O ~ N M
00 O~ O~ O~ O~ ~ O~ O~ O~ a1 a1 O O O O O
.~ ,-.. ,~ ~ N N N N N
N M ~ ~ N_ 00 O M ~' ~ N O M N O
3 d ° a cM G=°, ~ H d °° O O w >
xxxx~xx~' xxaa a as ~a x3~~xx~ xx~x ~3 ~3~ 33 xx x x x xx N~ ~~ ~ ~~~~''~ ~d'~~~~p ~O O O O~ ~ 00 O N ~ d' d1 O
M Q1 O 00 M ~ O Q1 O M M O~ ~ M ~D (~ M
01 M ~ N M l~ O1 ~ '"" .--mp ~ M (N ~_ ~ M ~ M ~ p ~O p ~MV~d'0~~.~NM~_~ OO~ OOOOO
xN ~ d~ ~~~d ~ d~
M ~ E"a \O ~h ~ ~ ~ ~ ~ M ~ ~ ~ ~ ~' ~ ~' ~ M 00 O ~ O~O
O M ~ ~ M ~ l~ dN' N O1 ~h ~ N M ~ M D1 ~_O M ~ ~ ~_ ~ ~ ~ M
~p ~ V N p~ N O Q\ N N ~ I~ ~ r' '-' N ~ ~
U d ~ .-.
~~dd~dd~~~~°~~~~~d~~~d~d~~
d .o ~.,; ~ N ~; ° ~ ~ ~ ~_ 00 ow o t~ ~ ~n ~ ~ ~ O O ~ ~ ~ ~ ~ N ~ ~ O ~ ~ ~ ~ I~ ~ M ~
~ 00 00 ~ N N ~ ~' N h ~O ~ I~ ~ N N N N o0 ~O t~ N O
M N l~ ~ ~n O ~ ~ ~ ~ N ~ ~ M N ~!1 ~ 00 ~ I~ d' ~ p O O
N dw0 v7 M M l~ M 01 ~
~O ~aO~~~M 3 3Mp~~
dMd~~daHo z~~~~~~~~Qd~~~~
-., p vp ~ 00 ,"' ~ N N ~_ d ~ ~ ~ 00 0~0 ~ ~ M N O_1 ~ ~ I~ pip N ~O
~ O~ tn ~ ~ ~ pp ~ ~ ,~_, \O 00 ~ ~ ~ ~ O l~ ~ M ~ ~ ~ V7 O O ~ ~ p 01 ~ ~ 00 ~O ~ ~ VWO O ~ 01 ,_, I~ ~ d' ~ M 01 O
O O O M O l~ t~ 00 v~ I~ ~ ~n O O v~ ~ .~ ~ ~ N m ~n oho O ,~ O
~x~~~~~~H~~~~~~~~~~~~~d~a i i N l~ v~

i N~
N
I~ 00 ~h M
M O l~
N
O O O
N N N
~n M l~
aaa xxx 00 ~ ~ ~p ~ ~ ,--. ~ ~ ~t ~--~ t~ M ~D N ~; o ~ ~i ri ,--.
M ~ ~O N M M .~ N ~ l~ 00 " Q\ M l~ O~ ~ M 00 ~D M
N ~ ~' M ~ ~ ~D ~ ~ OM N ~_ ~ '-' O M O ~ ~ ~ ~ O
p M O p p ~ O O O ~. p O o0 N O O O p O p O O ~ p O O O p O
U ~o U U U o U N U ~ o U U '~ ~ U U U o U a ,..~ U U U U M
dUdddUdo~,dtVUddo~ddd°~dd~d~ddddN
~ ,~_, 01 d ~ '~ M 00 ~ M V7 ~ N ~ 00 ~O ~ 00 pip O 00 ~ (~ O ,~ \O ~ 01 N ,.~ M O ~ ~O M ~ ~ 00 ~ ~ ~n ~ ~D ~ O ~ O '~ M ~n l~ 00 O 01 ~D
a\ N O ~ ~O O 00 '~ l~ M ~ ,--r 00 O N ~ M M ~n ~ ~n ~n N ,~ O ~ O O M
O O O p O d' O O v~ M M O M O O O ~ O O O O N N O O O O N
O O O t~ O oo O O ~n O O ~ M O O O ~ O N O O O ,~ p O O O O
U ~ U (~ o U M U C~ o ,~ o ~ U U U N U U U ,~ ,~ U
d.~d~d~dNddUd~pNddd~d~dddd~d~~d nj t~ oo p~ N ~ 00 d N '~ ~ ~ d1 ~ 00 M ,--; M N oo p~ O O ~p d' O ~O
01 0o M d M ~ ~ oo ~n N ~ N M N oo N N ~ ~ .-mp I~ p~ ~ M ~D
t!~ ~p O~ 01 00 .~ M ~ M ~O I~ M N ~ ~ l~ ~ ~O M O ~ ~ V' 01 l0 ~ ~D V~ d' d' O v~ p~ N ~ ~ ~ O ~ l~ ~ ,~ ~ 00 ~ Q1 ~h N 01 ~ ~ l~ O ~ V~ N ~O O
N O O M O O O O O .~ O O O O O O O N O O O O O O O O O O
O O .~ ,~ O O O O O ~~-~ O ~p O O O ~ O .-. .-r O O O O O O O O O
-1 o ,~ U W o, U U oo ,~ U o U a U a U a a U U U W U U U U
d~~dd~d~d~~~dUddddddddddddddd~
o~ ~ o .~ t~ ,--. ~-~ ~ oo wi os o0 os d t~ os ri ~ ~~ v-i a\ ~t oo ~." vo a;
ri d~ o0 os 00 O ~ ~. ~O M N I~ O U ~ Q1 M 00 ~O Q\ M Q1 l~ ~ Ov ~ M
N ~ N ~ ~O l~ ~' ~,, ~ v~ d' ~_O O ~ M_ O_ ~ O ,--~ O O ,--m~ 00 ~ v~ op o0 I
M ~ O M O 01 O p O O O M O ~ N N O O O O O O O ~ O O ~ O p O
O .~ O O O O O O O O O ~ O O O O O O O O O O O O O O ,~ O
~Nd~~~~~~~~ddNd~~d~d~~~~~d~ddd ~j ri ~O vD t~ ,-i O" ~ p~ ~ ~ O O ~ ~ ~ M
M
~
~ N t!1 N l~ ~' ~ M O~ V7 pp (~ rr ~p ~

_ _ _ 01 ~; ~ 01 N_ O O ~ I~ Ov ~ ~ 01 ~M,, ~t \O ~ ~ N N
N

~

' 0 ~O
N_ OOMONM~ ~MM~N 000 O O O O O O O ~ O O ~p O ~O O O O
~
U
U

~ U ,~ U ,-~ ,..a ~..~ o ,~ a o~ ,~ ~ U U o U
o c ~
ddod~hodadNd ~o~d~dddd~o~d~~
., ~
, V~ vp l~ .~ O 00 I~ p ~' 00 ~ M ~ p ~ ~ ~, Im! O d d ~ N M

N ~ ~ ~ ~ ~ ~ O
N ~ ~ O N ~ ~ ~
M N O ~ ~ ~ ~ ~ ~ ~
O
~

~
~
~
d V~OOOr'ON ~'0000 OO~O~ OOOO,~O
N
~

O \p O O O
--~ O O p ~' O O ~ ~ O O O O ~ O O
O O N
U o U a a U,~ ~ o U U U U U U o U a N ~ U o U U U ~n ~~
~
' MOdddd do dddd M
ddodd NdUddd~

a, ; ~ t~ oo ~" M .~ ~ ~ W 7 ~ as vi ~ o ~ ~ av ~ yo 0o d os ~ -: M
-i ._ o p .
v r _ M

O O ~ N ~ ~ ~ ~ ~ ~ ~ 01 ~ d ~ vp ~ D\ ~n ~ O ~ ~ v~
pip pip O O p p O
D
~
~
O

O N O ,~
O O O b O O
O ~ O O 0 N O O
~
,~ N U U U U U U U ~ oo U ~l ~l U M U U ,~ a U ~ ~ ~
o U

d ~
daddddddd~lNdddd dd~.,dd U

~
~
~n a; p~ cri ri ri ~n ~ ~ N oo I~ ~n N vo vo p" O cri N O oo ~ ov ri N ri ' vp oo l~ ~p p~ ~D I~ M M ~ l~ N o0 v0 N ~n M ~ O N d ~n N ~ ~n ~ ~ ~ O N M ~ ~ ~ ~ O l~ O ~ 00 ~
~ ~ ~ ~ ~ ~ O
~

M
,~ I~ ~ ~D vD p~ 01 O N D1 d' o p O ~
O O ~ ~ p O
O O O ~ O O O O O O O O O
O O

~
N
p ~Nddddd~dddddddNd~dddNdd~d~dd ~n a, a1 N

a x x ls~

y0 O; N ~ ~ 000 01 ~p m O .~ y0 ~ ~p ~i' ~O ~ ~ ~ O~p M ~ ~O
M 00 ~-' O M O ~ ~ ~ M ~ N O N ~ ~ O ,_, .~ M 00 \p d' Q O ~~ O r~ ~ N ~' ~ M N p ~ ~ ~ ~ N ~ NO
00 N ~ ~ ~ ~ ~ Ov M N O pp ~ ~ ~ oho ~ x ~ ono N ~n ~ "., O ~r ,~ ,~ ~ ~ M O ~
~ M ~ p~ I~ ~ op M d ~ ~ N N o0 ~ M ~ ~ ~ ~ N
0o Qv O O O oo t~ ~ p~ l~ o0 op N Z ~ ~ ~ I~ M ~ v~ ~n 00 M ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ N ~ ~ p 0M0 ~--~ ~ ~O
v7 O1 d d d 01 ~ ~ ~ ~ N ~
°1 ~ ° ,~ ,err, ~,~; o °' " a~ ,r; ~ ~ M N d ~r ~ o ,~ p v'i pip M ~ M ~ N v~ N ~' N ~ ,~ ''.'' ~ M ~ O N M ~O ~
t~ ~p N ~p '-' O N M ~ ~ ~ M ~' ~ ~ ~' 0~0 ~ x ~ ~ ~ 00 ~ V
I~ 00 O N O O ~ 00 ~' '~ 00 ~O ~ ~ M ~ M ~ N M ~p ~ l~ ~ M V7 M N
l~ \p ~ ~ ~ O N '~ ~ r~.~ 01 ~ ~ N N ~ ~ ~O M
3 3~od~~M~~d~~Nd~~,~~~°~o~Q~H~
c~i U a\ ~ ,~ o ,--~ ~ r~ o ~t Z d oo ~ Wo c~i pp ~O l~ .-r l~ 00 .~ ~ M N N 01 M ~ \O .~ 00 ~ l~ v1 O 00 ~ l~ ~ l~ ~ O M ~ ~' ~ N M ~t ~ I~ 00 ~ M ~ ~ O d1 ~n N O 01 ~ 00 ~ ~ O ~ ~O N .~ O ~ ~ N o0 M v1 ~ O l~ v~ M N N .-~ ~O ~O N
V) r' 01 ~' ~ O ~' ~ ~ Q1 M ~ ~ ~O M 00 01 O O O M ~') O 00 O1 ~
Q, 'o d ~ ~ ~ ~ 'b pd ~ ~d Wa o ~ d ° d ~ ~ ~ ~ M ~ ~ ~
d3dd~d~d~~d~~~Hdddd~x~~w~d~~dd N
V7 ~ M
i i ~ N
O o0 I
d1 ~
v7 M N

N O ~-~

O
N N N
~_ a a x xx .-: vv ~ o °~ c~ W o 00 ~ o Q1 ~t ~ O pMp ~ ~!1 ~ ~ ~ Ov O ~ ~O ~O ~. M ~ ~ I~ N
V1 '-' O 01 ~ in d1 M ~p ~ .-i v~ ~ M N O ~n Vj ~ ~-~ ~ p N ~ IW _O ~.,.j I~
~t °pO~p_M~O~~p~ONO~~NO~MO~N~MM
,.°.a U U ~ ° U U a a~ o U ~ M o ~p-1 U x.0-1 U o'n'o U o U No a U ° a o ~ .0-1 aaaN~add~pdp~~adda~d~a~ad~ap~a v~; ~ v c~; ~ I~ pp o ~t ~ ~ ~ ~ N ~O o; Q\ vi ~--~ ~,~ c~ v-i d oo ~''~ ~ ~p ~ N M
tn O ~O ~ ~ N M 00 V1 00 ~ V7 ~ V1 V7 M d1 00 M V~ ~ M N
V~oO~~~°0~~~~001N~o~oO~°DN~~~~N01~~M0_~
M O O p O .~ ,-, ,~ O M ~ O O O O O O ° O N p '~ r., M O O
O O O p O O r., O O ~ ~ ~ ,~ O O O O O O N O O v~ O O .~ p ~ O O
,~ U U U U U U U M a ov M W U U U U ~ U U o U U ,~ ,~ ~ U W
addddd~ddad~aa~dddadddUddaaNdd 00 ~ ~ cri M I~ ,~ oo N a ~ oo d M O o0 ~ N O M N O d op N M ~n ~D O ~t ~n O O~ M ~n O N ~O t~ N ~ v~ N N O~ ~n M ~O ~ ~ ., ,~ ~t v~ O ~ (~ O
\O O ~p 00 y~ .~ 00 ~ N [w N 00 ,~ ~p \O M l~ 00 O ~ ,-, ~ N M M V1 ~ M
O v~ v~ N .~ M M .-. a\ op ~ Q1 00 ~ ~ Ov ~ N v~ O ~ ~ N M ~ M d1 l~ N l~
O O O O N N M ~ ~ ~ O O M '~ O O O O O N O O M M O M I~ O O O
O O O O .~ p O .~ O ~ O r. ~ O O .~ O O O O O O ,-, O O .~ O O O O
W U U U W ,~ a oo U ,~ o U U W U U U r~ U U o, U a a U U U
ddadd~d~dNddUddddda~dd~~addadd I~ N t~ ~ ~ os p ~n W o ~~ ~n yo WO oo t~ os o '~ Wo ,~ o wi oo v~ wi wi ~O ~O ~O ~p l~ ~ ~ ~-m~ .--, ~p ~ t~ O N ~ OWE O ~ M N o0 v~ I~ v~ O v~ ~p ~n N ~ ,--~ ~ ~ p~ ~~ O N o0 N ~ oo ~ N O M v~ ~ ~ N O ~ M M ~t O O O
l~ I~ V~ .-W~ ~O ~p 00 l~ '~h ~f M ~ ~ ~O l~ M .-~ .--~ ~ V~ ~ ~O p~ N N I~ r, I~ M
.~ O O ,~ O O p~ O O O O M M O O O N N N M O O O ~ O O O M O O
O O O ,~ O O .~ O O O O ~ ~ O O O O O O O O O O O O O O O O O
~~~d~d~~~d~d~~dddQa~d~dddd~ddd ~
~ ~ M ~ M
~

V~ O 00 ~ ~ 00 O~
N ~ W ! M
M \O O ~ ~
M ~.,~ ~ I~ M d1 M ~ '-' ~ O O~ ~--~ O N N
O M M O ~
N
~
~ ~

N O ~ 00 O O
~ N ~ O M ~D
p 01 l~ ~ M ~
N ,-, t~ ~ ~ ~ N --~
O N ~ O O ~''~ O .~ O O O O O O N O O o0 ~' ~' l~
O O M

O N M O V~ O ~ O O O O pp O O O O O O pp '- ~ M N
M O O ~1~ ~ oo U oo N a ~ U ~ U U U U .-.~ ~7 U U U U ~ ~l ~ ono O U ,~
d ~
a a d a ~...a a U d d ~ .n~ ~
d N O a ~ Q N d d ~
d d (V M ~ ~ O p\ ~j Ui O ~ O O ~ 00 ~--~ I~ N ~
~ trj p a uj ~ N O
~

00 ~O ~,, v~ ~ l~ v~ O N O ~ o0 ~ l~ ~--' ~ ~t~
~ 01 0o N N ~
~O M ~ M ~ ~ ~ M ~ N I~ ~ O I~ ~ N ~ ~ O d1 ~ ~ ~ V7 ~

O

O O ~ O O I~ l~ O O O O O p O O O M O O ~ O
~ O
O

O O ~ O ~ N ~-, N
O O O O O O O O p O O O O O O ~. O O U o o0 U U p U o U ,...a ~l U U U U U U U U ,~ U U a oo U ,..~ d ~d ~
~

ddUd~ddd U ~a dddddddaadd~7d v v7 ~ ~ ~] N '~ O M ~ O ~ t~ ~ ~D O ~ ~ O O ~'N
I~ v~ p ~
~

N M
00 00 M ~ N ~ t~ 00 ~ I~ 01 M O ~ M p O N
~ ~O ~ M ,~ d1 O O ~ p~ N ~ 01 00 N ,-, ~ O ~ ~ I~
~ O o0 O p _ l~ N l~ M O p M ~W O ~n ~ ~n N N a\ ~ .- ~ ~ ~
O N O ~
00~0 p0 y0000000000 ~~p 0.~00~ pav ~OC 1 _ a dadad ~dNd~d~d~d~~d~ddd~d~~~d p \O ~ 01 M M W N o0 .-~ 00 .-r ~D ~rj ~D N ~ M O D
O V7 ~ \O ~''~ ~ <t ~

00 Wit' v~ ~ ~ O a1 ~ ~n ~ ~ I~ ~ a1 01 N N
01 ~' 00 V~ 00 01 N pOp 01 00 N o0 ~ N N --~ N ~ M ~' ~ N ~ ~ M O O d W O ~O ~
-r V7 V7 N o0 N N
00 N \O I~ M ~
~ ~
p1 ~ 'ct ~D ~ M Q

1 Q ~p (~
O . ~ Op , -..s N
O O O O M ~ O O O O M O O M O O N O M O M N

,_, O O O O O O O O O O O .~ O p O O O O O O O N
O ,~ ~
N
~~
~~
~

ddaddd aadd ddddd dddadd d t~o M .~

N ~

O

i N N

Q1~

N M

N N

~ U

U

a x x l~ 00 t~ ~ V7 ~D N l~
Q1 wj ~!j '-r O ~ N l~ ~ M V7 ~ 01 M O 01 ~ .-~ l~
N O ~D ~ ~ N N N N Q\ N N ~ ~ ~ ~ M v0 ~ ~ ~ ~ ,~ ~O ~
O 0o ~ O~ N O ~ ,~ N O ,~ ~ p~ .-. I~
00 M '~ ~ \p O ~ ~D ~ M M '"~ O ~ D1 00 pip ~ ~ ~ i--i O ~ D1 M O
,~~.~~,~~~~d~~~~,p ~~~d d ~~~N~d~~~~~~d~~~~d~dd ~ M ~r N ~ ~ ~ o; ~ ~ d d ~ ~ ~ ~ os N vo ~ o0 O~~~O~~ON~~oNO~~~M~'NONNp'-'p~MO~Oa\~M~
N o0 Ov ~ ~ M ~n Ov oo ~ ~ O Ov ~ I~ ~ v~ ~ .~ M .-. ~ ~--~ O~ N I~ O l~
O .~ 00 00 ~ oo ~ ~ O O l~ ~ O N ~ ~n O ~ 00 p~ ~ ~p p~ ~ l~ ~n N O
t~ Q\ O O d O ~ 01 ~ ~ M 01 d d' r3 M O M d' 00 t~ M ,~ V7 Ov N
~~~d~~ ~da~~~~~~~~,~,~~, o~~dd~Mdo d~~N~o~,~a;ood~~ddr;~dddd M
~n ~ ~ ~ ~ 01 ~ ~ ~ ~n ~ ~ ~, d' N_ ~ ~_~ l~ ~_ ~' M ~_ ~O 01 00 01 ~O ~t ~O
o0 M M d~ O O a\ ~ '~ ~ ~ 01 OM N o0 ~ ~n M O ~ ~ Ov M N ~ ~ O ~
M ~ V7 M M ~ 01 ~ ~ 00 Q~ ~~ r-, .~ N p~ l0 00 p ~ ~D ~ ~ r-r O ~ 00 M 01 d' ~ pp r, l~ N ~ t~ ',~ M ~ 3 O N d' I~ .~ ~ .~ \O a ~ 01 01 (~ M
d~~~~d~~d~~~dd~d~~~~~~d~ddd~dd ~ ~i O O~ OW t ~ N ~ v-i oo a1 Ov M
O N ~O ~ ~j d' ~ ~p M ~ O OWp ~ ~' N ~D p l~ ~ (~j M ~ Q1 ~ l~ ~ ~ ~O M
O I~ ~n ,-r .-~ 00 N ~p V1 O .-~ .--~ ~ O ~ ~ ~1 M N O N Q\ N ,~ ~ M o0 N
v~ 00 00 ~ p 00 O~ ~p l~ M 00 l~ .-r N 41 ~ ~ p ~ ,-, ~ ~ M I~ M 00 V'1 N N 00 ~D I~ 00 pp N ~O N O o0 O N ~O ,~ ~ N o0 v~ N N N ~ ~ I~ ~ ~ M l~ O~ ~O O~
N O O ~ ,~ ~ ~~ v~ O ~ l~ M p~ N ~' O O M O ,~ v~ O 01 ~ O N M v~ 00 v~
dddd~dddddddddddddd~~ddddd~d~~

,~ o --~ ~ ~: o ~_ ri °° ~
00 N o0 ~ ~O M O 01 ~ ~ l~
N ~.,~ 0o O ~' ~,~ ~ O N M M M ~ N ~-~ N N ~ ~ ,~ I~ ~ M O wt O ~ ~ ~ l~ ~ N V~ p N M ~ ~ O O ~ O ~ ~ vp O ~' v~ oo ~ M l~ ono O Ov ~ ~ M d ~ ~p oo ~t ~ O N ~ 00 t~ p~ '"' ~--~ .~ ,~ M o0 ,~
2r ° d~ ~~~~~~ ~~d~~~~dd~; 33°M~
d~;do~ do, ddd °dd ~ ~NV,~Ndd~~nd " N " M ~ ~ O ,~ I~ 00 0~1~~ ~~o~O~~p~OVN~OV~N~~ON~~o~o~~~~~xM
N ~ M N ~ 01 ~~ ,~-, ~ ~ ~ d1 ~ ~ l~ M I~ N ~ 00 N M O M ~' d1 ~
O W --i ~ ~ ~ l~ ~ M Q1 N ~ .~ ~ M ~~ 00 ",~ ~O M ~
°,d.~o dd~d~~~~3d~~~~~3~3~d~,',°,~o°~°~C
~M~~~;od~~~d~d~dd~;~;~dd~_~c,NO~~~'-' 0o p ~ O Q\ ~ '-' 00 .-. ~ v'i p ~ 00 ~O '"'' ~ ,~ N .-i M N ~ M v0 O .-i ~ O ~ N M ~ N N M ~ M l~ ~n M N ~O ~D ~ ,~ ~ ~ ~ ~ O ~n 00 ~p ~ 01 O pp ~ l~ .~ ~ l~ 00 W O O O ~ ~ M o0 ,-, ~O N O ~. N 00 ~ M
O N o0 0o O ~' 00 ~ ~ p O ~ M N '~ ~ ~ l~ M N ~ O O ~ N ~ N M
M o0 ,~ pp O O M r3 O I~ ~ ~D N M N ~' ~ 00 00 W O ','~ ',~ N o0 ~O N ,~
yo vo d ~ o~o ~-, O M ~ ~ °, ~ d a ~ t~ 3 d d ~ 'r' oo ~
dN~~dd~~dddd~MdddNdd~~d~.,»d~~p 00 ~ ~D l~ ~O W O N pip M ~ ~ .W p N ~ I~ N M d' O~ M N ~ ~ Ov pOp t~ ~ d' N oo N N '-' O o0 ~.,~ O O ~ ~n OW O ~ O M ~~ 00 00 ~ ~ O 41 ~-~ O
O r'' v7 O O t~ ~ N N M ~ M o0 ~ .-, ~ pp ~-'' l~ O N M r.r M M M l~ l~ ~ M
.~ V1 O d' ~ ~ M O l~ O ~ l~ Q\ I~ M ~ M Q1 N 01 ~ ~ 01 l~ ~
M ~ l~ ~ ~ V) ~D '~ M ~ ,M_, M ~D O 00 V7 00 ~' M I~ 00 (~ 00 '" O 00 l~ ~D
r-~~ r~-irk-i~~~~~~~r~-~~~~r-~1~ ~~~~O~O O~O~ O,....~
dddddd~dddddddddddddd~ddddd~~~

M ~ ~j O N
M h0 ON1 N ~
_h ~ ~ ~ ~ ~ ~ O ~ ,N-,, 01 O ~_ N .--, ~n r' ~ ~ M ~ ~ N ~ '-' N o°~O '-"' N M ono ~ N x ~ O
.-M, ~p ~ p '-' d' N ~ N o~0 ~ O ~ p y0 O ~ M ~' Wit' O N ~ ~ ~ ~ ~ M O
O ~ ~ a N ~ v~ ~ d' h ~ ~ ~) O o0 N .-. M
~ °° ~ o N ~ d o ~ ~ a ~ o rr~ ~ ~ ~ o d ~~ ~ ~ ~ ~ o o~oo~
° i°
O ~ oo ~ ~ W ~ ~ ~ ~ N ~ o0 ~
o ~ ~ ~ .~ d u; ri c~ o; a ~ ~ ,~ ~ ~ x o M d o0 ~ N ~ M \O N ~ M h ~p ~ p M M ~ ~' p a1 O ~ ~ 00 ~ h O p N O~ ~ M ~n N M ~ '-' ~n M
M M ~.,~ h ~" ~D O ~, 00 h O h ~ ~O O M ~ h N O ~ N h ~O O '-"
M M O ~ ~ h 00 ~ O M ~ ~-~ .~ ~D ~' M M M .--i ~t a oo ~ ~ o. ~ r-~ ~ '° N
a °; a w oMO d ° a o d ~ o '"'' o x ° a d ~ a oy ~~-1 i dNd~oo~dd~~d~~~~dodd~dadNao oa°~_.-,°
h M Q1 DI~h~O h~~M.~~Opd~Np~pNho~OOMO~~php~d.~~d0 O h O M_ ~t O~ op oo O ~ v~ ~p N .~ N O ~ oo ~O ~ a\ ~ ~' r,j ~n d' vj o0 °~-~° U N ,~ M O ~ O M O ~ ~ '~ ~ M ~1 ~ ° ~ ~ o M ~ v~~, ~ ° o a1 -' ~ d d ~ d ° d rn ~ a d rr~ oo ~ ~C ~ d o ~ ~ rr~ d N d ~ ~ ~ ~ d ~; 0 00 ~° ~ ~ ~ oo t~ a ~ o; ~ o ~ ~ o ~ ~ o r; ci ~ oo d ~ c~i a o0 N h N ~~ ~ 01 ~ M 01 01 ~ O O N ~ 00 ~ h M
N ~ ~ N O~ N M h ~ ,~ ~ ~ O ~ p ,-. ~ ~ v~ ,-~ h ~ ~n ,-~ ~ O ~ ri N
NpN~~M~U~M_oO~~O~d'~~NM'ON_NO~N~pOp~~O
o ~ p ~ o ~ o ° a ° o ° U ~ ° ~ h ° ~ p a ~p W~~aw~a~d~d~aw~~N~~~ddd~~aaad~

M 00 I~ ~ ° ~ ~ V7 V7 d1 00 ~ ~ M a; ~ N ~ p ° a\ 0~1 N O 01 O I~
--.~ 00 ~ O 01 'p ~, O N ~ ~p N lp ° ~ N N v7 l~ ~ ~ O ~ ~ 00 a1 ~ ~'~~ ~ O_ ° ~ 0~0 ~ O ~ 000 .~ a1 ~ M N o0 N'-'vjd~~0~1~°00N~-~~~NM~~~~ ~'G°
d cN~W ~ ~ ~ o ~~., o ~ ~ ~ ~ ~ ~ ~ x d a; o o b U
rivo;°d°do°~~~d~~-'a~',~~oN~Md~ ~ ~ay~~
00 pp ~ ° '° v0 av ° ~--' ~ M ~) ''' oo O O ~ ~ M
N ~ ~ ~ ~D N ~ N N ~ d '~ ."' ~ ~' ~ ON ON ~ ~ N O
C/~ d x 41 d N 01 ~ ~ ~ ~ ~ r-W--i pp ~ ~ OM1 pip V~ r~ ~ r~ ~ ~ v~ M
~t ~ ~ a1 ~ c~j ,~ <.,j ,~~~~ ~ yp ~ ~ ~ ~ ~ N N o0 ~
_l_~Q\M Nl_ N '-' ~-' l~ I~ N O~ ~ ~' ~ M oNO ° ~ ~ ~ O ~ l~ O M ~ pip O ~ N M
M O
N a N l~ 00 l~ Gz., ~..~ O ~O ~O ~ ~ N v~ ~ ~ M U ~ p M ~n N v~ V~ ~ ~O
~d~d~x~d~wx~do~xdwaddo°°
00 N 01 tn t~ V'1 ~ 00 M ~ ~ d1 ~ ~ ~ Q1 00 d ~ ~ ~ ~ M d ~ d ~ x M ~ ~ N O gyp ~p v~ .~ O~ O M ~p ~ N N I~ '~t pp ~~rj O N p~ ~, ~j O p~
N W O I~ M I~ M .-r l~ ~ M N 00 ~ N t~ I~ .-~r ~. I~ ,-, ~ ~ O
O O O M M M ~ ~ M .--~ ~ N O d~ ~ ~-~ M ~ ~p M ° N M 00 pp ~ ,-, [w O O O .-~ ,~ ~ O ,~ ,~ .--~ .-~~~ ,--~ .~ d1 N ~ .~ ,~ [w ,~ O N O O M v7 M o0 a~~~dd~.~d~~d'~~°w~~~dad°wxd ~~~x~
a1 ~n o M M ~O
~ i N N
O ~ ~O
M M lp i i ~ O I~
00 N Ov O V~ 01 ~D ~O O
N N N
N
a x cri -~ ~t o M t~ ,~ o; o wi ~
pp N ~ ~ ,-..i ~ ~ N f~ ~ M M M
N ~ ~ N ~ p°~ ~ ~ ° ~ M l~ M ~O d' O d' N N
M l~ ~ v~ ~ ,~ ~D ~ ,~ v~ ° ° o0 ,-, ~ ~ O o0 ,w0 N ~ a\
xNN ~ ' N~OM~ M °
rd~'~d' ~ZdO°°ZdO~~ ~ ~aa,N~~~~pz _ '-. cd ,~.
rr N ~ ~ p~ ~ M N ~ pip ~ p~ 'Z' 'b M ~ O M M N
01 N pp N ~ p~ ~ ~ N ~ ~ ~ N ~ N o0 ~ ~ b ~ M ~ 00 ~ M
Z ~n ~ ~ N ~ l~ M ~ N oo N .~ O ~ .~ t~ M ~ M ° M N O
N ~ ~ z ~ ~ ~ O
pp ~ ~ ~ ~ ~ ~ ~ ~ ~ H Q. ~ ~ ~ ~ ~ ~ ~ -~-~ ~ ~' ~~--~ I~ ~ d' N 0_O pp N O_1 ~D p ~ 00 M 01 v7 ~ N
xd'~~ MI~~~NNONO~N~°p~p~~01 ~R~ v~'lM~~dlO~!
° ~ ~ ~.,~ .~ M M O ~~ N M tn ~ ~D M N 00 H 00 .~ ~ N ~ ..r N
00 Q1 M O ~O N ~ M ~ I~ V7 ~ ~') ~ ~t O o0 M ~.,~ ~ \p .-r Q1 ~ N ~ l~
N N ~ ~ ~ ~ ~ ~' p~ Q Ov O ° N ~ ~ N ~ v~ oo ~ M ~
~~dd~~d~~~~~~~° ~~~~~~ddx ~i ~' o o ~ o; .W o ri oo ° ~-!
x d l~ N ~n ~ Ov O N oo ,w0 v0 M N O ~ v~ ~ .-, ~ O t~ N W O o0 V~ M [~ ~ O~ M O~ ~ a1 I~ l~ ~ ~ 00 V~ ° O M v~ 00 ~ a1 O M
M M ~ p~ ~ ~ O O p~ N d' 00 ~ N d- ~ .~ N D1 ~ I~ ~ '-' ~ ~ ~O d1 O
M l~ 00 N ~ ~""' ~ O ~ \p ~ M ~D M 00 ~ O~ M ~O ~ ~O ~' 01 00 ~ l~ ~O '~
O M~ ~ ~O ~ O 00 00 00 O 00 ~ Q1 ~ O ~ 01 ~O ~ t/7 "'r 01 ~~z~~d~~~~~~~~~~~H~~x~H3dd~d~~~
, , , o o ~ °
M I~ 00 d' , , , O 01 v~ l~
Q\
l0 l~ 00 O
I~ 00 O~ O
--i .-r ,-.., N
N N N N
oa a°aa xx xx~

N
~-r V7 ~ l~ ~ Ov M
M ~ ~ ~ d' ~ ~ ~ ~ ~ ~ M N
N ~ ~ N ~ ~ o x ~ N
M z p ~ ~ ~ ov d ~ V; 3 ~ N d ~ .-; vo ~ o; ~, .b o 00 ~n t~ oo ~
N ~ I~ O .~ ~ N cd ~ ~ O N ~ M O
p~ M N (~ ~ M V~ ~ p~ ~ ~ ~ V ~O O ~ O
N N ~O ~ ~ d' O V~ ~n d N ~ .~ M
O 00 ~ ~ O ~ 00 '-'~ ~ M O M
d N O ~ z ~ ~ ~ ~O ~ O oo ,-, O1d N~~ NNp~p ~~~ '~ rj~ O
~~oxx~~_o~~°~ ~M~~ ~~~ ~~ z "~ N N N ~ O 0o ~ N 01 M x ~ ~ ~ O ~, ~ d O ~ ~ O
N N ~ ~ ~ r' ~ ~ M ~ d- ~p ~D op ~ ~ N ~ ~ ~ p Q1 ~ M ~ rN-~ d d l~ ~ ~ O~O ~ V'7 ~ ~ ,~ ~ M p ~ O O
~,d~3xdddzx ~o ~~~Q~obN ~ad~x~o .p ri ,~ vs ~ ~ ~ oo E-~ '"" ~ °° ~ ~ c<; a; ~ t~ a; t~ Wn ~~~d~~,~~~M~xMz xNOH M,~~N~~p M N ~n o, a, ~ ~ oo ~ o, o ~ ~ o ~ ,~ ~ ~ ~ °1 ~' ° o U ~
0O ,~ 0O ~ N M 01 I~ M ~ ~O ~ ~ ~ O pip ~ U N ~ N M N ~ ~' .~ ~ N
~o V~ ~ ~p ~ ~ ~h d' M N ~ ~ N O ~ O O ~ .~ ~ ~n ~ ~ N M d ~ ~ v~ d' ~t M ~ ~O O d O N
t~ d d d d °° ~n ~ °° o, ~ 'b ~ N ~"' d a ~ '~ 'b o0 dd~z~d~dd~~cxd~~xd ~Hx~d~d ~~~ ~~ri' N ~~ N I~ ~O ~D o0 '~fi o0 O ~n o0 O d\ O '~ O O ~ N ~ N ~O ~~
~O l~ ti' ~ ~ l~ M t~ M V1 .~ 'ch i i i i ~ ~ i ~ i i ~ i In V1 V~ ~ N V~ t!7 t!7 ~ V~ V~ In 00 I~ 00 M N N d' O ~ ~D ~-~ d' 00 l~ 00 M Q1 01 M .~ 00 O V~ O
V7 l~ M V' ~ ~ M I~ M ~1 i i ~ ~ i i i i i ~ i ~
Q1 M ~O v~ Own N O1 O ~-W O
N Wn N o0 00 M o0 ,~ ,~ Q1 V7 O ~n ~n o0 ~D ~O v~ I~ l~ ~D l~ ~
I~ ~-~ l~ ~t ~ 00 N N v~ 01 v~
O~ 01 ~O O I~ 00 01 ~n 00 ~--m0 N
N
N M ~ ~n ~O l~ o0 a1 O ~-r N
N N N N N N N N N M M M
N N N N N N N N N N N N
~O .~ ~p V~ Ov o0 01 00 ~ .W O .--~
aoa o'aa ~ ~ °
~33 3 3 ~ 3 xx xxx x x x xx xx ri b p ~ O M
O
O
N ~ ~ ~ ~O N o0 -d ~ O
--W' N v~o ~ ~ ~ ~ cd I~ t~
O ~ O ~ W V1 N
r., ~ ~j O
~,~MO~~O~'',N_,x0 pOp ~N MO
d' ~ p ~ 00 O ~ r, ~ O ~
M ~ d ~ o ~° ° ~ ~ o ~ o, ~; ~ o 0 M ~--' ~ ~ 01 M 01 N O ., O~ ~ l~ O~
t~ ~ N ~ l~ v~ Q\ M N ~ l~ Q\ v7 ,b ~n ~ ,b l~ ~ N ~''~ ~ N ~ ~ ~ O oo a; 0~0 "C7 ~ fir' p N_ O ~ I~ ~ ~ E-i ~ ~ ~ 00 01 ~ o°°, ~ ~ '~ o d Is, ~ ~ x ~ ~ N ~ ~ N
~ ~ N x t~ o ~ ~ ~ oo ~n ~ ~ 00 o c~i ri M ~ d t~ ri ~.-i N .~ ~ 'p o0 O ~ ~ ~O V~ l~ N O v~ N O ~ O o0 ~O
M M ~ O ~ ~ QI ~ ~ I~ O ~ N M ~ I~ O <h Q1 00 ~D ~.; ~D 0 01 ~
~ .~ M ,--. ~ ~ p p .-r I~ v~ pp ~ N ~ ~ o0 ~n o0 00 l~ ~ o0 ~n ~ N 0o O
~O ~D ,n M M Q~ ~. M M M N ~ N I~ ~ ~ ~ ~ l~ l~ M ~,, l~ l~ O ~
l~ l~ ~ O ~ ~D l~ d1 .-~ M O Q\ '~ N ~ ~ 00 ~ I~
N ~ ~ N i-N--i ~ rN-i ~ ~ ~ ~O ~ b ~ a. O
z~dza~~z~~~~~~~xx~~~~~~z~~~xx~
M
.~ M Q1 lGt O o0 N o0 v7 N O o0 00 00 M M O
N ~O M l~ ~ ~ O C1 00 (~ \O M Q1 .~ M V~
M ~D C~ ~1 ~ ~ ~--~ 00 M ~-~ l~ ~' 01 l~ 01 ~
i i i ~ i ~ ~ ~ ~ i ~ i i ~ ~ i i l!7 N ~ In V7 ~ ~ In N ~ V~ VWl ~ N ~ h, l~ Q1 V7 N ~ ~ O ~ ~~ 00 ~O ~ '~ ~ ~ 01 ~O
O ~ N ~O 01 O O o0 I~ ~n ~ N o0 O ~ M 01 M ~D l~ ~ ~ ~ ~ 00 M ~ l~ ~ 01 l~ Q1 ~ M
i ~ ~ ~ i ~ i ~ ~ ~ i ~ i i i i i I~ t~ O O N ~-~ ~1 M 01 00 00 Ov 00 N N O ~h .~ O ~O ~O ~ O M o0 00 I~ v~ v~ ~t O ~ N N
N WO d1 \O N l~ ~ ,~ O ~ 01 00 N ~ O
.-, M ,~ ~ .-r V7 .~ ~ V7 '~t ~ M 01 O~ O O~ M
00 I~ ~O ~ ~ ~ ~ N ~D N M ~O ~O ~~ t~ l~ M
~ I~ ~ l0 l~ Q1 Q1 01 01 I~ I~ O~ l~ ~ Q1 l~ l~
M ~ N lp l~ 00 ~ O ~ N M ~ N ~O l~ 00 01 MM M M MMM <t ~ ~~ ~~~d d'd' N N N N N N N N N N N N N N N N N
.~ M o0 r,, p l~ O ~O N v0 N
N I~ ~ O ~D .~ O O ,-, ~ ~ ,~ l~ O 00 00 V7 x ~ d~~ ~ ~ 3 a~x~
3 ~ ~ ~33~3 xx x x xxx x x xx xxxx xx o cn vo a; 'b Q' cri ~ ~-M ~ d' O M ~_ Wn N N
N O M ~ ~ N ~ ~ 0~1 ~ 01 ~
'-, ~ pv M
00 ~ ~' ~ ,~ Ov ~ ~ ~ O 'd N ~
N oo O ,~ d OyV 01 z N O
M x~~N~~N~~xNM ~ ~ x~
N M M ~ O pip O ~ ~ ~ M ~ ~ ~ M ~ \O ~ Np o0 d1 ~ N pp x N 01 M ~O ~ N ~ ~ M y0 Ov ~ M ~ ~ ~ N ~p N ~ ~ M
00 O ~n ~ t~ ~ N I~ ~ ~ ~. M M ~_-' 'p ~_' ~ M ~ p ~O O M o0 ~ ~ ~ ~ o N d ~ .b ~ d ~ d d ~ ~ d 00 O l~ b ~ ~ ~ ~ ~ ~ ~ ~ M V7 01 M ~
~
-..i ~ M V~ ~ cad ~ ~ N O ~ ~ M N ~ M M M ~ ~ ~ N N
~' a\ M N ~ ., oo ~ p M ~ N ~ ~ O~ O~ a\ a1 ~ ~ ~ N ~ 00 ~~~o_ boo ~'~'~°~d'"'dd~~~d~ d~Ca~~~
NNN~M ~~~~~dd~°Nd~N°~°o~,cw M o0 00 ~ ~ ~ ~ vi O N ~D ~ O~ N ~n ,-: ~ N ~ ~ ~n M ~-' ,.-~ ~''~ ~ ~ M ~i N
01 O O ~ ~ O 01 v'1 ~ N ~ N I~ Q1 N v7 ~ I~ ~ ~ ~ 01 '~' ~ ~' ~p O 01 Q1 M
O ~ N 0 ~ ~ N ~ ~ M O ~ ,~ M ,~ ~ ~ .~ O ~ '~' '-, ~ O ~ M
d'~~~~x~~'~~x~x~~~~~~d~~~d~~d~~x O ~ ~ ~O M N 01 N
i i ~n v~ ~n ~n v~ v~ ~n v~
~ ~ ~ ~ ~ ~ ~ .-.
I~ ~ N O N 01 00 N d' N ~ ~ ~ 01 O
OW D ~ d1 ~O v~ ~D
~ ~ ~
~ ~ ~ ~ ,~ ~ ~ ~
N ~ 00 01 M ~ oo ~n 01 ~t ~ ~~ M ~~ oo v~
~O ~ ~ O 01 M M 01 ~n v~ v~ I~ ~n V~ N 01 N 01 ~ ~ .~ ~ 00 O~ l~ l~ 01 01 01 01 00 O ~~ N M ~ ~ ~D l~
N N N N N N N N
I~ . ~' V~ N .~ M I~ V1 '~x x~~
~x x~x x N
N V7 ,_, 0~0 ~ h 00 M ~ N p ~ ~ 00 \O O ,.~ ~' N O wj '-'' ~ N O Q1 O h .N,~ N x M z \p" ~ ~ O ., ~! '-' ~ 00 N ~ 00 ~ ~ ~ 00 0~0 M '-' N ~ ~ O N ~ ~ vp M O N h p~ ~n o0 O ~ M O N ~ M O ~O h ~ h M O_O ~ O O ~ ~ ~ 00 0o i-..-i °N M N a, ~ o ~ ~ a, ~n 't a, °.~° ~ ~ d d M o ~ p ~ ~ ~ ~ °.~° ~ ~ ~i C~ ~ o d M x h ~ ~ v; ~ ~ ~ ~ x o0 00 ~ z "'' 00 00 " ~O M ~ 00 O N ~O ~ O h o0 h N d' 3 O ~ ~n ..-r N h ~ ~ o0 ~ O ~ Q1 O_ ~ 0o O O ~n ~ ~O h N ~ O ~t ~ ~ h \O N M ~ Ov h ~ M ~ M ,,3 ~ D1 00 ~ ~ ~t O
~~OO~~°°,~hhxh~p~o~M~~~~ MMA ppd O M ~ ~ N ~ M ~ M ~ 00 ~ ~ ~ ~ ~ M ~ ~ ~ d ~ f~1 M x ~ M
h M M ~ N ~
01 N h O pp x ~ ~ ~ M z ~ M ~ M ~
O~ ~n M O O N O ~ v~ h N N ~O ~ ~ W p~ p_p N ~ N N N ~t ,~ ~ M N h O
~ d' O ~ ~ ~O O N .~ ~ d ~ tn ~ N ~ \O M ~ M '-' ~t ~ O r-h.~ ,"~ M O
M~o~ochyMV~-,~dxha,doo~-.. d ~hd d~ddd~
N lCr ~O ~ V7 N h ~~ M M d d d ~ ~ ~ ,~ o ~ ., d xxr~~xxx~x~~NxxNdo~~o~~NN~~~~~~
~ N ~O M h 01 M Ov ~ h N ~ ~-' ,~ O ~,,~ O ~ ~ t~ ~ ~ .-~ N N O h ~' h 00 ,~ 00 .W p ~O o0 Q1 O ~ N N M o0 ~O ~' ~ ~t t~ O N N ~ ~p ~ v~ v~
00 ~ ,~ ~ O \O O M ,N-,, ~ M ~ ~ M '-' ~ ~ M ~ V1 h M N ~ pip 0 ~ ~ N
~O ~~ h ,~ ~! ~h ~ D1 M ~ ~p ~ ~ ~ ~ M \O a rM-ii xxxdxxx~H~~d3~d~ddddxx~~d~ddd~

0 00 ~n p ~n ~, .-~ M ~ a;
0~0~O~v~'1~~~~~~~ M N
M ~i ~ M ~ ~ O O~ ~ N I~ N ~ a1 Q\ ~ '-' v0 ~ 'n ,-:
M O ~ M Q\ d' ~ ~ ~ N M ~ ~ O ~
~ o °° o ~" ~ d ~ ° ~ ~ ~ ~ ~ d ~ M ~ ~ ~ M O
'-' ~° ~ M 3 ~ ~t ~ ~ N °~ ~o '° °° d ° ~' ~ w d ~ U M d "' d ~ ~ a' o M ~ ~ .°~ ~ ° ,.~ ~ o N ~ o O O N ~ D1 O ~ ~ ~ N ~ ~ ~ ~ ~ ,-, ~ ~ ~ Q1 N ~
m ~ ~ p N ~O ~ '~t d d d d O .~ t~ N ~ ~ Q OM O
~~~~,~N~~~d~~~~~~~o~~~o°,~° o ~o ~O~o~OdM rM.,~M''~~MOO~~NN~~~~w~ ~ N
'-' V7 p ~ O~ ~ l~ 00 ~O ~ O ~' pip ~' p~p ~ Q1 00 ~ 00 M ~ Ov O
N ~ t~ O N p~ O M O o0 ~ ~ ~ ~ M M b b N .~ M
x~~~~~az~~~~d~~~~~x~~~~ ~~~~x~
'° d ~ z ~' ~ ~ o wo ~ ~ o ~ ~ N ~ vp N ~ v~ ~ ~t O
~O v~ N I~ ~ M N oo ~ ~ ~p o0 v~ oo ~ ~ z M N N O~ d' N Owe N ~O ~h N ~ ~_D M d' O O N l~ O ~ ~D t~ ,~ 00 00 ,~j ~O ~ ,-I O ~O l~ ~ I~ '~Y
.wp ~ M ~ ~' ~ O O l~ .-1 ~ O~ l~ v> ~t ~ ~ l~ N N N l~
00 00 l~ ~ O ~ ~ ~ I~ 01 ~ O O~ N 00 M l~ 00 ~ N V7 ~ O M O O M O M
d \O D1 N d N d d ~ r~- ~ r-N-~ rN-~ d d ~D ~ N O U d M M d d~xz~zd~dd~~dd~~~d~xdw~dd~~~d~
vo ,-, ~ ,~ o l~ ~ O N
V7 ~ ~ ~ M
i i i i i In ~ V7 tn N I~ N I~

V7 <t ~ M M
i ~ i i i N Q~ 00 O M
~ O l~ M
I~ ~n N O v7 ~O N ~ ~D v~
V7M~01 l~
00 Q1 O ~~ N
N
N N N N N
M ~ O ~O d1 N o0 O~ O
r~xc~a xxxx x ~ o ~ as a; ~ o0 M

M _ M
~Y ~
~
~~~
~
~
~
N
~

~N O v0 V~M o ~NOo ~
~~
p p O

~U,~UU,,~Uo~,UUU~,..~~oU oU~oQ,oo od do~~

d dNdd ~
d N
o d dNd N
U
dd ''~ ~ oo y_o v~ ~ ~ a~ ' ~; ~ o N o; d a; c~i a;
~ ~ ~ ~t d i p ~ O
~ lp M N ~ M O ~ ~ ~ ~ 00 ~ V7 I~ ~ O M p d N
~

~O O
N ,-, ~ ~ N ~D ~p v~ ~t .~
D1 ~h ~ ~
M N O ~ p~ O ~ ~' M ~ ~ O N O M

a~ ~ U ,~ U ,~ U U ,~ U U ~ ,-a U ,off o ~0-1 ~ o~, U U oMO U U o ~

~,~d~ddddd dd~ddddddNdd M ~ cV N N O O O -V N ~ ,-; M ~D U M t~ M ~j O ~D
~ ~i ~ M

I~ ~ 00 ~
d 00 M
~

M N
~ ~
~
O ~
pp ~ V~
lp ~ ~
p p ' ~ ~' ~ N ~ ~
~
O
~

v~
N M
~
~ ~
~ d , -, ~ V
l~
,~ N O p O O O O O O M ~ O O ~ O O O O O ~
~

ono ~ U U U U U U U ~l ,~ U U N U U U ~p-1 ~ r; yo dddd~d d d~d~ddo '~d~~d do M
d ~ ~
d ~ O M

0 ~ ~
~
~
\
~
~

v t t v ~ O
N I ~O
M O ~j O .~ a N d vj ~n ~ O I
oo 01 M v~ ~ .-~ ,yn o0 l~ ~ N O ,~ D1 N 01 00 00 ~O o0 Q\ ,~

~D O [~ O M M N O M p~ M N M M ~ M ~ ~O M 01 I~l~
~ o O ~~
~000~0~0oo0p0pocN.~oo~o0o0~a o~

-, O O O O O O O O O O O O O O O O O O M M
zd~dd~~d~~~~~~~~N~dd~~~N~ ~ a~ ~~

l~M
O

i ~

O N a\

v7~O M
i l~~ M
' M

O O V~
N

M ~ N

N N N

a100 ~O

x x x M ~ N
V1 ~ ~ O M 00 p V7 \O ~ M .~ M .~ .-~ 00 ~t O ~ ~ ~ r' N '-' ~ vp M ~ ~''~ O ° ° ~ ~ M ~ Ov ~ ~ N
N ~p ~ O l~ M N ~n ~ ~O ~ 00 00 ~ ~ N N M M ~ N v~ ,~ N ~ ~D O
M ~D O ° M ~ ~ N ~ v7 ~O M '-' ~D ~ O~ I~ 'ct O ~ ~ O , M y--m--~ M O~ M N (V M r-v r-i ~ N r-i ~ .~ N N ~D ~ I~
~~ddd~O3ry 3° ~dd~~nd~oo~o,~o~d Mdd d oo ri ~' ~ ~ oo N d ° ~ d .~ oo d ~ ~ d d d d d ,~ N W.; o M O ~ M 01 ~ d .~ w~j I~ 00 ~ M O pp ~ \O l~ N '"~ ~ O~
41Opo0O~~o0v~~~OO~W t'~~~,~-,p~pOOp°ppmpM
v1 ~ N .-, v7 N ~ ,~ l~ ,_.., ° .mn p~ p°p N M ~ O M ~~ l~ O
r3 p\ ~ ° pp ~ ~ pp 00 O\ M 01 ~' ~ M
°o ~ d d d ~ i d 3 ~ N oo ~ "' d ~ °° ~ ~ 'n "-, vo ~ ~ 3 d "' d d ci~d'°r.;d~3~ ~V;d~~~~~d~~d~d ,~ o N oo ~ ~ ~ ~o v-i ~ d ~ ~ ~ cri M ~ oy" ~ ~-~ ~t ~o ~ ~ o M
UjdN~~~~NO~Op~p,~~,p°p~~~~N~0~0 r-.i 00 ° 00 I~ ~ 00 ~O ~ ~ 00 O ° ~D ~ N Q1 V~ M ~ M M M ~ N
~C1 ~O ~n N O N ~ M ono ~ ,~ .-~ ~ ~ ono t~ ~ ~ N ° ~..~ oo N N ~D O ~
O
°o d ~ ~ o M '~.,° d °° '~ t~ 3 ~ z ~ ~ ~ 00 3 ~
~~d~ddd~~~~~~~d~~~3dd~ddd~d~~d v-i l~ ~n ~t O~ o0 ~ v'i O ~ ~i N ~ N ~ ~ ~n ~ ~ M O~ ~ d~ N N a; M
01 M ~ ~ O ~ ~n p~ N M ~ M 01 O ~ O ""~ l~ ~ ~O M 00 I~ I~ I~
O d1 d' ~-~ O N O ~ M .~ N N ~ .~ N M p~ N N ~ N ~ O ~' ~T ~n oo ~ 01 v~
O I~ \O V7 01 00 ~ O M V7 ~ N M N O N V1 00 M ~ N N ~D ~ ~ D1 O M D1 ,..w ,~ 00 l~ O M \O M ° O ~ ~O ~ M ~ '-' 00 .-r ~O V~ N ° O ~-' ~ .~ 00 Q~ 00 \O
M N N oo .~ oo M ~ I~ ,~ ~ ~p oo ~n 01 00 ~n ~ I~ ~ oo O~ oo v0 v0 ~t ~~~d~~~Hd~~w~~dd~~~ddd~~d~d~a~
N N
'~h M
M
M
N O
O N
Q\ N
~ Q\
N N
~t O
xx ~n ~ t~ ~--~ ~ N t~ M ~ ~i N ~ .--~ ~ O ~ d1 00 ~ ~ ~ N .-.~ ~_ O N ~ ~ 00 N 0~0 M ~ ~ d1 ~ Ov 01 ,~-, Ov No 00 ~ ~ ~ M ~ v7 O ~ ~ ~ ON M vp O l~ Q\ M O ~ ~t ~ O
~d~~~dNd~"~° ~~~~~d3~ o°~~d~~d~d~
~~MMdN~~N~~~~dd~~~dddd~~~oM~~M
0~0 O ~ ~ ~ Ov ~ O ~ ~ v0 O N ~ 0~1 vp N O ~ ~ m N ~ ~ '-' vp V~ ~ ~ ~ N l~ ~ V~ pyn l~ ~ ~p ~ ~ O ~t O~ '~ ~ 00 ~
01 .~ ,_, O~ 00 I~ ~ V7 ~ N ~ ~ 00 ~ O N ~ ~O ~ ~O M ~
y--m....~ ~ r~
Q~~~~~~~~~~~~~~~~~~~~~d~~~N~~~
I~ O ~O I~ M o0 ~j d' ~ " ~ M ~ 00 O N ~ M 01 V~ N 01 O V7 O ~ 01 ~ .~ ~ ..-r M I~ N l~ N N O M N
00 I~ ~D M I~ ~ N .~ O ~ N N ~ ~ ~ N ~ M l~ a1 01 00 O v~ N
00 00 M O ~ ~ t~ ~ O ~ 00 M O~ 01 ~ ~ ~ M t~ ~ N ~O r-' 00 M
00 ~O M N v~ oo ~ N ~ N o0 p~ l~ ~ M O 01 00 l~ ~ O ~ O ~ ~. pp N
dM' ~ ~ ~ M O ~ N V1 M N ~ M lp M ~O \O ~ ~ 00 ~~~_~_d~~~~~~d~~d~d~~dd~ddd~~dd~
N ~-wl' ~ ~ 00 O N ~ O 0N0 M \O ~ O ~ ~ ~ N M ~ ~ ~ O ~ ~n O_ ~ N
M O ~ 00 ~ M ,~ ~ ~ t~ 01 N O~ ~ ~ ~ ~ ~ ~ 00 N pip ~ ~~ M M ~ M
N o0 0o N p~ ~h N O ~ O ~ Q1 00 M Ov O p ~ M Ov M O o0 ~ ~ 01 00 01 M O O O M ~~ O M ~ 01 \O ~ O l~ ~ '"" 00 '~ M ~O 00 I~ N I~ ~ I~ 00 ~ '~ M
~3~~~~33~~~~~~.~~ d dddd~ddddd~dd~dd~ddddd~~~d~~dd 0 o ri ~O v~ d1 ~j l~ N ,-. ~ ~ N ~ M M ~ ~O
' v7 v0 O d' ~ M ~ O ~ l~ O ~ ~ ~ N ~ ° N ~ ~ ~D N O ~ ~ M
W M .~ l~ ~' M M ,~ ~ ~ M ~ N '""' M °N a O ~D ~ N 01 'O O ~ M
d d ~ ° ~C o '-' '-' a~ ~ o '~ ~ v~ M_ ~ o °° ° d o ° ~ ~ o ~ o N ~, o O ~, ~ N Q ~ N O ~"~ ~ a M
O ~ ~ ~ d ~ ~ x ~ ~ .~ oo ,..~ M ~ ~ ~ ~ ~ ~ ° N a\
d. ~ a ~ .-. ~ ~ ~ N o ~ ~ a\ ~ o a~ ~ w ~ o ~ ~, a ~ ri os ~ ° Sri d 00 ° p~ ~ ~ ~ ~ M N 00 ~ .-r ,~ I~ \O M ~ ~O d ~ p1 00 pp M V~ ~p ,~ ~ N ~ M O ~O N V~ M O Q1 d ,,~ O 00 ~ \O
00 ~ O M vD O v~ t~ ~ I~ O O M C/~ ° ~ O~ '"" ~ ~ O .~ N N ~ .-1 ' I~ M ..~ ~ W l~ M p~ lp W M V~ '~ W ~ ~ ~ ~ M N l~ N
~ ono ~ d ~ ~ ~ ~ ° d ~ ~ cri Do ~ ~ ~ ~° ~ wi ~ ~ ~ ~ ~ ~ ~ ~ o l~ M ~' ~ 01 M ,-, l~ O op ~ oo M ~O oo ,<j ~ t~ N M ° ~ ~ N ~ ~ N a ~ I~
~O ~p 41 pp ~ [~ 00 O ~ M ~ Q~ .-r d V~ .-r ~ O ~ M d ~ l~ M .~ 00 M N V7 ~ ~ 01 ~.,~ I~ I~ ~ 00 N ~p d1 M M O ~ ~ I~ M 00 O
M_ ~ ~"' op ~_ O o0 p~ M O O ~ ~ ~_ p oo O ~ M ~ ,~ o_0 ~' I~ O O
d ~ ,-, ~ ~ N .-. M oo d v~ d ~ ~ ~C ~ ~-' W d ~ °° ~) M o, ~ ~
° 00 M O 01 N_ ~"~
N o0 ~ ~ .~ d d ~ d v~ ~D ~ Q\ 00 ~ ~ ~ l~ N ~D N ~n pip M o0 ~ ~ ~ W
M N ~ r-~ 01 \O Q1 01 ~ <h ~ ~ N N ~ ~D r-i N O N M p1 N M O ~rj ~ ~ ~' O ~ M M ~O ~O 00 O ~ ~ N N O V7 .~ O O pp" ~j t~
I~ ~ ~ N l~ ~ ~ l~ ~~ I~ M I~ 00 M ~ M O l~ ~ M d1 V1 O O M ,n N .~
N O M .~ ~p ,~ M Q1 M 00 ~ .~ M l~ ~p M M ~~ M O M I~ N V7 V7 M N ~ \p ~ N O ~--~ N O M o0 ~ O ~ ~ ~~ O M ° ~-' ,~ ,~ O .~ 01 ~O O O ~~ ~ ~ O
d ~l a, ~ ~ o p o a w r~, a w N ~ a ~ ~ U ~l ~ ~ a a ~ o w ~d~C~~x~~dd~d~ddddddddd~~~~dNwd l~ N
Ov O~ Q; oo vp N ~ O N
O ~ ~p ~ ~ cn N ~O ~ pip ~ ,-; ~ V~ ~ ~ M 01 V7 ~--W~ ~ O ~, .~ ~O l~ ~ M l~ M ~ ~ M p~
0~1 ~ ~ ~ pMp ~ v~ ~ ~ O ~ ~ ~ O N O N ~n '_"' O ~ M O O M O b ~,,~ N ~ l~ ,~ N r, ~O ,~ ~O a M V~ ,-, W N .-. l~ 00 l~ ~ W O a ~pNo~M~~~x~~xM~~~~~~~a~~d~Qd~~;
~ ~ oo W y"' ~ ~ ~_ c~i Q: ~ o ° ~ ~ ~ .~ ~C ,-: ~ d M ~C M ~ ~ ~ ~o °' ~ O oo ~ I~ ~ WO p N ~ 'd ~ ~ ~ ri ~ Ov N W O ~ O~ ~' ~n ~ 00 O
M ~h' ~ ~ ~t N ~ N O t~ O 00 l~ 00 l~ O 00 ~ O lp ~ M ~ O O
~~ M ~ M N Q1 O M 00 V7 N ~ M ~ M \O V'1 O N d' ~ ~D ~ O M ~ '~h p O~ M
O ~ ~ O o0 W ~ r"' O O o0 O O '"_' C/~ ~ ~--~ .~ O O N ~ M '-' ~ O
~x~~~~~°~~~d~~~~~~~'~~~~~~d~~'~d tn o0 M ~ ~ ~ N Wn pp O p~ ~O C/~ l~ ~ ~' ~ 00 O ~ ~ ~, 00 N ~ 00 (wj ~O O M ~ ~n N O N ~ ~ ~ ., 00 ~ ~' ~ v~ M
l~ N ~O ~ M o0 '~ O ~~ N M pp ct' O N O ~ O O ~ ~ v7 ~ l~ ~ O N ~t Ov O ~D ~ .--r tn ~ \O 01 O I~ N Q\ \O ~ Q. N ~ 00 01 ~ \O o0 M M M l~ I~ O
t~ O I~ N O N .. ~ O r.r ~ ~ ~ v7 N N 01 ~ N ~~ ~O ~j M M p 01 M
O O O ~ M ~ ~ l~ ~ ,~ O N O M ~ N l~ O N ~~ ~' l~ N 00 ~~ p ,/~ ,~ O
M .~ .~ .~ M a M 01 M ~O M tn lp U I~ a ~~~d~~Na~x~~a~~~d~~~
~°o°~,d~~d~~~o°o~c"'v~
°~dr°~~,aM,ocM~,~~~M~'r-.°o~ ~
t~ ~ ~ W p d O ~D N ~t ~O ~ 00 ~ M M d' O N V~ ~p ~ N M O ~ OW O O
I~ O ~ ~ ,~ ~O ~ 00 ~ O I~ I~ O N O M \O 00 M M (~ ~p N I~ O~ M l~ .~ N 01 I~ O ~ N a1 I~ <T ~p O ,...., M .-~ 00 N 00 N [w O M ~ M (w V7 M ~' M .~ M
,~ O O ~ 00 O O\ 00 ~ p ~' ~~ ~~ ~O O 00 O O ~ .-. .-, N 00 ~' O ~' ~' O
~~~~~~~~~~~~~xd~~~~~~~~~dda~~~
N
N
N
~n N
O

O p°p ~ N
~ c'y0 0o O r' _~ v_j N
~ tn ~,~ ~ ~ \O 00 00 ~ O
v7 V1 ,-, M ~ 00 C~ ~p N N ° ~ ~ M r.rj ~ ~O O v7 '~ M
O O~ ~ d' ~ M ~ O ~,N,~ ~ ~ ~ 01 00 ~ p~ ~ ~ 01 0o M ~ N
~p ~ O ~ O ""~ N ~ I~ ° M M M N .~ I~ 01 ~ ~ M t~
y0 N oMO . 00 O o0 N ~ ~ . l~ ~M N ~ ~O
~d ~ 3x~~~x °~ ~ °~:M ~~ N
~" d ~ ,~ N N ~_ ~ v°NO d ~ o ~ o ''' ~ ~ ~ o ~_ cr; ~
M ~ ~ ~') ~ 00 M 00 M ~ ~ l~ ~D d M ~ ~ ~ N ~ 00 M N
N pW p ~ N v~ ~ N ~' o0 O~ r.,~ ~n ~ N M O N ,~ l~ ~ O
d ° ~ ~ ~ ~ H ~ ~ ~ ~ d ~ M ~ ~ O p1 ~Q; ° ~''' ~ ~ ~r ~ ~ d o 1N~~O ~~ ~~ M~ °p~p ~~O _~ N
O~ N ~ ~ M O 01 N °N ~ O ~''~ M d N ~ ~ v~ ° r' ~O M o0 ~

'-. ~O. N ~ ~ ~n ~n ~ ~D ~ ~ ~ N ~n ~ ~ ~ Ov 0 ~ ~ ~ ~ M ~ OW O
~;dd~~ N ',~°~ o~,~~~ d ~~ d M o ~: oo t~ °; M oMO ~ ~ cri c~i ~ ~ ci x d N o ri d ~ ~ ~ c~i ~
~ d a\ t~ a\ oo ~ ~, ~t o0 0, ~ .~ ° op ~n Q, ~ N t~
Q1 O ~ ~ M M ~ ~ ~ ~ 00 '--~ ~ ~p ~ M ~ ~ ~ O d' 00 Q1 M ~ ~ ~ O
M M_ O oo vp ~ ~ M ° ~n O N ~ M ~ l~ M ~ 00 ~' ~ M ~ ~ O~ ~_D o0 N iN ~ N o0 ~' M l~ N ~ 01 v~ l~ ~ ~ N o~o p ~ O ~ Q1 a1 N 01 0~1 °
~O o00 O
~~~~~~d~~x~~~x~zw~~~~N~z~ad~x~
00 o a~ .~ o a1 V7 M d' M
~ i i ~ i, N N
V~
O I~ O
V7 M d' M
i M M ~D ~ 00 M .~ ~ 41 N
N l~ ~ ~ ~D
O V) 00 ~ 00 ~ Q1 ~ ~ Q\
Q1 O ~ N M
N N N N N
.~ ~ O M N
dd x x ~i N WO ~ ~ v~ O O o~0 ,ri '~ ~ opo N ~p N o0 O V7 ~ M O
M N M ~ ~ ~t N ~ 01 M M O O N op 00 ~ ~ ~ ,-, N
00 ~' O N ~ ~ ~ ~ ~ O O
_ ~ _ ~_'~d~w~~a, ~~~~ dd 0o d o0 N ~ ~' t' N ""' O .-.~ ~ p p p ~ ~ "'~ ~ 01 '_-' O
01 N .~ .~ M ~ O vp ~ l~ lc' ~ d M ~ ~ d '~ ~--~ d~ M ~' ,~ d1 I~ N N ~ O yn M '"' t~ N .~ ,_, '--, '-' pp N M M 41 a\
.--i M M ~. ~ .~ N ~ l~ O ~ ~ O ~ 0o N_ pp M O N ~ ~_ ~:
d a~~~~~M ~ ~~ ~r~d~~ X0.3 o; a ~ No N ~ ~ d d ~ ~ d ~ d ~; ~ '~~'' E~-~ ~ d ~, d c~i .--~ 00 a; ~
~ d N o .-~ .-.. o ~ ~ ~.,, o ~ 0 00 ~-i Z o ~, '-' o .-, o; M
01 l~ ~n ~ V1 O ~ ~.,~ M M N M ~ ~ ,-; '~ ~n d ~n Q1 ~ N ~ O
~ 01 ~ M ~ p~ N O l~ r' '~t ~ ~ ~ N O d1 ~ ~,~ 01 N ~' .~ M ~ .~ ~ O ~ ~ ~ l~ 00 O ~O N O O ~p O O ~ N 01 N O1 a v0 "C ~ ~n p~ ~ I~ N ~t ~ N 01 0o v~ ~ ~. '~ oo I~
d~dddd '~ °~d~dd~'~ ~'' due' O O~ ~Y ~T' M ~h ~ ~ ~ ~ ~ ~ 01 ~ ~n M
a1 01 01 ~ oo l~ 00 O ~ ~ v»p ~ 01 t~ d p p ~ 00 N V7 t!7 ~ M ~O ~ ~ [~ M O M N lp M 01 ~ 00 ~ ~ N l~ I~
N 01 01 N ~ 01 N ~ 01 M o0 M ,-, O M pp ~,,~ N o0 M l~ O ~O
M ~ ,~ ,~ ~ ~ O M V7 ~ ~ M M 00 01 O ~ ~ M '~ ~O O ~ 00 y~ O ~ ~ 00 M .~ O .~ .~ O O O ~ ~D l~ l~ ~ oo ,~ I~ M d1 Iyp ~O ,~ py0 ~ i-r~ ~ ~ ~', 00 3aaaaa d~ ~~°;~°°°3~°d ~~N ~ ~ 3~
dddddd~~Z~~ddddddZd~rxd~~7~~rx~dd N 01 N I~
N ~ M ~ ~ ~t i i ~ i ~ N
00 V7 00 M l~ O
O ~ ~ M ~ M
N
i i ~
Q\ I~ N M ~1' M
Q1 l~ \O M ~-~ O~
O O N N ~ a\
M \O O~ .~ ~ 00 I~ ~O N oo M O
p1 I~ I~ 00 W~ 01 V7 lp l~ ' 00 01 N N N N N N
d~
x x 0o a;
M

00 ~ 00 O 01 ~"-' M l~ ~ ~ ~ ~ O O '-' v0 p ~ ~ ~ M U N ~ O p Q\
cd ~t O ~' 0,~0 ~ o~-ro 0~1 M
N ~ ~ ~ ~ ~ O ~ ~ O
M ~ ~D M M O ~ ~ O N ~ ~ ~ N Cn M O 00 ,-1 M o0 N ~ ~ a N ~ ~ O ~ ~ 00 p1 ~ ~ ~ M ~ O O ~D 0o ~ O M ~O
N v'i ~ ~ d pp ~ N O ~i ~ ~'' ~ N ~
~D \O ' M r. ~ ~ 00 01 t~ Z N
V M ~ ~' t~ N ~ tYi ~ ~W p N v0 0o M N
r-~ M ~ ~ c0 ~ opo '-' ~1 ~ N p o00 ~ O~ p ON ~n0 O
~ M M ~ p ~n ~D ~ O O Q~, l~ 00 [w ~ v7 pp M ~ 00 O ~_p Q1 b 00 'b (~ O M "~ ~' M "C~ l~ 00 M
b ~ d ~ ~ ~ ~ ~ O ~ ~ ~ O d M
N O~ l~ 00 00 ~ ~n l~ ~ M ~ l~ I~ N ~ ~i ~ ~ M
00 ~ N O ~ N N d' 00 ~ ~ 'n ~ N M ~n ~O N N
~O O 01 ,~ .~ M N oo N O ~ ~ ~ ~ '-' ~ cn ~ 00 ~O l~ ~ v~ ~ r; 01 O~ V~ M O~ Cn ~ ~ M M O ~h l~ M oo ~ N O I~ O
N V7 N I~ l~ O ~ O ~ ~ DO ~' ~ '-' N M ~ N D1 ~p o0 ~ N 00 ~ ~ 00 M M O ~ O O '-~ ~ ~ N M ~ O N ~ Q1 ~
d~~~~~~°H~ z ~~dddazxd~~~H~zz~
M 00 O~ O V'1 ~ N M M ~ C~ l~ 00 O ~ ~O
t~ N O ~ N M ~ ~' ~O .~ 0o 'O O l~ ~n ~
I~ M ~ ~ ~ ~ M M \O V7 N M
i i i i ~ i i ~ i i i ~ i i V'1 In N Vl In tn Vl N N ~ In N ~ V~ ~ ~
Q1 ~' V1 lp .~ O 00 ~ O1 l~ M M ~ ~O O N
V1 .~ 01 01 ~ N 01 N ~t 01 ~ ~1 01 ~ d' v l~ M M M ~O ~~ N M \O ~ N M M
i i ~ ~ ~ i ~ i i ~ i i i i i l~ l~ 01 O M V~ oo 'O ~ v~ oo ~n N l~ ~O a1 N o0 N ~~ l~ N M ~ 00 ~ M Q1 ~ .-i .~
N ~ pv .~ .~ M O o0 ~n N d' ~I' ~-~ ~ ~t ~
M 00 Q1 O ~ ~--~ ~ 00 .~ t/~ lc, N I~ N ~ N
0o I~ ~n ~D ~O ~n N 01 a1 N N O~ I~ l~ t~ ~O
O ~~ N M ~ ~ ~O l~ 00 01 O ~ N M ~
00 00 00 00 00 00 00 00 00 00 O~ 01 01 N N N N N N N N N N N N N N N N
-, M l~ O N M O~ N M 00 l~ ~' 00 ~O ~ l~ I~ l~ v7 M 01 O V7 Q1 .-r l~ N O
C~ U ~ ~ ~ ~ ~1 ~ U H vy; ~ p4' O O O O O O O O O O O O
xx xxxxxxxx xxx xxx 0 os ~ o o ~t vo ~ ~ V' v'i ~ ~ ~ ~ ~ ~ 01 Op O ~ N ~ ~ ~ v7 ~ .~ M ~ N v~ N ~.,~ 00 ~ N N ~ O
V7 ~ N I~ .~ O ~.~ d' [w ~ ~ .--r M I~ ~ ~ O I~ 00 00 ~ O O\
~-~ N N N N ~n op ~O ,~ ~n ~ N N "~ M v~ C~ ~
N O M ~ ~ ~ ~' 00 ~ ~ M d' ~ ~ O ~ N M ~ N N
O O ~ ~ ~ ~ ~ ~ ~ M ~ ~ ~ 00 d~°°~~ d ~~n dd ~ ~ ~a~ d o M_ Q1 ~ M d ~ ~ N M ,~ N 'G ~ ~ N d ~ ~ .-i O ~ M ~ oo ~ .~ p t~ O ,~ ~ N ,-. t~ N M O ~j ~ ~ ~ N
~O l~ O ~ ~ ~ N ~ ~ p ~ ~ 00 ~ p M 01 p ~ ~ ~ ~ ~ ~ ~ M ~ M ~ ~ ~ ~D 'fit d' d O
xZ~ ~ ~d~xdz ~N~d ~E-M~i ~~xvod.n I~ O oo ~ ~ ~ ~ oo ~ .~ ~ N ~ '-: ~ O
o~o~Or-'~~~F''oMO~~o~O~M~~ O O~~N ~O N NN
0 0o M ~o ~ '~' ~' ~ ...., ~r v. o~ ~ ~ oo Z ~ ~ ~ a\ .~ oo ~t ~r ~ ~ M t~ ~ N ~" ~ ~ O c~ oo N ,~ ° c~ ~ ~ ~ ~n a1 ,~ '~ w oo .~ p v~ d O O ~D ~n ~ ~ N M ~ ~ '-' N ~O oo ~ 00 ~ ~W O ~ ~D M ~ O ~n Q\
\O 0o I~ ~ N ~ ~ O ~ N o0 ~ ~ ~ ~ O N M pip ~ ~ ~ ~ ~ ~ O
d ~ ~ ~ d d z ~ ~ ~ °; ~ ~ N w ~ ~ ~ N
M .-~ ~O ~ ~ v~ oo d' N M ~ I~ I~ ~ v~ N ~ ~ N o0 ~t c~j a1 00 ,~ vj d~ d ~ ~ 00 0o v0 M ,-, ~' ~. 01 O N O l~ N ~ ~ O oo a\ Z
~t N I~ ~ ~ °~ ~O v~ ~ 00 O N l~ O I~ ~ ~O ~ Qv ~ ~ 00 00 ~ O
M O l~ V~ M N O ~. ~ M N N N V~ M .-~ 01 \p ~ lp O~ 00 00 l~ V7 O ~ ~ tn ~t ° d °' d d d M ~ c.~~, N M N a, ~ p ~ ~d '° M M
d~d~ddZd~ddddd~d~d~~~~~HH~~~~w ~O M
i i I~ ~ M
M
i i O M M
N M I
V7 M ~O
N 00 l~
O~ Q\
N N N
x c~
xx x ' ~ N
00 V~ p ~ 01 M ~ M ~ N t~ M '~ M .-~
-; N pip N ~ ~ N ~ l~ ~ M ~ v7 ~t O p p ~ ~ ~ ~ ~ ~ ~ ~t o0 N [wM'~~ppV1p0M01 '~N~~tM~.-~..~NM~M'-'Q
01 ~ 01 ~~ O 01 .~ O ~n ~ ~ l~ a\ ~ O '-' V~ M O ~ M ~ N
M ~ N_ M v~ l~ N ~ ~O ~ 00 ~D N ~n ~n M 3 ~ oo N o0 ~ ~ ~ N d ~ M N H E'' ~ ~ ° N ~ ~ 3 d ~ ~ ~ d ~~~~~;~~dN~NOO~~~~a~0~~~~_~l~~z~~N0~1~
M M oo Ov ~ ~ 0~0 ~t ~ l~ ~ ~ ~ N ~ '~ ~ ~ ~ p ~n I~ ~ ~t N
~' 01 N ~ ~ M M ~ 00 ~O ~ M pp ~' '~ \O p~ ~ ~ ~--, 00 ~ O 01 l_~ O I_~ M
~ M_ M ~ O ~O O M M M ~ 00 M ~ 00 ~ ~ M ~ ~ ~ ~ V~ O~ M ~ l~ N
N
M ~ O 'ch (s., ~O 'n f.~ ~ Q '~ ~ d ~ ~ O
d~ ~~Mdd~3 o _ M
O ~_ ~ ~ ~-' ono ~ WO ~ ~ ~ ~ ~ N o0 vD
~ ~O O_ ~ M O ~O O ~ l~ N ~ 00 '"" M N M ~ ~ ~' ~ M O ~ pp ~ ~ N
~ 00 \p ~ V'1 M ~ r'~~., N M ~ ~ ~ 00 ~ ~ ~d ° 00 00 ~ ~ 00 M ~ ~O 01 V~
d~d~H~~wZd~dd~~dww~~~Z
~O O ~ N ~ N ~O o0 O o0 00 '-' c* ~ 01 vD O ~ 00 d M 00 pp I~ ~ ~ V~ ~p .~ M ~ ~~ l~ N ~ 00 O 00 00 ~ I~ ~ ~' 00 N ~ l~ ~~ 01 01 ~ 00 00 M ~ ~ ~ 00 M O O 00 00 d' O Q 00 M ,~
O M '~ op ~ l~ ~.,~ .~ O WO ~ W ~ N ~ M O d' O M ~ l~ ~ M .~
M l~ ~ ~ ~ ~ ~ O I~ ~p ~p M ~D V~ v7 ~ v~ W n '~ 00 ~O V~ v~ ~O 00 ~D 00 xUdw~d~wd~~~~~~d~~~d~d~~~~dd~~

~
~ ~ ~ ° '-' ~ ~
~p O O op ~n O ~' M N o0 M O ~ ~ M ~ ~ r' N N
N O ~ ~ t~ ~ M O ~ .~ ~ ,~ ~ ~O 00 01 N M ~O ~' d' O
Ooo3O~N~~ O~N~M O O~O~ M~~UOp~pMO
~ o o ~ rM, ~ ~ ~ U Q n-1 ~ N U d d U ~1 N °o o ~ '~ ~ o d v, oy o o ~ "., ~.; ~ N d r~ d ~ ~ d ,~ ~ d r~; o M ~ l~ ~ O ~ O~ U U V~ l~ V'1 p .~ M ~ N t~ O v~ M
0 0 °~ ~ oo ~ o'n'o c~~y~,, ~ N d d ~ ~C o a o ~ °o ~ ~ ,Mr, ~
°; ~ ~ ° "~ o N M ~ ~ ~ N ~ ~ 00 00 ~ N r.; ~ 01 O O .~ ~ O v0 p O p ~ ~ O
~ i c U ~ o d o 0 0 ~ U o U i U M
~° d ~ d ~ ~C ~ ~ d o ~ ° ~ o ~ o ~ d o d ~ ,.-~ ~ d o o_; ~; o; ~ i ~ u_i ~ ~ M ° N ~ ~ d '° ~ ~ ~ d N o ~ ~ ~
N V7 ~ 00 ~ ~ ~ M M .~ \p ~ M d O N '-' ~ O M N ~ ,-; ~ M ~ M
v7 M ~ M ~ ~ ,~ M 01 v7 O N .---~ ~ v7 O ~ ~ ~ 00 ~' ,-, N 01 ~ 00 M ,~ ~ N ~ ~ ~' V1 O V~ l~ ~ ~ ~ ~p ~ ~p Op O ~ ~ M O O
N ,~ ,~ U o ~ a ,~ ~ O d' Hdd~~r.~~~G"''~~~~°o~~d~~~~odo~da~~NddU
~ ~ o'''o ~ d ~_ c~i ~ ~ ~ 'o ~ ~ ~ o ~~, d ~_ c~ ~i ~ v, d ~ M ~ .-~-~ O 00 M O_ M_ ~ ~O M ~ ~ O O ~ pM 00 O 00 01 vp O N ~ ~ ~ O Ov ~ O l~ O O I~ O ~ O ~ O I~ O O ~ M O O O ~ O
.N-rr ONO o0 00 O
O ~ O O ~ O ~ O N O ~ O O ~. O O O O
d dd~~~~~~~~~~~N~~ddN~~ddaN

00 ~''~ ~ ~ wi ~,-; t~ '~ ° oWp ~t o 00 ~ ., ~ [~ 00 [~ M 00 ,~ ~ V7 M
M~~~ ~00~0~ oMONM~oNON ~0 ° N ° ~ ° O O Ov M O O N ~ oo ~ N O N ~ ~ N
N ~ °° M W ,~ ~ ~ U o U ~ ° o ~ o °o o ° ° a, o ~ ~ o o ° o ~.~,, o ~ ~ ~ ~ o ~ ° o d U d N d d ° d o°_o U d ~ o ~ ~ °
'" ~ ~ d a o ~ o ~ ~ °M° °o °o ~ ~ d o ~ o c,; d ~ ~ d "' ~ d ~
oo N ~ oo ci oo d o " 00 N pp ~O h ~ ~~ ~ o U U M ~." t~ N ~ ~t vi ~' N ~' ,~ ° ~n d ~ ~ ~ o ~''' ~
cri ~_ ~ d d ~ ~ ~ oo ~ o ° ~ ° o M ~ ~ ~n o o_o ~ o ~ 00 ~_M~MOO~O~OMOp°~°N~'OOM°ON~OO°N
O
~ ~ ~ Q' v ° ~C ~ U ~ a ~ ~ N d N U d d a ~ ~ o U d ~ ~ U ~
v "' w' ,~-~ o '-' t~ '-' d °° d o ~ M ~i M d t~ oo d .-. ,~ o d ~ ~ a; d ~
w ~C ~ M oo ,~, ~ ~ .~ ~wo ~ o ~ U M ~ ~
N ~ ° ~ ,~ d ~ o ~ ~ ~ ~ vN-, °° ° ~ v ~ ~ ~
~M., ~ ~ d o n ° N ~ ~
00 ° .-r o ,~ o ° o ° ~ 0 0 0 0 0 0 0 o N o, o ~ o ~0 0 0 ~ ~ o ~ N ~ ° o ~ o o ° 0 0 0 0 0 0 0 ~ ° o ~ ~ ,~ o o ~
~
~ M U ~ o a' o U oo U oo U U U U ~ U U o ,..~ U o °° U U
°° °o ~~d~~~W~d~dNd~ddd~ddU~d~~~ddUU
ri .-. ,-~ t~ o ~ .-. ~ ri ~ ~ ~ ~ ,~ ~ v-i ~ oo c~ ~ ~ d d N ~ V'1 00 N N o0 01 00 ~ ~ ~D ~ ~ ~ M d ~
M ~ ,.-~ 00 N ,~ ~ ,~ ° O \O ~ .-.WO vp 01 ~O N 01 O o0 p~ M v~ N ~
O_1 ~rj O O O ~ M ~ ~O Ov ° O O O N p O ~ p O M O ~ O O ~ O p p O 01 O O O ~ O N O O ° O O O ,-, O O ..~ ~ O .~ O ~ O O M O p °
O I~ N
~r~~~d°w~~~~~d~~~~~~~dN~d~d~~~~~

N O ~ ~O
O M N M ° O
O .-r pp l~ l~ I~ ~_ ~ ~D ~ ~d ~, O '~ r''> '~ pp ~ ,'~ N ~ ~ cd N N ~ O~ ° ~ O U d °x~. d vo o d ~ ~ ~ o°~, 0 0 0 'p ~ ~ ~n .~ E-~ ~ ° d os N a;
M M ~ ~ O ~ ~ .~ V' ~ a ~ p O
M I~ 00 00 00 ~ l~ O N ,~ ~ N d ,_; ~D ~ M
vW O ~ ~ ,~ ,_, O ~ ~ ~ ,_.., N N 01 pp O
O O M O rte- ~ U O ~ ~ ~ 01 ,b ,~ ~' ~ ~ 01 N p O N
'r' d 'o ~_~, ° M N "., N N M ° U G~) d d U ~ M ~ ate. ~ ~ ° p,,' d ° N '~ ~ ~ ~ ~ o ° d d oo~dooo~a,o~~ N~or~,, o~ ~~do~~°~N
~ o o r, ~ ~C ~ ~ ~ ° ~ U ~ U o °°°°~~o;o;~o; ~°d ~ d~ ~~~''~oo~ooo U U ~ U N OWE N ~ 01 ~ U b ~ ~ ~ ~ ~ °; o p°., U U
ddNd°~ o~°~~~ ~d ~~ '~~ x~xd~addd ~ ~ a, x ~ ~C ~C ~ yo N ~ ~ o o"'o o , ~ ~ o oMO N d ~ ~ °
N V1 N rr d' O N M N M ~ ~ ~ ~ l~ '-' ~ ~--~ 01 ~ M ~ 01 M O
p O N O M M Q\ 01 01 ~O O ~ O O ~ M ~ ~' ~ M ~ M ~n 00 ~ O O
O O O O pp o0 v~ v~ v~ '~t N ~ O O ~ O l~ IW p O ~D l~ ~ l~ ~ O O O
0.. U U U .~-. .-~ ~n ~n ~n .-. O ~ U a., ,~d d ~ ,~ ~ '° ~ U U U
dddd~~~C~C~C~~ ~d~~~d~z~d~~~~!ddd ~ 01 ~D N 00 01 ~ O
O O ~ ~O o0 N ~O 01 l0 M ~ M M
i i i ~ ~ ~ ~ i ~ N ~
O V~ N 00 ~ v~ O ~O
01 d1 M ~ I~ ~ V7 I~
~ N ~ M M
i i ~ i i i ~ i 00 O M 00 ~ .~-i 00 O
~ M a1 O ~ ~ N d' 00 Q1 01 0o O~ O ~D ~n O M Q1 I~ 00 ~ M
.~ l~ ~n v~ v~ ~D in N
Q\ ~
d1 O ~ N M d' ~ ~O
O~ O O O O O O O
N M M M M M M M
O O V'1 41 00 .~ 00 O ~l ~ d xxx x xxxx v-i ~_ vi .~ '~ M N <.j av ~W p ~p ~ as o t~ N o0 00 r.; O N Ov ,.~ V~ ~ ~ O N ~ O ~; ~O pp M N O ~ ~ M v~
M ~ M ~ O ~ ~ I~ ~p ~O O N ~ ~ ~ I~ 00 O ~ OWE 00 N ~ 01 N
O O ~ O M O pp ~ O ~ ~ O 00 O O ~,,~ p O ~n O ~ O 01 p p ~ ~ p p O
O V~ ~ O .-r O ~ V7 O ~ O O O O O ~, ~p O ~ O ~ V7 O 00 U o o U U o p U M ,.~ U ,~ U U o0 o U o o U °p U U o a a o dUUd~d~~d°o~ddddNUdU~UdoddUd~dU
o°oddovooMNdd°di ~ n~~odod~d n~~od~~% od 00 O O pp O~ M ~-, ~t ~O N ~ ~ M ~,.j v~ ~ M ~ O N l0 pp O ~ p~ ~ ~p ~D ~ ,~. ~ v~ ~' ~n O ~ ~ ~ I~ "' l~ o0 ~p y> ~ V~ N tn ~
O ~ M O O ~ N ,~ O ~ M O O O O ~ ~ O M .~ ~ O O O M ~ O O M
UNOUU~o-lo~U~~UUUU~o-l~U~~,...~~U~U~~-lo'~'oUUao d~UdddU~dN~ddddd~d~C~Nd°oddNdddU
~ ~ d °o o, ~ d o ~ ~ ~ ~ ~ ~ o't, ° °; ~ o ono ~ ~ d N N
°~ N ~ M d O ~n V_~ O "~ ~ ,~ 00 v~ M_ ~O O a1 l~ ~ t~ ~O .~ .~ V~ 00 ~ O~
O O ~ O M O ~ O O p O M M O O O ~ N O O p O ~ O Q\ O O O
O ~ ~ O O O ~ O O O O O O O O O O O O O O O ~ O O O O O O N
U M o U a~~., o U U U U ~,~ U U U ~ U U U U o ,~ U U U °p daUdd~Uddddd~dddd~ddddo~~d~ddp '~ d d oo v~ .~ d ~ t~ M o 00 .-: r; o c-~i ri o .-: .o 0o c~ ~ t~ a; tW o c_ri ~
\p N O ~ M I~ ~ M Vl '-r .~ 00 l~ O O ~r O ~ O O <f ~ ~ 00 O Uj N .-. .~ ~ v~ ~' M oo O~ N oo ,wp .~ .-a ,~ p O M ~ t~ ~ O pW O ~O ~ v~ d' ~ l~ ~O ~D ,.-. ~_O ~ ~n ~t I~ ~ ~ N N
~ O O O N O O ~ p O O O O O O O O O O O ,~ O O O O O O ,M_, ~NNdd~Ndd~ddddddddddddNdddddd~

N Vj N ~ v'i O ~ ~ yn 00 p; '".' 00 N M N N
I~ v~ op N ~ O O ~ M O ~ N
.b ~ ov ° o ° M o U ~
U o o U d U ~ d o°~, '~ ~ '° d .b ~ '~ ~ d '° d ~ ,~ o '~ ~ o w M N O U N ~ O M ~
o d~ t~ o ° ° 0 0 0 o d 3 00 o d U ~ ~ ~ o ° U o U 0 ~ M
d ~ o o°~o ~ ~ ~ ~ G4 U d U d d ~ M d b ~ ~ . ., N
003,.." o .b~d~da;ooo <°°,,a; ~o,--~yo 0o N ~ "' ~ o ~ ~ ~ ~ ~ ~ z °; ~ ~ o 0 00 p~pNpN~~~ O N MO
O ,~ ~ ~ ~ U ~ O ~p O M ~ N O O pp N o0 M
d "'O~d~ ~ o~o3~d~d~~ Md N
M .-~ d ~ °~ d °
0 o ri x o ~ ~ ~ ~ ,~ ~ N ~ ~ cx ~ o d vo x ~ ~
~O 01 N ~ v~ r-~ O O N . ~ r,., 00 ,~ v~
M ~ N ~ ~ V1 N ~; ~",~ ~ ~ .~ .~ M ~ l~ M N M ,-, ~ O~ ~ M 01 O O O ~ ~ ~ ~ ~ N M O N O O O N O ~ N M pMp ~ 00 O M
U U 0 0°°0 ~ d ~ ~ ~ °, ~ ~ U U U ~0-1 U ~ M ~
° ~ o M M ~
dd ~~zddr~zwdwddddd33Q~~~x~d ~O I~ M O o0 Qv ~ v~ 00 ~n .~ N l~ ~O o0 Ov N ~ ~ I~ ~f' l~ M ~-~ ~ O ~--~ v7 M ~D M 01 N N M M V1 M ~O ~ M ~ t~ ~ ~ M M
i i i ~ ~ i i i ~ i i i ~ i ~ i V~ V'1 V~ V'1 ~!1 V~ V7 V'1 V7 ~ V7 N N
N M 01 ~O d' ~ l~ .~ ~ ,~ I~ 00 M N d' ~
~ V1 M ~D ~~ O M 01 01 M N V> N 00 N N M M N M ~O ~ M M ~D ~ ~O M M
~ i i i ~ i i i i i i i i i ~ i ~ l~ N ~~ I~ M O O l~ O ~O v~ 00 O 01 l~
00 01 ~ M ~ ~O N ~~ ~ ~ O V7 N ~ M lr ~O ~O N ~ ~ I~ O M O o0 M 01 v7 00 M ~
.-r Q1 v7 N I~ ~~ <t t~ M 01 01 M ~O ~~ l~ N
~O oo ~ N oo ~ M ~n N O I~ oo .-~ Own .~
~ a1 ~ ~ a\
I~ oo a1 O ~~ N M ~ ~n ~O (~ oo ~ O ~~ N
O O O ~~ ~ ~ ~ .~ ~-, .-r .-r ~ ~ N N N
M M M M M M M M M M M M M M M M
M .--i M .~ O M O ~ M ~ N y ~ M O_ M
dd~~WOO ~~~~ ~~ WdC7 xxxx ~x ~x x x ~~x _ ~''~ N ~ N M O D
~ ° O N N M Wit' O .--r cYi p N ~ ° v~ d-M ~ ~ .~ O ~p ~O v~ M ~ ~ ~ p~ ° ~ O ~ 00 v~ v~
0~0 .~ M N ~ p O ~ ° ° ~ M ~ p ~ ~ ~ ~n O p O N O ~
o n ~~~VUMUU~°U°o°o~oUU°,°~N,°~
~_~.°',, dddoddvyd~VodUdd~d~d d 01 M ~ ~ \O U ~' O~ V1 00 ~ ~ ~ M d ~ M '<h ~ N .~
~ O ~ d O ~ M ~W d ~ N M ~ ~ p ~ ~ ~ N ~ W O ~ ~ N
M ,-~ ~.,~ ~ ~n M pp ° ~O ,~ ° M v~ O ° O ~ M O I~ O
O O O M N
v~; ~ N ~ U o ~ V o o U V o °~° o U o U U U ° ,°-1 ,_0..1 .~xd~ H~r~ 3~d~,~d~~dd~oododdd~dd M ~n N v0 Q\ N ".~ (, U
~ 'd ° ~ O M .~ d M N '"" ~O ~O ~ nj N p ~ ° d o0 ~ 01 O v'i Ov N ~ O ~ ~ ~ M oo .~ ,..~ O N '~ oo N M d ~ o0 N ~ O O ~n l0 ~ ~ O N N ~ d °M I~ ~ ~ d' O Oy''~ ~ N ~ ~ O ~t o0 00 Ov ~ Q°1 ~ ~n ~ ~' ~ v~ ~ .N-~ O ~ O O ° M O O 00 ~ O ~ O
° N M O O
~ ~ ~ -~s x ~ ~ -d ~ N ~ d U N U U U ~0-1 U ° o ~ U o~', 0 U 0pq .°-1 U U
0 0 ~ ~ ., o adz ~d~ddddd~Vad~~d~ddd O c~i ~ ~ ~n N ~n N N N N ~ ,~ O~ oo ~ d ~ 00 ~~ 00 ~ M
00 ~ ~ ~ M 00 '~ ~ ~ N ~ 01 00 ~ ~ M ~ O ~ ,-~.i ~ ° ~ ~ V'1 O ° ~ ~ V1 M ~D ~ I~ ~ ~ I~ v7 .-~ ~ 01 'n O N ~ ~ N ~ O M ~t p ~' v7 O
° M O ~ M ~ O ~O 00 M o0 O 01 ° ~ O O O M .~ pp O O O O O O M O
pp 01 01 ~ N ~ I~ ~ ~ ~ O l~ .~ ° O O O p M M O O O O ° O
° O
x~~dfxdadd~~d d~d~d~d~~Nd~~d~dd~

O ~ N M
~D d' i ~
~n ~n ~mn 00 00 ~n 0o N O N
M ~D ~h v~
~ i O ~O ~ 00 O 00 ~O d' I~ M O
~ O1 N
~ P
M ~' ~ ~O
N N N N
M M M M
'-r .~ \p M
O ~O O~ O
rx a z r~
x xx x ~n ~ .~ O ~ M ~-' ~ .. N ~ N .~ ~ ~rj o0 M
00 N I~ O I_~ .~ pp .~ ~ ~"'' N O .~ O N ~; O 'n o0 ~ O ~ O ~ d' M ~ ~ O N ~ ~ N M ~ ~ ,~ ~D N o0 O ~ ~ 01 ~
O M M O 01 ~ O O N O O O N ~ p O O O ~ O ° M ° "' O N ~ O
O O O ° O O O O M ° O O O O O O .-r O ° ~ .~ ~n O
U ,~ U ,~ U U U M N U U ° a ~ U U o oM, N o U ~ a ~
°° U ,...~ M U
d~dddddd~~'dd~d~dd°No°d~dNUd~od ~ ~ ~ <t o a; a; o d vo 'o r.; M 0N0 ~ N d ~ d d oNO o ° ° d ~
o°°o ~ o~°o 00 ~p oo M v, ~n N ~ U ~' N ~ U M
_~n N oo M d' N_ N M O ~ ~n N ~' ,---~ 00 O ~t WO
O ~ M O O O O M d ~ O N ° O O M O M O O M O O O d ~n °p O ~
O
O ° O O O O O O ~ ,N-, O O O O O r., O O ~ ,-, O O O O N O O ~ O
U ..-a a U U U a M M a ° U a U o U ~° o~ U U U °
° U N U
ddddd~dd~°~d~d~ddUd°o~ddd~_U~dNd i M ~ t~ ~ 00 I~ ~ N O M d' ~-i .~ ,~ d V~ ~ 00 \O ~"' r' LV d N ~''~
~n N '-' ~ 01 N N ~ O M ~ oo N ,~ ~n ~n ~ ~ ~ .. ,~ N v~ 00 O N ~ ~ 01 ~ ,~ N ~ ~; ~ d' O ~O O ~ M ~; N 1~ 01 ~ ~ ~ '"'' N N M M
~n N ~ ~n O ~n l~ O1 ~ ~ 01 N ° ~n ~n N '-, ~ v~ ~ Ov N ~ ~ l~ M O ~' O
~n N ° O ~ O ~ ,~ O O O p~ V7 O O O O
O O O O D O O O ~ ~ O O '~' O O N M ~' ° N ° O ° O
o0 N O O ° O
U U U U U U U ~t o~ U U ,.~ U U ,~ a ~ U a U U U N ° U U
dddd~dddaadddddddNdodddd~U~d~d -. ,-. Q\ ~' ~ N ~ M d ~ ~ vW O o0 t~ ,--~ N N N ~ ~ ~j l~ ~~ ~t d O M
00 ~ N .-, ,~ v~ ~ O 01 v~ p~ 00 ~ 00 I~ Q1 N ~D p O d' N Q\ l~ 00 01 ~ N ~ \O ~ ~ ~ N N ~' ~ ~ ~O N o~o oM0 ~ d' '~ N p pM ~ ~ ~ V' 0N0 ~O ~
O M O O O O ~ (~ M ~ O O M O O (~ O O O M O ° O O O ~n O O O O
O ,-, O O O O ,-~ .~ ,-, ~ O O O O O O O O O M O O O O O M O O O O
~~~dd~~~~~~~~dddd~~Nd~a~~~~~~~

ri V~ N V~ ~ d' 00 '-' ~ ~O ~ v1 O ~O ~f' 00 00 ~ 00 N ~ ~ 00 00 ~ ~O p~ 01 ~ M ~ N ~ 01 v~ Q\ ~ O
00 ~ ~ ~ ~' ,~ M M pp 01 M l~ N N N N 01 N v1 o U °; ~n oo M ~ '-' o°~, '° ~_ ~ ~_ o ~ 0 0 0 ~ °
o M ~ ~ .~ ~ ~ .-r ~ r"'i o: d a " M " o, N ~h ~ d '-' ~ ~ oo O~ .~ ~
M M ~ N N ~-i .~ v>
'n ~ M ~D p~ '~ O p~ ~ p~ ~O M '~ ~ ~ d' V~ ~ O V1 °
M ,-, I~ ~ ~ V~ ,_, ,--r d1 ~ a V ~' ~ ~ O~ M N ~
Op l~ O O ~ N ~ ~.-~ a .~ d' ~ d M ~ ~ .M-r ,~ O O O
U ~do,'''~~~~d~~ao;~aoaaa~..~
ddd ~~~~ o adM~daddd~~
N ~O o0 N ~ ~' ,~-, M ~ N M d ,Wp M M d o0 N N M ~O ~ V7 00 O ~n oo ~ ~D O ~ ~~ ~p N M ~t ~t oo M .-~ d' .-~ O M D1 N
~ t ~ '~ ~ oo ~n ~ ~ a N ~ ~ ~ M ~ ~ ~n p ~ ~n a\ o d' d~ N
N M O ~ ,~ V~ Q\ 01 d' V7 ~ N 01 M ~ M
°o°oo ~~d'-'~~~bb~'3a~y~~,..~~~,,~oMO~ooo~~
b~d~~N ~ ~~dd~~ o~~~dd~~~°~~
o 'b c~j t~ 01 wi ~ ~D ~D ~ ~ ~ oo ~ ~D ~ N d ~''~ ~ ~ N ~ 00 ~n O Ov ~ ~ ~ ~ 01 p p pMp ~ ~ ~ l~ M D1 ~ 00 01 M <f N ~' M O ~ V~ V'i ~ Q\ 01 O d' .-i ~
p ° ~ o0 ~t ~''~ M ~ O 01 ~ oo l~ ~' ~ 'n ~ ~ ~ M ~ ~ ~ ~ l~ 01 ~t N
° O O Op ~ ~ ~--W~ O ° 01 N ,~ M M M ~ M ~ ~ M ~ ° O O ~~
~ N
ac» ~ d~~Md d 33~a~aa~a,...~ aaaao~~
ddd~H~dd~H~d~dd~d~dd~d~ddddd~~
o\
00 0, o ~ a, l~ ~ M ~ M ~--, ~ M .--i V~
i i i ~ i i V7 ~ ~ N V~ V~
~D M ~n N
~O M ~ ~~ N O
,~ d~ M .-i ~ ~ i i i O v~ O V~ d' ~O
~' 01 ,~ M ~ V7 I~ v7 O .
N O I~ ~D

l~ 00 01 O ~-~ N
N N N M M M
M M M M M M
~ C7 ~, P~ a.. O
xx xx x x N ~ .r ~ rM,, ~ ~ O cd Q\ ~
o ~o~o ~ ~~ U~p ">~o ~ ~d~ d~
d °' ~ "' N ~ ~ .D b ,-.
O M '~ O M ~ r~-~
°' oo ~ 3 c~i ~ ~ o o r~, ~ o~ n o " d °"-,° ago ~ c o o ~~~o o ~-1 cd ,.~ ~ ~t U
d d wi ~ ~ ~ ~ ~ ~ ~ ~ b ~
c~i ,~ ., c~ t~ r..~ ~ ~ r; .~~. M ,~ ~ z N ~
x~o~bM~
d . d ° ~ ~ . ~ d ci o d o; ~ .. ~ ~ '~ ~ ~ d oo ~ ~" ~ ~ W o 00 ° a; r~ ~ ~.., "., ~ .~ ~ ~ oo d ,~ d z ~r vO N N O \p 0o v~ N o0 pp yp oo p M M op ~_ N M M M p~
(~ v0 oo M vp p~ ~ ~ ~ M ~ l~ ~ l~ ~ ~ ~ ~p O~ °N O ~ ° ~ v> d' O
N [w o0 ~ v~ O '-r ~ ~"' M o0 N .~ d1 l~ t~ 00 p~ d ~ O O
o_~ ~_ 'o ~_ ~ ~n o o_o -b ~ M ~ ~ o ~ ~ ~ ~ oo ~ U
dddxd~x~x~~ d ~~ddHr~~HN d~~ ~3dd M M M I~ l~ M ~ 01 M V7 .~ M (~ M M N
O I~ 00 V7 M d' 00 00 I~ ~ 01 l~ ~ 00 r-r \O
M M ~O V7 \O V7 ~ M \O V~ M M d' ~~ ~ M l~
i ~ i ~ ~ i i i i ~ ~ ~ ~ i N N ~ ~ ~ N ~ ~ N ~ N ~ ~ N N
l~ O~ 01 01 M N Q\ l~ ~ 01 .-, l~ 01 M d1 O~ 00 N o0 ~n ~O ~ N N ~O l~ v~ M I~ ~ M ~ a1 M N ~O V7 ~O V7 ~ M ~ V7 M , M V' ~~-~ ~ N
i ~ i i i i i ~ i ~ ~ ~ i ~ i i ~t M M V~ O V~ 00 ~O ~h ~~ O v~ N ~--~ 00 O
d1 01 l~ ~ N V7 .--r ~ 01 ~ M ~ v~ M 00 O 00 l~ N O o0 N l~ .~ d' l~ ~' O o0 M 0o ~t M
00 O d' 00 41 N ~~ ~ ~ .~ 01 ~ O M o0 01 I~
M l~ ~ l~ ,~ N o0 ,~ l~ 00 00 N ~ l~ 00 I
M <h V1 ~O l~ 00 01 O ~~ N M ~ V7 ~O l~ 00 M M M M M M M ~ ~ ~' ~ ~t ~t ~ ~t d' M M M M M M M M M M M M M M M M M
N o0 N ~n N vp M ~ N M ~ ,_, O ~ O N
00 ~D N O N N ~O N oo ~ ~ ~ ~ Ov Q\ I~
U U U U U U ~U ~U U U U ~ ~ ~ ~ O' x x ~~x ~xx ~~~ xx xx xx o t~
M wj M ~ M ~ V1 ''.~ .r M ~ ~ p~ ~O M V7 v~ 00 ~ ~' ~ ~ N
'-' O °N O ° ~ ,~1 ~"~ O ~.]
'b~~~N O~~ ono ~~~Nd~d w~
I~ ~n f~j pp ~ N 01 x ~ ~ 00 ~ ~ E-. d ~ ~ ~ d ~ . ~, o; a, o ~ ~.~i ~n ~n N
a~ .~ ~t yp '-' ~ cM~~ ~ ~ ~ '~' 'n ~ ~ p o0 000~z~~~M~ ° ~O ~~~~°v ONON
N N oo O ~' M ~ ~ N ~ ~ O l~
o ~ ...~ ~ N ~ o ~ N ~ ~r d d d x~-~ ~ ~~3 ~dd~~
d -~ .-: d H
dv;~~z~~°NO~o~~ ~ . ~ ~ ~N~
M O
M~ O O ~ ~ ONN~~o~o~~~D
N OO~_ ~MM~ ~ ~ ~ M~~~~N
N~~~Md~~~N ~ 'd ~~~ d~~d~~~ddd MO~~~''~'~00 ~ ~O ~ ONO~_O~_~M_a;~pO~M_~
p v> ~ pp N O~ ~ ~ oMO o0o O ~ ~ can o'~o U ~ ~ O O ~ ~ D N ~ ~ oo D
0 O 01 ~ O Q\ M ,-, ,~ N ~ O ° D1 ~ ~ 00 ~ 0 d' N ~ I~ ~ N
~ N M ~ ~ p M O ~dt ~ O N ~ O ~ ~ ~ M M N ~ ~ ~ N
dQd~r~da;d~~ ~wdHd~ ~d~~~~~~~d ~ ~t t~ O v~ N v~ 'O o0 M l~ l~ ~n a\ t~ .-~ O O O v~ ~ M ~~
V7 l~ ~ V7 M ~ M ~Y ~ N
i ~ ~ i i i ~ i i ~ i i ~
N ~ ~ ~ N
O O M \O ~ 00 ~ N ~ 01 M M
00 \O O 00 01 00 ~' V7 N Q1 M M M
V1 V'1 ~D M ~ M ~' M M ~ N
~ i i ~ ~ i i ~ ~ i i i i N 00 ~ N U1 00 N M ~ ~ a1 00 Qv ~ l~ '~?' Q1 v~ N Ov N N o0 O O
~ O ~n N M I~ O ~ l~ ~ ~O ~ M
M N t~ M Q\ ~O ~O M ~O ~ O V~ V~
O 01 00 l~ M ~ l~ v7 .-~ M ~D N V~
~ N ~
O ~ N M ~t v~ ~D t~ 00 O~ O ~--~ N
M M M M M M M M M M M M M
M ~ M 01 .-, ,-, t~ ~ ~O ~O M pp N ~.a°~°xA~
www~www w~w ww xwww ww~
xx x xx~ xxx xx M N ~p l~ ~n p~ ~ O ~ v7 N O ~ ~ Q\ O ~ pp I~ N 0_O ~ V1 l~ ~ ~ M
O O o~o ~ ~ N ~ ~ due' ~ ~ ~ ~h ~ N O N ~
ONO N N O t~ O ~ 01 ~ M O O '~ O 00 ~ d' ,~-, O O .~ O O O
N O O O ~ v~ ~. N p O O O ~ o0 l~ ~. O O ~. O O ~ O op N ~ d ~ ~ d a x O O ~ Q,' d d O O ~ ~ d x d d d O
N o d d ~ "' U ~ ~ ~ a; o ~
~ ~-~ oNO r.-, ate., U ~ 0 00 ,MO" o ~ ~ ~r vNo d ~ ~ a, 0 00 ~ ~ ~ o, I~ v7 ~ M Nl~ M M ~ ~ O O N ~n 00 M ~ ~' ~ '-' O ~p ~ M
N ~ O ~ O ~w_n O ~n ..~ ~ t~ N O N v~ ,~ oo M N I~ N ~n .~ O ~ M
O O O O O O ~ N O O O O ~ N O O O O O p p ~ O
o~o U N ,~ a U a U ~ oNO ~l U '-' N ,~ U U U U U U o °~~dMOd~ddddNN~dd~~a~dddd~d~~U
wo ~' ~ o ~ ~ ~ ~ r; ~_ ~ ~; ov ~-: 00 a; d r; ~_ o ~ ~ ~ c~i ~ ~ d 0~1 ~O ~j U ~ I~ ~ N d' O 00 ~O ~ O N O ~O ~ M O a\ O ~ M M ~ ~j 0 d O O O p M O O O O O O ~' N ~ ~ O O O O O ~ p O p ~' U ~ ° ° 0 0 0 0 .-~ 0 0 0 0 0 ~ ~ p o 0 0 0 0 0 0 o M
M ~., ~ a U U U ,~ U ,~ U U U U o ,~ U U U U U a p.., U o 'ad~d~~ddd~dd~d~ddd~UdddddddddU
N uj N N ~ 00 ,-.yj o0 M ~ l~ ~ 00 0o O~ ~ ~j d .~ O O M ~ N ~ ~j O Q~' ~O 00 M p ~O ~ ~ ~ ~ ~ p M ~ O l~ Q\ M ~' N ~-a p1 M
~." ~n .--, 01 O ~ O M .-~ r-.~ O v~ ~ ~O ~ l~ v~ N ~ O Q1 00 O o0 O ~y o0 p"
'-' ~ pip N ~ O a1 M w0 ~n v~ oo OWO ~ ~ ~D 'n ~ ~n v~ vi ~n ~ ~_O
pMp v1 O ~ ~ ~ O O O ~ ..M~ O O ~ O O O O N ~j p O O O O O O p O
x~~~~Na~~~ddd~d~aa~~~~~d~dddd~

N ~ 00 Vj wj 00 Q1 I~ Q~ O '-'~ ~O N M M ~ Vj M

00 00 N ~ v~ ~O l~ ~ 00 ~ v~ ~ N N ~O N ~ N ~O
vi O ~ N ~ ~t ~ O ~ oo '~ I~ O ~ ~ 01 O M

O p ~ ,~ 0 ~O o0 ~ oo '-' ~ ~ ~ O pMp O p ~ O O O
O
~
~ O ~ O O 0 p O p M
N p O
~
~

p O , O O ~ N .
O O O O
oUU ,-a UUUo~,~oi ~'~'oU ~oUMUUUUUU'n d~
~
d ~

~dodd N
ddd~
oodd~d dd~UddUa N ~ U p N
d ~
~
d ~ O
M ~

O lp OO
~O
M ~ ~ Q1 l~ V7 00 M
O
yp M v0 O N O d' N ~j ~ vW D p ~D ~ ~ c~ p1 r. O
N ~
o ' ~

~n v-, ~ d ", ,~ ~ ~ ~ "-, d ' N ov o N ~ ~ ,~ v-, ~ ~ ~ o, '~ M ~
O ~ O O ~ O O ~ O O M

O O ~ O O O O ~ O ~ ~ O O M O O ~ O ~ O O O O
ono o a <.N., U U a U U o U a o U U ~ U U ~ U N U
U U U ,.~ ~
~
~

dddddUddU~ddt~ddNd d NUd dddd~

~
o o d o a; o r~ o o wi ~ ~ ci ~ d ~o ~ ~." 00 o M ~
~i N vo o a~ c~; ~ N N

00 l~ ~ U 000 t~ ~n ~ N N " O ~ ~ ~ ~O 00 O t~ v~
~ N v1 N .-, N .~ M M O l~ v7 ,-, v7 ~ .~ .~ ~' N pp ~n <.j M '~
'~ ~ M
M v~ v~ d M v'1 O~ 00 ~ pip N ~ v~ ~' ~ ,~ V ~D l~
.-r M l~ M N v~ O O O M

M o0 N O ~ O O ~ O O .~ N O N O O O N N ~O N O O
O N O O
O O ~ O O O 0 0 ~ O O M O 0 O O 0 O O o0 O O O
O N

a ~ U U ,~ U ,,~ U U U o ~-7 U ,..a U U U ~ ~ U U
d~ddo'~odd~dddddddddd~d~C~dd~~~o N v-i ,~ v> r; ~n ~ W p ~ O oo ~j ~j N a1 M O ~ OW
~ O vp ~
N N M I~ p~ ,~ ~ ~ ~ ~n l~ ~ l~ 00 ~O ~ d' N ~ M O
d' N ~ ~ v~

00 N O d1 pp N O ~' ~ ~ 'p" ~D N ~p M M 01 ~ ~ O ~
I~ O O N d1 v~ N ~D
v~ l~ v~ ~p M .~ O N ~ t~ .-r .--. ~. N w .-i N M
~ ~ O M a1 V~ .~ ~O .

O O O O ~ M O M O ~ M M ~ O O M Q~ O N M O O O O
O N O N
O O O O ~ O O p O ~ O O p~ O O .-a O O O O O O O
O p O .~ N

dddd~C~~dddNdd~ddd~d~dddddd~ddN

~° ~ o O N ~ O 'ct ~O O
N O~ ~ ~ ~O ~'~ O
p O ,--~ O~ ~ ~ ~O ~% ~ v~ O ~ ~ 00 ~ ~ 00 N
t''i O M ~,M,~, M ~ ~ N ~ v~ ~ d OW ~ t_~
d ~ O O ° yo d ~ ° ~ d oo d N ~ ~ ~ o 0 M~~ ~l~ ~oM0~~0~1 d~ ~ OM O
~Qv~~N~NOp~p00 N,M~ O~
~ ~O ~O ,~ ~ ~ ~ ~ p ~ O d ~D ~O v'1 t~ '-' O 00 ~' '~ M O O O_ ~ ~b .~ M U I~ N 'b I~ I~ ~' 'b ~ N M o ~ ~ 3 d ~ ~ ~ b d 3 '~ ~ ~ o ~~z~3~d~ ~d~ ~ ~~M dd ~ ~NM ~;d O~ I~ ~O 00 M N
ONp~p~p 0~1~~ Nit N ~N O
p O N N ~ ~ M ~ ~ M ~ N ~ p O M ~ ~ O p ~ O ~ O
O N N I~ ~ ~--~ ~ O O ~n ~ a\ ~n "~ O M
~~~Nx~x~N~~~'~ ~ ~~ ~,~xd dd ~d ~Hb d I~ t~ t~ N .-: ~p O oho N ~ ,-. t~ M oo ~n d O ~ ~ ' '' O o0 M .~ N M M
l~ p r-, 00 N ~ ~ N ~ O N ~ ~ O O I~ ON1 N ~ N
O w ~ t~ ,-m0 ~1 ~O ~ ~ ~ ~ N ~ O O ~O O 00 O ~ M N ~ .~ N r-.
N ..~ ~--y~ ~n ,'~_, 00 ~O pip O O O ~ O I~ .-m_-~ M ~ v~ O
p O ~ ~ O O O ~ ~ O N O O I~ ~ ~ d' ~ ~ ~_ N ~ ~ ~ ~ O ~O
~dxd~~~~d~~dddddd Nod H~xH ~~d Q\ Q\ O O ~ O a1 O~ 'O I~ ~n ~ ~ N
Wit' 01 ~~ ~ Q1 N N O M l~ I~ l~ ~ ~ N
~O N M ~ M M M ~ V~ M N
i i i i ~ ~ i ~ i i ~ i i ~ i V1 ' l/~ In ~!1 In In V7 V7 V'1 V~ V~ V7 In In V7 ~n ~w0 ~ O ~D ~n ~n N M ~-r N O o0 M o0 01 N ~ ~ O Q1 N ~ ~D \O ~ O O
~ N M <t M M M .~ ~ M N
i i i ~ ~ ~ ~ i i i ~ i ~ i .~ rr .-~
00 ~ O M ~~ N N ~D l~ ~t .~ ~O .~
M N ~ 01 M N l~ M ~ M M .~ l~ ~ 01 ~O ~~ O ~n l~ ~ ~ V'1 ~ ~n W t~ ~p .~ V~
O N ~ ~ 00 t~ N ~ 01 v~ v~ M ~ O~ ~O
O N ~-~ O ~ M l~ l~ M ~n ~n I~ ~n N Q\
t~ t~ Own v~ ~n ~ O~ n o0 0o a1 ~ v~ t~
M ~ ~n ~O l~ o0 01 O ~-~ N M ~ ~n ~ l~
M M M M M M M M M M M M M M M
v1 ~n I~ ~ ~ N ~ WO N ~D
d d d ~' '~' z z N .-~ ° ~ ov x ~ ~~ xxx xx xxxxxx O ~ ~ ~ ~ ~ ~ N p ~ N
~O OMO ~ 0~1 ~ ~ ~O ~ ~ ~O ~ ~ ~~., ~ ~ ~ N 00 l~ ~ ~ ~ 000 O ~ ~ O N M M ~ ~ ,~ ~ ~ ~ O ~ l~ N ~' V1 O M p~ M
O ~ l~ .~ ~, 00 ~ M ~n N ~ l~ ~O O M O .~ ~ M p"
'ch O ~ Q\ 00 N ~' N '-'' v0 '° ',~° ~ N ~, M
U d d ~ r, "; ~ ~ ~ ~ ~ d x N ~ ~ ~ ',r,° ~ ~ d o N TWO ~ ~ ~-i cue; ~ r; ~ ~r d~~' °~ '~°'o°'~°
o~dN°~~~'~°Mo~~d~dao~o:~
~D ~ ~ M ~ ~ M N ~j ~t v1 ~ ~ ~' ~p V~ O ~ ~ N o0 O~ O d' O oMO 000 O O . N ~ ~ ~ I~ M ~ ~' ~ O ~ ~ .-, .-~ I~ ~n ~O O N ~ ~t vWD
O ~ b d Q\ l~ v~ ~D ~ O M M_ ~ v~ ~ a\ ~O ,~ O ~ v7 N
° ~ d '~ d ~ d N ~ ~ ~ ~ ~ ~ Q ~ ~ ~ ~ ~ o ~ ~ ~ ~ d o, d °° ~ ° o 'r,° o °~t o d oo d ~ u; M ~ ~
oo ~ ~ d ~ '-: ~ '°
vp t~ O .. N O oo ~ 00 ., ~ ~ N ~ ~ O x Q> N U M ~ ~ l~ M M ~ l~ ,_, ~D N ~ l~
~ v~ ~O O O N O O oo ~ M N O o0 O N N M
0 O 00 d ~ M ~ ~ M ~ M t!~ V'1 ~ ~ pNp M .~ ~ \O ~' 00 ~ 00 00 p ~ ~ .-~, O O b O I~ M ~ M ~ ~.~ t~ d ~O
d~~d d~dx~~xdddd~~dHdHdN~d~
d ~ o; v; ~ oo Q: M a; N o ~ cri ~ ~ ~ ~ ri ~ ~; ~ o d N ° ~ d t~ O ~ M 01 ~ d' N ~p ~' 00 N .~ M o0 M o~0 ~' ~ ~D ~ M ~ O N N
p00~'oNO~~~~~O~~N~~,N-,N~O~~~MN~MOM
.-a O 01 ~ ~ M 00 O O ~ M .~ 00 N \O M V7 ~ ~D '~ O I~ d~ d' ~ V~ 01 Nddd~~dd~d~ddd~d~~~d ~dd~~4~d~r~;
N Ov M
O~ t~ O
M
i , In ~ In ~ D1 M
M .~
~O ~1' N N
~ Q\
0o Ov O
M M M
O .-~ v7 x xx ~ oM_o 'r' '-' ~ oo ~ ~ ~ ~ o p ~ ~ o N ~ os ~ O
t~N~~N~N ~~'-' M~O~p~~N ~M~'N~
O M ~ O O O O ~ ~ N ~ pp ~ M O O O O ~ O N O ,.M-~ O O p ~ O ~ 00 ~~~dd~d°°~NNo~°, N° U addc ddddddddN
~a~d~d 00 O ~n ~ cri ~ ~ ~ ~ N ~ oM,~ ~ a ° ~ ~ ~i ~ o ~ o "~ ~ N ~ ~ ~ ~ M N ~
M~~~~'~°,°,°o"~'OO°odMOOp~~~~N
N ~ ~ O ~ O O ~ O O O ~ O ~ O O O O ~ O ~ O O O O O ~ O O O
~ ~ ° U ,°~ U ,~ o U ,~ U ~ U o U (> ,..a U ,°~ ~ ,..a a U U ~..a r~., U U U
d ~ddd~Ud~dodUddddd~oddddd~ddd l~ N o0 0o d ~ O M O~ d ~-' M M O1 op" .-r N l~ v'i ~ O~ ~ O y0 ~ t~ O pp d1 ~ ~ 'p N N I~ .--~ N ~ ~ .~ N N l~ D1 I~ N o0 01 ~O ~ N N v'1 ~ ~ M ~ N O N ~ ~ ~ ~ ~ ~ ~ ~ ~ V7 O ~' ~h ~ v~ ~ v~
pOO~,M-,,OpOOtn0000~0~000000000 U U U o, ,~ U U o U ~ U a U U ,~ U ~ U U U U U U U U
~~~ddd~ddd°d~dddddd~7ddddddd~d ~i .~ ~ oo wi oo ~ I~ ~ U p~ t~ 0 00 00 0o p .~ 00 00 ~ ri o ~-! wi oo Vi os 01 ~ M p ~ ~p ~D Q~ 00 N d ~ O~ 00 V7 U M ~ d' M .-r d' 00 V7 O N p~ M N
M N ~O oo y0 oo ~ ~ N l~ ~tW p O ~O ~ O ~n O m N O ~n op ~ N ~-~
O t~ M l~ t~ V ~ ~ M ~' ~ O ~ ~ ~ O O O O O O O N p O O
O O O .-~ O M O O O O O
N ~dO p O O O ,~ O p O O O O O O O O O O O O O O .-~ .-~ O O
ddd~~ddddddNdddddd~dddddddd~dd 0M0 M ~ ~ ~ N ~p ~ O o0 N ~ N M
V7 ~ l~ O N O I~ ~ ~ ~ <''~ 0 ~ ,~ ~ ~ ~ ~ ~ ~ N N ~ ~ h .-i O ~ D o0 O O O N O ~ O pip O p O O O O ~ O N ~ ~ ~ ~ ~ p vp odoo~ndddod~dNd~d~dd~~d~o~dd~NN
d M d ~ N ~ N N d ~ M p ~ N ~ N ~ ~ ~ .--' ~ ~ O U
M O~ N O oo O ~ ~ ~ ~ O ~ ~ M ~ M O~ N U ~' N v0 ~
~ o, ~.-, .-~ ~ ~n M .~ ° ~n d 0 0 .-, d N ~ ,--~ ~n ~ ,~, o°~°o~~°o°o °°o n°p~°o°°°o°o°oo~~oa;'~o.degree .o~ooo N U p ~ U U M U ~ U ,~ U U U ~ U ~o M U U ,~ U U ,~
adU~N~ddaddd~d~dd~~~~d~~dddddd ~' vD ~ '-' ~ N d ~ v'i ~D o0 t~ N M ~ N M ~ ,~ Ov ~ N uj .~ t~ ~ oo ~p [w ~ M ~ pp V1 ~ y~ N O M .~ N ~D l~ V7 .~ ~ ~ V~ ~ ~ M
~ 0~1 ~ ~ ~ N ~ ~ ~ ~ ~ ~ N O ~ ~ O O O ~ N o0 p OOp~MOpO00000~~~pO1~00000~00p o U U U U a o U a U U U U "' U ~ .-~ Cs, ,~
Udo~ddddUd~dddd~d~~M~d°,~,'~dd~dd~
~' l~ ~ O N_ O ~ 01 d M ~ O N N p~ ~ ~ 00 N ~ N M N M ~D ,-, ~ ~_ 00 pp m ~ ~ p ~ ~ y0 00 ~ ~ ~ O ~ ~ ~ ~ O N ~ ~ ~ ~O r-. ~O
M ,~_, M O O p N ~ O M O ~ O ~ O M O O ~ O O ~ O O M O O O M
N O N O O O O O ~ O O O O O O O .~ O O ~ O O ~ O O O O O O O
NdNddd~~NddddddddddNddNddddddd vo o_o vo °,.,.,° ~ ~ ~ o of Wo ~ o ~ ,~ o MO ~~O M ~Oo~OONO~~N00O
O M ~ ~ 00 01 ~ ,~ 01 t~ ~ 41 V7 ~ ~ O N
O ~ O O f~ ~ ~ o°o ~ ~ ~ O O O O O p p p o0 00 ~ O
U o ,..~ U ~ '~ d r"'., ,~ ,...a U U U U ° o oMO U °o dU~d°°~ N ~ ~°pdd~dddd~UNd°
O N ~ ~ ~V ~ 01 ~ ~ ~ N ~ ~ '-' d N N
M pp" O Ov U ~ I~ N O ~ ~ ~ O M Wit' ~ Ov ~ vj ~' t~ .~
O~~d ~M~ U ~~OOOOMM~'-'~pOO~p O ~ O p O ~b ~ ~ O p O O O O O O .-. 0 ~ O O ,n a o U ,~ ~ ~ ~ ~ "-~ d 3 M U U a ,.~ a o U U o dUddN~ ~ddo~ dNd~d~d~~d~ddU
N d Q1 0~0 ~ ~ ~ ~O '~t ~ ~ ~ M ~ 0~0 N OWO ~ N M ~ O~1 ~O d (~ ~ O M a\ l~ ~n M ~ 00 ~ ~ M O O ~ V~ N ~ O O N
00000 pNOOM~N ~~~O~OOpp00~~00 0~00~0 ~~d O pNp~0000000~NOON
a, U ,.,~ U ~ d d d ~ ~-~ r~-, U U U U ,~ U a ~ U U o dt~~ddd "d~.~x d~~dddd~ddNddU
d~ a; oo ~ ~ vi N o t~ ~ o_ ~i ,-. M ,-: ~ o v-i ~ cri ~ o ~-: oo d cNyp~~ p1~ ~~~ ~ ~ ~ N~O~~o~oo~o~ ~OM~~pp V1 ~p ~ O ~ .--. M ~ ~ v7 ~ M ~ N M ~ M N ~ ~ l~ ~' ~ N ~ v7 O p~ O p ~ N p M ~ ~ N ~ N N ~ ~ M ~-' O O M O O O O M O O O N
O ,-, O p ~ O ~ ~ N ~ I~ ~ Q\ O ~ ~ v~ N O O D O O O O ~ 0 0 0 N
d~d~ddx~dddH~dHx~dddddddddddd~

\O M a1 W O O
00 ~O O M .~ V7 d\ O
I~ ~ ~ M ~ ~ M V7 ~ i ~ i i i i i V~ V7 ~ In V~ N N ~
O .~ V'1 O~ ~ ~~ N ~O
I~ ~ O ~ O ~ 00 00 I~ t/7 ~ M ~ ~ M
i i i i i i i i M ~n N ~ O~ oo O O
l~ l~ l~ d' ~ ~~ N I~
V~ d' ~ ~' v~ ~n O M
t~ O ~' 00 Q~ 00 M O
O o0 ~n N M I~ a1 N
l~ 00 ~ t~ (~ ~ l~ ~
N M ~ ~n ~D (~ 00 M M M M M M M M
t~ ~O ~ M 01 ~ ~ ~O
M ~~ O ~ ~ N ~ N
xxx xxxxx N M
~ p 0 ~
~

N N O o0 O O l~ ~
l~ ~~ O v0 0o O o0 Q~ Q\
'p O

vp ,~~. ~n ~ N ~ I~ I~ a\ O v~ ,~ .~ N
v7 op O O o0 pp .-. 00 d1 O O M ~ O O O O O

o ~'', '~' o o .~ o o ~ U M ~ N
.-~ o ~ o ,~ N o U U ~ ,~ U ,..~ U d M U ~ ~ M
~
~~do ~
dd~

d b ~ l~ ~ oo O
dd ~
~o O U N

N ~ oMO ~ ~N
~O
N~~~

o ~ ~ ~ ~oNO"
O
od~"~~cM~io~~ao~U

.-. 0 0 0 ,~ 0 0 0 0 ~ o ~ oMO N ~. N ~ o .~
d O oo ,~ ~n ~
N p p O .-. O O O O 01 O ~ ~ O
a ~
U U U
'b ~

U U N ~ "-, ,.. d o ,.a ~~
~
M

oddd ~ -d ~ ~ ~ M
~'d ~ ~ ~ .; o do ~ ,.a dd O
o, U ~ ~

~ ~ N ~ ~ r N ~ p~ --~ O M ~ V7 01 .~ t~ 00 l~
N d w' d ' a, o N M ~ ~
.-V o0 0 ~ owt ~r o~ ,~ ~

N O ~ l~ 01 O O ~ p N ~ y p N O O N
~O ~ I~ l~ N ~ O O V~ I~
N ~O M Q1 ~ ~n ~ N r" M O
N I~ ~ ~ O ~ 01 .-~
~ V7 '~ M
O ~ O M O O O O O O ~ O M M

o~opoooooo~o a, '~ b oo3'N
o , ~ ~ a, "' U a U U U U U U o U ~ '~ ~ ~ ~ a U ov N ~ ~ d d d~o d Z
dddddddN
~

d M a d ' M N ~ ~ N
j ~
~
~
~
' ~

c~ ~ N ~ 0 01 l 0 M
M
n ~~ d ~ M
N N
~O ~n 00 00 ~p v0 N oo l~ O oo v0 ~ ~ '~t l~oo l~ O M
Ov o0 yp d1 00 ~D oo ,~ ~ N ~ M oo l~
~ p~ .~ l~N00 Q\ N N ~ t~
M d' ~ ~ V1 ~ V7 M ~ ~ ~!1 ~p d1 M M
~

O O ~ ~~ ,-, M l~M ~n N ~ N
M O O O O O O O ~O N ~ O o0ppN O M
O O ~ O O O O O O O O ~
O O

d~N~~~~ddd~~~~ x ~H z ~ ~ d~d~ d~d~

N

l~I~00~ ~ I~N ~ O l~

N N~ Wit'O I~~ ~ N

M ~ ~ l~
i ~~ i ~ i ~ i ~ N ~ N ~ ~ N

N M~ O O M o0N ~OM

M O~~DN ~ O M

M ~ N l~rr V~~
i i~ i ~ ~ ~ i M MN -r~ ~ t~I~ N '~t N l~l~00M N V1M M N

M0101~ ~ ~ ~ .~l~

I~I~~ .~M V~~ M M O

00N~Ov~l~~O~ I~ l~M

Q, ~ D\

O~ N M ~ ~ ~ t~o0 00d101Q\01D1D1D1 OvO~

M MM M M M M M M M

00V7O I~O ~ ~ ~ l~
~'O ,~ O ' ~iO

H

b a; a; ~
M a1 .-r ~ r; Vi ~ o o ~ ono M ~' Q M ~ .-. ~ 00 N
N t~ .~ M ~p ~ ~ V7 M
~ ~ a M a a ~ ~ ~ M O ~ ~ N ~
o apt, ~' ~ oo ri ri o ~ ,~ o t~ ~ ~ oo .~
~ ~n Om vp '~ M oMO N ~ 0~~0 M ~ N N N ~ ~ N
~ 00 ~ M N ~ 01 ~ M d ~' ~ pp H ~ ~ N ~ M
O ~ V7 O O d1 'b p ~ ~ 00 iM b -i ~t ~D d ~ O d ~ ~' N v~ O1 l~
~ O M M O ~ b d ~ O~ O ~ N ~n ,-. 01 ~O N O
~0~1~~~0~10 v~1 ~ ~ Od' M~d'-' ~~p ~n ~O M ~ ~ ~ ~ ~O ~ ~ ~ ~n M M d1 N ~ ,-: ~ f~j ,~ ~ t~ N
~ ~ ~ a ~ a a ~ ~ ~ '' ~
N d d d ,-. N d M .~ M oo ~
° ~ c~ .-.; ~ cu ch yo d~ ~° ~--~ d N .--~ ~i ~ oo ~
°° d ''' ,d N ° oo N ~~~, N~ M~,MH.~ ~o~,~~~~xdH~"~~~Q
~ M v7 ~O Vj ~ ~ M N ~ O_ M 01 ~_O I~ ~ O ~ ~ O
OW1 ~ 01 0~1 ~ N ~O M M O ~ N d N M QN o0 ~ M ~ N ~
.-r .~ ~ ,~ O ~ O O N ~ ~ '"_' 00 d M O M O M ~ l~ O ~ M ~ M d Q
~ oo ,o ° 'b d ° M °~° b d ~ d 'b a, d~~~~dd~~~dH~Nd ~~~~~~H~cz! ~ddd vo vo ~ t~ ~ ~ .-r ~ o O t~ l~ O ~~ ~O N 01 M o0 00 ~O ~ N ~O ~ ~n ~ Vwp i ~ i i i ~ i i ~n ~n ~n v~ ~n ~n ~mn ~n ~n N N N M M O t~ N ~O l~
01 ~ ~ ~ O V7 O 00 .-r ~O
I~ ~O ~' ~ ~O ~ ~ ~ V~ ~O
i i i i ~ i i i 01 ~ O ~ \O V7 I~ I~ N
l~ 00 Q1 \O d' I~ V7 01 M \O
l~ O 01 v~ O O~ N ~' ~ I~
O v7 I~ ~ V7 N Ov ~ ~D O
M ~D v~ O o0 O N I~ ~ ~O
Q\ Ov Q\ v> ~O I~ 0o Ov ~ D1 O ~--~ N M ~ ~ ~O l~ 00 Q\ O O O O O O O O O
M ~ ~ ~Y d' ~ ~ ~ '~h 0o M N M ~ ~ N O
° ~ ~ ~~ 3~ ~3 a x xx xx xx x ~i n ,--~ o°o vo o\ ono c~i a;
a1 ~' .~ ~~ ' wi ~O ° ~ N vD
[~ 00 ~ N ~ 00 ~ M N O N ~ l~ M M
N l~ .~ Qv M .-r .-r ON1 ~ O ~ l~ ~ ~ M ~ N M ~ ~O o0 N N o0 d ~ N M H ~ N
N ~ d N 01 N Q\ ~ ,-; ~O
°M° ~ o ,N ~ r' 'o ~ ,~ b N d ri M ° ri d ~
Wn ° o ~n oo ~ ~ ° ~ ~ ~' ~o d ° oo E-~
.-. yo ° ° ~ ,~ ~ o '~ b o ~ ~ ono ~ -d M O~ U ~ O .~ ~ M v7 ~p ~ I~ ~' ~
d ~ ~ 00 ~ M ~ ~ ~ M '-' ~ ~O O cd d ~-~ N M M ~ O ~ ~ ~ 00 ~ 00 d d N o M z ~ ~ ~ ~ ~ ~~ x ~ ° d ''' o ,-. ~ ° ° N "' o 0 O_ .~ N ~ oN_o O ~O O N ~D ~D ~
c~ d N ~ ~ ° U ~ ~ ~ M ~ ~ M O ~ ~ ~ Q~ ~
d M
z ~d~ ~~dM N zd~~ d dd ~d~'N
M M
as '~ooo°°cN°i ~~o~~'' x ~o'~~oooo~o~
~O oo I~ Ov WO W O M N O~ ~ I~ 0o N ~ N l~ .~ ' N
00 ~O ° M 00 ~ ~ V7 tn ~ M .-, 00 00 N ~, ~,~ O \O ~ ~ ~ ,~ ~O O~ ~
a~o~ooN oo~o~da.°~N°~~~o°o~o0oa\,o~o 00 00 ~ oo ~ ~ oo .-. ,-. -d d o'r'o ~~~~dx~dd~d~xxd~~zxH~d~ddd~Hxa 00 01 M M ~ ~ M ~O ~ 01 N 'O O O M ~1' N N ~ M l~ ~ N ~n M ~ ~n v~ ~n o0 00 ~n M V7 ~ ~ lCt ~ ~ V1 N M ~ V~ 00 M V~
~ i i i i i i i i ~ i ~ i i ~ i N N N ~ ~ ~ N N ~ N
V> 01 01 .~ O 01 N l~ ~ 00 N ~ ~O ~ O
N .~ ~p O O ~ ~~ O M ~ M ~O ~O ~' M V7 ~' ~ ~O ~ ~ V7 V7 M ~ V'1 00 M V1 ~ i ~ ~ i ~ i i i ~ i i ~ ~ i i I~ ~ ~-r l~ N v~ ~ N o0 O1 t~ l~ V~ ,~ I~ I~
M I~ ,~ .~ ~ \O M 00 V7 N
~ O N ~~ N ~ ~ ~O Q\ M ~~ N I~ ~ N M
In In ~1 ~ I~ N N M ~ M ~ 01 V1 N ~ ~ I~ ~O ~
00 ~ Q\ l~ l~ l~ l~ 00 l~ 00 l~ l~ l~ ~O 00 ~O
Q\ O ~~ N M ~ v~ ~D l~ o0 01 O .~ N M ~
O .-r ~ .~ .~ .~ ,~ ,~ ,~ ,~ .~ N N N N N
~ ~t ~t ~ ~ ~t ~ d d' ~ ~ ~ ~ ~ ~t ~
M N M ~h ~ 41 O N M M ~' ° ~ N ~O N
d ~ ~ ~ M ~ ~ ~ d x ~ C7 . C7 C7 dd xx~x xxx xxxx xxx x ~x o ~ oo vo ,~ M v-i ~i ~ ~ 'n O a1 ~t ~ 0 ~ Vi ~o~o ~~OOO~NM~O~~N N O~~~o~o "
00 ~ ~' ~O y~ ~ l~ .~ 00 0_O ~ O_O O\ N O ~ ~ M N ° ~ N ~O
0o WO N M ~ ~D O ~ ~ ~ ~ t~ ~ oho N ~ ~ " oo ~ v7 ~ O
d d d1 M d1 ,~ ~ O 00 00 ~ M ~ ~ ~, d ~ ~ O ~ ,~ M M
° l~ ~ 00 ~ ~ ~ V~ ~ ~--' O pip ~ ~,~ " ~ V .-~ M N
~ o0 ~ O O N ~ a1 00 ~ I~ N ~ l~ ~ ,~ M Q\ 00 OMN~~ O°llxp~M~d~'~~~~,~_,N~NM~_Nd~~~N~M
i",~~ M ~ ~ O ~ O ° ~ M ~ ~ \O 00 ~ ~ ~O 00 N l~ ~
0 d .~~~~~~d~~~~~~~~M_dMx~~~~~
o; ~ d ~ .-~ ~ ~ os d ~ ri d oo ~ ~ d ~ ~ d ~ ~ ~ ~ o 0o d ~ ~ d ~°-1 '~ ~ o°o ~ M o ° o ~ "' ° d .~ ~ o ~ ~ ~0 00 ~ o ~ ~ d O ~h O O N ~ ~M ~ 0000 ~ o~O ~ dO' ~ ~ ' " ~n ~ p~ N~ ~ O N
N l~ d' N o0 ~W O t~ ~n ~D ,_, t~ o_0 .~ ~ ~ d' v'i ~_ ~ ~ ~O
-."MAO N
N ~ I~ d M ~O ,-, d o a; ~oo 00 ~_ ~ ,-. ~o N °° ~ o v d a; r-, o ~ ~ ~
°° o ~ ~n o d O ON1 ~ O ~' 00 O ~ ~ ~ 0M0 ~ d' ~ I~ N O_ '-' 00 ~ 0M0 ~D ~ '-' p ~ ~'~ ~-' pNp l~
N ~ ~ N ~ l~ O 'p l~ v~ ~ N ~ ~ ~ ~ ~~' ~ ~ M ~ N ~ ~ N ~ N M
00 01 M l~ ~ 00 ~ O O (w ~ D1 .~ V7 ._~ ~_ M M
~~~~dNH~~~dd~d~~' d x~~dddZ
HdHdH
N ~ ~
N O l~
i 00 .~ N
O Q\ ~O
V7 M ~D
i i ~ I~ O
l~ M 00 ~O O 00 N
N N N
d' ~ d w N
p°p ~ N O o0 M M O ~--i N
N N N N ~ ~ ~ O N N ~ ~ M N N o~0 h o0 h ~ p ~i N Oh1 ~ N
O h ~O O O o0 00 v~ O
~ N ~ ~ ~ ~ ~ ~ Z h o~0 p~ ~ ~ N ~ U U N a ~ ~ ~ O\
~ h o~ M '° ~ d° ~ d °1 ~ ~..Mo-1 N ° ,°~ d d O d M o ~~3 ~;~3 ~ °~n° ci ono ° '~' d o d vo d ~, N d M ~ N °
~M v-, N ~ o U
oho ~ N ~ M ,~ ~ O N ~ ~ N M M ~ d1 ~ ~ ~-~--~ o0 Ov ~
N'"'~~~~OV~o~oO~ohod~'v ~NON~OO~ONp~NO
d M o ~ N N ° o U U ~ U M U ~ M ~
NM~o~~dd ~~~~°~Na~dN~~d~°d d " oo O oo d ,-, N M ~ t~ ~"~ ~ N
h h ~ 00 h_ '-' V7 h .~ M_ 00 h ,~ ~ O1 ~ ~ ~°p ~ h ~ N ~ ~ O ~ ~ h ~ ° M ~ 00 O h O N
00 ~ ~ V7 00 ~D M h ~ 00 ~ ~ ~ M M h M ~ O .N-~ M O O ~ N pp O N O O
ooo°~~M o~ '°~ooooo~oNO°~o VU°UoMO°c,~U
~~~~x~~~d~~~~~dd~N~~Ndd~dN~a~d ~ h oo _ ~ ~ 00 0o r~i M d ri ~
00 ~ o ~ '-' 00 oMO °~ °~° Vi ~_ oo ~ ~ o N .--. o O O ~ ~ ,~ ~ 01 ~ ~ ~ ~ ~ oMO 0o v°p ~ ~ O M N ~ oho ~ ~ ~ O M ~ O
d' M ~O d' ~O ~O v7 Q~ ~O 'd' ~ ~ O~ O D1 00 ,n '~h .~ M M N M N ~ ~p O d' O O O 00 ° O O O O ~' 01 M O '~h N N M
ONM~~NOM~.hvp~.-,~p,~00°OOOOOOOOON
~' ~n ° ~ ~ 3 d ~ d d d ~ '~° M ~ U U U U U a a a U .o ddZd~wddddddddd~dddddddddd~~~d p ~ I~ ~ ~ ,-~ c~ ~ oo ~ ~i p~ t~
l~ ,n O_ M ~ O O O 01 M [~ N
O O ~n v~ ~ ~ ~ (~ M ~ O r.. O ~ ~O N 00 ~ .-W p ° Q1 _ ~ M 00 °N O O O M O ~ ~ ~ ~°. ~ p ~ cV 0 0 0 ~ 0 0 0 o N U U U "~ U o ~ U ° o U o a' U U o ° U o 0 o U
° ~ ~ .~ ~ '~ d d d N d ° ,...a d o o d °
°.~° ~ d ° ° d o 0 o d U o0 0o r-, ~ ° U U U d U W U ~ U
d ~ ° a N ~ ~.'°,, o ~ U ° d o ~ d ~ N d a ~ ~ N d d ~ ~
d °' d 00 N ~ N ~ d O oo ~ ~ M O ~ ~ d p ~ ~ N ~ ~t N
N ~ ° ~ O M ° M N I~ O t~ ° O M M .~ M ° I~ ~
° M O
w ~ ~ O ~ O O O l~ O ~ r.. O l~ O ° O~ ~ O ° O ~n ~' ° m op ° .
°o ° o 0o U ~ U U a o U ~ ~..a W o o U N ~ U a W ~ o, U o ov U
ov U
or~.,~UNd~dddoodr'ddUod,~~ddd~~dUNd~d ~dd,°NN°~o°o~°M°~~~d~~~N~N~o"',~~do°o o~o \O ~ ~ d N .~ ~ V7 ~ N ~ l~ O O ~ N M ~ ~ ~ ~h o0 N ~ l~ pp" ~ M M M
n N ~ .~ N ~ N o0 O v0 ~ O v1 N ~ ~ ~n .~ ~O ~ O~ M ~.~ O O ,-, N ~ ~ W ° ~ ° O ° p~ ° ~ O O ~ p D .~ M D O p ~ O p ~ O O p O
00 ~ o o U o ,~ U U o U o U a ~ U a r-1 a o a a ,...~
~f~U~dU~~dNdU~dN°dd~ddd~~dUdd~~
~ ~ d N ~ d v~ ri vs o ~ d ~" ~ ri ~ ~t ~''~ ri ~ ~ oo ni ~ ~i d t~ ~ ,~ ~
~p ~' ., ~p N ~ O N l~ a\ ~ ~ Ov ~' N ~n O t~ N I~ v~ ~ ~' N l~
~O ~ r.; '-' O 00 ~ ~ ~ 01 ~ ~p M .~ M 'p O ~ l~ ~ O V7 N M ~O ~t v7 M d' ~D ~D ,~ 'p N N Q1 ,~ ~ N N pp O N O ~ 00 'n M .~ ~O ~O V ~ ~ I~ O ~O
.~ O N ~' O N O O O N O M O p v~ ,-, O O ~ O O N O O O ~ O O m O
O O ~.,~ ~ O 00 ° O O O O ,-, O O O 00 O ° O °
° O O O O M O O O O
U ~ N oo U U U W U o~ U ,.~ N U U U ~l U U U ~ U U ~l U
~dN~~~ddddd~d~~~~d~ddddddNdddd N 01 N ~~rj ~D N_ N <t ~' M pp 01 .-i \O N O .-r ~ ~ N ~ ~ O O ~ ., O '"' O ~ N ~ ~ ~' ~j N
O O ~O v7 d' N ~ O ~ ~t ~ ~ ~ M ~ v~ r. ~O
O O O ., ~ O M ~ O O O 00 ~O ~ O M ~O ~ 00 O N O O ~ pip M
iU°ddd~d ~ ~~;N~°~ ~~ Ud~~dU~°o,Nd N ~ ~n ~' ~ ~ ~ 0N0 ~ ~ ~ 0 ., ~ ~ ~p p '~t d N ~
_ a1 Ov N ~_' l~ ~~ .~ r, ~t ,~
~O M O O1 l~ ~O l~ N pip O ~ N ~ ~ 0~0 0~1 00 O M ~ ~ N ~O
O O ,-, O O ~t ~ O O N ~ ~"a N p~ ~ d' ~O ~ O~ M M O O pp M O
U o U ~ 0 0 0.. o o U U a d ~ v'<'-, M a ~ '-' '~ a a ° U o ,°~ U
d~dN°°d~~ddd ~~x~~d~d~dd~d~7dd N 4i ~n ~-~ d d t~ ~ ~'' ~i v'i Ov ~ ~ ~ d O~ ~ ~ D\ ..-s ~O ~n ~n N
O ~ O ,-; ,-; ~ ~ ~ o~o ~ ~ O ~ N N ~ ~ M N ~ ~ M ~ ~ N N
~O N ~n O1 ~ ~ N 01 ~' N 01 01 ~ w ~ ~t ~ O~ ~ 00 l~ ~ ~O .~ ~ ~O ~n I~
O N O O ~ O ~ O O N O ~ ~ ,_; ~ N ~ .~ .~. O .~ O p~ M O O O O
O O O O O pp O ~-r O O ~-~ O ~t O ,~ p O O O O
U U a o ov U a U a a d ° ~ °r..° 3 c~ 3 ~ U ,~ U U U
U
d~dd°Nddd~~d ~N~33~~d~d~ddddd ~..; 00 o ri ~ v~ ~ .-. N N <r o~ a; ~n .~ Z ~ d d ~' ., y" v; ri ~ 4W o vp l~ N M ~_ p~ ~ ~ 00 O_ d1 p 00 M ~D O N M ~' V'1 N r-r \O 00 N ~ 0~0 ~ ~ ~ O N v'1 ~ ~ ~ M I~ N O O ~ ~ O ~ N ~ ~' ~ ~ ~ ~ v~ N
N O O O ~ O p O M O O O p l~ Q\ ,~_, ~ N ,~ ~ ~ ~ ~ O O M O O O O
0 0 0 0 ~., o 0 0 0 0 0 0 0 0 .~ N ~r ~., ~ o o '-' ~ U U a U U U U
ad~a~~~ddd~~~~~z~xxd~~~ddddddd M O
~M
M
N N
.~ O

xx N ., ~ ~ pip ~ O O p M M p ~ ~ O .-.
O ~ N py0 ~ O ~ oo ~ ~ ~ ~ ~ p N ~ ~ ~ ~ ~ yD
~°o~,~~°o~~o~°oooc~i°oo~o °°oo °~°°'~'~°o°'~°~wi~
dog,dNUodd°°~.,o~od~~~~d~°oodd°o°~do~,d.degr ee.~_°o oUo'r'oNdUcM.~~~No o~oM~~°'°odd~~~d~~N~dod d o ~O O~ .-~ v-i ~. N ~ ~ .~ ~p ,.-~ ~n ~ p ~n ~ ,-; O ~p ,~j M ~ gyp" l~ oo ., 00 d N N ~t ~ ~ ~' ~ .~ O ~ ~ ~ ~ ~ '~ N ~ ~n .~ ~ ~ N N ~n N
I~ M O n1 O~.--~OMMpO~~pMppOOpO~~OpN~pOMOM VSO
~ a M N U o r a U o U o o U ,...~ r'' o U U N U ~ ~ o, °d~~o~dUa~dd~op..,~dUUd~~Uddad~o~
°~N~~dd~i do~o~~~~~o~o"~dd°~'°~dy°odc"~.~NUd 00 ~ M ~O ~ N V~ M ~ ~ d' ~ ~ ~p p~ N 00 ~ M ~ ~ ~ ~ I~
p ~ '-' ~ O O O M v~ O O ~ ° O '-' ~ M ~ O ~ O O ~ O O ~ N
O M O p ~ ~ ° O ~ O O O O O I~ ° O N ~ O O ~ ~ p O ~ O O I~
,M-, U M U ~ U oo U ,...~ U U ~r p., U N o ,~ o o, U o U ,...a '~ ~
dad~~N~d~ld~~dd~ddaUd~oN~dUdd°
a, d O N ~ M O ~ 01 ~ O M ~ N v'i o 0o d d ~ ,-: ~ 00 0o t~ d ~t ~ N ~i 00 00 M ~ ,~ V7 ~' ~O 41 00 00 .~ l~ ~ M ~O ~ ~ O 00 ~ 00 O ~ \p O 00 M V' 00 .--~ M 00 O ~ ~ ~ O r, ~ M V7 ~ ~ O ~O
~ p1 V~ ~ p~ ~D ~ V7 ~D ~ p0 ~O ~ 00 M O O O M O O O
O ~ O O O O O O O O O O O O O .~ p M ~ O O ~ O O O M O O N O
dNdddddd~ddd~ddddNNd~NdddNddNd _ o ~t .~ N O ~n ~ O ,.~ l~ O O ~h M ~ N ~ p ~ ~ ~ O ~ O ~ ~ ~ ~ O d' Ov ~
-~ ~ ° U ° ° U '°° ° °
° U ~ ,°~ ~ ui U o ~ 'd° o~o ~d~dd~d~°o~d~~dNd~ d ob~~°pdd ~~~~~~o~Od~~~~~~o ~~ '~t~~ ~N
~ ~i ,~ ~ O ~ o0 ~n O N '~ N o0 O
V M N ~ M
ooooM°,_~°oo~o°oooNOO_ U
ddddd~~Ud~dd~d~ -c~~~ ~ o d ,-..
O ~p l~ O .-i ~j M .-i ~ 'p O~ O N ~wj ~ ~"~ ~ \O
00 "~ M N O ~.,~ I~ d ~t ~p t~ ~t ~, ~ N ~ ~.r; d ~O l~ ~O
O ~ Ov ~ ~ ~ a1 ~t '-' ~n M t~ oo ~ '~ O ~ O
~ v1 N v~ ~ ~' ~ ~ ~O N ~ l~ Q\ pp M E-~ ~ ~ N ~ N O~ p O M V~
oo~oooo°oM°o°'oooooo°
°M°~'°oNOO~ U
d~dddddod~~d~~~o N~°w~~
W o cri ~ .-~ cn ~ ~ 00 0o t~ O t~ ~i ~..~ ~ l~ ~, ~ ~ ~ ~ N ~ ~ v-i ~
~O N O ,~ ~n ~ 00 00 v~ ~n M l~ ~ a1 O Wit' O ~ O oo ~O
t~ ,~j v1 ~O l~ ~ 00 O~ ~ O p O ~ ~ ~p ~ l~ d. O~ t~ N M l~
N a\ w o o .~ ~n ~ ~n o a, ~ ~n ~ o U ~ .~ ,~ ~ ,~ o N ~ ~ ~ ~ ~ ~
ooo°ooo°o~°oo°o°o°o°od ~~~~~a,~o,~d~ °oo U ~l ~l U U oo ~l U U U '~ N o0 ov '~...~° ~ ~ ~ 'b t~ U
dddd~~dN~~dddd~~ ~~H~zdzdd~wd~
00 o c~ ~n ~ ~ .~ oo N o M V1 M ~ I~ O ~ ~-~ ~O 00 M ~n ~ ~i' ~ ~ ~1 M N
i i ~ i i i ~ i i i V7 V~ In ~ ~ ~ In Vl V~ In ~O M ~-~ l~ l~ l~ ~ 00 ~O
N M N ~ ~n o0 M O ~ 'O
M ~ ~ ~ M ~ V~ M N
~ i i i 00 l~ N l~ ~ 00 lp M ~
M N ~ ~O I~ ~ ~~-~ 00 .-~ N oo N ~ M ~D N
N I~ 01 00 ~ ~ O ~--W1 V7 l~ ~n v~ oo ~n v~ ~t M ~n N
Q\ ~ ~ ~ ~ ~ ~ ~ D1 ~D
O ~ N M ~ N ~ l~ 00 M M M M M M M M M M
M O pp O 01 V7 O ~ 00 E-i O C7 A U GA ~1 al ~
~xxxxxx xx~x a1 N 'C7 ~O N N ~ M ~ ~ ~ ri ~ ON1 00 l~ .~ ,~ ~ 00 ~ p ~ N ,_; Q\ p v~ O ~ ~h' ~ ~O ~ d' t~ ~ ~O
00 00 ~--~ d ,~ M ~ O ~ ~ N ~ ~ ~ [~ 3 ~ ~ ~ N N
vo d Q ~ Z o o .-. ,~ N d d ~ d o_o °o ri P; ~ ~ '~ N r.; wt 2r ~ ~ ~ ~ ~ ° o~°o ~ o d o p M ~ 00 OMO ONO N ~O ~ ~ d' ~_ N d N ~ 01 .~ ~ '"_' (~ .-r \O ~ M d1 N 00 ~ ~ ~ ~ O O ~ ~ ~ N Z z ~ ~ ~ ~ ~ ~ 0~1 ~ ~ ~ M
00 I~ _ ~_ ~ ~ M V7 ~, ~ _ 00 d ~ °°
d N o r; c~i d x ~ ~ o ~ ~ o r; N
00 p Ov N ~ ~rj O M O ~p O ~n N ~ ~ N_ ~_ .~ l~
~ D1 ~ N vp N Q1 ~ ~ M ~ p x x M Q1 M N l~
r~.~ O 01 M ~ M N N l~ M d1 ~p ~ M ~ ~n ,~ ~ O ~ N l~ ~ ~p M \O d' .~
,.d d1 00 ~ ~ l~ ~ ~ ~ ~ 00 ~1' O '~' O 00 ,-, ~ ~ ~ [w d.
~~~d ~~d~° ~x~°~d~~~~~3zxzx ° ~ M
O 00 pip V' 'fit ~ ~O ~ '~ M ~ V7 ~ ~ V7 M ~ ~ ~ O M 00 N ~ N 00 pip O I~ 01 'n ~ v~ ~ d' O .-. ~ 00 ~D l~ ~O ~ N o0 00 M M ~O Np [w 00 ~ t~ ~
~ M 01 O \O N M 00 01 00 M o0 N O ~ ~ N ~ ~ O ~ ~ ~p ~ \O O M
00 O .~ l~ ~ N 01 N '-' l~ N ~ p O t~ D1 .~ M I_~ r., 00 O 00 ~ ,N-,, M 01 M
~ ~ M r~-~ M ~ 00 O d ~ ~ r~-i ~ r~-~ d ddddddd~ddddZd~~dH~ddd~dd~ddd~
l~ l~ Q\ M
O ,-~D v~ I~ d' ~ i i Q1 O ~wh N N
i i i i O ~ OWD
00 ~ M 00 l~ Q~ ~ O
~t 01 O l0 O ~~ N M
d' v7 N o0 v~
W W W A

n ~-rj ~ ~p O~ ~ v~ N I~ M V'1 N M M M ~ M

yp N ~ N ~ 01 ~ .-mh N O ~ N o0 01 d' ~ N M

N ~ oo ~ N O o0 M ~O ~_O ~ 01 .--~
~ ~ N ~ d_' M l_~
N

N M N ~' M ~ ~ ~ M ~ ~' M ~ ~ V
N ~ ~ ~ ~ M ~
'~' ~ a, M
M ~t N "., oo ~ ~ ~ M Z o o o ,...~
~~d'~~~o ~n~i~ ,..~
~ ~~d~~~d~~~ddddd d M

'~' ,-.~

~ N oo IWO ~' N l~ ~n oo M ~
~. ~ I~ N
.. ~ M M M l~ 00 ~ I~ M M N l~
01 v'i v'i ~ N M ,~ ,b N ~ V7 N ~ p~ M 01 d y0 ~/ ~ ~ ~ M ~ N O V~ ~O ~ ~D M V~ M
M ~p ~ ~ ~O N O '~ ~ O M M O
' tn ~D ~ ~p M M ~ ~ 01 N ~ d ~ 00 ,-, ~ O ~ <f M I~ M I~
~ ~ ~ O
M

OO a a a a a a a a O N M ~ M o0 ~ l~ ~
~ ~

d d t~ yn cr;
~ N ' ~ z M ~
d ~
00 d v~ ~ U ~ ~ ~

pp N o0 N ~O v7 M M .-~ N ~O
~ ~
M v~ z O ~O D I
~ 01 ' M ~ M I~ ~ ~ ~ I~ ~ 00 ~n ~O ~
~h O O O
~

I I~ ~t ~ O .--, .--~ O O O
M d O ~O d' O .~
N a1 o0o N ~
~ ~
~ M
~

~

1 aa d' -1 ~M~N , Myi N W..
~x'-'d'~~ , ~'~ od"'~ ~
~dd~d~~~~dd~~d~

r ,~~ N I~ ~i O M Q\ N I~ ~ ~ v'i N r-i O M I~ 01 ~ ~ N d ~' ~ () ~' N ,~ M l~ V~ .~ 01 ~O ~
00 d1 ~h M I~ ~

N M ~ "' 0~0 D oho M ~ ~ ~ ~ ~O ~t ~ (~ N Wt o0 ~ oo ~ v'i .-~
00 O (~ l~ N I~ M M M M M
~

N N M ~' M ~ ~ ~ ~ ~ ~ ~h ~
O ~ ~ ~ ~t ~ O ~ ~ ~ O O O O O O O O O O O
~

~ d~~d~d~~~~~dddd '-' ~ ~
~~~~~~z~~~~z~zH

N

N

v7 N

N
O

O

d' N

x O v7 O O ~D ~ ~O N o0 00 I~ ~O M V~ ~n ~O ~n ~ O I~ N ~ N Ov '-' M

~ U1 t~ N ~ ~ 01 ~ d' ~ Ov M V7 00 V7 .~ N O
00 ~O ~ ~ N 00 ~

0o Q1 ~n O~ M l~ N ~ ~ O -~ O ~n N -~ ~D ~n ~ .~ O O N O ~ M ~ ~ ~ N
~ M ~n 01 00 ~ ~' O 01 .~ ~ O O O ~ d' -~ O ~ O M

~d'~~MMd'~'~~~~~~~T~'~'~~~~~~~'oo l~~n M
O O O O O O O O O O O O O O O O O O O
O O O O O

aaaa,..~,....~ a,..~ as aaaaaaaa ,...~,.~
M
dddd~~d~~dd~dddddddd~~~~ M~M~

p ~
~
~

~h ~ ~ M ~ ~ M O OW i o0 01 ~n N 01 0o d O ~D
l~ N o0 N ~ l l~ ~ ~' 'd' v7 Cn V7 M N 00 M ~ I~ I~
O~ M ~ ~ ~' ~ d' N ~ Q1 ~O V7 .--r 00 ~ O M M M N ~ M N ~1 N O ~ l~
'~h M N ~ M O~ 00 O ~ N

O O M O l~ ~ ~ O d' ~ ~ -r O ~ O ~ Q\
O I~ O ~ O 01 N d\ ~n v~
' ' ' ' ~ M
~t M ~ ~ ~ M ~
~ ~ ~ ~
d ~h ~t ~ ~ d' M ~ d O O O O O O O O ~ O O O O O O O O O O
O O O O O O N N ~ ~ M

aaaa,~ aaaa aaaaaa,~ ,..~,..~~..~ aaa~~
dddd~~~~~~ddd;
~
~

,~
Ma,~
dddd dd dddd _ N
~

~t ~O oo O~ N oo ~O 4i v'i O N oo v0 O
O~ ~ ~ ~n oo ~i ~i D; ~n [-~ d o N l~ -~ ~ ~ v7 OW ~ 00 N ~ O~ ~ M a1 ~
00 d' O l~ M ~n N l~
O
~
l~
l~
~
~

~ N l ,-;
~ M

N O
n t N N
N
~

~ N

_ _ _ _ _ _ M
~

M ~t M ~ ~' ~ ~ ~ ~ ~' M
N ~ ~
N

O O O O O O O O O O O O O O O O O O O
O O O O O O ~
aaaaa,.~,~,..~ aaaaaaaa a ,~,~ aaaa~~MN~
dddd~d~~~dddd o l d~~~dd aa ~~
dddd dd M o0 ~ V7 M ~ N Qv ~ 00 O 00 M o0 I~ I~
~ N DO N o0 I~ M O 00 ~' ~' N l~
N ~O Q1 M ~O l~ l~ 01 ~' V1 l~ N M l~
l~ N ~ M ~ ~n I~ VO ~ N M M

V'1 l~ D1 .~ .~ ~p N .~-i M ~ M ~ N o0 N M I~ '~t -i ~O N o0 N N M ~ pp" ~j O O

~ N .~ ~n ~ ~ ~ O O O r, O~ .~ ~O D1 a\
O a1 ,~ ~n O .-. w ,~ N ,n ~ O

d' ~ ~ ~ ~ ~ ~ I~ ~ ~ M M ~ l~ ~' ~ ~
~ ~ p~ O M ~' 00 O O O O O O O O O O O O O O O O O O O
O O O O O O M ,~ op O O
aaaa a a s ,..~,~
N....N
aaaaaaa,~ a aaaaa ddddddd~dd~ddddddddd~~~~dHHN~d ~_ r. _ M 01 I~ ~ (~ N ~ N ~ ~ N O O M pp ~ M M M ~ 00 ., p M ,~ p ,~ ~ O V'1 r., M Qv M .-, O ~1' ~ ~ ~ l~
00 ~ O ~ O M O ~p ~!1 ~ O ~ d1 00 ~p y~ d 00 ~
O M ~
a; ~ ~ W O ~ O d d ~ p d ~ ~ U ~ d ~ ~ '-' ~ °° '%~ ~ d d ~ ~o ~ M ~ ,~ .~ "' o r-i ~ ~
d ~ ~"'~ N l~ l~ CV N ~' M I~ ~ r.r ~ 0 ~ N \O N
M ~ ~ V~ ~ O 0~1 iM ~ M U ~ 00 ~ ~ 0 d ~ ~ ~ ~ O ~ l~ ., 01 M O
,~ ~ M ~ ~ .~ ... ~ ~', r' ~ o, " ~ ~ ~ ~.; d N o '° o d ~ d d d r~
o ~ ~ ~n ~r M ~n '~° ~ ~, o o d o ~ ~ ~ ~ d ~n ~ _° o ~ ~o d V7 ~!1 V'1 d ~O tn d V7 ~ ~ ~D d' 00 '-' Q M
N rr 01 ~ M O l~ 00 M ~ l~ O~, o0 00 N \O
~~~~.~~,,~a,~o~o~0°1,~ o~~~oM~o°o°°o~
W~'°"o~~~dd~~
01 ~ N ~ N o0 M I~ OW1 00 ~ ~ O ,_, r-i " ~O r., ~ ~ W
O ~ N ~. ~ \O v7 I~ ~O ~ l~ ~rj M M ~ ~ ~' D1 ~ O M 00 ~ N_ M O l_~ M
~_MI~O~_O~_~~~~~~~~Or~-~ NoMO~ ~o~o~I~~ d'~ON
d H ~ ~ U d ~ ~ ~ A ~ ~ ~ d d d ~ ° ° ~ ~ ~ ~ '""~ o ~ °
o°°, o ~
Udo ~°d~;d~-'~odNddddd'-' as ri ~ O ~ os ~n oo I~ ~ ~t ~ vi "., ~ ~ r, O~_~~~~o~ooNOO~I~~M~~~_NQv00~''~NNM~~~o~OMMN
N N N M <t <t ~ ~ N N ~ ~ ~ M M N ~ Q\ ~ 00 M O ~O O ,~ ~ .~ 00 V1 V'I ~' V~ V7 V7 ~ t!1 N Q ~ ~ ~ p V7 p ~O O N M p O M p O 00 M 00 N
m--.i .y--i ~ W N ~ r-~ '-r W ~ \O h 01 01 ,~ ~ ~ O O
ddHd~d~~~d~~~ddd~ddd~~~~ddddd~

~ o vo ~ ~ M M ~D
M M N ~ ~--~ 00 00 ""' O ~t N ~ M ~ d1 M ~ ~ ~ ~ ~ ~ c~j N ~
M M ~ v~ N V~ ,,.N.., O~ ~ ~ p I~ ~ M N ~ p ~ O~
N cri M
O d ~ pip O ~ N ~' N d o0 ~ O t~ gyp" N Wn ~ ° vo ~ M ~ d o C~~~Md~dd~~~.;~~o~~~N~00~ON~N oMO; , ~j o0 ~O ~ M ~ N ~ ~ d ~ ~ x M ~' ~ O ~ ~ ~ N M
oho ~ ~'~ ~ ~ 0 N ~ ~ ,y-~- d ~j ~ ~ N M t~ r"' ~ ~ N O
d d O ~ N ~ ~ ~ ~ ~ ~ N ~ ~ 0o ~ ~ ~O
N ~O ~--m, d' ~ Ov oho ~ ~ N O y--, M
0 '"" d ~ ~ N d ~ N ~p Wit' dwn ~; ~ M N "~ oo ~ ~ ,n ri ~p 01 0~0 ~ w ,--~ .-i w ~O ~ O ~--m0 ~ p ~ o0 N ,~ M a1 ~ ~j r,~ N ~ ~D
'-' M M ~ O O ~ vM1 O o0 N ~ M ~ ~ Q-i ~ O ~' ~ O O N pp O c,~ ~ 01 pv~p 0o ~ '_'_' I~ M ~ ~ M ~n ~ ~; ~ ~ ~ d ~ ~ ~ ~~., N O ~ Ov N O
O d x o~0 O N ~ ~ ~ O 0 ~ N ~ ~''~ ~ Own O ~ ''~ N ~ w ~ N ~ d d N
O O~ N ~n ~; ~ d ~ °d d ~ o ~ ~ d vo o ~°,a ~ ° ~ ~ c~,~ o ~ ~ N ~
~ ~, N oo v~ op Q\ N O O
O ,-; ~ ~ pp ~ ~O ~ ~" ~ ~ ~ ~ O M ~ ~ N M ~ ~ p N ~ O ~n .~
M ~ d ~ M ~ ~'' ~ '~ °M ~n N "' '° °M° '~
c°1~W' d '' p ~ d °M d ~ N av M o 00 ,~ ° ~r o, o 0 0 ~ d ~ ~ d d oo d M
M
,r d ,n ° w ~ °o ~ ~ d N o W ~ ~ ~ ~ c~i ,err, o; ~ o ~ ~ ~ ~ ~' ~
00 M ~' N M '"' in N '""' r-i r-i O O O ~ O ~ ~ ~ 00 Ov 0 'n o0 t~ ~ N M oo y0 M o0 ~n 1~ ~ V~ ~ ~ a\ 0 ~--~ ~ o~0 000 0 01 M [w M m .W O N p~ ~p I~ a1 N ~p 00 M ~. 00 ~,~ 'n M ~n M O N v~ M M ~
r, M O ~ O o0 N M O ~~ ~p ~ 01 01 O ~ O N M N O N N N I~ O O N . 00 0o V1 ~ N 01 .-, N r.r ~p O ~p ,~ 00 ~, h .-r ~ ~~ P~ h N
w°~d~dd~°~dwd°~w °~d°d~dwd~~ld~d°

'-' ~ ~ N ., ~ ~ N ~ N M c~C I~
M O 00 pp 01 ,-.-~ 00 M N ~ ""' O
N O ~1 °b goo°o°ooU °o°o,n~o~ ~vN~, ~dddd°dd-d dd N
M ~ .~ ~0 ~; l~ ~ N ,~ ~ I~ O d ,~ ~ d1 01 p M l~ c~ ~ N M ~ 00 N o0 v7 Q\ N ~O ~ N ~ M p ~ ~O ~ ° ~ ~ ~t p ~ ., O
O M ~ O O ~ O O O O O O D1 O O M O O l~ \O '-r N
~'~i°dd ~tdddd~ddo ~dddo d~
x ~Ly ~ ~~ p~ 0l N ~p v~ vi 0l p~ O d' ~O ~ ~ ~ M ~ d 'O ~ N vp v0 ~ N N M ~ 00 M cG O ~ ~ N ~ ~ V~ ~ ~ ~ 01 N 01 p z ~n ~ O d ,n M N N 01 ~ N ~ N 01 ~ l~ ~ M "' M O ° ~ 00 ° M ~ N ~ ~ N ~. 0 0 ~ O° ~ ° ~ N
ri~~UU x ~U°U~~o-100- °M° U~UUo~ °~'M°
o°o ~ood~ ~ ~d~ddd~do ddddWo M d z ~ ~ 00 00 ~ x ~ ~ o ~ ~ o M ~ ~ o o ~; d o vo d °°
M p~ ~ V7 ~ ~ ~p ~ M .~ N ~ 00 M tn N
OO ~ ~ ~ ~ ~O O ~ ~ 4I M ~ O ~ O ~ ~' OO OO N_ c~Mn ~
O D N M ~ ,.M~ p O 0 0 p 0 ~ ~ O O ~ ~
M o o d ~ ~q ~ o a r~., U U U a., U a\ d U U a oo '~ M ~ o, °d~~d~~z w~~ddd~d~ ~ddd~ ~~dr~!~x ~n O t~ N O O t~ ~O O N
O ~D 00 v~ ~ ~ .--~ 00 01 l~
M ~ ~' ~h ~t ~ ~ M l~ N
~ i ~ ~ i ~ ~ i i N N N ~ ~ N N N ~ N
~O M 00 ~D \O M N ~ 00 d1 ~ I~ M N ~ O l~ l~ v~
N ~ ~t d' ~ ~ ~~ M I~ ~n i ~ ~ i i i ~ i O 01 O I~ 0o O O N N ~O
O O Wit' M I~ ~D O 00 ~ ~O
O ~ M v~ N ~ l~ M v~ N
Q1 41 M O ~ 01 M M N
dw0 M lw0 N ~-, oo ~n 01 Q\ ~ ~ ~ ~ D\
~n ~ ~ 00 ~ O ~~ N M
d' ~ ~ ~ ~ v~ ~n ~n ~n ~n d' ~ ~ d' ~
° ~O,P- o_o _~~_ N~ v1 A ~ ~ A A

os ~ o ~
yo '~ ri N a\ cri ~ ~ ~n r1' ~ t~ O~
~~~~~~,~'~°°~'os°oo'-'°oo~~o N ~ c.~ ~° °o ° ~ ~ °M ~ ~ o ~ U U ~ U U
,°~ M
~ MNM~~~-°~U~dd,~ ddd0~
V7 ,N-, d ~O .-i ~ ~ w ~ ~ ~ d d' a 41 M ~ M O ~ ~ 00 wj ~.,.j M O .-; <t N O
00 o x ~ p ~ ~ M ~ p d ,-. p r, ,~ ~n ~ M ~n o~p d ~~N~'Op~N~~~O~NppO'"''oOM
r~ C, ~~., d ~ 3 d w d ~ N ~ ~ ~ ~ ~ ~~ M o; ~ oo "; ~ o M p1 v1 00 N .~ O .-r 00 01 'n ~ ~ O N ~ ~D O ~ 00 O O N ~ ~ N
~ M ,~ (~ ~ ~ ~ O O O ~ '-' O
M~OO~~pNp~00000~~0 ~ t~ ~ M d ~ o~ 00 o U U U U U ,~ ~ U
d~~ ZF"a~~~w~z~N~lUddddddNd M N ~O M ~ 00 O M ~ 00 .~ (~ .~ ,~ M ~ Vj o0 M
M oo ~-~ N ~ d o, ~ d ,~ ~ a\ ~ .~ ~ d .~ o ~. .--~ oo ~, oo N
M ~ 00 N N ~" ~ M O N ~ O V7 M 00 '~' M M ~ O N
~O l~ 00 00 ~ ~ O o0 ~ ~D O O ~ M ~O ~ v~ W p ~O N p ~n ~t M I~ l~ ~ N O O ~ V~ v~ O~ t~ O O N O O O O ~ p O O
00 ~ I~ v~ N ~ ~ ~n O M .~ o0 ~O V1 O ~ O O O O O O O O
N "' d "' d '° 'n U ~ U U U U U p.., U U
ddHd~w~~~d~dNdddddddd N ~ N ~ N ~O l~ v~
00 \O I~ 01 M V7 ~ M
~n d' M ~
i ~ i i.
N ~ N ~ ~ ~ N N
0o N oo N oo N M
~p ~ ~ 00 ~ ~ M N
~f ~ ~ ~ ~ ~h M ~
~ i i ~ i ~ i i M 00 l~ ~O ~O O l~ ~
~-~ N ~ ~ l~ N N O
01 ~D O ~ ~--~ ~ M
d1 v7 M N M l~ v7 O~
v0 ~ ~ M '~Y' ~n N ~n Q1 ~
~O l~ 00 01 O ~~ N
~h ~
N ~ N M ~ 00 ~D ~ ~ O ~O ~~ O O
U U U U GA G~l d N
~ A ~ A ~1 ~ ~ U
xxxxxxx O ~ O ~ oo ~ ~ p D ~ N ~ M ~ M v0 ~ ~ ~ ~ 00 M M ~ O N N ~ t%j OM '~ ~ V~ 00 ~ ~ 00 .~ ~ O O ~ O ~ O n 00 M ~ ~O
~O
N l~ O N N op ~n M vp Ov O ~ ~' ~O '"' N
N O p O O M O O p p O M N p O p p t~ N O p ~ .-~ p~ N O O
ppOOOOOppOO,npO VIpO~NOp~~M''"pO
UC~UUaUU UUaoUUoU~~ Uoo~aU M
~~ddddddd~dddUddUdad~d~U~dd~a~
MO~~M~I~~N~Np~OdN~dNdo~oo°'oopdd M Ov p v~ ~ ~_ ~ l~ ~n v1 W p ,--~ ~ ~n N ~ (~ N ~ v7 01 ~--~ ~.j ~ o0 ~t 0o ~

N ~ ~ ~ M N O ~ ~ ~ ~ M ~ M ~ ~ O '_-, ~O ~' d' 01 \p N ~ d' ~ p d1 O ~-~ D O O ~ p O r' ~ p p ~ O N O O O ~ ~ ~ ~' O O
U ~l U~IU~IUU oa ooU~nU,~Uo~oa UM~I
~~d~d~dddddd~pd~Nd~dddU°-~Ud~dpd ~ ,~; ~~ ~ ~; N cue; O o; ~ ~ ~ ~O ,~ ,--~ os ~ ~ "; a; d d d N ~~~i cn ~ O
O~ O_ M N Q1 M .-i M ~ ~ O ~ ~O 00 M O ~~-~ ~ Q1 V~ (~ ~ ~_O 00 V~
M O1 ~ ~ f~ ~ t! ~ ~ ~ p ~ ~ d' ~ ~ ~ ~ ~ ,~ ~ ~ ~ M ~ N Ov p O O O O O O O p O D p p M p O O O O D O ~ O N O p O O ~ O
~l U U U U U U U ,~ U U w U N U U U U U U ~l o0 00 o U ,~ o, dddddddddddddaddddddd~~Npd~~N~
,~ O ~O ~ O ~~ (~ O V7 I~ N t~ ~' ~ O I~ ~ ~ V7 M v) 00 O\ ~ ~, 'n M M ~ N
O O ~ ~ oNO p~1 ~ N ~ 00 ,~ v~ <''~ ~h N N ~n o0 .-.~ M .-~ ,~ I~ O N 00 0o v~
~n I~ ~n ~ ~ ~ ~ ~ p N O O ~ ~ ~ N ~ ~ M O O o~0 0~0 ~
O O O ~ O D O ~ p ~ p p O ~ D O O O ~ p O O O O ~ O O O O O
dddddd~d~dd~dNdddddd~~~d~ddddd '~ ~
o ~ ~ ~ _ ~ ~ ~
~
~

0 v~ ~ ~ ~ _, 00 0 ,. '_-' 0 0 , 00 ., N N 00 00 01 N M ~ ~ p Q~ ,b ~ ~ ~ ~ N ~
O

d '' ~ N ~dOO
~

dd d "; ~ ,'~',' ; a ~ H ~ d U
~ ~

t~ O . N N
~p d O ~
O ~

N ~; ~ d O ~ ~ oo .~ N o0 l~
~ U o0 ~O O O M
l~ N

O M O d N ~ N ~ ~, ~ M ~
o 00 N
.-. M op N p U U ~ U ~~-1 N o ~ ~ ' U ~ o U d ~, ' 3 ~ ~
dd d d dd-d Q r;
~ ~

~ v ci ~t ~ d c,; ~ ~ O -d~ o, ~ d ~
W ~ oNO
o _ , l~ 00 I~ 00~ ~ 00 ~ 00 l~ M ~
~O O ~ O M ~ N p '~h N \p V7 M ~ 00 N , M d ~ ~ ~ N 01 ~ O
M p M -, M ~
D O O D O N
~ M

O ~ ~ N M N '.~~M ~ d N z U ~ O O
U U U U oo ~ ~~
~

ddddd ~ x ~dd~.,; '~~ d~
N d~
o~~~

N M v'i O ~ N oov'i ~ ~Doox vi M d oo t~
I~ ~ ~ M ~ ~ N a\N M W ~ M O ~ d M
o0 O N ~ M oo O
'~t ~ ~-' O

n ~jv~ l~O O ~O O O ~ d' r,;O ,rj N o0 v~
O oo ~ N l~ M ~ 0000 00 0 ~ M O 00 O ~ 00 N
' ~

I~ ~ M ~ M ~ ~ N ~ M
.~ c? O 01 p ,-. ~ N -~~ ~n ~ pp ~ M N O O 0o I~ ~ r, 00 ~ ~n ~p ~D
O M O '".' ~
'"'' O O O O O z N O ~ O ~ ~ O z v1 O~ ~ M
O O n ~ ~ ~ ~ ~ O
~~~~dd~ ~~ ~d~~ d~ ~zdd~Nd~

\Oo0 N O o0 00lp~ M l~Q1 O .W p \ON O \Ol~~ N t~

V1 ~ ~ M V7~ ~DV7 M
i i ~ i i ~ i ~ i ~ i N ~ ~ N ~ N N ~ ~ N

N ~ 00~O~ ~ N I~

O~O ~ ~ ~ Q\v~~ N ~ ~D

V1 ~ ~ M ~ ~!1~D~ M
i ~ i i i ~ i ~ i i ~

O O~ ~ ~ oo M O l~~n N ~n M 01 010000 ~ O M O

O M M \O~O O ~O~ O \O~

~O d'M '~ d'O ~OD1 O O

O w 0 N -r N OvN

a1 ~ ~ a\ d1 M d' ~ ~ l~ 00O~O ~ N M

~t ~ ~ ~ ~ ~ ~t~ d'~

~ M O O M ~ l~~O O~M

U ~ ~ U ~

U U U U U U U d d x x x x x x x x x x x ~n ~--~ ~--~ t~ N '~ ~ ~~ a; ~i ri O i!1 I~ 00 ~ O ~ M
vp M ~n ~n ~ ~ O ~ ~ I~ N ~ OM ~ oho ~ ~ ~ O
O ~ O O ~ O v7 N ~ ~ ~' M
O ~ O O ~ O O O 0 M O M ~ ~ p ~ ~ M V~ ~ ~ M
U p U U (~ a a oo ~ °' °~ ,~ ~n °~ 'o d o d d ~ d d d ~ ° ~ o °~°° ~ ~ ~ ~ ~ N 3 U o 00 ~ ~' d ~ M ov o ~ d d M ~
N ~ ~ ~ o N oo vo ~ o ~ ~ ~ v-, ,~ N o ~" o ~ M ~ ~ Z M
U ~ O l~
N ~ v .-~~ ~ O ~ ~ ~ ~ 01 N ~n O N O O ~,,~ M O O p~ ~ U N M ~. O v7 N M O ~ d ~p O o~p N
U M U U o 0 0 0 "i ~ o d n o°~o ~ °; ~ ~ '-' ~ ~t ~ d a, ~
~ ° ~
x~zN~~dd~~M
O I~ ~ N ~ ~ O ~ N c.~ ~ d O O N M v~ M ~ '-' m N
M
O_ N N ~ ~ ~ U ~ ~ ,~-, d o0 p ct O ~ ~ ~ d' O ~ '~' ~ ~D o00 ONOOOOOOb~ ~~ OW'~~Ol~oN0OMIN O ~M
O O ,~ ,~ N ~_O N ~.,~ O d r-i r-i ~ U ~ pip ~"~ N
do~ddd~d~° °3~°x~~~d~dNdU~d~~
c~s d~ oo ~ c~i wi vi ~ x x d d Ca 'o v~ '~ o°°o yo ~ ~ v-i o 0 op ~-~ O op ~p .~ ~ ~ ~ N N ~n N ~O oo N ~
[~ 00 O~ M ~ ,-~ O M ~ ~ ,~ ,~ p~ ~ N ~O D1 ~ ~ M .~ l M M O O ~ O O M O pM1 ~ d1 <t ~ O ~ N ~ 00 N M M ~ ~ M M ~ O v0 M
.~ ,-, O O O O O O O O M O d' t~ M ~ ~ '_"' ~ '-' I~ V~ N ~~ V~
a ov U U ~ ,~ U l~ ~p ~ M 'n M ~ d 3 ~ d 'b °' d~ddd~~~dxHxxzz~~~dddd~~~~dddd N ~ ~O M ~ a1 ~D O
N 01 01 l~ V7 ~ M M ~ d' M ~ 00 i i ~ ~ ~ i i ~
~ V~ ~ N N N V~ V7 00 ~ N Ov O ~n N ~D
O o0 00 v~ d' M O N
01 M M V' ~' M V7 00 i i ~ i i i i i ~' ~O Q~ M 00 ~O '~' \O
~ N ~O N ~ oo ~n O
00 M ~ 00 00 00 O O
~ O ~O N ~ ~D 01 M
~ d, ~
Q\ ~ ~
~t VW O l~ 00 Q\ O
V
M O ~p N ~ ,~ ~ ~O
dd xxxxx xxx I~ M
~ I~ 00 N p ~ ~ ~ O ~ O
O
O M ~ M ~ ~ ~ l~ 0~0 0~0 N N ~ ~O o0 V'1 ~ ~ M O~ ~ ~ ~ ., 01 01 ~O O ~ ~ M
O ~ d d d ~O ~ M N N p~ M ~ M ~ ~ ~ N N M l~ 00 '~~~~~~dw~~HQ' ~ o,~ "' O vp ~j pp ~O m N N ~ ~ ~ r' O a1 ~ ~ ~ ~ l~ O
M oo N ~ ~ ~ ~ ~n ~ x ~ p~ ~' op o0 v~ pp ~ N O v~
~ N M ~ oo M o0 N O ~ M ~''W p M ~ l~ M ~ ~ p ~ ~ O
O N ~ ~ N ~ ~ ~." ~ O O
~ ~ °° o ~ ~ d ~ ~ ~ ~ o d ~, d ~ o N ~ x ~ ~ ~ ~ ~ ci o 0 00 ~p M ~ ~ O M M ~ '~' N V7 00 O ~ U r~r O l~ ~ V7 V~ 00 I~ 00 ~ C
N ~ v7 00 ~ M pp I~ 00 t~ ~ O
'p N ~ ~-.~ N ~ ~ N .~ N d 'p" ~ ~' 01 p~ O O 01 00 d N
p1 d i"'i d ~ ~ '"'' 00 ~ ~ pMp d ~ ~ ~ ~ d ~ ~ 00 ~~ddddz~ddd N d ~~"~ ''' M
o, c~i t~ ~ t~ os t~ ~ ~ t~ ~ M ~ oo ,~ Z c~i ~ ~ N --~ ~ ~ z ~ os o, t~ ~ Q\ ..-~ oo t~ t~ o t~ 0 00 .~ ~i cri ~ ~ ,.., ~.
-.~ O N ~p ~ M N .-~ 00 ~ l~ ~O ~ N ~ ~h ~ " ~ ~ ~' Q1 ~ M ~ N
V7 O d1 ~ ~ O~ ~ N ~~ 00 l~ M M ~ ~ r"' 00 ~ ~ ~ ~ N ~'-' lp ~ 00 I~ ~
N ~ I~ l~ ~ v~ ~ ~ 00 ~ v~ O N oo ~ N p~ ~ ~ 00 ~ 00 O ~ ~ p~ p~ O
~n ~ d' l~ O ,~ 00 N V v~ M 01 O o0 ~ M v~ O O ~ ~ O ,~ N N N N ~ M
~~~~~~~~~~~~~~~~~dz~z~~~d~~~ x M I~ O
O
i 01 V) M
O N
N

N M ~h' Wit' ~t ~t O "'~ M
U U
x x O o0 N I~ M
~ M l~ 00 M M N M
00 ~ ~-~ 00 v7 t~ O N ~ I~ pp N O '-. M
M ~ I~ N M N M ~ ~ N ~' ,~-, ~. ~ .-~ M ~ l~ ~ N
r-~ N N ~ ~ M N ~ O ~ M ~.ij O .~ O ~ N ~ ~D
~ N p ~ 0 t~ oo N O ~O .~ lmn ~ ''.' ~", 000 .-, ~ O ~ ~ ~ O N O
O ~ ~ F-pp-. ~ ~'"T'"' N ~ M d d ~ ~, M d1 ~ V7 N ,-i l~
p0l~o~Od~_~~~01~~~M~~~,M_, ~ ~N
ono '~ ~ t~ N r' ~ ~ t~ ~ '"'' ~ ~' ~ N r" ~ ~ ~ v_~
rN, cd O l~ ~ '~ O Ov ~ M ~ '-' ~ N Q1 N vN'1 ~ M N N ~ O U
~ M ~ ~ M 00 00 ~"~ M ~ ~ ~ 00 N ~ pp d3~~3~d~ x°~ d p '~ ~ ~''~ l~ M N O d' d oo ~ ~ ~ ~n M ~ ~ oo " N O
O ~ py0 N d N M M ~ .~ I~ ~ V~ V~ ~ r, ~' 00 V7 M M p~ O ,-, ~wj 01 00 N M I~ ~ M M M oo O ~' O~ p ~ O ~ ,--~ M Ov p N ~ N vp ~ d ~ d ~ ~ ~ oo d ~ x ~ ~ ~ d ~ ~ ~ M ~ ~n 3 ~dH~~~d ~.~~~~~
Mxp~p~ dNO~ht~ON N~~oM001N~NO~ N ~Nx ~ ~O M l~ ~ 01 ~ 00 ~ O
M M ,~~. ,~ yn 01 l_~ N .~ I~ ~O ~O l~ I~ ~ N '-~ r' N l~ pp ono O
N M'~~ N~M~pWOO~o~M~o~OO~o~O~N~O~ ~~t~
N '_'.' ~ ~fi '-' d- d ~ .~ O d1 N v0 vD d d ~ ~ M ~ ~ O
~~x~~ x~d~dd~r~d~~d~~wddddH~ ~d~
~n t~ N O N

~1' ~n v~
~ i ~ i i ~n ~mn ~n ~n ,~ M 00 ~D 00 N V' i i i i O N l~
I~ V7 O 00 00 ~ M O~ Q1 M

Op 00 OO 00 00 d' d' N M M l~
~x dd d dd xx x xx N
M
N
,~ N
c~
N
N N
Q\
. d °' ~~d ~~z d' N
O l~
M l~
-~ N
N I
M
O N
d xx Code Description Tissue Orean Cell DiseaseVector Line AR022 a Heart a Heart AR023 a Liver a Liver AR024 a mammary glanda mammary gland AR025 a Prostate a Prostate AR026 a small intestinea small intestine AR027 a Stomach a Stomach AR028 Blood B cells Blood B cells AR029 Blood B cells Blood B cells activated activated AR030 Blood B cells Blood B cells resting restin AR031 Blood T cells Blood T cells activated activated AR032 Blood T cells Blood T cells resting resting AR033 brain brain AR034 breast breast AR035 breast cancer breast cancer AR036 Cell Line CAOV3Cell Line AR037 cell line PA-1cellline AR038 cell linetransformedcell line transformed AR039 colon colon AR040 colon 9808co65R)colon (9808co65R

AR041 colon 9809co15)colon 9809co15) AR042 colon cancer colon cancer AR043 colon cancer colon cancer (9808co64R) (9808co64R) AR044 colon cancer colon cancer 9809co14 9809co14 AR045 corn clone corn clone AR046 corn clone corn clone AR047 corn clone2 corn clone2 AR048 corn clone3 corn clone3 AR049 Corn Clone4 Com Clone4 ARO50 Donor 11 B Donor II
Cells 24hrs B Cells 24hrs ARO51 Donor 11 B Donor II
Cells 72hrs B Cells 72hrs AR052 Donor 11 B-CellsDonor II
24 hrs. B-Cells hrs.

AR053 Donor II B-CellsDonor II
72hrs B-Cells 72hrs AR054 Donor II RestingDonor II
B Cells Resting B
Cells ARO55 Heart Heart AR056 Human Lung Human Lung (clonetech (clonetech AR057 Human Mammary Human Mammary (clontech) (clontech) AR058 Human Thymus Human Thymus (clonetech) clonetech AR059 Jurkat (unstimulated)Jurkat unstimulated AR060 Kidne Kidne AR061 Liver Liver AR062 Liver Clontech)Liver Clontech) AR063 Lymphocytes Lymphocytes chronic lymphocytic chronic lymphocytic leukaemia leukaemia AR064 Lymphocytes Lymphocytes diffuse large B cell diffuse large lymphoma B cell lym homa AR065 Lymphocytes Lymphocytes follicular 1 homa follicular 1 homa AR066 normal breast normal breast AR067 Normal OvarianNormal Ovarian AR068 Normal Ovary Normal Ovary AR069 Normal Ovary Normal Ovary AR070 Normal Ovary Normal Ovary AR071 Ovarian CancerOvarian Cancer AR072 Ovarian CancerOvarian Cancer 97026001) 97026001 AR073 Ovarian CancerOvarian Cancer (97076029) (97076029 AR074 Ovarian CancerOvarian Cancer (98046011 (98046011 ) AR075 Ovarian CancerOvarian Cancer 98066019) (98066019) AR076 Ovarian CancerOvarian Cancer 98076017 (98076017 AR077 Ovarian CancerOvarian Cancer (98096001 (98096001 AR078 ovarian cancerovarian cancer AR079 Ovarian CancerOvarian Cancer AR080 Ovarian CancerOvarian Cancer AR081 Ovarian CancerOvarian Cancer AR082 ovarian cancerovarian cancer AR083 Ovarian CancerOvarian Cancer AR084 Ovarian CancerOvarian Cancer AR085 Ovarian CancerOvarian Cancer AR086 ovarian cancerovarian cancer AR087 Ovarian CancerOvarian Cancer AR088 Ovarian cancerOvarian cancer C00 3rd C00 3rd AR089 Prostate Prostate AR090 Prostate clonetechProstate (clonetech AR091 rostate cancerrostate cancer AR092 prostate cancerprostate #15176 cancer #15176 AR093 prostate cancerprostate #15509 cancer #15509 AR094 prostate cancerprostate #15673 cancer #15673 AR095SmalllntestineSmalllntestine (Clontech) (Clontech) AR096S leen S Teen AR097Thymus T cellsThymus T
cells activated activated AR098Thymus T cellsThymus T
resting cells restin AR099Tonsil Tonsil AR100Tonsil geminalTonsil geminal center centroblast center centroblast ARI01Tonsil germinalTonsil germinal center B

cell center B
cell AR102Tonsil 1 h Tonsil 1 node h node AR103Tonsil memory Tonsil memory B cell B

cell AR104Whole Brain Whole Brain AR105Xeno raft ES-2Xenograft AR106Xeno raft SW626Xeno raft H0068Human Skin Human Skin Skin diseaseUni-ZAP
Tumor Tumor XR

H0081Human Fetal Human Fetal Skin Uni-ZAP
Epithelium Skin Skin XR

H0086Human epithelioidEpithelioid Sk Muscle diseaseUni-ZAP

sarcoma Sarcoma, XR
muscle H0344Adipose tissueAdipose - Uni-ZAP
(human) 6825A

(human) XR

H0345SK1N Skin - 4000868HSkin Uni-ZAP

XR

H0427Human Adipose Human Adipose, pSportl left hi 1i oma H0443H. Adipose, Human Adipose, pSportl subtracted left hi 1i oma H0494Keratinocyte Keratinocyte pCMVSport 2.0 H0540Skin, burned Skin, leg Skin S ortl burned H0548Human Skin Human Skin pBluescript Fibroblasts, normal Fibroblasts H0586Healing groin healing groingroin diseasepCMVSport wound, 6.5 hours postwound, 6.5 3.0 incision hours ost incision H0587Healing groin Groin-2/19/97groin diseasepCMVSport wound;

7.5 hours ost 3.0 incision H0592Healing~groin HGS wound diseasepCMVSport wound - healing zero hr post-incisionproject; 3.0 abdomen (control) H0593Olfactory Olfactory pCMVSport epithelium epithelium;nasalcavityfrom roof 3.0 of left nasal cacit H0600Healing AbdomenAbdomen diseasepCMVSport wound;70&90 3.0 min post incision H0601Healing AbdomenAbdomen diseasepCMVSport Wound;l5 days 3.0 post incision H0602Healing AbdomenAbdomen diseasepCMVSport Wound;21 &29 3.0 days post incision 50040Adipocytes Human Adipocytes Uni-ZAP

from Osteoclastoma XR

50280Human Adipose Human Adipose Uni-ZAP
Tissue, re-excision Tissue XR

50342 Adipocytes;re-excisionHuman Adipocytes Uni-ZAP

from Osteoclastoma XR

50348 Cheek CarcinomaCheek Carcinoma diseaseS ortl S6022 H. Adipose Human Adipose Uni-ZAP
Tissue Tissue XR

T0001 Human Brown Brown Fat pBluescript Fat SK-T0004 Human White Human White pBluescript Fat Fat SK-T0060 Human White Human White pBluescript Adipose Fat SK-L0005 Clontech human aorta of A+ mRNA
#6572 L0021 Human adult K.Okubo L0060 Human th mus NSTH II

L0361 Stratagene ovary Bluescript ovary #937217) SK

L0362 Stratagene Bluescript ovarian cancer(#937219) SK-L0363 NCI CGAP GC2 germ cell Bluescript tumor S K-L0365 NCI CLAP Phel pheochromocytoma Bluescript SK-L0366 Stratagene schizophrenic Bluescript schizo brain brain S11 S-11 frontal SK-lobe L0369 NCI CGAP AA1 adrenal adenomaadrenal Bluescript gland S K-L0375 NCI CGAP_Kid6 kidney tumorkidney Bluescript SK-L0376 NCI CGAP Larl larynx larynx Bluescript S K-L0385 NCI CGAP Gasl gastric tumorstomach Bluescript S K-L0388 NCI CGAP HN6 normal gingiva Bluescript (cell l ine from S K-immortalized kerati L0435 Infant brain, lafmid LLNL BA

array of Dr.
M. Soares L0438 normalized total brain brain lafmid infant brain BA

cDNA

L0439 Soares infant whole Lafmid brain brain BA

L0455 Human retina retina eye lambda cDNA gtl0 randomly primed sublibrary L0471 Human fetal Lambda heart, Lambda ZAP ZAP
Express Ex ress L0483 Human pancreatic Lambda islet ZAPII

L0517 NCI CGAP Prl AMP10 L0518 NCI CGAP Pr2 AMP 10 L0519 NCI CGAP Pr3 AMP 10 L0520 NCI CGAP_Alvl alveolar pAMPlO

rhabdom osarcoma L0521 NCI CGAP Ewl Ewin "s sarcoma AMP10 L0527 NCI CGAP Ov2 ovary AMP10 L0532 NCI CGAP Th th oid AMP10 L0539 Chromosome placenta pAMPlO

Placental cDNA
Libra L0545 NCI CGAP Pr4.1prostatic prostate pAMPlO

intraepithelial neoplasia - high ade L0562 Chromosome HeLa pAMPlO
7 HeLa cell cDNA Library line;

ATCC

L0581 Stratagene liver pBluescript liver #937224 SK

L0589 Stratagene pBluescript fetal retina L0591 Stratagene pBluescript HeLa cell s3 L0592 Stratagene pBluescript hNT neuron #937233) SK-L0593 Stratagene pBluescript neuroepithelium SK-#937231) L0594 Stratagene pBluescript neuroepithelium SK-L0595 Stratagene neuroepithelialbrain pBluescript NT2 cells neuronal precursor SK-L0596 Stratagene colon pBluescript colon (#937204 SK-L0598 Morton Fetal cochlea ear pBluescript Cochlea SK-L0599 Stratagene lung pBluescript lung #937210) SK-L0600 Weizmann Olfactoryolfactory nose pBluescript epithelium E ithelium SK-L0601 Stratagene pancreas pBluescript pancreas (#937208) SK-L0602 Pancreatic pancreatic pancreas pBluescript Islet islet SK-L0603 Stratagene placenta pBluescript placenta (#937225 SK-L0604 Stratagene muscle skeletal pBluescript muscle 937209 muscle SK-L0605 Stratagene fetal spleenspleen pBluescript fetal spleen (#937205 SK-L0608 Stratagene lung carcinomalung NCI-H69 pBluescript lung carcinoma 937218 SK-L0616 Chromosome pBluescriptI
21 exon IKS+

L0637 NCI CGAP Brn53three pooledbrain pCMV-menin iomas SPORT6 L0638 NCI CGAP Brn35tumor, 5 brain pCMV-pooled (see d escri tion SPORT6 L0653 NCI CGAP Lu28 two pooled lung pCMV-squamous SPORT6 cell carcinomas L0655 NCI CGAP_Lyml2lymphoma, lymph pCMV-node f ollicular SPORT6 mixed small and large cell L0659 NCI CGAP Panl adenocarcinomapancreas pCMV-L0662 NC1 CGAP_Gas4 poorly differentiatedstomach pCMV-adenocarcinoma SPORT6 with si et r L0663 NCI CGAP Ut2 moderately- uterus pCMV-differentiated SPORT6 endometrial adenocarcino L0664 NCI CGAP-Ut3 poorly-differentiateduterus pCMV-endometrial SPORTG

adenocarcinoma, L0666 NCI CGAP Utl well-differentiateduterus pCMV-endometrial SPORT6 adenocarcinoma, L0667 NCI CGAP CML1 myeloid cells,whole pCMV-18 blood C ML cases, SPORT6 pooled BCR/ABL rearra L0717 Gessler Wilms SPORT1 tumor L0731 Soares-pregnant uterus pT7T3-Pac uterus NbHPU

L0740 Soares melanocytemelanocyte pT7T3D

2NbHM (Pharmacia) with a modified of linker L0741 Soares adult brain pT7T3D
brain N2b4HB55Y (Pharmacia) with a modified of linker L0742 Soares adult brain pT7T3D
brain N2b5HB55Y (Pharmacia) with a modified of linker L0743 Soares breast breast pT7T3D
2NbHBst (Pharmacia) with a modified polylinker L0744 Soares breast breast pT7T3D
3NbHBst (Pharmacia) with a modified olylinker L0745 Soares retina retina eye pT7T3D
N2b4HR

(Pharmacia) with a modified olylinker L0746 Soares retina retina eye pT7T3D
N2b5HR

(Pharmacia) with a modified of linker L0747 Soares_fetal heart pT7T3D
heart_NbH

H 19W (Pharmacia) with a modified of linker L0748 Soares fetal Liver pT7T3D
liver spleen and 1 NFLS Spleen (Pharmacia) with a modified of linker L0749 Soares fetal_liver_splee Liver pT7T3D
~ and n 1NFLS SI Spleen (Pharmacia) with a modified olylinker L0750 Soares_fetal . lung pT7T3D
lung NbH

L19W (Pharmacia) with a modified olylinker L0751 Soares ovary ovarian tumorovary pT7T3D
tumor NbHOT (Pharmacia) with a modified of linker L0752 Soares-parathyroidparathyroid parathyroid pT7T3D
tum tumor or_NbHPA gland (Pharmacia) with a modified of linker L0754 Soares placenta placenta pT7T3D
Nb2HP

(Pharmacia) with a modified olylinker L0755 Soares-placenta placenta pT7T3D
8to9we eks_2NbHP8to9W (Pharmacia) with a modified of linker L0756 Soares_multiplemultiple pT7T3D
scleros sclerosis is 2NbHMSP lesions (Pharmacia) with a modified polylinker V TYPE

L0757 Soares_senescent_fibrobsenescent pT7T3D
fibroblast lasts NbHSF (Pharmacia) with a modified polylinker V TYPE

L0758 Soares testis_NHT pT7T3D-Pac (Pharmacia) with a modified olylinker L0759 Soares_total pT7T3D-fetus Nb2 HF8_9w Pac (Pharmacia) with a modified olylinker L0763 NCI CGAP_Br2 breast pT7T3D-Pac (Pharmacia) with a modified olylinker L0764 NCI CGAP-Co3 colon pT7T3D-Pac (Pharmacia) with a modified of linker L0766 NCI CGAP GCB1 germinal pT7T3D-center B

cell Pac (Pharmacia) with a modified olylinker L0768 NCI CGAP_GC4 pooled germ pT7T3D-cell tumors Pac (Pharmacia) with a modified of linker L0769 NCI CGAP-Brn25anaplastic brain pT7T3D-oligodendroglioma Pac (Pharmacia) with a modified of linker L0770 NCI CGAP_Brn23glioblastomabrain pT7T3D-( pooled) Pac (Pharmacia) with a modified polyl in ker L0772 NCI CGAP ColO colon tumor colon pT7T3D-RER+

Pac (Pharmacia) with a modified polyl i nker L0773 NCI CGAP Co9 colon tumor colon pT7T3D-RER+

Pac (Pharmacia) with a modified of linker L0774 NCI CGAP Kid3 kidney pT7T3D-P ac (Pharmacia) with a modified polylinker L0775 NCI CGAP Kids 2 pooled kidney pT7T3D-tumors (clear cell Pac type) (Pharmacia) with a modified olylinker L0776 NC1 CGAP_Lu5 carcinoid lung pT7T3D-Pac (Pharmacia) with a modified of linker L0777 Soares_NhHMPu Pooled humanmixed pT7T3D-S1 (see melanocyte, below) Pac fetal heart, and (Pharmacia) pregnant with a modified of linker L0779 Soares NFL_T_GBC pooled pT7T3D-S

1 Pac (Pharmacia) with a modified of linker L0780 Soares_NSF_F8 pooled pT7T3D-9Wy0 T_PA_P_S1 Pac (Pharmacia) with a modified of linker L0781 Barstead prostate prostate pT7T3D-BPH

HPLRB4 Pac (Pharmacia) with a modified polylinker L0783 NCI CGAP_Pr22 normal prostateprostate pT7T3D-Pac (Pharmacia) with a modified polylinker L0789 NCI CGAP Sub3 pT7T3D-Pac (Pharmacia) with a modified olylinker L0791 NCI CLAP Subs pT7T3D-Pac (Pharmacia) with a modified olylinker L0792 NCI CGAP_Sub6 pT7T3D-Pac (Pharmacia) with a modified polylinker L0794 NCI CGAP GC6 pooled germ pT7T3D-cell tumors Pac (Pharmacia) with a modified of linker L0800 NCI CGAP Col6 colon tumor,colon pT7T3D-RER+

P ac (Pharmacia) with a modified of linker L0803 NCI CGAP_Kid kidney pT7T3D-Pac (Pharmacia) with a modified polylinker L0804 NCI CGAP_Kidl22 pooled kidney pT7T3D-tumors ( clear cell Pac type) (Pharmacia) with a modified of linker L0805NCI CGAP_Lu24 carcinoid lung pT7T3D-Pac (Pharmacia) with a modified of linker L0806NCI CGAP Lul9 squamous celllung pT7T3D-carcinoma, Pac poorly differentiated (Pharmacia) (4 with a modified of linker L0809NCI CGAP Pr28 prostate pT7T3D-P ac (Pharmacia) with a modified of linker L2251Human fetal Fetal lung ~ lung I

OMIM Description Reference 102200 Somatotrophinoma 106100 Angioedema, hereditary 107680 ApoA-I and apoC-III deficiency, combined 107680 Corneal clouding, autosomal recessive 107680 Amyloidosis, 3 or more types 107680 Hypertriglyceridemia, one form 107680 Hypoalphalipoproteinemia 107720 Hypertriglyceridemia 108725 Atherosclerosis, susceptibility to 120550 Clq deficiency, type A

120570 Clq deficiency, type B

120575 Clq deficiency, type C

120700 C3 deficiency 120950 C8 deficiency, .type I

120960 C8 deficiency, type II

130500 Elliptocytosis-1 131100 Multiple endocrine neoplasia I

131100 Prolactinoma, hyperparathyroidism, carcinoid syndrome 131100 Carcinoid tumor of lung 133171 [Erythrocytosis, familial], 133100 133200 Erythrokeratodermia variabilis 133780 Vitreoretinopathy, exudative, familial 134934 Thanatophoric dysplasia, types I and II, 187600 134934 Achondroplasia, 100800 134934 Craniosynostosis, nonsyndromic 134934 Crouzon syndrome with acanthosis nigricans 134934 Hypochondroplasia, 146000 136836 Fucosyltransferase-6 deficiency 138140 Glucose transport defect, blood-brain barrier 143100 Huntington disease 145981 Hypocalciuric hypercalcemia, type II

147050 Atopy 147141 Leukemia, acute lymphoblastic 147791 Jacobsen syndrome 153454 Ehlers-Danlos syndrome, type VI, 225400 153700 Macular dystrophy, vitelliform type 159555 Leukemia, myeloid/lymphoid or mixed-lineage 161015 Mitochondrial complex I deficiency, 252010 164009 Leukemia, acute promyelocytic, NL1MA/RARA type 164953 Liposarcoma 167410 Rhabdomyosarcoma, alveolar, 268220 168000 Paraganglioma, familial nonchromaffin, 1 168360 Paraneoplastic sensory neuropathy 168461 Multiple myeloma, 254250 168461 Parathyroid adenomatosis 1 168461 Centrocytic lymphoma 171760 Hypophosphatasia, adult, 146300 171760 Hypophosphatasia, infantile, 241500 176100 Porphyria cutanea tarda 176100 Porphyria, hepatoerythropoietic 177070 Spherocytosis, hereditary, Japanese type 177070 Hermansky-Pudlak syndrome, 203300 178300 Ptosis, hereditary congenital, 1 180072 Night blindness, congenital stationary, type 3, 180072 Retinitis pigmentosa, autosomal recessive 180721 Retinitis pigmentosa, digenic 180840 Susceptibility to B7DM

182500 Cataract, congenital 186740 hnmunodeficiency due to defect in CD3-gamma 186830 Immunodeficiency, T-cell receptor/CD3 complex 187040 Leukemia-1, T-cell acute lymphoblastic 188025 Thrombocytopenia, Paris-Trousseau a 188070 Bleeding disorder due to defective thromboxane A2 receptor 191181 Cervical carcinoma 193235 Vitreoretinopathy, neovascular inflammatory 194190 Wolf Hirschhorn syndrome 203750 3-ketothiolase deficiency 209901 Bardet-Biedl syndrome 1 218000 Andermann syndrome 227220 [Eye color, brown]

230000 Fucosidosis 232600 McArdle disease 243500 Isovalericacidemia 252800 Muco olysaccharidosis Ih 252800 Mucopolysaccharidosis Ih/s 252800 Mucopolysaccharidosis Is 255800 Schwanz-Jampel syndrome 256700 Neuroblastoma 259700 Osteopetrosis, recessive 259770 Osteoporosis-pseudoglioma syndrome 261640 Phenylketonuria due to PTS deficiency 300011 Menkes disease, 309400 300011 Occipital horn syndrome, 304150 300011 Cutis laxa, neonatal 300127 Mental retardation, X-linked, 60 305450 FG syndrome 313700 Perineal hypospadias 313700 Prostate cancer 313700 Spinal and bulbar muscular atrophy of Kennedy, 313700 Breast cancer, male, with Reifenstein syndrome -313700 Androgen insensitivity, several forms 600045 Xeroderma pigmentosum, group E, subtype 2 600048 Breast cancer-3 600101 Deafness, autosomal dominant 2 600319 Diabetes mellitus, insulin-dependent, 4 600528 CPT deficiency, hepatic, type I, 255120 600650 Myopathy due to CPT II deficiency, 255110 600650 CPT deficiency, hepatic, type II, 600649 600722 Ceroid lipofuscinosis, neuronal, variant juvenile type, with granular osmiophilic deposits 600722 Ceroid lipofuscinosis, neuronal-l, infantile, 256730 600839 Banter syndrome, 241200 600957 Persistent Mullerian duct syndrome, type I, 261550 600965 Deafness, autosomal dominant 6 600975 Glaucoma 3, primary infantile, B

601238 Cerebellar ataxia, Cayman type 601382 Charcot-Marie-Tooth neuropathy-4B

601800 [Hair color, brown]

601846 Muscular dystrophy with rimmed vacuoles 601884 [High bone mass]

602216 Peutz-Jeghers syndrome, 175200 602477 Febrile convulsions, familial, 2 60 Deafness, autosomal dominant 12, 601842 _ eafness, autosomal dominant 8, 601543 602574 ~

Polynucleotide and Polypeptide Variants [095] The present invention is also directed to variants of the connective tissue associated polynucleotide sequence disclosed in SEQ >D NO:X or the complementary strand thereto, nucleotide sequences encoding the polypeptide of SEQ ID NO:Y, the nucleotide sequence of SEQ ID NO:X encoding the polypeptide sequence as defined in column 6 of Table 1A, nucleotide sequences encoding the polypeptide as defined, in column 6 of Table 1A, the nucleotide sequence as defined in columns 8 and 9 of Table 2, nucleotide sequences encoding the polypeptide encoded by the nucleotide sequence as defined in columns 8 and 9 of Table 2, the nucleotide sequence as defined in column 6 of Table 1B, nucleotide sequences encoding the polypeptide encoded by the nucleotide sequence as defined in column 6 of Table 1B, the cDNA sequence contained in Clone ID
NO:Z, and/or nucleotide sequences encoding a polypeptide encoded by the cDNA
sequence contained in Clone >D NO:Z.
[096] The present invention also encompasses variants of the polypeptide sequence disclosed in SEQ m NO:Y, a polypeptide sequence as defined in column 6 of Table 1A, a polypeptide sequence encoded by the polynucleotide sequence in SEQ ID NO:X, a polypeptide sequence encoded by the nucleotide sequence as defined in columns 8 and 9 of Table 2, a polypeptide sequence encoded by the nucleotide sequence as defined in column 6 of Table 1B, a polypeptide sequence encoded by the complement of the polynucleotide sequence in SEQ )D NO:X, and/or a polypeptide sequence encoded by the cDNA sequence contained in Clone m NO:Z.
[097] "Variant" refers to a polynucleotide or polypeptide differing from the polynucleotide or polypeptide of the present invention, but retaining essential properties thereof. Generally, variants are overall closely similar, and, in many regions, identical to the polynucleotide or polypeptide of the present invention.
[098] Thus, one aspect of the invention provides an isolated nucleic acid molecule comprising, or alternatively consisting of, a polynucleotide having a nucleotide sequence selected from the group consisting of (a) a nucleotide sequence described in SEQ >D
NO:X or contained in the cDNA sequence of Clone )17 NO:Z; (b) a nucleotide sequence in SEQ >I7 NO:X or the cDNA in Clone >D NO:Z which encodes a mature connective tissue associated polypeptide; (c) a nucleotide sequence in SEQ >Z7 NO:X or the cDNA

sequence of Clone >D NO:Z, which encodes a biologically active fragment of a connective tissue associated polypeptide; (d) a nucleotide sequence in SEQ >D
NO:X or the cDNA sequence of Clone >D NO:Z, which encodes an antigenic fragment of a connective tissue associated polypeptide; (e) a nucleotide sequence encoding a connective tissue associated polypeptide having the complete amino acid sequence of SEQ >D NO:Y or the complete amino acid sequence encoded by the cDNA in Clone >D
NO:Z; (f) a nucleotide sequence encoding a mature connective tissue associated polypeptide of the amino acid sequence of SEQ >D NO:Y or the amino acid sequence encoded by the cDNA in Clone >D NO:Z; (g) a nucleotide sequence encoding a biologically active fragment of a connective tissue associated polypeptide having the complete amino acid sequence of SEQ >D NO:Y or the complete amino acid sequence encoded by the cDNA in Clone )D NO:Z; (h) a nucleotide sequence encoding an antigenic fragment of a connective tissue associated polypeptide having the complete amino acid sequence of SEQ >D NO:Y or the complete amino acid sequence encoded by the cDNA in Clone >I7 NO:Z; and (i) a nucleotide sequence complementary to any of the nucleotide sequences in (a), (b), (c), (d), (e); (f), (g), or (h), above.
[099] The present invention is also directed to nucleic acid molecules which comprise, or alternatively consist of, a nucleotide sequence which is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100%, identical to, for example, any of the nucleotide sequences in (a), (b), (c), (d), (e), (f), (g), (h), or (i) above, the nucleotide coding sequence in SEQ
>D NO:X or the complementary strand thereto, the nucleotide coding sequence of the cDNA contained in Clone >D NO:Z or the complementary strand thereto, a nucleotide sequence encoding the polypeptide of SEQ m NO:Y, a nucleotide sequence encoding a polypeptide sequence encoded by the nucleotide sequence in SEQ )D NO:X, a polypeptide sequence encoded by the complement of the polynucleotide sequence in SEQ
>D NO:X, a nucleotide sequence encoding the polypeptide encoded by the cDNA
contained in Clone >D NO:Z, the nucleotide coding sequence in SEQ )D NO:X as defined in columns 8 and 9 of Table 2 or the complementary strand thereto, a nucleotide sequence encoding the polypeptide encoded by the nucleotide sequence in SEQ >D NO:X as defined in columns 8 and 9 of Table 2 or the complementary strand thereto, the nucleotide coding sequence in SEQ >D NO:B as defined in column 6 of Table 1B or the complementary strand thereto, a nucleotide sequence encoding the polypeptide encoded by the nucleotide sequence in SEQ ID NO:B as defined in column 6 of Table 1B
or the complementary strand thereto, the nucleotide sequence in SEQ ID NO:X encoding the polypeptide sequence as defined in column 6 of Table 1A or the complementary strand thereto, nucleotide sequences encoding a polypeptide as defined in column 6 of Table 1A
or the complementary strand thereto, and/or polynucleotide fragments of any of these nucleic acid molecules (e.g., those fragments described herein).
Polynucleotides which hybridize to the complement of these nucleic acid molecules under stringent hybridization conditions or alternatively, under lower stringency conditions, are also encompassed by the invention, as are polypeptides encoded by these polynucleotides and nucleic acids.
[0100] In a preferred embodiment, the invention encompasses nucleic acid molecules which comprise, or alternatively, consist of a polynucleotide which hybridizes under stringent hybridization conditions, or alternatively, under lower stringency conditions, to a polynucleotide in (a), (b), (c), (d), (e), (f), (g), (h), or (i) above, as are polypeptides encoded by these polynucleotides. In another preferred embodiment, polynucleotides which hybridize to the complement of these nucleic acid molecules under stringent hybridization conditions or alternatively, under lower stringency conditions, are also encompassed by the invention, as are polypeptides encoded by these polynucleotides.
[0101] In another embodiment, the invention provides a purified protein comprising, or alternatively consisting of, a polypeptide having an amino acid sequence selected from the group consisting of-. (a) the complete amino acid sequence of SEQ ID NO:Y
or the complete amino acid sequence encoded by the cDNA in Clone ID NO:Z; (b) the amino acid sequence of a mature connective tissue associated polypeptide having the amino acid sequence of SEQ >D NO:Y or the amino acid sequence encoded by the cDNA in Clone ID NO:Z; (c) the amino acid sequence of a biologically active fragment of a connective tissue associated polypeptide having the complete amino acid sequence of SEQ
ID NO:Y
or the complete amino acid sequence encoded by the cDNA in Clone ID NO:Z; and (d) the amino acid sequence of an antigenic fragment of a connective tissue associated polypeptide having the complete amino acid sequence of SEQ ID NO:Y or the complete amino acid sequence encoded by the cDNA in Clone ID NO:Z.
[0102] The present invention is also directed to proteins which comprise, or alternatively consist of, an amino acid sequence which is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100%, identical to, for example, any of the amino acid sequences in (a), (b), (c), or (d), above, the amino acid sequence shown in SEQ ID NO:Y, the amino acid sequence encoded by the cDNA contained in Clone m NO:Z, the amino acid sequence of the polypeptide encoded by the nucleotide sequence in SEQ m NO:X
as defined in columns 8 and 9 of Table 2, the amino acid sequence of the polypeptide encoded by the nucleotide sequence in SEQ ~ NO:B as defined in column 6 of Table 1B, the amino acid sequence as defined in column 6 of Table 1A, an amino acid sequence encoded by the nucleotide sequence in SEQ ~ NO:X, and an amino acid sequence encoded by the complement of the polynucleotide sequence in SEQ ID NO:X.
Fragments of these polypeptides are also provided (e.g., those fragments described herein). Further proteins encoded by polynucleotides which hybridize to the complement of the nucleic acid molecules encoding these amino acid sequences under stringent hybridization conditions or alternatively, under lower stringency conditions, are also encompassed by the invention, as are the polynucleotides encoding these proteins.
[0103] By a nucleic acid having a nucleotide sequence at least, for example, 95%
"identical" to a reference nucleotide sequence of the present invention, it is intended that the nucleotide sequence of the nucleic acid is identical to the reference sequence except that the nucleotide sequence may include up to five point mutations per each nucleotides of the reference nucleotide sequence encoding the polypeptide. In other words, to obtain a nucleic acid having a nucleotide sequence at least 95%
identical to a reference nucleotide sequence, up to 5% of the nucleotides in the reference sequence may be deleted or substituted with another nucleotide, or a number of nucleotides up to 5% of the total nucleotides in the reference sequence may be inserted into the reference sequence. The query sequence may be an entire sequence referred to in Table 1A
or 2 as the ORF (open reading frame), or any fragment specified, as described herein.
[0104] As a practical matter, whether any particular nucleic acid molecule or polypeptide is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to a nucleotide sequence of the present invention can be determined conventionally using known computer programs. A preferred method for determining the best overall match between a query sequence (a sequence of the present invention) and a subj ect sequence, also referred to as a global sequence alignment, can be determined using the FASTDB
computer program based on the algorithm of Brutlag et al. (Comp. App. Biosci.
6:237-245 (1990)). In a sequence alignment the query and subject sequences are both DNA

sequences. An RNA sequence can be compared by converting U's to T's. The result of said global sequence alignment is expressed as percent identity. Preferred parameters used in a FASTDB alignment of DNA sequences to calculate percent identity are:
Matrix=Unitary, k-tuple=4, Mismatch Penalty=1, Joining Penalty=30, Randomization Group Length=0, Cutoff Score=1, Gap Penalty=5, Gap Size Penalty 0.05, Window Size=500 or the length of the subject nucleotide sequence, whichever is shorter.
[0105] If the subject sequence is shorter than the query sequence because of 5' or 3' deletions, not because of internal deletions, a manual correction must be made to the results. This is because the FASTDB program does not account for 5' and 3' truncations of the subject sequence when calculating percent identity. For subject sequences truncated at the 5' or 3' ends, relative to the query sequence, the percent identity is corrected by calculating the number of bases of the query sequence that are S' and 3' of the subject sequence, which are not matched/aligned, as a percent of the total bases of the query sequence. Whether a nucleotide is matched/aligned is determined by results of the FASTDB sequence alignment. This percentage is then subtracted from the percent identity, calculated by the above FASTDB program using the specified parameters, to arnve at a final percent identity score. This corrected score is what is used for the purposes of the present invention. Only bases outside the 5' and 3' bases of the subject sequence, as displayed by the FASTDB alignment, which are not matched/aligned with the query sequence, are calculated for the purposes of manually adjusting the percent identity score.
[0106] For example, a 90 base subject sequence is aligned to a 100 base query sequence to determine percent identity. The deletions occur at the 5' end of the subject sequence and therefore, the FASTDB alignment does not show a matched/alignment of the first 10 bases at 5' end. The 10 unpaired bases represent 10% of the sequence (number of bases at the 5' and 3' ends not matched/total number of bases in the query sequence) so 10% is subtracted from the percent identity score calculated by the FASTDB program. If the remaining 90 bases were perfectly matched the final percent identity would be 90%. In another example, a 90 base subject sequence is compared with a 100 base query sequence. This time the deletions are internal deletions so that there are no bases on the 5' or 3' of the subject sequence which are not matched/aligned with the query. In this case the percent identity calculated by FASTDB is not manually corrected.

Once again, only bases 5' and 3' of the subject sequence which are not matched/aligned with the query sequence are manually corrected for. No other manual corrections are to be made for the purposes of the present invention.
[0107] By a polypeptide having an amino acid sequence at least, for example, 95%
"identical" to a query amino acid sequence of the present invention, it is intended that the amino acid sequence of the subject polypeptide is identical to the query sequence except that the subject polypeptide sequence may include up to five amino acid alterations per each 100 amino acids of the query amino acid sequence. In other words, to obtain a polypeptide having an amino acid sequence at least 95% identical to a query amino acid sequence, up to 5% of the amino acid residues in the subject sequence may be inserted, deleted, (indels) or substituted with another amino acid. These alterations of the reference sequence may occur at the amino or carboxy terminal positions of the reference amino acid sequence or anywhere between those terminal positions, interspersed either individually among residues in the reference sequence or in one or more contiguous groups within the reference sequence.
[0108] As a practical matter, whether any particular polypeptide is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to, for instance, the amino acid sequence of a polypeptide referred to in Table 1A (e.g., an amino acid sequence identified in columns or 6) or Table 2 (e.g., the amino acid sequence of the polypeptide encoded by the polynucleotide sequence defined in columns 8 and 9 of Table 2) or a fragment thereof, the amino acid sequence of the polypeptide encoded by the polynucleotide sequence in SEQ
ID NO:B as defined in column 6 of Table 1B or a fragment thereof, the amino acid sequence of the polypeptide encoded by the nucleotide sequence in SEQ >D NO:X
or a fragment thereof, or an amino acid sequence of the polypeptide encoded by cDNA
contained in Clone ID NO:Z, or a fragment thereof, can be determined conventionally using known computer programs. A preferred method for determining the best overall match between a query sequence (a sequence of the present invention) and a subject sequence, also referred to as a global sequence alignment, can be determined using the FASTDB computer program based on the algorithm of Brutlag et al. (Comp. App.
Biosci.6:237-245 (1990)). In a sequence alignment the query and subject sequences are either both nucleotide sequences or both amino acid sequences. The result of said global sequence alignment is expressed as percent identity. Preferred parameters used in a FASTDB amino acid alignment are: Matrix=PAM 0, k-tuple=2, Mismatch Penalty=1, Joining Penalty=20, Randomization Group Length=0, Cutoff Score=1, Window Size=sequence length, Gap Penalty=5, Gap Size Penalty=0.05, Window Size=500 or the length of the subject amino acid sequence, whichever is shorter.
[0109] If the subject sequence is shorter than the query sequence due to N- or C-terminal deletions, not because of internal deletions, a manual correction must be made to the results. This is because the FASTDB program does not account for N- and C-terminal truncations of the subject sequence when calculating global percent identity.
For subject sequences truncated at the N- and C-termini, relative to the query sequence, the percent identity is corrected by calculating the number of residues of the query sequence that are N- and C-terminal of the subject sequence, which are not matched/aligned with a corresponding subject residue, as a percent of the total bases of the query sequence.
Whether a residue is matched/aligned is determined by results of the FASTDB
sequence alignment. This percentage is then subtracted from the percent identity, calculated by the above FASTDB program using the specified parameters, to arrive at a final percent identity score. This final percent identity score is what is used for the purposes of the present invention. Only residues to the N- and C-termini of the subject sequence, which are not matched/aligned with the query sequence, are considered for the purposes of manually adjusting the percent identity score. That is, only query residue positions outside the farthest N- and C- terminal residues of the subject sequence.
[0110] For example, a 90 amino acid residue subject sequence is aligned with a residue query sequence to determine percent identity. The deletion occurs at the N-terminus of the subject sequence and therefore, the FASTDB alignment does not show a matching/alignment of the first 10 residues at the N-terminus. The 10 unpaired residues represent 10% of the sequence (number of residues at the N- and C- termini not matched/total number of residues in the query sequence) so 10% is subtracted from the percent identity score calculated by the FASTDB program. If the remaining 90 residues were perfectly matched the final percent identity would be 90%. In another example, a 90 residue subject sequence is compared with a 100 residue query sequence.
This time the deletions are internal deletions so there are no residues at the N- or C-termini of the subject sequence which are not matched/aligned with the query. In this case the percent identity calculated by FASTDB is not manually corrected. Once again, only residue positions outside the N- and C-terminal ends of the subject sequence, as displayed in 'the FASTDB alignment, which are not matched/aligned with the query sequence are manually corrected for. No other manual corrections are to be made for the purposes of the present invention.
[0111] The polynucleotide variants of the invention may contain alterations in the coding regions, non-coding regions, or both. Especially preferred are polynucleotide variants containing alterations, which produce silent substitutions, additions, or deletions, but do not alter the properties or activities of the encoded polypeptide.
Nucleotide variants produced by silent substitutions due to the degeneracy of the genetic code are preferred. Moreover, polypeptide variants in which less than 50, less than 40, less than 30, less than 20, less than 10, or 5-50, 5-25, 5-10, 1-5, or 1-2 amino acids are substituted, deleted, or added in any combination are also preferred. Polynucleotide variants can be produced for a variety of reasons, e.g., to optimize codon expression for a particular host (change codons in the human mRNA to those preferred by a bacterial host such as E.
coli).
[0112] Naturally occurnng variants are called "allelic variants," and refer to one of several alternate forms of a gene occupying a given locus on a chromosome of an organism. (Genes II, Lewin, B., ed., John Wiley & Sons, New York (1985).) These allelic variants can vary at either the polynucleotide and/or polypeptide level and are included in the present invention. Alternatively, non-naturally occurring variants may be produced by mutagenesis techniques or by direct synthesis.
[0113] Using known methods of protein engineering and recombinant DNA
technology, variants may be generated to improve or alter the characteristics of the polypeptides of the present invention. For instance, one or more amino acids can be deleted from the N-terminus or C-terminus of the polypeptides of the present invention without substantial loss of biological function. As an example, the authors of Ron et al., J. Biol. Chem. 268: 2984-2988 (1993), reported variant KGF proteins having heparin binding activity even after deleting 3, 8, or 27 amino-terminal amino acid residues.
Similarly, Interferon gamma exhibited up to ten times higher activity after deleting 8-10 amino acid residues from the carboxy terminus of this protein. (Dobeli et al., J.
Biotechnology 7:199-216 (1988).) [0114] Moreover, ample evidence demonstrates that variants often retain a biological activity similar to that of the naturally occurring protein. For example, Gayle and coworkers (J. Biol. Chem. 268:22105-22111 (1993)) conducted extensive mutational analysis of human cytokine IL-la. They used random mutagenesis to generate over 3,500 individual IL-la mutants that averaged 2.5 amino acid changes per variant over the entire length of the molecule. Multiple mutations were examined at every possible amino acid position. The investigators found that "[m]ost of the molecule could be altered with little effect on either [binding or biological activity]." In fact, only 23 unique amino acid sequences, out of more than 3,500 nucleotide sequences examined, produced a protein that significantly differed in activity from wild-type.
[0115] Furthermore, even if deleting one or more amino acids from the N-terminus or C-terminus of a polypeptide results in modification or loss of one or more biological functions, other biological activities may still be retained. For example, the ability of a deletion variant to induce and/or to bind antibodies which recognize the secreted form will likely be retained when less than the majority of the residues of the secreted form are removed from the N-terminus or C-terminus. Whether a particular polypeptide lacking N- or C-terminal residues of a protein retains such immunogenic activities can readily be determined by routine methods described herein and otherwise known in the art.
[0116] Thus, the invention further includes polypeptide variants which show a functional activity (e.g., biological activity) of the polypeptides of the invention. Such variants include deletions, insertions, inversions, repeats, and substitutions selected according to general rules known in the art so as have little effect on activity.
[0117] The present application is directed to nucleic acid molecules at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequences disclosed herein, (e.g., encoding a polypeptide having the amino acid sequence of an N
and/or C terminal deletion), irrespective of whether they encode a polypeptide having functional activity. This is because even where a particular nucleic acid molecule does not encode a polypeptide having functional activity, one of skill in the art would still know how to use the nucleic acid molecule, for instance, as a hybridization probe or a polymerase chain reaction (PCR) primer. Uses of the nucleic acid molecules of the present invention that do not encode a polypeptide having functional activity include, inter alia, (1) isolating a gene or allelic or splice variants thereof in a cDNA library; (2) in situ hybridization (e.g., "FISH") to metaphase chromosomal spreads to provide precise chromosomal location of the gene, as described in Verma et al., Human Chromosomes: A
Manual of Basic Techniques, Pergamon Press, New York (1988); (3) Northern Blot analysis for detecting mRNA expression in specific tissues (e.g., normal connective tissues or diseased connective tissues); and (4) in situ hybridization (e.g., histochemistry) for detecting mRNA expression in specific tissues (e.g., normal connective tissue or diseased connective tissues).
[0118] Preferred, however, are nucleic acid molecules having sequences at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequences disclosed herein, which do, in fact, encode a polypeptide having functional activity. By a polypeptide having "functional activity" is meant, a polypeptide capable of displaying one or more known functional activities associated with a full-length (complete) protein of the invention. Such functional activities include, but are not limited to, biological activity, antigenicity [ability to bind (or compete with a polypeptide of the invention for binding) to an anti-polypeptide of the invention antibody], immunogenicity (ability to generate antibody which binds to a specific polypeptide of the invention), ability to form multimers with polypeptides of the invention, and ability to bind to a receptor or ligand for a polypeptide of the invention.
[0119] The functional activity of the polypeptides, and fragments, variants and derivatives of the invention, can be assayed by various methods.
[0120] For example, in one embodiment where one is assaying for the ability to bind or compete with full-length polypeptide of the present invention for binding to an anti-polypeptide of the invention antibody, various immunoassays known in the art can be used, including but not limited to, competitive and non-competitive assay systems using techniques such as radioimmunoassays, ELISA (enzyme linked immunosorbent assay), "sandwich" immunoassays, immunoradiometric assays, gel diffusion precipitation reactions, immunodiffusion assays, in situ immunoassays (using colloidal gold, enzyme or radioisotope labels, for example), western blots, precipitation reactions, agglutination assays (e.g., gel agglutination assays, hemagglutination assays), complement fixation assays, immunofluorescence assays, protein A assays, and immunoelectrophoresis assays, etc. In one embodiment, antibody binding is detected by detecting a label on the primary antibody. In another embodiment, the primary antibody is detected by detecting binding of a secondary antibody or reagent to the primary antibody. In a further embodiment, the secondary antibody is labeled. Many means are known in the art for detecting binding in an immunoassay and are within the scope of the present invention.
[0121] In another embodiment, where a ligand is identified, or the ability of a polypeptide fragment, variant or derivative of the invention to multimerize is being evaluated, binding can be assayed, e.g., by means well-known in the art, such as, for example, reducing and non-reducing gel chromatography, protein affinity chromatography, and affinity blotting. See generally, Phizicky et al., Microbiol. Rev.
59:94-123 (1995). In another embodiment, the ability of physiological correlates of a polypeptide of the present invention to bind to a substrates) of the polypeptide of the invention can be routinely assayed using techniques known in the art.
[0122] In addition, assays described herein (see Examples) and otherwise known in the art may routinely be applied to measure the ability of polypeptides of the present invention and fragments, variants and derivatives thereof to elicit polypeptide related biological activity (either in vitro or in vivo). Other methods will be known to the skilled artisan and are within the scope of the invention.
[0123] Of course, due to the degeneracy of the genetic code, one of ordinary skill in the art will immediately recognize that a large number of the nucleic acid molecules having a sequence at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100%
identical to, for example, the nucleic acid sequence of the cDNA contained in Clone >D
NO:Z, a nucleic acid sequence referred to in Table 1A (e.g., SEQ ID NO:X), a nucleic acid sequence disclosed in Table 2 (e.g., the nucleic acid sequence delineated in columns 8 and 9) or fragments thereof, will encode polypeptides "having functional activity." In fact, since degenerate variants of any of these nucleotide sequences all encode the same polypeptide, in many instances, this will be clear to the skilled artisan even without performing the above described comparison assay. It will be further recognized in the art that, for such nucleic acid molecules that are not degenerate variants, a reasonable number will also encode a polypeptide having functional activity. This is because the skilled artisan is fully aware of amino acid substitutions that are either less likely or not likely to significantly effect protein function (e.g., replacing one aliphatic amino acid with a second aliphatic amino acid), as further described below.

[0124] . For example, guidance concerning how to make phenotypically silent amino acid substitutions is provided in Bowie et al., "Deciphering the Message in Protein Sequences: Tolerance to Amino Acid Substitutions," Science 247:1306-1310 (1990), wherein the authors indicate that there are two main strategies for studying the tolerance of an amino acid sequence to change.
[0125] The first strategy exploits the tolerance of amino acid substitutions by natural selection during the process of evolution. By comparing amino acid sequences in different species, conserved amino acids can be identified. These conserved amino acids are likely important for protein function. In contrast, the amino acid positions where substitutions have been tolerated by natural selection indicates that these positions are not critical for protein function. Thus, positions tolerating amino acid substitution could be modified while still maintaining biological activity of the protein.
[0126] The second strategy uses genetic engineering to introduce amino acid changes at specific positions of a cloned gene to identify regions critical for protein function. For example, site directed mutagenesis or alanine-scanning mutagenesis (introduction of single alanine mutations at every residue in the molecule) can be used. See Cunningham et al., Science 244:1081-1085 (1989). The resulting mutant molecules can then be tested for biological activity.
[0127] As the authors state, these two strategies have revealed that proteins are surprisingly tolerant of amino acid substitutions. The authors further indicate which amino acid changes are likely to be permissive at certain amino acid positions in the protein. For example, most buried (within the tertiary structure of the protein) amino acid residues require nonpolar side chains, whereas few features of surface side chains are generally conserved. Moreover, tolerated conservative amino acid substitutions involve replacement of the aliphatic or hydrophobic amino acids Ala, Val, Leu and Ile;
replacement of the hydroxyl residues Ser and Thr; replacement of the acidic residues Asp and Glu; replacement of the amide residues Asn and Gln, replacement of the basic residues Lys, Arg, and His; replacement of the aromatic residues Phe, Tyr, and Trp, and replacement of the small-sized amino acids Ala, Ser, Thr, Met, and Gly.
Besides conservative amino acid substitutions, variants of the present invention include (i) substitutions with one or more of the non-conserved amino acid residues, where the substituted amino acid residues may or may not be one encoded by the genetic code, or (ii) substitutions with one or more of the amino acid residues having a substituent group, or (iii) fusion of the mature polypeptide with another compound, such as a compound to increase the stability and/or solubility of the polypeptide (for example, polyethylene glycol), or (iv) fusion of the polypeptide with additional amino acids, such as, for example, an IgG Fc fusion region peptide, serum albumin (preferably human serum albumin) or a fragment or variant thereof, or leader or secretory sequence, or a sequence facilitating purification. Such variant polypeptides are deemed to be within the scope of those skilled in the art from the teachings herein.
[0128] For example, polypeptide variants containing amino acid substitutions of charged amino acids with other charged or neutral amino acids may produce proteins with improved characteristics, such as less aggregation. Aggregation of pharmaceutical formulations both reduces activity and increases clearance due to the aggregate's immunogenic activity. See Pinckard et al., Clin. Exp. Immunol. 2:331-340 (1967);
Robbins et al., Diabetes 36: 838-845 (1987); Cleland et al., Crit. Rev.
Therapeutic Drug Carner Systems 10:307-377 (1993).
[0129] A further embodiment of the invention relates to polypeptides which comprise the amino acid sequence of a polypeptide having an amino acid sequence which contains at least one amino acid substitution, but not more than 50 amino acid substitutions, even more preferably, not more than 40 amino acid substitutions, still more preferably, not more than 30 amino acid substitutions, and still even more preferably, not more than 20 amino acid substitutions from a polypeptide sequence disclosed herein. Of course it is highly preferable for a polypeptide to have an amino acid sequence which comprises the amino acid sequence of a polypeptide of SEQ ID NO:Y, an amino acid sequence encoded by SEQ ID NO:X, an amino acid sequence encoded by the portion of SEQ ID NO:X
as defined in columns 8 and 9 of Table 2, an amino acid sequence encoded by the complement of SEQ ID NO:X, and/or the amino acid sequence encoded by cDNA
contained in Clone ID NO:Z which contains, in order of ever-increasing preference, at least one, but not more than 10, 9, 8, 7, 6, S, 4, 3, 2 or 1 amino acid substitutions.
[0130] In specific embodiments, the polypeptides of the invention comprise, .or alternatively, consist of, fragments or variants of a reference amino acid sequence selected from: (a) the amino acid sequence of SEQ ID NO:Y or fragments thereof (e.g., the mature form and/or other fragments described herein); (b) the amino acid sequence encoded by SEQ m NO:X or fragments thereof; (c) the amino acid sequence encoded by the complement of SEQ m NO:X or fragments thereof; (d) the amino acid sequence encoded by the portion of SEQ m NO:X as defined in columns 8 and 9 of Table 2 or fragments thereof; and (e) the amino acid sequence encoded by cDNA contained in Clone >D NO:Z or fragments thereof; wherein the fragments or variants have 1-5, 5-10, 5-25, 5-50, 10-50 or 50-150, amino acid residue additions, substitutions, and/or deletions when compared to the reference amino acid sequence. In preferred embodiments, the amino acid substitutions are conservative. Polynucleotides encoding these polypeptides are also encompassed by the invention.
Polyuucleotide and Polypeptide Fragments [0131] The present invention is also directed to polynucleotide fragments of the polynucleotides (nucleic acids) of the invention. In the present invention, a "polynucleotide fragment" refers to a polynucleotide having a nucleic acid sequence which, for example: is a portion of the cDNA contained in Clone m NO:Z or the complementary strand thereto; is a portion of the polynucleotide sequence encoding the polypeptide encoded by the cDNA contained in Clone >D NO:Z or the complementary strand thereto; is a portion of a polynucleotide sequence encoding the amino acid sequence encoded by the region of SEQ m NO:X as defined in columns 8 and 9 of Table 2 or the complementary strand thereto; is a portion of the polynucleotide sequence of SEQ )D NO:X as defined in columns 8 and 9 of Table 2 or the complementary strand thereto; is a portion of the polynucleotide sequence in SEQ m NO:X or the complementary strand thereto; is a polynucleotide sequence encoding a portion of the polypeptide of SEQ m NO:Y; is a polynucleotide sequence encoding a portion of a polypeptide encoded by SEQ m NO:X; is a polynucleotide sequence encoding a portion of a polypeptide encoded by the complement of the polynucleotide sequence in SEQ m NO:X; is a portion of a polynucleotide sequence encoding the amino acid sequence encoded by the region of SEQ >D NO:B as defined in column 6 of Table 1 B or the complementary strand thereto; or is a portion of the polynucleotide sequence of SEQ >D
NO:B as defined in column 6 of Table 1B or the complementary strand thereto.
[0132] The polynucleotide fragments of the invention are preferably at least about 15 nt, and more preferably at least about 20 nt, still more preferably at least about 30 nt, and even more preferably, at least about 40 nt, at least about 50 nt, at least about 75 nt, or at least about 150 nt in length. A fragment "at least 20 nt in length," for example, is intended to include 20 or more contiguous bases from the cDNA sequence contained in Clone m NO:Z, or the nucleotide sequence shown in SEQ >D NO:X or the complementary stand thereto. In this context "about" includes the particularly recited value or a value larger or smaller by several (5, 4, 3, 2, or 1) nucleotides, at either terminus or at both termini. These nucleotide fragments have uses that include, but are not limited to, as diagnostic probes and primers as discussed herein. Of course, larger fragments (e.g., at least 160, 170, 180, 190, 200, 250, 500, 600, 1000, or 2000 nucleotides in length) are also encompassed by the invention.
[0133] Moreover, representative examples of polynucleotide fragments of the invention, comprise, or alternatively consist of, a sequence from about nucleotide number 1-50, 51-100, 101-150, 151-200, 201-250, 251-300, 301-350, 351-400, 401-450, 451-500, 501-550, 551-600, 651-700, 701-750, 751-800, 800-850, 851-900, 901-950, 951-1000, 1001-1050, 1051-1100, 1101-1150, 1151-1200, 1201-1250, 1251-1300, 1301-1350, 1351-1400, 1401-1450, 1451-1500, 1501-1550, 1551-1600, 1601-1650, 1651-1700, 1701-1750, 1751-1800, 1801-1850, 1851-1900, 1901-1950, 1951-2000, 2001-2050, 2051-2100, 2101-2150, 2151-2200, 2201-2250, 2251-2300, 2301-2350, 2351-2400, 2401-2450, 2451-2500, 2501-2550, 2551-2600, 2601-2650, 2651-2700, 2701-2750, 2751-2800, 2801-2850, 2851-2900, 2901-2950, 2951-3000, 3001-3050, 3051-3100, 3101-3150, 3151-3200, 3201-3250, 3251-3300, 3301-3350, 3351-3400, 3401-3450, 3451-3500, 3501-3550, 3551-3600, 3601-3650, 3651-3700, 3701-3750, 3751-3800, 3801-3850, 3851-3900, 3901-3950, 3951-4000, 4001-4050, 4051-4100, 4101-4150, 4151-4200, 4201-4250, 4251-4300, 4301-4350, 4351-4400, 4401-4450, 4451-4500, 4501-4550, 4551-4600, 4601-4650, 4651-4700, 4701-4750, 4751-4800, 4801-4850, 4851-4900, 4901-4950, 4951-5000, 5001-5050, 5051-5100, 5101-5150, 5151-5200, 5201-5250, 5251-5300, 5301-5350, 5351-5400, 5401-5450, 5451-5500, 5501-5550, 5551-5600, 5601-5650, 5651-5700, 5701-5750, 5751-5800, 5801-5850, 5851-5900, 5901-5950, 5951-6000, 6001-6050, 6051-6100, 6101-6150, 6151-6200, 6201-6250, 6251-6300, 6301-6350, 6351-6400, 6401-6450, 6451-6500, 6501-65.50, 6551-6600, 6601-6650, 6651-6700, 6701-6750, 6751-6800, 6801-6850, 6851-6900, 6901-6950, 6951-7000, 7001-7050, 7051-7100, 7101-7150, 7151-7200, 7201-7250, 7251-7300 or 7301 to the end of SEQ ID NO:X, or the complementary strand thereto. In this context "about" includes the particularly recited range or a range larger or smaller by several (5, 4, 3, 2, or 1) nucleotides, at either terminus or at both termini. Preferably, these fragments encode a polypeptide, which has a functional activity (e.g., biological activity). More preferably, these polynucleotides can be used as probes or primers as discussed herein.
Polynucleotides which hybridize to one or more of these polynucleotides under stringent hybridization conditions or alternatively, under lower stringency conditions are also encompassed by the invention, as are polypeptides encoded by these polynucleotides.
[0134] Further representative examples of polynucleotide fragments of the invention, comprise, or alternatively consist of, a sequence from about nucleotide number 1-50, 51-100, 101-150, 151-200, 201-250, 251-300, 301-350, 351-400, 401-450, 451-500, 550, 551-600, 651-700, 701-750, 751-800, 800-850, 851-900, 901-950, 951-1000, 1050, 1051-1100, 1101-1150, 1151-1200, 1201-1250, 1251-1300, 1301-1350, 1351-1400, 1401-1450, 1451-1500, 1501-1550, 1551-1600, 1601-1650, 1651-1700, 1701-1750, 1751-1800, 1801-1850, 1851-1900, 1901-1950, 1951-2000, 2001-2050, 2051-2100, 2101-2150, 2151-2200, 2201-2250, 2251-2300, 2301-2350, 2351-2400, 2401-2450, 2451-2500, 2501-2550, 2551-2600, 2601-2650, 2651-2700, 2701-2750, 2751-2800, 2801-2850, 2851-2900, 2901-2950, 2951-3000, 3001-3050, 3051-3100, 3101-3150, 3151-3200, 3201-3250, 3251-3300, 3301-3350, 3351-3400, 3401-3450, 3451-3500, 3501-3550, 3551-3600, 3601-3650, 3651-3700, 3701-3750, 3751-3800, 3801-3850, 3851-3900, 3901-3950, 3951-4000, 4001-4050, 4051-4100, 4101-4150, 4151-4200, 4201-4250, 4251-4300, 4301-4350, 4351-4400, 4401-4450, 4451-4500, 4501-4550, 4551-4600, 4601-4650, 4651-4700, 4701-4750, 4751-4800, 4801-4850, 4851-4900, 4901-4950, 4951-5000, 5001-5050, 5051-5100, 5101-5150, 5151-5200, 5201-5250, 5251-5300, 5301-5350, 5351-5400, 5401-5450, 5451-5500, 5501-5550, 5551-5600, 5601-5650, 5651-5700, 5701-5750, 5751-5800, 5801-5850, 5851-5900, 5901-5950, 5951-6000, 6001-6050, 6051-6100, 6101-6150, 6151-6200, 6201-6250, 6251-6300, 6301-6350, 6351-6400, 6401-6450, 6451-6500, 6501-6550, 6551-6600, 6601-6650, 6651-6700, 6701-6750, 6751-6800, 6801-6850, 6851-6900, 6901-6950, 6951-7000, 7001-7050, 7051-7100, 7101-7150, 7151-7200, 7201-7250, 7251-7300 or 7301 to the end of the cDNA sequence contained in Clone ID NO:Z, or the complementary strand thereto.
In this context "about" includes the particularly recited range or a range larger or smaller by several (5, 4, 3, 2, or 1) nucleotides, at either terminus or at both termini. Preferably, these fragments encode a polypeptide, which has a functional activity (e.g., biological activity). More preferably, these polynucleotides can be used as probes or primers as discussed herein. Polynucleotides which hybridize to one or more of these polynucleotides under stringent hybridization conditions or alternatively, under lower stringency conditions are also encompassed by the invention, as are polypeptides encoded by these polynucleotides.
[0135] Moreover, representative examples of polynucleotide fragments of the invention comprise, or alternatively consist of, a nucleic acid sequence comprising one, two, three, four, five, six, seven, eight, nine, ten, or more of the above described polynucleotide fragments of the invention in combination with a polynucleotide sequence delineated in Table 1B column 6. Additional, representative examples of polynucleotide fragments of the invention comprise, or alternatively consist of, a nucleic acid sequence comprising one, two, three, four, five, six, seven, eight, nine, ten, or more of the above described polynucleotide fragments of the invention in combination with a polynucleotide sequence that is the complementary strand of a sequence delineated in column 6 of Table 1B. In further embodiments, the above-described polynucleotide fragments of the invention comprise, or alternatively consist of, sequences delineated in Table 1B, column 6, and have a nucleic acid sequence which is different from that of the BAC
fragment having the sequence disclosed in SEQ ID NO:B (see Table 1B, column S). In additional embodiments, the above-described polynucleotide fragments of the invention comprise, or alternatively consist of, sequences delineated in Table 1B, column 6, and have a nucleic acid sequence which is different from that published for the BAC clone identified as BAC >D NO:A (see Table 1B, column 4). In additional embodiments, the above-described polynucleotides of the invention comprise, or alternatively consist of, sequences delineated Table 1B, column 6, and have a nucleic acid sequence which is different from that contained in the BAC clone identified as BAC m NO:A (see Table 1B, column 4). Polypeptides encoded by these polynucleotides, other polynucleotides that encode these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides and polypeptides are also encompassed by the invention.

[0136] In additional specific embodiments, polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more fragments of the sequences delineated in column 6 of Table 1B, and the polynucleotide sequence of SEQ m NO:X (e.g., as defined in Table 1B, column 2) or fragments or variants thereof. Polypeptides encoded by these polynucleotides, other polynucleotides that encode these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention.
[0137] In additional specific embodiments, polynucleotides of. the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more fragments of the sequences delineated in column 6 of Table 1B
which correspond to the same Clone m NO:Z (see Table 1B, column 1), and the polynucleotide sequence of SEQ >D NO:X (e.g., as defined in Table 1A or 1B) or fragments or variants thereof. Polypeptides encoded by these polynucleotides, other polynucleotides that encode these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention.
[0138] In further specific embodiments, polynucleotides of the invention comprise, or alternatively consist of, one, two, three, four, five, six, seven, eight, nine, ten, or more fragments of the sequences delineated in the same row of column 6 of Table 1B, and the polynucleotide sequence of SEQ ID NO:X (e.g., as defined in Table 1A or 1B) or fragments or variants thereof. Polypeptides encoded by these polynucleotides, other polynucleotides that encode these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention.
[0139] In additional specific embodiments, polynucleotides of the invention comprise, or alternatively consist of a polynucleotide sequence in which the 3' 10 polynucleotides of one of the sequences delineated in column 6 of Table 1B and the 5' 10 polynucleotides of the sequence of SEQ m NO:X are directly contiguous. Nucleic acids which hybridize to the complement of these 20 contiguous polynucleotides under stringent hybridization conditions or alternatively, under lower stringency conditions, are also encompassed by the invention. Polypeptides encoded by these polynucleotides and/or nucleic acids, other polynucleotides and/or nucleic acids that encode these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides, nucleic acids, and polypeptides are also encompassed by the invention.
[0140] In additional specific embodiments, polynucleotides of the invention comprise, or alternatively consist of a polynucleotide sequence in which the 3' 10 polynucleotides of one of the sequences delineated in column 6 of Table 1B and the 5' 10 polynucleotides of a fragment or variant of the sequence of SEQ ID NO:X (e.g., as described herein) are directly contiguous Nucleic acids which hybridize to the complement of these 20 contiguous polynucleotides under stringent hybridization conditions or alternatively, under lower stringency conditions, are also encompassed by the invention. Polypeptides encoded by these polynucleotides and/or nucleic acids, other polynucleotides and/or nucleic acids encoding these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides, nucleic acids, and polypeptides are also encompassed by the invention.
[0141] In further specific embodiments, polynucleotides of the invention comprise, or alternatively consist of a polynucleotide sequence in which the 3' 10 polynucleotides of a fragment or variant of the sequence of SEQ ID NO:X and the 5' 10 polynucleotides of the sequence of one of the sequences delineated in column 6 of Table 1B are directly contiguous. Nucleic acids which hybridize to the complement of these 20 contiguous polynucleotides under stringent hybridization conditions or alternatively, under lower stringency conditions, are also encompassed by the invention. Polypeptides encoded by these polynucleotides and/or nucleic acids, other polynucleotides and/or nucleic acids encoding these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides, nucleic acids, and polypeptides are also encompassed by the invention.
[0142] In specific embodiments, polynucleotides of the invention comprise, or alternatively consist of a polynucleotide sequence in which the 3' 10 polynucleotides of one of the sequences delineated in column 6 of Table 1B and the 5' 10 polynucleotides of another sequence in column 6 are directly contiguous. In preferred embodiments, the 3' polynucleotides of one of the sequences delineated in column 6 of Table 1B is directly contiguous with the 5' 10 polynucleotides of the next sequential exon delineated in Table 1B, column 6. Nucleic acids which hybridize to the complement of these 20 contiguous polynucleotides under stringent hybridization conditions or alternatively, under lower stringency conditions, are also encompassed by the invention. Polypeptides encoded by these polynucleotides and/or nucleic acids, other polynucleotides and/or nucleic acids encoding these polypeptides, and antibodies that bind these polypeptides are also encompassed by the invention. Additionally, fragments and variants of the above-described polynucleotides, nucleic acids, and polypeptides are also encompassed by the invention.
[0143] In the present invention, a "polypeptide fragment" refers to an amino acid sequence which is a portion of that contained in SEQ ID NO:Y, a portion of an amino acid sequence encoded by the portion of SEQ ID NO:X as defined in columns 8 and 9 of Table 2, a portion of an amino acid sequence encoded by the polynucleotide sequence of SEQ ID NO:X, a portion of an amino acid sequence encoded by the complement of the polynucleotide sequence in SEQ JD NO:X, and/or a portion of an amino acid sequence encoded by the cDNA contained in Clone ID NO:Z. Protein (polypeptide) fragments may be "free-standing," or comprised within a larger polypeptide of which the fragment forms a part or region, most preferably as a single continuous region.
Representative examples of polypeptide fragments of the invention, include, for example, fragments comprising, or alternatively consisting of, from about amino acid number 1-20, 21-40, 41-60, 61-80, 81-100, 102-120, 121-140, 141-160, 161-180, 181-200, 201-220, 221-240, 241-260, 280, 281-300, 301-320, 321-340, 341-360, 361-380, 381-400, 401-420, 421-440, 460, 461-480, 481-500, 501-520, 521-540, 541-560, 561-580, 581-600, 601-620, 640, 641-660, 661-680, 681-700, 701-720, 721-740, 741-760, 761-780, 781-800, 820, 821-840, 841-860, 861-880, 881-900, 901-920, 921-940, 941-960, 961-980, 1000, 1001-1020, 1021-1040, 1041-1060, 1061-1080, 1081-1100, 1101-1120, 1121-1140, 1141-1160, 1161-1180, 1181-1200, 1201-1220, 1221-1240, 1241-1260, 1261-1280, 1281-1300, 1301-1320, 1321-1340, 1341-1360, 1361-1380, 1381-1400, 1401-1420, 1421-1440, or 1441 to the end of the coding region. In a preferred embodiment, polypeptide fragments of the invention include, for example, fragments comprising, or alternatively consisting of, from about amino acid number 1-20, 21-40, 41-60, 61-80, 81-100, 102-120, 121-140, 141-160, 161-180, 181-200, 201-220, 221-240, 241-260, 280, 281-300, 301-320, 321-340, 341-360, 361-380, 381-400, 401-420, 421-440, 460, 461-480, 481-500, SO1-520, 521-540, 541-560, 561-580, 581-600, 601-620, 640, 641-660, 661-680, 681-700, 701-720, 721-740, 741-760, 761-780, 781-800, 820, 821-840, 841-860, 861-880, 881-900, 901-920, 921-940, 941-960, 961-980, 1000, 1001-1020, 1021-1040, 1041-1060, 1061-1080, 1081-1100, 1101-1120, 1121-1140, 1141-1160,' 1161-1180, 1181-1200, 1201-1220, 1221-1240, 1241-1260, 1261-1280, 1281-1300, 1301-1320, 1321-1340, 1341-1360, 1361-1380, 1381-1400, 1401-1420, 1421-1440, or 1441 to the end of the coding region of SEQ >D NO:Y. Moreover, polypeptide fragments of the invention may be at least about 10, 15, 20, 25, 30, 35, 40, 45, S0, 55, 60, 65, 70, 75, 80, 85, 90, 100, 110, 120, 130, 140, or 150 amino acids in length. In this context "about" includes the particularly recited ranges or values, or ranges or values larger or smaller by several (S, 4, 3, 2, or 1) amino acids, at either extreme or at both extremes. Polynucleotides encoding these polypeptide fragments are also encompassed by the invention.
[0144] Even if deletion of one or more amino acids from the N-terminus of a protein results in modification of loss of one or more biological functions of the protein, other functional activities (e.g., biological activities, ability to multimerize, ability to bind a ligand) may still be retained. For example, the ability of shortened muteins to induce and/or bind to antibodies which recognize the complete or mature forms of the polypeptides generally will be retained when less than the majority of the residues of the complete or mature polypeptide are removed from the N-terminus. Whether a particular polypeptide lacking N-terminal residues of a complete polypeptide retains such immunologic activities can readily be determined by routine methods described herein and otherwise known in the art. It is not unlikely that a mutein with a large number of deleted N-terminal amino acid residues may retain some biological or immunogenic activities. In fact, peptides composed of as few as six amino acid residues may often evoke an immune response.
[0145] Accordingly, polypeptide fragments include the secreted protein as well as the mature form. Further preferred polypeptide fragments include the secreted protein or the mature form having a continuous series of deleted residues from the amino or the carboxy terminus, or both. For example, any number of amino acids, ranging from 1-60, can be deleted from the amino terminus of either the secreted polypeptide or the mature form.
Similarly, any number of amino acids, ranging from 1-30, can be deleted from the carboxy terminus of the secreted protein or mature form. Furthermore, any combination of the above amino and carboxy terminus deletions is preferred. Similarly, polynucleotides encoding these polypeptide fragments are also preferred.
[0146] The present invention further provides polypeptides having one or more residues deleted from the amino terminus of the amino acid sequence of a polypeptide disclosed herein (e.g., a polypeptide of SEQ ll~ NO:Y, a polypeptide encoded by the polynucleotide sequence contained in SEQ >D NO:X or the complement thereof, a polypeptide encoded by the portion of SEQ ID NO:X as defined in columns 8 and 9 of Table 2, a polypeptide encoded by the portion of SEQ ID NO:B as defined in column 6 of Table 1B, and/or a polypeptide encoded by the cDNA contained in Clone ID
NO:Z). In particular, N-terminal deletions may be described by the general formula m-q, where q is a whole integer representing the total number of amino acid residues in a polypeptide of the invention (e.g., the polypeptide disclosed in SEQ ID NO:Y, or the polypeptide encoded by the portion of SEQ ID NO:X as defined in columns 8 and 9 of Table 2), and m is defined as any integer ranging from 2 to q-6. Polynucleotides encoding these polypeptides are also encompassed by the invention.
[0147] The present invention further provides polypeptides having one or more residues from the carboxy terminus of the amino acid sequence of a polypeptide disclosed herein (e.g., a polypeptide of SEQ ID NO:Y, a polypeptide encoded by the polynucleotide sequence contained in SEQ ID NO:X, a polypeptide encoded by the portion of SEQ
ID
NO:X as defined in columns 8 and 9 of Table 2, and/or a polypeptide encoded by the cDNA contained in Clone ID NO:Z). In particular, C-terminal deletions may be described by the general formula 1-n, where n is any whole integer ranging from 6 to q-1, and where n corresponds to the position of amino acid residue in a polypeptide of the invention. Polynucleotides encoding these polypeptides are also encompassed by the invention.
[0148] In addition, any of the above described N- or C-terminal deletions can be combined to produce a N- and C-terminal deleted polypeptide. The invention also provides polypeptides having one or more amino acids deleted from both the amino and the carboxyl termini, which may be described generally as having residues m-n of a polypeptide encoded by SEQ ID NO:X (e.g., including, but not limited to, the preferred polypeptide disclosed as SEQ >D NO:Y and the polypeptide encoded by the portion of SEQ ID NO:X as defined in columns 8 and 9 of Table 2), the cDNA contained in Clone ID NO:Z, and/or the complement thereof, where n and m are integers as described above.
Polynucleotides encoding these polypeptides are also encompassed by the invention.
[0149] Also as mentioned above, even if deletion of one or more amino acids from the C-terminus of a protein results in modification of loss of one or more biological functions of the protein, other functional activities (e.g., biological activities, ability to multimerize, ability to bind a ligand) may still be retained. For example the ability of the shortened mutein to induce and/or bind to antibodies which recognize the complete or mature forms of the polypeptide generally will be retained when less than the majority of the residues of the complete or mature polypeptide are removed from the C-terminus.
Whether a particular polypeptide lacking C-terminal residues of a complete polypeptide retains such immunologic activities can readily be determined by routine methods described herein and otherwise known in the art. It is not unlikely that a mutein with a large number of deleted C-terminal amino acid residues may retain some biological or immunogenic activities. In fact, peptides composed of as few as six amino acid residues may often evoke an immune response.
[0150] The present application is also directed to proteins containing polypeptides at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to a polypeptide sequence set forth herein. In preferred embodiments, the application is directed to proteins containing polypeptides at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%
identical to polypeptides having the amino acid sequence of the specific N- and C-terminal deletions. Polynucleotides encoding these polypeptides are also encompassed by the invention.
[0151] Any polypeptide sequence encoded by, for example, the polynucleotide sequences set forth as SEQ ID NO:X or the complement thereof, (presented, for example, in Tables 1A and 2), the cDNA contained in Clone ID NO:Z, or the polynucleotide sequence as defined in column 6 of Table 1B, may be analyzed to determine certain preferred regions of the polypeptide. For example, the amino acid sequence of a polypeptide encoded by a polynucleotide sequence of SEQ ID NO:X (e.g., the polypeptide of SEQ ID NO:Y and the polypeptide encoded by the portion of SEQ
ID
NO:X as defined in columns 8 and 9 of Table 2) or the cDNA contained in Clone ID
NO:Z may be analyzed using the default parameters of the DNASTAR computer algorithm (DNASTAR, Inc., 1228 S. Park St., Madison, WI 53715 USA;
http://www.dnastar.com~.
[0152] Polypeptide regions that may be routinely obtained using the DNASTAR
computer algorithm include, but are not limited to, Gamier-Robson alpha-regions, beta-regions, turn-regions, and coil-regions; Chou-Fasman alpha-regions, beta-regions, and turn-regions; Kyte-Doolittle hydrophilic regions and hydrophobic regions;
Eisenberg alpha- and beta-amphipathic regions; Karplus-Schulz flexible regions; Emini surface-forming regions; and Jameson-Wolf regions of high antigenic index.
Among highly preferred polynucleotides of the invention in this regard are those that encode polypeptides comprising regions that combine several structural features, such as several (e.g., 1, 2, 3 or 4) of the features set out above.
[0153] Additionally, Kyte-Doolittle hydrophilic regions and hydrophobic regions, Emini surface-forming regions, and Jameson-Wolf regions of high antigenic index (i.e., containing four or more contiguous amino acids having an antigenic index of greater than or equal to 1.5, as identified using the default parameters of the Jameson-Wolf program) can routinely be used to determine polypeptide regions that exhibit a high degree of potential for antigenicity. Regions of high antigenicity are determined from data by DNASTAR analysis by choosing values which represent regions of the polypeptide which are likely to be exposed on the surface of the polypeptide in an environment in which antigen recognition may occur in the process of initiation of an immune response.
[0154] Preferred polypeptide fragments of the invention are fragments comprising, or alternatively, consisting of, an amino acid sequence that displays a functional activity (e.g. biological activity) of the polypeptide sequence of which the amino acid sequence is a fragment. By a polypeptide displaying a "functional activity" is meant a polypeptide capable of one or more known functional activities associated with a full-length protein, such as, for example, biological activity, antigenicity, immunogenicity, and/or multimerization, as described herein.
[0155] Other preferred polypeptide fragments are biologically active fragments.
Biologically active fragments are those exhibiting activity similar, but not necessarily identical, to an activity of the polypeptide of the present invention. The biological activity of the fragments may include an improved desired activity, or a decreased undesirable activity.

[0156] In preferred embodiments, polypeptides of the invention comprise, or alternatively consist of, one, two, three, four, five or more of the antigenic fragments of the polypeptide of SEQ ID NO:Y, or portions thereof. Polynucleotides encoding these polypeptides are also encompassed by the invention.
[0157] The present invention encompasses polypeptides comprising, or alternatively consisting of, an epitope of: the polypeptide sequence shown in SEQ >D NO:Y; a polypeptide sequence encoded by SEQ m NO:X or the complementary strand thereto; the polypeptide sequence encoded by the portion of SEQ m NO:X as defined in columns 8 and 9 of Table 2; the polypeptide sequence encoded by the portion of SEQ ~
NO:B as defined in column 6 of Table 1B or the complement thereto; the polypeptide sequence encoded by the cDNA contained in Clone )D NO:Z; or the polypeptide sequence encoded by a polynucleotide that hybridizes to the sequence of SEQ >D NO:X, the complement of the sequence of SEQ >D NO:X, the complement of a portion of SEQ m NO:X as defined in columns 8 and 9 of Table 2, or the cDNA sequence contained in Clone ID NO:Z
under stringent hybridization conditions or alternatively, under lower stringency hybridization as defined supra. The present invention further encompasses polynucleotide sequences encoding an epitope of a polypeptide sequence of the invention (such as, for example, the sequence disclosed in SEQ ID NO:X, or a fragment thereof), polynucleotide sequences of the complementary strand of a polynucleotide sequence encoding an epitope of the invention, and polynucleotide sequences which hybridize to the complementary strand under stringent hybridization conditions or alternatively, under lower stringency hybridization conditions defined supra.
[0158] The term "epitopes," as used herein, refers to portions of a polypeptide having antigenic or immunogenic activity in an animal, preferably a mammal, and most preferably in a human. In a preferred embodiment, the present invention encompasses a polypeptide comprising an epitope, as well as the polynucleotide encoding this polypeptide. An "immunogenic epitope," as used herein, is defined as a portion of a protein that elicits an antibody response in an animal, as determined by any method known in the art, for example, by the methods for generating antibodies described infra.
(See, for example, Geysen et al., Proc. Natl. Acad. Sci. USA 81:3998- 4002 (1983)). The term "antigenic epitope," as used herein, is defined as a portion of a protein to which an antibody can immunospecifically bind its antigen as determined by any method well known in the art, for example, by the immunoassays described herein.
Immunospecific binding excludes non-specific binding but does not necessarily exclude cross-reactivity with other antigens. Antigenic epitopes need not necessarily be immunogenic.
[0159] Fragments, which function as epitopes may be produced by any conventional means. (See, e.g., Houghten, R. A., Proc. Natl. Acad. Sci. USA 82:5131-5135 (1985) further described in U.S. Patent No. 4,631,211.) [0160] In the present invention, antigenic epitopes preferably contain a sequence of at least 4, at least 5, at least 6, at least 7, more preferably at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 20, at least 25, at least 30, at least 40, at least 50, and, most preferably, between about 15 to about 30 amino acids.
Preferred polypeptides comprising immunogenic or antigenic epitopes are at least 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 100 amino acid residues in length. Additional non-exclusive preferred antigenic epitopes include the antigenic epitopes disclosed herein, as well as portions thereof. Antigenic epitopes are useful, for example, to raise antibodies, including monoclonal antibodies, that specifically bind the epitope. Preferred antigenic epitopes include the antigenic epitopes disclosed herein, as well as any combination of two, three, four, five or more of these antigenic epitopes.
Antigenic epitopes can be used as the target molecules in immunoassays. (See, for instance, Wilson et al., Cell 37:767-778 (1984); Sutcliffe et al., Science 219:660-666 (1983)).
[0161] Non-limiting examples of epitopes of polypeptides that can be used to generate antibodies of the invention include a polypeptide comprising, or alternatively consisting of, at least one, two, three, four, five, six or more of the portions) of SEQ m NO:Y specified in column 6 of Table 1A. These polypeptide fragments have been determined to bear antigenic epitopes of the proteins of the invention by the analysis of the Jameson-Wolf antigenic index, which is included in the DNAStar suite of computer programs. By "comprise" it is intended that a polypeptide contains at least one, two, three, four, five, six or more of the portions) of SEQ m NO:Y shown in column 6 of Table 1A, but it may contain additional flanking residues on either the amino or carboxyl termini of the recited portion. Such additional flanking sequences are preferably sequences naturally found adjacent to the portion; i.e., contiguous sequence shown in SEQ )D NO:Y. The flanking sequence may, however, be sequences from a heterolgous polypeptide, such as from another protein described herein or from a heterologous polypeptide not described herein. In particular embodiments, epitope portions of a polypeptide of the invention comprise one, two, three, or more of the portions of SEQ ID
NO:Y shown in column 6 of Table 1A. Polynucleotides encoding these polypeptides are also encompassed by the invention.
[0162] Similarly, immunogenic epitopes can be used, for example, to induce antibodies according to methods well known in the art. See, for instance, Sutcliffe et al., supra; Wilson et al., supra; Chow et al., Proc. Natl. Acad. Sci. USA 82:910-914; and Bittle et al., J. Gen. Virol. 66:2347-2354 (1985). Preferred immunogenic epitopes include the immunogenic epitopes disclosed herein, as well as any combination of two, three, four, five or more of these immunogenic epitopes. The polypeptides comprising one or more immunogenic epitopes may be presented for eliciting an antibody response together with a carrier protein, such as an albumin, to an animal system (such as rabbit or mouse), or, if the polypeptide is of sufficient length (at least about 25 amino acids), the polypeptide may be presented without a Garner. However, immunogenic epitopes comprising as few as 8 to 10 amino acids have been shown to be sufficient to raise antibodies capable of binding to, at the very least, linear epitopes in a denatured polypeptide (e.g., in Western blotting).
[0163] Epitope-bearing polypeptides of the present invention may be used to induce antibodies according to methods well known in the art including, but not limited to, in vivo immunization, in vitro immunization, and phage display methods. See, e.g., Sutcliffe et al., supra; Wilson et al., supra, and Bittle et al., J. Gen.
Virol., 66:2347-2354 (1985). If in vivo immunization is used, animals may be immunized with free peptide;
however, anti-peptide antibody titer may be boosted by coupling the peptide to a macromolecular Garner, such as keyhole limpet hemacyanin (KLH) or tetanus toxoid.
For instance, peptides containing cysteine residues may be coupled to a carrier using a linker such as maleimidobenzoyl- N-hydroxysuccinimide ester (MBS), while other peptides may be coupled to carriers using a more general linking agent such as glutaraldehyde. Animals such as rabbits, rats and mice are immunized with either free or carrier- coupled peptides, for instance, by intraperitoneal and/or intradermal injection of emulsions containing about 100 pg of peptide or carrier protein and Freund's adjuvant or any other adjuvant known for stimulating an immune response. Several booster t injections may be needed, for instance, at intervals of about two weeks, to provide a useful titer of anti-peptide antibody which can be detected, for example, by ELISA assay using free peptide adsorbed to a solid surface. The titer of anti-peptide antibodies in serum from an immunized animal may be increased by selection of anti-peptide antibodies, for instance, by adsorption to the peptide on a solid support and elution of the selected antibodies according to methods well known in the art.
[0164] As one of skill in the art will appreciate, and as discussed above, the polypeptides of the present invention (e.g., those comprising an immunogenic or antigenic epitope) can be fused to heterologous polypeptide sequences. For example, polypeptides of the present invention (including fragments or variants thereof), may be fused with the constant domain of immunoglobulins (IgA, IgE, IgG, IgM), or portions thereof (CHl, CH2, CH3, or any combination thereof and portions thereof, resulting in chimeric polypeptides. By way of another non-limiting example, polypeptides and/or antibodies of the present invention (including fragments or variants thereof) may be fused with albumin (including but not limited to recombinant human serum albumin or fragments or variants thereof (see, e.g., U.S. Patent No. 5,876,969, issued March 2, 1999, EP Patent 0 413 622, and U.S. Patent No. 5,766,883, issued June 16, 1998, herein incorporated by reference in their entirety)). In a preferred embodiment, polypeptides and/or antibodies of the present invention (including fragments or variants thereof) are fused with the mature form of human serum albumin (i.e., amino acids 1 - 585 of human serum albumin as shown in Figures 1 and 2 of EP Patent 0 322 094) which is herein incorporated by reference in its entirety. In another preferred embodiment, polypeptides and/or antibodies of the present invention (including fragments or variants thereof) are fused with polypeptide fragments comprising, or alternatively consisting of, amino acid residues 1-z of human serum albumin, where z is an integer from 369 to 419, as described in U.S. Patent 5,766,883 herein incorporated by reference in its entirety.
Polypeptides and/or antibodies of the present invention (including fragments or variants thereof) may be fused to either the N- or C-terminal end of the heterologous protein (e.g., immunoglobulin Fc polypeptide or human serum albumin polypeptide).
Polynucleotides encoding fusion proteins of the invention are also encompassed by the invention.
[0165] Such fusion proteins as those described above may facilitate purification and may increase half life in vivo. This has been shown for chimeric proteins consisting of the first two domains of the human CD4-polypeptide and various domains of the constant regions of the heavy or light chains of mammalian immunoglobulins. See, e.g., EP
394,827; Traunecker et al., Nature, 331:84-86 (1988). Enhanced delivery of an antigen across the epithelial barrier to the immune system has been demonstrated for antigens (e.g., insulin) conjugated to an FcRn binding partner such as IgG or Fc fragments (see, e.g., PCT Publications WO 96/22024 and WO 99/04813). IgG Fusion proteins that have a disulfide-linked dimeric structure due to the IgG portion desulfide bonds have also been found to be more efficient in binding and neutralizing other molecules than monomeric polypeptides or fragments thereof alone. See, e.g., Fountoulakis et al., J.
Biochem., 270:3958-3964 (1995). Nucleic acids encoding the above epitopes can also be recombined with a gene of interest as an epitope tag (e.g., the hemagglutinin (HA) tag or flag tag) to aid in detection and purification of the expressed polypeptide.
For example, a system described by Janknecht et al. allows for the ready purification of non-denatured fusion proteins expressed in human cell lines (Janknecht et al., 1991, Proc.
Natl. Acad.
Sci. USA 88:8972- 897). In this system, the gene of interest is subcloned into a vaccinia recombination plasmid such that the open reading frame of the gene is translationally fused to an amino-terminal tag consisting of six histidine residues. The tag serves as a matrix binding domain for the fusion protein. Extracts from cells infected with the recombinant vaccinia virus are loaded onto Ni2+ nitriloacetic acid-agarose column and histidine-tagged proteins can be selectively eluted with imidazole-containing buffers.
Fusion Proteins [0166] Any polypeptide of the present invention can be used to generate fusion proteins. For example, the polypeptide of the present invention, when fused to a second protein, can be used as an antigenic tag. Antibodies raised against the polypeptide of the present invention can be used to indirectly detect the second protein by binding to the polypeptide. Moreover, because secreted proteins target cellular locations based on trafficking signals, polypeptides of the present invention which are shown to be secreted can be used as targeting molecules once fused to other proteins.
[0167] Examples of domains that can be fused to polypeptides of the present J
invention include not only heterologous signal sequences, but also other heterologous functional regions. The fusion does not necessarily need to be direct, but may occur through linker sequences.
[0168] In certain preferred embodiments, proteins of the invention are fusion proteins comprising an amino acid sequence that is an N and/or C- terminal deletion of a polypeptide of the invention. In preferred embodiments, the invention is directed to a fusion protein comprising an amino acid sequence that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical to a polypeptide sequence of the invention.
Polynucleotides encoding these proteins are also encompassed by the invention.
[0169] Moreover, fusion proteins may also be engineered to improve characteristics of the polypeptide of the present invention. For instance, a region of additional amino acids, particularly charged amino acids, may be added to the N-terminus of the polypeptide to improve stability and persistence during purification from the host cell or subsequent handling and storage. Also, peptide moieties may be added to the polypeptide to facilitate purification. Such regions may be removed prior to final preparation of the polypeptide. The addition of peptide moieties to facilitate handling of polypeptides is familiar and routine techniques in the art.
[0170] As one of skill in the art will appreciate that, as discussed above, polypeptides of the present invention, and epitope-bearing fragments thereof, can be combined with heterologous polypeptide sequences. For example, the polypeptides of the present invention may be fused with heterologous polypeptide sequences, for example, the polypeptides of the present invention may be fused with the constant domain of immunoglobulins (IgA, IgE, IgG, IgM) or portions thereof (CH1, CH2, CH3, and any combination thereof, including both entire domains and portions thereof), or albumin (including, but not limited to, native or recombinant human albumin or fragments or variants thereof (see, e.g., U.S. Patent No. 5,876,969, issued March 2, 1999, EP Patent 0 413 622, and U.S. Patent No. 5,766,883, issued June 16, 1998, herein incorporated by reference in their entirety)), resulting in chimeric polypeptides. For example, EP-A-O
464 533 (Canadian counterpart 2045869) discloses fusion proteins comprising various portions of constant region of immunoglobulin molecules together with another human protein or part thereof. In many cases, the Fc part in a fusion protein is beneficial in therapy and diagnosis, and thus can result in, for example, improved pharmacokinetic properties (EP-A 0232 262). Alternatively, deleting the Fc part after the fusion protein has been expressed, detected, and purified, would be desired. For example, the Fc portion may hinder therapy and diagnosis if the fusion protein is used as an antigen for immunizations. In drug discovery, for example, human proteins, such as hIL,-5, have been fused with Fc portions for the purpose of high-throughput screening assays to identify antagonists of hIL-5. See, D. Bennett et al., J. Molecular Recognition 8:52-58 (1995); K. Johanson et al., J. Biol. Chem. 270:9459-9471 (1995).
[0171] Moreover, the polypeptides of the present invention can be fused to marker sequences, such as a polypeptide, which facilitates purification of the fused polypeptide.
In preferred embodiments, the marker amino acid sequence is a hexa-histidine peptide, such as the tag provided in a pQE vector (QIAGEN, Inc., 9259 Eton Avenue, Chatsworth, CA, 91311), among others, many of which are commercially available. As described in Gentz et al., Proc. Natl. Acad. Sci. USA 86:821-824 (1989), for instance, hexa-histidine provides for convenient purification of the fusion protein. Another peptide tag useful for purification, the "HA" tag, corresponds to an epitope derived from the influenza hemagglutinin protein (Wilson et al., Cell 37:767 (1984).) [0172] Additional fusion proteins of the invention may be generated through the techniques of gene-shuffling, motif shuffling, exon-shuffling, and/or codon-shuffling (collectively referred to as "DNA shuffling"), briefly described below, and further described herein. DNA shuffling may be employed to modulate the activities of polypeptides of the invention, such methods can be used to generate polypeptides with altered activity, as well as agonists and antagonists of the polypeptides.
See, generally, U.S. Patent Nos. 5,605,793; 5,811,238; 5,830,721; 5,834,252; and 5,837,458, and Patten et al., Curr. Opinion Biotechnol. 8:724-33 (1997); Harayama, Trends Biotechnol.
16(2):76-82 (1998); Hansson et al., J. Mol. Biol. 287:265-76 (1999); and Lorenzo and Blasco, Biotechniques 24(2):308-13 (1998); each of these patents and publications are hereby incorporated by reference in its entirety). In a preferred embodiment, one or more components, motifs, sections, parts, domains, fragments, etc., of a polynucleotide encoding a polypeptide of the invention may be recombined with one or more components, motifs, sections, parts, domains, fragments, etc., of one or more heterologous molecules encoding a heterologous polypeptide.
[0173] Thus, any of these above fusions can be engineered using the polynucleotides or the polypeptides of the present invention.

Recombinant and Synthetic Production of Polypeptides of the Invention [0174] The present invention also relates to vectors containing the polynucleotide of the present invention, host cells, and the production of polypeptides by synthetic and recombinant techniques. The vector may be, for example, a phage, plasmid, viral, or retroviral vector. Retroviral vectors may be replication competent or replication defective. In the latter case, viral propagatic5~n generally will occur only in complementing host cells.
[0175] The polynucleotides of the invention may be joined to a vector containing a selectable marker for propagation in a host. Generally, a plasmid vector is introduced in a precipitate, such as a calcium phosphate precipitate, or in a complex with a charged lipid.
If the vector is a virus, it may be packaged in vitro using an appropriate packaging cell line and then transduced into host cells.
[0176] The polynucleotide insert should be operatively linked to an appropriate promoter, such as the phage lambda PL promoter, the E. coli lac, trp, phoA and tac promoters, the SV40 early and late promoters and promoters of retroviral LTRs, to name a few. Other suitable promoters will be known to the skilled artisan. The expression constructs will further contain sites for transcription initiation, termination, and, in the transcribed region, a ribosome binding site for translation. The coding portion of the transcripts expressed by the constructs will preferably include a translation initiating codon at the beginning and a termination codon (UAA, UGA or UAG) appropriately positioned at the end of the polypeptide to be translated.
[0177] As indicated, the expression vectors will preferably include at least one selectable marker. Such markers include dihydrofolate reductase, 6418 or neomycin resistance, glutamine synthase, for eukaryotic cell culture and tetracycline, kanamycin or ampicillin resistance genes for culturing in E. coli and other bacteria.
Representative examples of appropriate hosts include, but are not limited to, bacterial cells, such as E.
coli, Streptomyces and Salmonella typhimurium cells; fungal cells, such as yeast cells (e.g., Saccharomyces cerevisiae or Pichia pastoris (ATCC Accession No.
201178));
insect cells such as Drosophila S2 and Spodoptera Sfi7 cells; animal cells such as CHO, COS, 293, NSO and Bowes melanoma cells; and plant cells. Appropriate culture mediums and conditions for the above-described host cells are known in the art.

[0178] Among vectors preferred for use in bacteria include pQE70, pQE60 and pQE-9, available from QIAGEN, Inc.; pBluescript vectors, Phagescript vectors, pNHBA, pNHl6a, pNHl8A, pNH46A, available from Stratagene Cloning Systems, Inc.; and ptrc99a, pKK223-3, pKK233-3, pDR540, pRITS available from Pharmacia Biotech, Inc.
Among preferred eukaryotic vectors are pWLNEO, pSV2CAT, pOG44, pXTl and pSG
available from Stratagene; and pSVK3, pBPV, pMSG and pSVL available from Pharmacia. Preferred expression vectors for use in yeast systems include, but are not limited to pYES2, pYDl, pTEFl/Zeo, pYES2/GS, pPICZ, pGAPZ, pGAPZaIph, pPIC9, pPIC3.5, pHIL-D2, pHIL-S1, pPIC3.5K, pPIC9K, and PA0815 (all available from Invitrogen, Carlsbad, CA). Other suitable vectors will be readily apparent to the skilled artisan.
[0179] Vectors which use glutamine synthase (GS) or DHFR as the selectable markers can be amplified in the presence of the drugs methionine sulphoximine or methotrexate, respectively. An advantage of glutamine synthase based vectors is the availabilty of cell lines (e.g., the murine myeloma cell line, NSO) which are glutamine synthase negative. Glutamine synthase expression systems can also function in glutamine synthase expressing cells (e.g., Chinese Hamster Ovary (CHO) cells) by providing additional inhibitor to prevent the functioning of the endogenous gene. A
glutamine synthase expression system and components thereof are detailed in PCT
publications:
W087/04462; W086/05807; W089/01036; W089/10404; and W091/06657 which are hereby incorporated in their entireties by reference herein. Additionally, glutamine synthase expression vectors can be obtained from Lonza Biologics, Inc.
(Portsmouth, NH). Expression and production of monoclonal antibodies using a GS expression system in murine myeloma cells is described in Bebbington et al., Bioltechnology 10:169(1992) and in Biblia and Robinson Biotechnol. Prog. 11:1 (1995) which are herein incorporated by reference.
[0180] The present invention also relates to host cells containing the above-described vector constructs described herein, and additionally encompasses host cells containing nucleotide sequences of the invention that are operably associated with one or more heterologous control regions (e.g., promoter and/or enhancer) using techniques known of in the art. The host cell can be a higher eukaryotic cell, such as a mammalian cell (e.g., a human derived cell), or a lower eukaryotic cell, such as a yeast cell, or the host cell can be a prokaryotic cell, such as a bacterial cell. A host strain may be chosen, which modulates the expression of the inserted gene sequences, or modifies and processes the gene product in the specific fashion desired. Expression from certain promoters can be elevated in the presence of certain inducers; thus expression of the genetically engineered polypeptide may be controlled. Furthermore, different host cells have characteristics and specific mechanisms for the translational and post-translational processing and modification (e.g., phosphorylation, cleavage) of proteins. Appropriate cell lines can be chosen to ensure the desired modifications and processing of the foreign protein expressed.
[0181] Introduction of the nucleic acids and nucleic acid constructs of the invention into the host cell can be effected by calcium phosphate transfection, DEAF-dextran mediated transfection, cationic lipid-mediated transfection, electroporation, transduction, infection, or other methods. Such methods are described in many standard laboratory manuals, such as Davis et al., Basic Methods In Molecular Biology (1986). It is specifically contemplated that the polypeptides of the present invention may in fact be expressed by a host cell lacking a recombinant vector.
[0182] In addition to encompassing host cells containing the vector constructs discussed herein, the invention also encompasses primary, secondary, and immortalized host cells of vertebrate origin, particularly mammalian origin, that have been engineered to delete or replace endogenous genetic material (e.g., connective tissue antigen coding sequence), and/or to include genetic material (e.g., heterologous polynucleotide sequences) that is operably associated with connective tissue associated polynucleotides of the invention, and which activates, alters, and/or amplifies endogenous connective tissue associated polynucleotides. For example, techniques known in the art may be used to operably associate heterologous control regions (e.g., promoter and/or enhancer) and endogenous connective tissue associated polynucleotide sequences via homologous recombination (see, e.g., US Patent Number 5,641,670, issued June 24, 1997;
International Publication Number WO 96/29411; International Publication Number WO
94/12650; Koller et al., Proc. Natl. Acad. Sci. USA 86:8932-8935 (1989); and Zijlstra et al., Nature 342:435-438 (1989), the disclosures of each of which are incorporated by reference in their entireties).
[0183] Polypeptides of the present invention can also be recovered from:
products . purified from natural sources, including bodily fluids, tissues and cells, whether directly isolated or cultured; products of chemical synthetic procedures; and products produced by recombinant techniques from a prokaryotic or eukaryotic host, including, for example, bacterial, yeast, higher plant, insect, and mammalian cells. Depending upon the host employed in a recombinant production procedure, the polypeptides of the present invention may be glycosylated or may be non-glycosylated. In addition, polypeptides of the invention may also include an initial modified methionine residue, in some cases as a result of host-mediated processes. Thus, it is well known in the art that the N-terminal methionine encoded by the translation initiation codon generally is removed with high efficiency from any protein after translation in all eukaryotic cells. While the N-terminal methionine on most proteins also is efficiently removed in most prokaryotes, for some proteins, this prokaryotic removal process is inefficient, depending on the nature of the amino acid to which the N-terminal methionine is covalently linked.
[0184] In one embodiment, the yeast Pichia pastoris is used to express polypeptides of the invention in a eukaryotic system. Pichia pastoris is a methylotrophic yeast which can metabolize methanol as its sole carbon source. A main step in the methanol metabolization pathway is the oxidation of methanol to formaldehyde using OZ.
This reaction is catalyzed by the enzyme alcohol oxidase. In order to metabolize methanol as its sole carbon source, Pichia pastoris must generate high levels of alcohol oxidase due, in part, to the relatively low affinity of alcohol oxidase for OZ.
Consequently, in a growth medium depending on methanol as a main carbon source, the promoter region of one of the two alcohol oxidase genes (AOXI) is highly active. In the presence of methanol, alcohol oxidase produced from the AOXI gene comprises up to approximately 30%
of the total soluble protein in Pichia pastoris. See, Ellis, S.B., et al., Mol. Cell.
Biol. 5:1111-21 (1985); Koutz, P.J, et al., Yeast 5:167-77 (1989); Tschopp, J.F., et al., Nucl. Acids Res.
15:3859-76 (1987). Thus, a heterologous coding sequence, such as, for example, a polynucleotide of the present invention, under the transcriptional regulation of all or part of the AOXI regulatory sequence is expressed at exceptionally high levels in Pichia yeast grown in the presence of methanol.
[0185] In one example, the plasmid vector pPIC9K is used to express DNA
encoding a polypeptide of the invention, as set forth herein, in a Pichea yeast system essentially as described in "Pichia Protocols: Methods in Molecular Biology," D.R. Higgins and J.

Cregg, eds. The Humana Press, Totowa, NJ, 1998. This expression vector allows expression and secretion of a polypeptide of the invention by virtue of the strong AOXI
promoter linked to the Pichia pastoris alkaline phosphatase (PHO) secretory signal peptide (i.e., leader) located upstream of a multiple cloning site.
[0186] Many other yeast vectors could be used in place of pPIC9K, such as, pYES2, pYDl, pTEFl/Zeo, pYES2/GS, pPICZ, pGAPZ, pGAPZalpha, pPIC9, pPIC3.5, pHIL-D2, pHIL-S1, pPIC3.5K, and PA0815, as one skilled in the art would readily appreciate, as long as the proposed expression construct provides appropriately located signals for transcription, translation, secretion (if desired), and the like, including an in-frame AUG
as required.
[0187] In another embodiment, high-level expression of a heterologous coding sequence, such as, for example, a polynucleotide of the present invention, may be achieved by cloning the heterologous polynucleotide of the invention into an expression vector such as, for example, pGAPZ or pGAPZalpha, and growing the yeast culture in the absence of methanol.
[0188] In addition to encompassing host cells containing the vector constructs discussed herein, the invention also encompasses primary, secondary, and immortalized host cells of vertebrate origin, particularly mammalian origin, that have been engineered to delete or replace endogenous genetic material (e.g., coding sequence), and/or to include genetic material (e.g., heterologous polynucleotide sequences) that is operably associated with polynucleotides of the invention, and which activates, alters, and/or amplifies endogenous polynucleotides. For example, techniques known in the art may be used to operably associate heterologous control regions (e.g., promoter and/or enhancer) and endogenous polynucleotide sequences via homologous recombination (see, e.g., U.S.
Patent No. 5,641,670, issued June 24, 1997; International Publication No. WO
96/29411, published September 26, 1996; International Publication No. WO 94/12650, published August 4, 1994; Koller et al., Proc. Natl. Acad. Sci. USA 86:8932-8935 (1989);
and Zijlstra et al., Nature 342:435-438 (1989), the disclosures of each of which are incorporated by reference in their entireties).
[0189] In addition, polypeptides of the invention can be chemically synthesized using techniques known in the art (e.g., see Creighton, 1983, Proteins: Structures and Molecular Principles, W.H. Freeman & Co., N.Y., and Hunkapiller et al., Nature, 310:105-111 (1984)). For example, a polypeptide corresponding to a fragment of a polypeptide can be synthesized by use of a peptide synthesizer. Furthermore, if desired, nonclassical amino acids or chemical amino acid analogs can be introduced as a substitution or addition into the polypeptide sequence. Non-classical amino acids include, but are not limited to, to the D-isomers of the common amino acids, 2,4-diaminobutyric acid, a-amino isobutyric acid, 4-aminobutyric acid, Abu, 2-amino butyric acid, g-Abu, e-Ahx, 6-amino hexanoic acid, Aib, 2-amino isobutyric acid, 3-amino propionic acid, ornithine, norleucine, norvaline, hydroxyproline, sarcosine, citrulline, homocitrulline, cysteic acid, t-butylglycine, t-butylalanine, phenylglycine, cyclohexylalanine, b-alanine, fluoro-amino acids, designer amino acids such as b-methyl amino acids, Ca-methyl amino acids, Na-methyl amino acids, and amino acid analogs in general. Furthermore, the amino acid can be D (dextrorotary) or L
(levorotary).
[0190] The invention encompasses polypeptides of the present invention which are differentially modified during or after translation, e.g., by glycosylation, acetylation, phosphorylation, amidation, derivatization by known protecting/blocking groups, proteolytic cleavage, linkage to an antibody molecule or other cellular ligand, etc. Any of numerous chemical modifications may be carried out by known techniques, including but not limited, to specific chemical cleavage by cyanogen bromide, trypsin, chymotrypsin, papain, V8 protease, NaBH4; acetylation, formylation, oxidation, reduction;
metabolic synthesis in the presence of tunicamycin; etc.
[0191] Additional post-translational modifications encompassed by the invention include, for example, e.g., N-linked or O-linked carbohydrate chains, processing of N-terminal or C-terminal ends), attachment of chemical moieties to the amino acid backbone, chemical modifications of N-linked or O-linked carbohydrate chains, and addition or deletion of an N-terminal methionine residue as a result of procaryotic host cell expression. The polypeptides may also be modified with a detectable label, such as an enzymatic, fluorescent, isotopic or affinity label to allow for detection and isolation of the protein.
[0192] Examples of suitable enzymes include horseradish peroxidase, alkaline phosphatase, beta-galactosidase, or acetylcholinesterase; examples of suitable prosthetic group complexes include streptavidinlbiotin and avidin/biotin; examples of suitable fluorescent materials include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride or phycoerythrin; an example of a luminescent material includes luminol; examples of bioluminescent materials include luciferase, luciferin, and aequorin; and examples of suitable radioactive material include iodine ('Z11ya3h 12s1 131~~ carbon ('4C), sulfur (35S), tritium (3H), indium (I11~' 112' 113mIn' 115m~)' technetium (99TC,99mTc), thallium (2°'Ti), gallium (~BGa, 6'Ga) alladium '°3Pd mol bdenum 99Mo xenon ('33Xe) fluorine ('$F) 'S3Sm "'Lu P ( )~ Y ( )> > > > >
159Gd' 149Pm' 140La' 175' 166HO' 90Y' 47SC' laurel lBaRe~ 142Pr, 105~~ and 9'Ru.
[0193] In specific embodiments, a polypeptide of the present invention or fragment or variant thereof is attached to macrocyclic chelators that associate with radiometal ions, including but not limited to, ' "Lu, ~°Y, 166Ho, and ' S3Sm, to polypeptides. In a preferred embodiment, the radiometal ion associated with the macrocyclic chelators is "'In. In another preferred embodiment, the radiometal ion associated with the macrocyclic chelator is 9°Y. In specific embodiments, the macrocyclic chelator is 1,4,7,10-tetraazacyclododecane-N,N',N",N"'-tetraacetic acid (DOTA). In other specific embodiments, DOTA is attached to an antibody of the invention or fragment thereof via a linker molecule. Examples of linker molecules useful for conjugating DOTA to a polypeptide are commonly known in the art - see, for example, DeNardo et al., Clin Cancer Res. 4(10):2483-90 (1998); Peterson et al., Bioconjug. Chem. 10(4):553-7 (1999);
and Zimmerman et al, Nucl. Med. Biol. 26(8):943-50 (1999); which are hereby incorporated by reference in their entirety.
[0194] As mentioned, the connective tissue associated proteins of the invention may be modified by either natural processes, such as posttranslational processing, or by chemical modification techniques which are well known in the art. It will be appreciated that the same type of modification may be present in the same or varying degrees at several sites in a given connective tissue associated polypeptide. Connective tissue associated polypeptides may be branched, for example, as a result of ubiquitination, and they may be cyclic, with or without branching. Cyclic, branched, and branched cyclic connective tissue associated polypeptides may result from posttranslation natural processes or may be made by synthetic methods. Modifications include acetylation, acylation, ADP-ribosylation, amidation, covalent attachment of flavin, covalent attachment of a heme moiety, covalent attachment of a nucleotide or nucleotide derivative, covalent attachment of a lipid or lipid derivative, covalent attachment of phosphotidylinositol, cross-linking, cyclization, disulfide bond formation, demethylation, formation of covalent cross-links, formation of cysteine, formation of pyroglutamate, formylation, gamma-carboxylation, glycosylation, GPI anchor formation, hydroxylation, iodination, methylation, myristoylation, oxidation, pegylation, proteolytic processing, phosphorylation, prenylation, racemization, selenoylation, sulfation, transfer-RNA
mediated addition of amino acids to proteins such as arginylation, and ubiquitination.
(See, for instance, PROTEINS - STRUCTURE AND MOLECULAR PROPERTIES, 2nd Ed., T. E. Creighton, W. H. Freeman and Company, New York (1993);
POSTTRANSLATIONAL COVALENT MODIFICATION OF PROTEINS, B. C.
Johnson, Ed., Academic Press, New York, pgs. 1-12 (1983); Seifter et al., Meth.
Enzymol. 182:626-646 (1990); Rattan et al., Ann. N.Y. Acad. Sci. 663:48-62 (1992)).
[0195] Also provided by the invention are chemically modified derivatives of the polypeptides of the invention which may provide additional advantages such as increased solubility, stability and circulating time of the polypeptide, or decreased immunogenicity (see U.S. Patent No. 4,179,337). The chemical moieties for derivitization may be selected from water soluble polymers such as polyethylene glycol, ethylene glycol/propylene glycol copolymers, carboxymethylcellulose, dextran, polyvinyl alcohol and the like. The polypeptides may be modified at random positions within the molecule, or at predetermined positions within the molecule and may include one, two, three or more attached chemical moieties.
[0196] The polymer may be of any molecular weight, and may be branched or unbranched. For polyethylene glycol, the preferred molecular weight is between about 1 kDa and about 100 kDa (the term "about" indicating that in preparations of polyethylene glycol, some molecules will weigh more, some less, than the stated molecular weight) for ease in handling and manufacturing. Other sizes may be used, depending on the desired therapeutic profile (e.g., the duration of sustained release desired, the effects, if any on biological activity, the ease in handling, the degree or lack of antigenicity and other known effects of the polyethylene glycol to a therapeutic protein or analog).
For example, the polyethylene glycol may have an average molecular weight of about 200, 500, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, 5000, 5500, 6000, 6500, 7000, 7500, 8000, 8500, 9000, 9500, 10,000, 10,500, 11,000, 11,500, 12,000, 12,500, 13,000, 13,500, 14,000, 14,500, 15,000, 15,500, 16,000, 16,500, 17,000, 17,500, 18,000, 18,500, 19,000, 19,500, 20,000, 25,000, 30,000, 35,000, 40,000, 50,000, 55,000, 60,000,

65,000, 70,000, 75,000, 80,000, 85,000, 90,000, 95,000, or 100,000 kDa.
[0197] As noted above, the polyethylene glycol may have a branched structure.
Branched polyethylene glycols are described, for example, in U.S. Patent No.
5,643,575;
Morpurgo et al., Appl. Biochem. Biotechnol. 56:59-72 (1996); Vorobjev et al., Nucleosides Nucleotides 18:2745-2750 (1999); and Caliceti et al., Bioconjug.
Chem.
10:638-646 (1999), the disclosures of each of which are incorporated herein by reference.
[0198] The polyethylene glycol molecules (or other chemical moieties) should be attached to the protein with consideration of effects on functional or antigenic domains of the protein. There are a number of attachment methods available to those skilled in the art, such as, for example, the method disclosed in EP 0 401 384 (coupling PEG
to G-CSF), herein incorporated by reference; see also Malik et al., Exp. Hematol.
20:1028-1035 (1992), reporting pegylation of GM-CSF using tresyl chloride. For example, polyethylene glycol may be covalently bound through amino acid residues via a reactive group, such as a free amino or carboxyl group. Reactive groups are those to which an activated polyethylene glycol molecule may be bound. The amino acid residues having a free amino group may include lysine residues and the N-terminal amino acid residues; those having a free carboxyl group may include aspartic acid residues glutamic acid residues and the C-terminal amino acid residue. Sulfhydryl groups may also be used as a reactive group for attaching the polyethylene glycol molecules. Preferred for therapeutic purposes is attachment at an amino group, such as attachment at the N-terminus or lysine group.
[0199] As suggested above, polyethylene glycol may be attached to proteins via linkage to any of a number of amino acid residues. For example, polyethylene glycol can be linked to proteins via covalent bonds to lysine, histidine, aspartic acid, glutamic acid, or cysteine residues. One or more reaction chemistries may be employed to attach polyethylene glycol to specific amino acid residues (e.g., lysine, histidine, aspartic acid, glutamic acid, or cysteine) of the protein or to more than one type of amino acid residue (e.g., lysine, histidine, aspartic acid, glutamic acid, cysteine and combinations thereof) of the protein.

[0200] One may specifically desire proteins chemically modified at the N-terminus.
Using polyethylene glycol as an illustration of the present composition, one may select from a variety of polyethylene glycol molecules (by molecular weight, branching, etc.), the proportion of polyethylene glycol molecules to protein (polypeptide) molecules in the reaction mix, the type of pegylation reaction to be performed, and the method of obtaining the selected N-terminally pegylated protein. The method of obtaining the N-terminally pegylated preparation (i.e., separating this moiety from other monopegylated moieties if necessary) may be by purification of the N-terminally pegylated material from a population of pegylated protein molecules. Selective proteins chemically modified at the N-terminus modification may be accomplished by reductive alkylation which exploits differential reactivity of different types of primary amino groups (lysine versus the N-terminal) available for derivatization in a particular protein.
Under the appropriate reaction conditions, substantially selective derivatization of the protein at the N-terminus with a carbonyl group containing polymer is achieved.
[0201] As indicated above, pegylation of the proteins of the invention may be accomplished by any number of means. For example, polyethylene glycol may be attached to the protein either directly or by an intervening linker.
Linkerless systems for attaching polyethylene glycol to proteins are described in Delgado et al., Crit. Rev. Thera.
Drug Carner Sys. 9:249-304 (1992); Francis et al., Intern. J. of Hematol. 68:1-18 (1998);
U.S. Patent No. 4,002,531; U.S. Patent No. 5,349,052; WO 95/06058; and WO
98/32466, the disclosures of each of which are incorporated herein by reference.
[0202] One system for attaching polyethylene glycol directly to amino acid residues of proteins without an intervening linker employs tresylated MPEG, which is produced by the modification of monmethoxy polyethylene glycol (MPEG) using tresylchloride (C1SOZCHZCF3). Upon reaction of protein with tresylated MPEG, polyethylene glycol is directly attached to amine groups of the protein. Thus, the invention includes protein-polyethylene glycol conjugates produced by reacting proteins of the invention with a polyethylene glycol molecule having a 2,2,2-trifluoreothane sulphonyl group.
[0203] Polyethylene glycol can also be attached to proteins using a number of different intervening linkers. For example, U.S. Patent No. 5,612,460, the entire disclosure of which is incorporated herein by reference, discloses urethane linkers for connecting polyethylene glycol to proteins. Protein-polyethylene glycol conjugates wherein the polyethylene glycol is attached to the protein by a linker can also be produced by reaction of proteins with compounds such as MPEG-succinimidylsuccinate, MPEG activated with l,l'-carbonyldiimidazole, MPEG-2,4,5-trichloropenylcarbonate, MPEG-p-nitrophenolcarbonate, and various MPEG-succinate derivatives. A number of additional polyethylene glycol derivatives and reaction chemistries for attaching polyethylene glycol to proteins are described in International Publication No.

66, the entire disclosure of which is incorporated herein by reference.
Pegylated protein products produced using the reaction chemistries set out herein are included within the scope of the invention.
[0204] The number of polyethylene glycol moieties attached to each protein of the invention (i.e., the degree of substitution) may also vary. For example, the pegylated proteins of the invention may be linked, on average, to 1, 2, 3, 4, S, 6, 7, 8, 9, 10, 12, 15, 17, 20, or more polyethylene glycol molecules. Similarly, the average degree of substitution within ranges such as 1-3, 2-4, 3-5, 4-6, 5-7, 6-8, 7-9, 8-10, 9-11, 10-12, 11-13, 12-14, 13-15, 14-16, 15-17, 16-18, 17-19, or 18-20 polyethylene glycol moieties per protein molecule. Methods for determining the degree of substitution are discussed, for example, in Delgado et al., Crit. Rev. Thera. Drug Carner Sys. 9:249-304 (1992).
[0205] The connective tissue associated polypeptides of the invention can be recovered and purified from chemical synthesis and recombinant cell cultures by standard methods which include, but are not limited to, ammonium sulfate or ethanol precipitation, acid extraction, anion or canon exchange chromatography, phosphocellulose chromatography, hydrophobic interaction chromatography, affinity chromatography, hydroxylapatite chromatography and lectin chromatography. Most preferably, high performance liquid chromatography ("HPLC") is employed for purification. Well known techniques for refolding protein may be employed to regenerate active conformation when the polypeptide is denatured during isolation and/or purification.
[0206] Connective tissue associated polynucleotides and polypeptides may be used in accordance with the present invention for a variety of applications, particularly those that make use of the chemical and biological properties of connective tissue associated antigens. Among these are applications in the detection, prevention, diagnosis and/or treatment of diseases associated with connective tissues, such as e.g., cancer, tumors, rheumatoid arthritis, psoriatic arthritis, discoid lupus erythematosus, systemic lupus erythematosus, scleroderma, CREST syndrome, Sjogren's syndrome, polymyositis, dermatomyositis, mixed connective tissue disease, relapsing polychondritis, vasculitis, Henoch-Schonlein syndrome, erythema nodosum, polyarteritis nodosa, temporal (giant cell) arteritis, Takayasu's arteritis, Wegener's granulomatosis, Reiter's syndrome, Behcet's syndrome, ankylosing spondylitis, cellulitis, keloids, Ehler Danlos syndrome, Marfan syndrome, pseudoxantoma elasticum, osteogenese imperfecta, chondrodysplasias, epidermolysis bullosa, Alport syndrome, cubs laxa, genetic disorders affecting skeleton, skin and muscles; formation of excessive scar tissue; deposition of pathological amounts of connective tissue in body organs, including kidney, intestines and heart, and in liver by liver cirrhosis, in skin by scleroderma, in lung by lung fibrosis, in bone marrow by leukemia, in blood vessels by atherosclerosis, and in joints by rheumatic diseases.
Additional applications relate to diagnosis and to treatment of disorders of cells, tissues and organisms. These aspects of the invention are discussed further below.
[0207] In a preferred embodiment, polynucleotides expressed in a particular tissue type are used to detect, diagnose, treat, prevent and/or prognose disorders associated with the tissue type.
[0208] The polypeptides of the invention may be in monomers or multimers (i.e., dimers, trimers, tetramers and higher multimers). Accordingly, the present invention relates to monomers and multimers of the polypeptides of the invention, their preparation, and compositions (preferably, Therapeutics) containing them. In specific embodiments, the polypeptides of the invention are monomers, dimers, trimers or tetramers.
In additional embodiments, the multimers of the invention are at least dimers, at least trimers, or at least tetramers.
[0209] Multimers encompassed by the invention may be homomers or heteromers.
As used herein, the term homomer refers to a multimer containing only polypeptides corresponding to a protein of the invention (e.g., the amino acid sequence of SEQ ID
NO:Y, an amino acid sequence encoded by SEQ >D NO:X or the complement of SEQ
ID
NO:X, the amino acid sequence encoded by the portion of SEQ ID NO:X as defined in columns 8 and 9 of Table 2, and/or an amino acid sequence encoded by cDNA
contained in Clone )D NO:Z (including fragments, variants, splice variants, and fusion proteins, corresponding to these as described herein)). These homomers may contain polypeptides having identical or different amino acid sequences. In a specific embodiment, a homomer of the invention is a multimer containing only polypeptides having an identical amino acid sequence. In another specific embodiment, a homomer of the invention is a multimer containing polypeptides having different amino acid sequences. In specific embodiments, the multimer of the invention is a homodimer (e.g., containing two polypeptides having identical or different amino acid sequences) or a homotrimer (e.g., containing three polypeptides having identical and/or different amino acid sequences). In additional embodiments, the homomeric multimer of the invention is at least a homodimer, at least a homotrimer, or at least a homotetramer.
[0210] As used herein, the term heteromer refers to a multimer containing two or more heterologous polypeptides (i.e., polypeptides of different proteins) in addition to the polypeptides of the invention. In a specific embodiment, the multimer of the invention is a heterodimer, a heterotrimer, or a heterotetramer. In additional embodiments, the heteromeric multimer of the invention is at least a heterodimer, at least a heterotrimer, or at least a heterotetramer.
[0211] Multimers of the invention may be the result of hydrophobic, hydrophilic, ionic and/or covalent associations and/or may be indirectly linked by, for example, liposome formation. Thus, in one embodiment, multimers of the invention, such as, for example, homodimers or homotrimers, are formed when polypeptides of the invention contact one another in solution. In another embodiment, heteromultimers of the invention, such as, for example, heterotrimers or heterotetramers, are formed when polypeptides of the invention contact antibodies to the polypeptides of the invention (including antibodies to the heterologous polypeptide sequence in a fusion protein of the invention) in solution. In other embodiments, multimers of the invention are formed by covalent associations with and/or between the polypeptides of the invention.
Such covalent associations may involve one or more amino acid residues contained in the polypeptide sequence (e.g., that recited in SEQ ID NO:Y, encoded by the portion of SEQ
ID NO:X as defined in columns 8 and 9 of Table 2, and/or encoded by the cDNA
contained in Clone ID NO:Z). In one instance, the covalent associations are cross-linking between cysteine residues located within the polypeptide sequences which interact in the native (i.e., naturally occurring) polypeptide. In another instance, the covalent associations are the consequence of chemical or recombinant manipulation.

Alternatively, such covalent associations may involve one or more amino acid residues contained in the heterologous polypeptide sequence in a fusion protein. In one example, covalent associations are between the heterologous sequence contained in a fusion protein of the invention (see, e.g., U.S. Patent Number 5,478,925). In a specific example, the covalent associations are between the heterologous sequence contained in a Fc fusion protein of the invention (as described herein). In another specific example, covalent associations of fusion proteins of the invention are between heterologous polypeptide sequence from another protein that is capable of forming covalently associated multimers, such as for example, osteoprotegerin (see, e.g., International Publication NO:
WO
98/49305, the contents of which are herein incorporated by reference in its entirety). In another embodiment, two or more polypeptides of the invention are joined through peptide linkers. Examples include those peptide linkers described in U.S. Pat.
No.
5,073,627 (hereby incorporated by reference). Proteins comprising multiple polypeptides of the invention separated by peptide linkers may be produced using conventional recombinant DNA technology.
[0212] Another method for preparing multimer polypeptides of the invention involves use of polypeptides of the invention fused to a leucine zipper or isoleucine zipper polypeptide sequence. Leucine zipper and isoleucine zipper domains are polypeptides that promote multimerization of the proteins in which they are found. Leucine zippers were originally identified in several DNA-binding proteins (Landschulz et al., Science 240:1759, (1988)), and have since been found in a variety of different proteins. Among the known leucine zippers are naturally occurring peptides and derivatives thereof that dimerize or trimerize. Examples of leucine zipper domains suitable for producing soluble multimeric proteins of the invention are those described in PCT application WO
94/10308, hereby incorporated by reference. Recombinant fusion proteins comprising a polypeptide of the invention fused to a polypeptide sequence that dimerizes or trimerizes in solution are expressed in suitable host cells, and the resulting soluble multimeric fusion protein is recovered from the culture supernatant using techniques known in the art.
[0213] Trimeric polypeptides of the invention may offer the advantage of enhanced biological activity. Preferred leucine zipper moieties and isoleucine moieties are those that preferentially form trimers. One example is a leucine zipper derived from lung surfactant protein D (SPD), as described in Hoppe et al. (FEBS Letters 344:191, (1994)) and in U.S. patent application Ser. No. 08/446,922, hereby incorporated by reference.
Other peptides derived from naturally occurring trimeric proteins may be employed in preparing trimeric polypeptides of the invention.
[0214] In another example, proteins of the invention are associated by interactions between Flag~ polypeptide sequence contained in fusion proteins of the invention containing Flag~ polypeptide sequence. In a further embodiment, proteins of the invention are associated by interactions between heterologous polypeptide sequence contained in Flag~ fusion proteins of the invention and anti-Flag~ antibody.
[0215] The multimers of the invention may be generated using chemical techniques known in the art. For example, polypeptides desired to be contained in the multimers of the invention may be chemically cross-linked using linker molecules and linker molecule length optimization techniques known in the art (see, e.g., U.S. Patent Number 5,478,925, which is herein incorporated by reference in its entirety). Additionally, multimers of the invention may be generated using techniques known in the art to form one or more inter-molecule cross-links between the cysteine residues located within the sequence of the polypeptides desired to be contained in the multimer (see, e.g., U.S. Patent Number 5,478,925, which is herein incorporated by reference in its entirety).
Further, polypeptides of the invention may be routinely modified by the addition of cysteine or biotin to the C-terminus or N-terminus of the polypeptide and techniques known in the art may be applied to generate multimers containing one or more of these modified polypeptides (see, e.g., U.S. Patent Number 5,478,925, which is herein incorporated by reference in its entirety). Additionally, techniques known in the art may be applied to generate liposomes containing the polypeptide components desired to be contained in the multimer of the invention (see, e.g., U.S. Patent Number 5,478,925, which is herein incorporated by reference in its entirety).
[0216] Alternatively, multimers of the invention may be generated using genetic engineering techniques known in the art. In one embodiment, polypeptides contained in multimers of the invention are produced recombinantly using fusion protein technology described herein or otherwise known in the art (see, e.g., U.S. Patent Number 5,478,925, which is herein incorporated by reference in its entirety). In a specific embodiment, polynucleotides coding for a homodimer of the invention are generated by ligating a polynucleotide sequence encoding a polypeptide of the invention to a sequence encoding a linker polypeptide and then further to a synthetic polynucleotide encoding the translated product of the polypeptide in the reverse orientation from the original C-terminus to the N-terminus (lacking the leader sequence) (see, e.g., U..S Patent Number 5,478,925, which is herein incorporated by reference in its entirety). In another embodiment, recombinant techniques described herein or otherwise known in the art are applied to generate recombinant polypeptides of the invention which contain a transmembrane domain (or hydrophobic or signal peptide) and which can be incorporated by membrane reconstitution techniques into liposomes (see, e.g., U.S. Patent Number 5,478,925, which is herein incorporated by reference in its entirety).
Antibodies [0217] Further polypeptides of the invention relate to antibodies and T-cell antigen receptors (TCR) which immunospecifically bind a polypeptide, polypeptide fragment, or variant of the invention (e.g., a polypeptide or fragment or variant of the amino acid sequence of SEQ ID NO:Y or a polypeptide encoded by the cDNA contained in Clone ID
NO:Z, and/or an epitope, of the present invention) as determined by immunoassays well known in the art for assaying specific antibody-antigen binding. Antibodies of the invention include, but are not limited to, polyclonal, monoclonal, multispecific, human, humanized or chimeric antibodies, single chain antibodies, Fab fragments, F(ab') fragments, fragments produced by a Fab expression library, anti-idiotypic (anti-Id) antibodies (including, e.g., anti-Id antibodies to antibodies of the invention), intracellularly-made antibodies (i.e., intrabodies), and epitope-binding fragments of any of the above. The term "antibody," as used herein, refers to immunoglobulin molecules and immunologically active portions of immunoglobulin molecules, i.e., molecules that contain an antigen binding site that immunospecifically binds an antigen. The immunoglobulin molecules of the invention can be of any type (e.g., IgG, IgE, IgM, IgD, IgA and IgY), class (e.g., IgGl, IgG2, IgG3, IgG4, IgAI and IgA2) or subclass of immunoglobulin molecule. In preferred embodiments, the immunoglobulin molecules of the invention are IgGI. In other preferred embodiments, the immunoglobulin molecules of the invention are IgG4.
[0218] Most preferably the antibodies are human antigen-binding antibody fragments of the present invention and include, but are not limited to, Fab, Fab' and F(ab')2, Fd, single-chain Fvs (scFv), single-chain antibodies, disulfide-linked Fvs (sdFv) and fragments comprising either a VL or VH domain. Antigen-binding antibody fragments, including single-chain antibodies, may comprise the variable regions) alone or in combination with the entirety or a portion of the following: hinge region, CHl, CH2, and CH3 domains. Also included in the invention are antigen-binding fragments also comprising any combination of variable regions) with a hinge region, CH1, CH2, and CH3 domains. The antibodies of the invention may be from any animal origin including birds and mammals. Preferably, the antibodies are human, murine (e.g., mouse and rat), donkey, ship rabbit, goat, guinea pig, camel, horse, or chicken. As used herein, "human"
antibodies include antibodies having the amino acid sequence of a human immunoglobulin and include antibodies isolated from human immunoglobulin libraries or from animals transgenic for one or more human immunoglobulin and that do not express endogenous immunoglobulins, as described infra and, for example in, U.S.
Patent No. 5,939,598 by Kucherlapati et al.
[0219] The antibodies of the present invention may be monospecific, bispecific, trispecific or of greater multispecificity. Multispecific antibodies may be specific for different epitopes of a polypeptide of the present invention or may be specific for both a polypeptide of the present invention as well as for a heterologous epitope, such as a heterologous polypeptide or solid support material. See, e.g., PCT
publications WO
93/17715; WO 92/08802; WO 91/00360; WO 92/05793; Tutt, et al., J. Immunol.
147:60-69 (1991); U.S. Patent Nos. 4,474,893; 4,714,681; 4,925,648; 5,573,920;
5,601,819;
Kostelny et al., J. Immunol. 148:1547-1553 (1992).
[0220] Antibodies of the present invention may be described or specified in terms of the epitope(s) or portions) of a polypeptide of the present invention which they recognize or specifically bind. The epitope(s) or polypeptide portions) may be specified as described herein, e.g., by N-terminal and C-terminal positions, or by size in contiguous amino acid residues, or listed in the Tables and Figures. Preferred epitopes of the invention include those shown in column 6 of Table 1A, as well as polynucleotides that encode these epitopes. Antibodies, which specifically bind any epitope or polypeptide of the present invention may also be excluded. Therefore, the present invention includes antibodies that specifically bind polypeptides of the present invention, and allows for the exclusion of the same.

[0221] Antibodies of the present invention may also be described or specified in terms of their cross-reactivity. Antibodies that do not bind any other analog, ortholog, or homolog of a polypeptide of the present invention are included. Antibodies that bind polypeptides with at least 95%, at least 90%, at least 85%, at least 80%, at least 75%, at least 70%, at least 65%, at least 60%, at least 55%, and at least 50% identity (as calculated using methods known in the art and described herein) to a polypeptide of the present invention are also included in the present invention. In specific embodiments, antibodies of the present invention cross-react with murine, rat and/or rabbit homologs of human proteins and the corresponding epitopes thereof. Antibodies that do not bind polypeptides with less than 95%, less than 90%, less than 85%, less than 80%, less than 75%, less than 70%, less than 65%, less than 60%, less than 55%, and less than 50%
identity (as calculated using methods known in the art and described herein) to a polypeptide of the present invention are also included in the present invention. In ~a specific embodiment, the above-described cross-reactivity is with respect to any single specific antigenic or immunogenic polypeptide, or combinations) of 2, 3, 4, 5, or more of the specific antigenic and/or immunogenic polypeptides disclosed herein.
Further included in the present invention are antibodies which bind polypeptides encoded by polynucleotides which hybridize to a polynucleotide of the present invention under stringent hybridization conditions (as described herein). Antibodies of the present invention may also be described or specified in terms of their binding affinity to a polypeptide of the invention. Preferred binding affinities include those with a dissociation constant or Kd less than 5 X 10-2 M, 10-Z M, 5 X 10-3 M, 10-3 M, 5 X 10-4 M, 10-4 M, 5 X 10-5 M, 10-5 M, 5 X 10-6 M, 10-6M, 5 X 10-' M, 10' M, 5 X 10-8 M, 10-g M, 5 X 10-9 M, 10-9 M, 5 X 10-' ° M, 10-' ° M, 5 X 10-" M, 10~" M, 5 X 10-' Z M, 10-' 2 M, 5 X
10-' 3 M, 10-' 3 M, 5 X 10-' 4 M, 10-'4 M, 5 X 10-' S M, or 10-' S M.
[0222] The invention also provides antibodies that competitively inhibit binding of an antibody to an epitope of the invention as determined by any method known in the art for determining competitive binding, for example, the immunoassays described herei-n. In preferred embodiments, the antibody competitively inhibits binding to the epitope by at least 95%, at least 90%, at least 85 %, at least 80%, at least 75%, at least 70%, at least 60%, or at least 50%.

[0223] Antibodies of the present invention may act as agonists or antagonists of the polypeptides of the present invention. For example, the present invention includes antibodies which disrupt the receptor/ligand interactions with the polypeptides of the invention either partially or fully. Preferably, antibodies of the present invention bind an antigenic epitope disclosed herein, or a portion thereof. The invention features both receptor-specific antibodies and ligand-specific antibodies. The invention also features receptor-specific antibodies, which do not prevent ligand binding but prevent receptor activation. Receptor activation (i.e., signaling) may be determined by techniques described herein or otherwise known in the art. For example, receptor activation can be determined by detecting the phosphorylation (e.g., tyrosine or serine/threonine) of the receptor or its substrate by immunoprecipitation followed by western blot analysis (for example, as described supra). In specific embodiments, antibodies are provided that inhibit ligand activity or receptor activity by at least 95%, at least 90%, at least 85%, at least 80%, at least 75%, at least 70%, at least 60%, or at least 50% of the activity in absence of the antibody.
[0224] The invention also features receptor-specific antibodies which both prevent ligand binding and receptor activation as well as antibodies that recognize the receptor-ligand complex, and, preferably, do not specifically recognize the unbound receptor or the unbound ligand. Likewise, included in the invention are neutralizing antibodies which bind the ligand and prevent binding of the ligand to the receptor, as well as antibodies which bind the ligand, thereby preventing receptor activation, but do not prevent the ligand from binding the receptor. Further included in the invention are antibodies, which activate the receptor. These antibodies may act as receptor agonists, i.e., potentiate or activate either all or a subset of the biological activities of the ligand-mediated receptor activation, for example, by inducing dimerization of the receptor. The antibodies may be specified as agonists, antagonists or inverse agonists for biological activities comprising the specific biological activities of the peptides of the invention disclosed herein. The above antibody agonists can be made using methods known in the art. See, e.g., PCT
publication WO 96/40281; U.S. Patent No. 5,811,097; Deng et al., Blood 92(6):1981-1988 (1998); Chen et al., Cancer Res. 58(16):3668-3678 (1998); Harrop et al., J.
Immunol. 161(4):1786-1794 (1998); Zhu et al., Cancer Res. 58(15):3209-3214 (1998);
Yoon et al., J. Immunol. 160(7):3170-3179 (1998); Prat et al., J. Cell. Sci.
111(Pt2):237-247 (1998); Pitard et al., J. Immunol. Methods 205(2):177-190 (1997); Liautard et al., Cytokine 9(4):233-241 (1997); Carlson et al., J. Biol. Chem. 272(17):11295-(1997); Taryman et al., Neuron 14(4):755-762 (1995); Muller et al., Structure 6(9):1153-1167 (1998); Bartunek et al., Cytokine 8(1):14-20 (1996) (which are all incorporated by reference herein in their entireties).
[0225] Antibodies of the present invention may be used, for example, to purify, detect, and target the polypeptides of the present invention, including both in vitro and in vivo diagnostic and therapeutic methods. For example, the antibodies have utility in immunoassays for qualitatively and quantitatively measuring levels of the polypeptides of the present invention in biological samples. See, e.g., Harlow et al., Antibodies: A
Laboratory Manual, (Cold Spring Harbor Laboratory Press, 2nd ed. 1988);
incorporated by reference herein in its entirety.
[0226] As discussed in more detail below, the antibodies of the present invention may be used either alone or in combination with other compositions. The antibodies may further be recombinantly fused to a heterologous polypeptide at the N- or C-terminus or chemically conjugated (including covalent and non-covalent conjugations) to polypeptides or other compositions. For example, antibodies of the present invention may be recombinantly fused or conjugated to molecules useful as labels in detection assays and effector molecules such as heterologous polypeptides, drugs, radionuclides, or toxins. See, e.g., PCT publications WO 92/08495; WO 91/14438; WO 89/12624;
U.S.
Patent No. 5,314,995; and EP 396,387; the disclosures of which are incorporated herein by reference in their entireties.
[0227] The antibodies of the invention include derivatives that are modified, i.e., by the covalent attachment of any type of molecule to the antibody such that covalent attachment does not prevent the antibody from generating an anti-idiotypic response. For example, but not by way of limitation, the antibody derivatives include antibodies that have been modified, e.g., by glycosylation, acetylation, pegylation, phosphylation, amidation, derivatization by known protecting/blocking groups, proteolytic cleavage, linkage to a cellular ligand or other protein, etc. Any of numerous chemical modifications may be carned out by known techniques, including, but not limited to specific chemical cleavage, acetylation, formylation, metabolic synthesis of tunicamycin, etc. Additionally, the derivative may contain one or more non-classical amino acids.

[0228] The antibodies of the present invention may be generated by any suitable method known in the art. Polyclonal antibodies to an antigen-of interest can be produced by various procedures well known in the art. For example, a polypeptide of the invention can be administered to various host animals including, but not limited to, rabbits, mice, rats, etc. to induce the production of sera containing polyclonal antibodies specific for the antigen. Various adjuvants may be used to increase the immunological response, depending on the host species, and include but are not limited to, Freund's (complete and incomplete), mineral gels such as aluminum hydroxide, surface active substances such as lysolecithin, pluronic polyols, polyanions, peptides, oil emulsions, keyhole limpet hemocyanins, dinitrophenol, and potentially useful human adjuvants such as BCG
(bacille Calmette-Guerin) and corynebacterium parvum. Such adjuvants are also well known in the art.
[0229] Monoclonal antibodies can be prepared using a wide variety of techniques known in the art including the use of hybridoma, recombinant, and phage display technologies, or a combination thereof. For example, monoclonal antibodies can be produced using hybridoma techniques including those known in the art and taught, for example, in Harlow et al., Antibodies: A Laboratory Manual, (Cold Spring Harbor Laboratory Press, 2nd ed. 1988); Hammerling, et al., in: Monoclonal Antibodies and T-Cell Hybridomas 563-681 (Elsevier, N.Y., 1981) (said references incorporated by reference in their entireties). The term "monoclonal antibody" as used herein is not limited to antibodies produced through hybridoma technology. The term "monoclonal antibody" refers to an antibody that is derived from a single clone, including any eukaryotic, prokaryotic, or phage clone, and not the method by which it is produced.
[0230] Methods for producing and screening for specific antibodies using hybridoma technology are routine and well known in the art and are discussed in detail in the Examples. In a non-limiting example, mice can be immunized with a polypeptide of the invention or a cell expressing such peptide. Once an immune response is detected, e.g., antibodies specific for the antigen are detected in the mouse serum, the mouse spleen is harvested and splenocytes isolated. The splenocytes are then fused by well known techniques to any suitable myeloma cells, for example cells from cell line SP20 available from the ATCC. Hybridomas are selected and cloned by limited dilution. The hybridoma clones are then assayed by methods known in the art for cells that secrete antibodies capable of binding a polypeptide of the invention. Ascites fluid, which generally contains high levels of antibodies, can be generated by immunizing mice with positive hybridoma clones.
[0231] Accordingly, the present invention provides methods of generating monoclonal antibodies as well as antibodies produced by the method comprising culturing a hybridoma cell secreting an antibody of the invention wherein, preferably, the hybridoma is generated by fusing splenocytes isolated from a mouse immunized with an antigen of the invention with myeloma cells and then screening the hybridomas resulting from the fusion for hybridoma clones that secrete an antibody able to bind a polypeptide of the invention.
[0232] Another well known method for producing both polyclonal and monoclonal human B cell lines is transformation using Epstein Barr Virus (EBV). Protocols for .
generating EBV-transformed B cell lines are commonly known in the art, such as, for example, the protocol outlined in Chapter 7.22 of Current Protocols in Immunology, Coligan et al., Eds., 1994, John Wiley & Sons, NY, which is hereby incorporated in its entirety by reference herein. The source of B cells for transformation is commonly human peripheral blood, but B cells for transformation may also be derived from other sources including, but not limited to, lymph nodes, tonsil, spleen, tumor tissue, and infected tissues. Tissues are generally made into single cell suspensions prior to EBV
transformation. Additionally, steps may be taken to either physically remove or inactivate T cells (e.g., by treatment with cyclosporin A) in B cell-containing samples, because T cells from individuals seropositive for anti-EBV antibodies can suppress B cell immortalization by EBV.
[0233] In general, the sample containing human B cells is innoculated with EBV, and cultured for 3-4 weeks. A typical source of EBV is the culture supernatant of the B95-8 cell line (ATCC #VR-1492). Physical signs of EBV transformation can generally be seen towards the end of the 3-4 week culture period. By phase-contrast microscopy, transformed cells may appear large, clear, hairy and tend to aggregate in tight clusters of cells. Initially, EBV lines are generally polyclonal. However, over prolonged periods of cell cultures, EBV lines may become monoclonal or polyclonal as a result of the selective outgrowth of particular B cell clones. Alternatively, polyclonal EBV
transformed lines may be subcloned (e.g., by limiting dilution culture) or fused with a suitable fusion partner and plated at limiting dilution to obtain monoclonal B cell lines.
Suitable fusion partners for EBV transformed cell lines include mouse myeloma cell lines (e.g., SP2/0, X63-Ag8.653), heteromyeloma cell lines (human x mouse; e.g, SPAM-8, SBC-H20, and CB-F7), and human cell lines (e.g., GM 1500, SKO-007, RPMI 8226, and KR-4).
Thus, the present invention also provides a method of generating polyclonal or monoclonal human antibodies against polypeptides of the invention or fragments thereof, comprising EBV-transformation of human B cells.
[0234] Antibody fragments, which recognize specific epitopes may be generated by known techniques. For example, Fab and F(ab')2 fragments of the invention may be produced by proteolytic cleavage of immunoglobulin molecules, using enzymes such as papain (to produce Fab fragments) or pepsin (to produce F(ab')2 fragments).
F(ab')2 fragments contain the variable region, the light chain constant region and the CH1 domain of the heavy chain. For example, the antibodies of the present invention can also be generated using various phage display methods known in the art and as discussed in detail in the Examples (e.g., Example 10). In phage display methods, functional antibody domains are displayed on the surface of phage particles, which carry the polynucleotide sequences encoding them. In a particular embodiment, such phage can be utilized to display antigen binding domains expressed from a repertoire or combinatorial antibody library (e.g., human or marine). Phage expressing an antigen binding domain that binds the antigen of interest can be selected or identified with antigen, e.g., using labeled antigen or antigen bound or captured to a solid surface or bead. Phage used in these methods are typically filamentous phage including fd and M13 binding domains expressed from phage with Fab, Fv or disulfide stabilized Fv antibody domains recombinantly fused to either the phage gene III or gene VIII protein.
Examples of phage display methods that can be used to make the antibodies of the present invention include those disclosed in Brinkman et al., J. Immunol. Methods 182:41-50 (1995); Ames et al., J.
Immunol. Methods 184:177-186 (1995); Kettleborough et al., Eur. J. Immunol.
24:952-958 (1994); Persic et al., Gene 187 9-18 (1997); Burton et al., Advances in Immunology 57:191-280 (1994); PCT application No. PCT/GB91/01134; PCT publications WO
90/02809; WO 91/10737; WO 92/01047; WO 92/18619; WO 93/11236; WO 95/15982;
WO 95/20401; and U.S. Patent Nos. 5,698,426; 5,223,409; 5,403,484; 5,580,717;
5,427,908; 5,750,753; 5,821,047; 5,571,698; 5,427,908; 5,516,637; 5,780,225;

5,658,727; 5,733,743 and 5,969,108; each of which is incorporated herein by reference in its entirety.
[0235] As described in the above references, after phage selection, the antibody coding regions from the phage can be isolated and used to generate whole antibodies, including human antibodies, or any other desired antigen binding fragment, and expressed in any desired host, including mammalian cells, insect cells, plant cells, yeast, and bacteria, e.g., as described in detail below. For example, techniques to recombinantly produce Fab, Fab' and F(ab')2 fragments can also be employed using methods known in the art such as those disclosed in PCT publication WO 92/22324; Mullinax et al., BioTechniques 12(6):864-869 (1992); and Sawai et al., AJRI 34:26-34 (1995);
and Better et al., Science 240:1041-1043 (1988) (said references incorporated by reference in their entireties).
[0236] Examples of techniques which can be used to produce single-chain Fvs and antibodies include those described in U.S. Patents 4,946,778 and 5,258,498;
Huston et al., Methods in Enzymology 203:46-88 (1991); Shu et al., PNAS 90:7995-7999 (1993);
and Skerra et al., Science 240:1038-1040 (1988). For some uses, including in vivo use of antibodies in humans and in vitro detection assays, it may be preferable to use chimeric, humanized, or human antibodies. A chimeric antibody is a molecule in which different portions of the antibody are derived from different animal species, such as antibodies having a variable region derived from a murine monoclonal antibody and a human immunoglobulin constant region. Methods for producing chimeric antibodies are known in the art. See e.g., Morrison, Science 229:1202 (1985); Oi et al., BioTechniques 4:214 (1986); Gillies et al., (1989) J. Immunol. Methods 125:191-202; U.S. Patent Nos.
5,807,715; 4,816,567; and 4,816397, which are incorporated herein by reference in their entirety. Humanized antibodies are antibody molecules from non-human species antibody that binds the desired antigen having one or more complementarity determining regions (CDRs) from the non-human species and a framework regions from a human immunoglobulin molecule. Often, framework residues in the human framework regions will be substituted with the corresponding residue from the CDR donor antibody to alter, preferably improve, antigen binding. These framework substitutions are identified by methods well known in the art, e.g., by modeling of the interactions of the CDR and framework residues to identify framework residues important for antigen binding and sequence comparison to identify unusual framework residues at particular positions.
(See, e.g., Queen et al., U.S. Patent No. 5,585,089; Riechmann et al., Nature 332:323 (1988), which are incorporated herein by reference in their entireties.) Antibodies can be humanized using a variety of techniques known in the art including, for example, CDR-grafting (EP 239,400; PCT publication WO 91/09967; U.S. Patent Nos. 5,225,539;
5,530,101; and 5,585,089), veneering or resurfacing (EP 592,106; EP 519,596;
Padlan, Molecular Immunology 28(4/5):489-498 (1991); Studnicka et al., Protein Engineering 7(6):805-814 (1994); Roguska. et al., PNAS 91:969-973 (1994)), and chain shuffling (U.5. Patent No. 5,565,332).
[0237] Completely human antibodies are particularly desirable for therapeutic treatment of human patients. Human antibodies can be made by a variety of methods known in the art including phage display methods described above using antibody libraries derived from human immunoglobulin sequences. See also, U.S. Patent Nos.
4,444,887 and 4,716,111; and PCT publications WO 98/46645, WO 98/50433, WO
98/24893, WO 98/16654, WO 96/34096, WO 96/33735, and WO 91/10741; each of which is incorporated herein by reference in its entirety.
[0238] Human antibodies can also be produced using transgenic mice which are incapable of expressing functional endogenous immunoglobulins, but which can express human immunoglobulin genes. For example, the human heavy and light chain immunoglobulin gene complexes may be introduced randomly or by homologous recombination into mouse embryonic stem cells. Alternatively, the human variable region, constant region, and diversity region may be introduced into mouse embryonic stem cells in addition to the human heavy and light chain genes. The mouse heavy and light chain immunoglobulin genes may be rendered non-functional separately or simultaneously with the introduction of human immunoglobulin loci by homologous recombination. In particular, homozygous deletion of the JH region prevents endogenous antibody production. The modified embryonic stem cells are expanded and microinjected into blastocysts to produce chimeric mice. The chimeric mice are then bred to produce homozygous offspring, which express human antibodies. The transgenic mice are immunized in the normal fashion with a selected antigen, e.g., all or a portion of a polypeptide of the invention. Monoclonal antibodies directed against the antigen can be obtained from the immunized, transgenic mice using conventional hybridoma technology.

The human immunoglobulin transgenes harbored by the transgenic mice rearrange during B cell differentiation, and subsequently undergo class switching and somatic mutation.
Thus, using such a technique, it is possible to produce therapeutically useful IgG, IgA, IgM and IgE antibodies. For an overview of this technology for producing human antibodies, see Lonberg and Huszar, Int. Rev. Immunol. 13:65-93 (1995). For a detailed discussion of this technology for producing human antibodies and human monoclonal antibodies and protocols for producing such antibodies, see, e.g., PCT
publications WO
98/24893; WO 92/01047; WO 96/34096; WO 96/33735; European Patent No. 0 598 877;
U.S. Patent Nos. 5,413,923; 5,625,126; 5,633,425; 5,569,825; 5,661,016;
5,545,806;
5,814,318; 5,885,793; 5,916,771; 5,939,598; 6,075,181 and 6,114,598, which are incorporated by reference herein in their entirety. In addition, companies such as Abgenix, Inc. (Freemont, CA) and Genpharm (San Jose, CA) can be engaged to provide human antibodies directed against a selected antigen using technology similar to that described above.
[0239] Completely human antibodies which recognize a selected epitope can be generated using a technique referred to as "guided selection." In this approach a selected non-human monoclonal antibody, e.g., a mouse antibody, is used to guide the selection of a completely human antibody recognizing the same epitope. (Jespers et al., Biotechnology 12:899-903 (1988)).
[0240] Further, antibodies to the polypeptides of the invention can, in turn, be utilized to generate anti-idiotype antibodies that "mimic" polypeptides of the invention using techniques well known to those skilled in the art. (See, e.g., Greenspan &
Bona, FASEB
J. 7(5):437-444; (1989) and Nissinoff, J. Immunol. 147(8):2429-2438 (1991)).
For example, antibodies which bind to and competitively inhibit polypeptide multimerization and/or binding of a polypeptide of the invention to a ligand can be used to generate anti-idiotypes that "mimic" the polypeptide multimerization and/or binding domain and, as a consequence, bind to and neutralize polypeptide and/or its ligand. Such neutralizing anti-idiotypes or Fab fragments of such anti-idiotypes can be used in therapeutic regimens to neutralize polypeptide ligand/receptor. For example, such anti-idiotypic antibodies can be used to bind a polypeptide of the invention and/or to bind its ligand(s)/receptor(s), and thereby block its biological activity. Alternatively, antibodies which bind to and enhance polypeptide multimerization and/or binding, and/or receptor/ligand multimerization, binding and/or signaling can be used to generate anti-idiotypes that function as agonists of a polypeptide of the invention and/or its ligand/receptor. Such agonistic anti-idiotypes or Fab fragments of such anti-idiotypes can be used in therapeutic regimens as agonists of the polypeptides of the invention or its ligand(s)/receptor(s). For example, such anti-idiotypic antibodies can be used to bind a polypeptide of the invention and/or to bind its ligand(s)/receptor(s), and thereby promote or enhance its biological activity.
[0241] Intrabodies of the invention can be produced using methods known in the art, such as those disclosed and reviewed in Chen et al., Hum. Gene Ther. 5:595-601 (1994);
Marasco, W.A., Gene Ther. 4:11-15 (1997); Rondon and Marasco, Annu. Rev.
Microbiol.
51:257-283 (1997); Proba et al., J. Mol. Biol. 275:245-253 (1998); Cohen et al., Oncogene 17:2445-2456 (1998); Ohage and Steipe, J. Mol. Biol. 291:1119-1128 (1999);
Ohage et al., J. Mol. Biol. 291:1129-1134 (1999); Wirtz and Steipe, Protein Sci. 8:2245-2250 (1999); Zhu et al., J. Immunol. Methods 231:207-222 (1999); and references cited therein.
Polynucleotides Encoding Antibodies [0242] The invention further provides polynucleotides comprising a nucleotide sequence encoding an antibody of the invention and fragments thereof. The invention also encompasses polynucleotides that hybridize under stringent or alternatively, under lower stringency hybridization conditions, e.g., as defined supra, to polynucleotides that encode an antibody, preferably, that specifically binds to a polypeptide of the invention, preferably, an antibody that binds to a polypeptide having the amino acid sequence of SEQ ID NO:Y, to a polypeptide encoded by a portion of SEQ m NO:X as defined in columns 8 and 9 of Table 2, and/or to a polypeptide encoded by the cDNA
contained in Clone ID NO:Z.
[0243] The polynucleotides may be obtained, and the nucleotide sequence of the polynucleotides determined, by any method known in the art. For example, if the nucleotide sequence of the antibody is known, a polynucleotide encoding the antibody may be assembled from chemically synthesized oligonucleotides (e.g., as described in Kutmeier et al., BioTechniques 17:242 (1994)), which, briefly, involves the synthesis of overlapping oligonucleotides containing portions of the sequence encoding the antibody, annealing and ligating of those oligonucleotides, and then amplification of the ligated oligonucleotides by PCR.
[0244] Alternatively, a polynucleotide encoding an antibody may be generated from nucleic acid from a suitable source. If a clone containing a nucleic acid encoding a particular antibody is not available, but the sequence of the antibody molecule is known, a nucleic acid encoding the immunoglobulin may be chemically synthesized or obtained from a suitable source (e.g., an antibody cDNA library, or a cDNA library generated from, or nucleic acid, preferably poly A+ RNA, isolated from, any tissue or cells expressing the antibody, such as hybridoma cells selected to express an antibody of the invention) by PCR amplification using synthetic primers hybridizable to the 3' and 5' ends of the sequence or by cloning using an oligonucleotide probe specific for the particular gene sequence to identify, e.g., a cDNA clone from a cDNA library that encodes the antibody. Amplified nucleic acids generated by PCR may then be cloned into replicable cloning vectors using any method well known in the art.
[0245] Once the nucleotide sequence and corresponding amino acid sequence of the antibody is determined, the nucleotide sequence of the antibody may be manipulated using methods well known in the art for the manipulation of nucleotide sequences, e.g., recombinant DNA techniques, site directed mutagenesis, PCR, etc. (see, for example, the techniques described in Sambrook et al., 1990, Molecular Cloning, A Laboratory Manual, 2d Ed., Cold Spring Harbor Laboratory, Cold Spring Harbor, NY and Ausubel et al., eds., 1998, Current Protocols in Molecular Biology, John Wiley & Sons, NY, which are both incorporated by reference herein in their entireties ), to generate antibodies having a different amino acid sequence, for example to create amino acid substitutions, deletions, and/or insertions.
[0246] In a specific embodiment, the amino acid sequence of the heavy and/or light chain variable domains may be inspected to identify the sequences of the complementarity determining regions (CDRs) by methods that are well know in the art, e.g., by comparison to known amino acid sequences of other heavy and light chain variable regions to determine the regions of sequence hypervariability. Using routine recombinant DNA techniques, one or more of the CDRs may be inserted within framework regions, e.g., into human framework regions to humanize a non-human antibody, as described supra. The framework regions may be naturally occurring or consensus framework regions, and preferably human framework regions (see, e.g., Chothia et al., J. Mol. Biol. 278: 457-479 (1998) for a listing of human framework regions). Preferably, the polynucleotide generated by the combination of the framework regions and CDRs encodes an antibody that specifically binds a polypeptide of the invention. Preferably, as discussed supra, one or more amino acid substitutions may be made within the framework regions, and, preferably, the amino acid substitutions improve binding of the antibody to its antigen. Additionally, such methods may be used to make amino acid substitutions or deletions of one or more variable region cysteine residues participating in an intrachain disulfide bond to generate antibody molecules lacking one or more intrachain disulfide bonds. Other alterations to the polynucleotide are encompassed by the present invention and within the skill of the art.
[0247] In addition, techniques developed for the production of "chimeric antibodies"
(Morrison et al., Proc. Natl. Acad. Sci. 81:851-855 (1984); Neuberger et al., Nature 312:604-608 (1984); Takeda et al., Nature 314:452-454 (1985)) by splicing genes from a mouse antibody molecule of appropriate antigen specificity together with genes from a human antibody molecule of appropriate biological activity can be used. As described supra, a chimeric antibody is a molecule in which different portions are derived from different animal species, such as those having a variable region derived from a murine mAb and a human immunoglobulin constant region, e.g., humanized antibodies.
[0248] Alternatively, techniques described for the production of single chain antibodies (U.5. Patent No. 4,946,778; Bird, Science 242:423- 42 (1988);
Huston et al., Proc. Natl. Acad. Sci. USA 85:5879-5883 (1988); and Ward et al., Nature 334:544-54 (1989)) can be adapted to produce single chain antibodies. Single chain antibodies are formed by linking the heavy and light chain fragments of the Fv region via an amino acid bridge, resulting in a single chain polypeptide. Techniques for the assembly of functional Fv fragments in E. coli may also be used (Skerra et al., Science 242:1038-1041 (1988)).
Methods of Producing Antibodies [0249] The antibodies of the invention can be produced by any method known in the art for the synthesis of antibodies, in particular, by chemical synthesis or preferably, by recombinant expression techniques. Methods of producing antibodies include, but are not limited to, hybridoma technology, EBV transformation, and other methods discussed herein as well as through the use recombinant DNA technology, as discussed below.
[0250] Recombinant expression of an antibody of the invention, or fragment, derivative or analog thereof, (e.g., a heavy or light chain of an antibody of the invention or a single chain antibody of the invention), requires construction of an expression vector containing a polynucleotide that encodes the antibody. Once a polynucleotide encoding an antibody molecule or a heavy or light chain of an antibody, or portion thereof (preferably containing the heavy or light chain variable domain), of the invention has been obtained, the vector for the production of the antibody molecule may be produced by recombinant DNA technology using techniques well known in the art. Thus, methods for preparing a protein by expressing a polynucleotide containing an antibody encoding nucleotide sequence are described herein. Methods which are well known to those skilled in the art can be used to construct expression vectors containing antibody coding sequences and appropriate transcriptional and translational control signals.
These methods include, for example, in vitro recombinant DNA techniques, synthetic techniques, and in vivo genetic recombination. The invention, thus, provides replicable vectors comprising a nucleotide sequence encoding an antibody molecule of the invention, or a heavy or light chain thereof, or a heavy or light chain variable domain, operably linked to a promoter. Such vectors may include the nucleotide sequence encoding the constant region of the antibody molecule (see, e.g., PCT
Publication WO
86/05807; PCT Publication WO 89/01036; and U.S. Patent No. 5,122,464) and the variable domain of the antibody may be cloned into such a vector for expression of the entire heavy or light chain.
[0251] The expression vector is transferred to a host cell by conventional techniques and the transfected cells are then cultured by conventional techniques to produce an antibody of the invention. Thus, the invention includes host cells containing a polynucleotide encoding an antibody of the invention, or a heavy or light chain thereof, or a single chain antibody of the invention, operably linked to a heterologous promoter. In preferred embodiments for the expression of double-chained antibodies, vectors encoding both the heavy and light chains may be co-expressed in the host cell for expression of the entire immunoglobulin molecule, as detailed below.

[0252] A variety of host-expression vector systems may be utilized to express the antibody molecules of the invention. Such host-expression systems represent vehicles by which the coding sequences of interest may be produced and subsequently purified, but also represent cells which may, when transformed or transfected with the appropriate nucleotide coding sequences, express an antibody molecule of the invention in situ.
These include but are not limited to microorganisms such as bacteria (e.g., E.
coli, B.
subtilis) transformed with recombinant bacteriophage DNA, plasmid DNA or cosmid DNA expression vectors containing antibody coding sequences; yeast (e.g., Saccharomyces, Pichia) transformed with recombinant yeast expression vectors containing antibody coding sequences; insect cell systems infected with recombinant virus expression vectors (e.g., baculovirus) containing antibody coding sequences; plant cell systems infected with recombinant virus expression vectors (e.g., cauliflower mosaic virus, CaMV; tobacco mosaic virus, TMV) or transformed with recombinant plasmid expression vectors (e.g., Ti plasmid) containing antibody coding sequences; or mammalian cell systems (e.g., COS, CHO, BHK, 293, 3T3 cells) harboring recombinant expression constructs containing promoters derived from the genome of mammalian cells (e.g., metallothionein promoter) or from mammalian viruses (e.g., the adenovirus late promoter; the vaccinia virus 7.5K promoter). Preferably, bacterial cells such as Escherichia coli, and more preferably, eukaryotic cells, especially for the expression of whole recombinant antibody molecule, are used for the expression of a recombinant antibody molecule. For example, mammalian cells such as Chinese hamster ovary cells (CHO), in conjunction with a vector such as the major intermediate early gene promoter element from human cytomegalovirus is an effective expression system for antibodies (Foecking et al., Gene 45:101 (1986); Cockett et al., Bio/Technology 8:2 (1990)).
[0253] In bacterial systems, a number of expression vectors may be advantageously selected depending upon the use intended for the antibody molecule being expressed. For example, when a large quantity of such a protein is to be produced, for the generation of pharmaceutical compositions of an antibody molecule, vectors which direct the expression of high levels of fusion protein products that are readily purified may be desirable. Such vectors include, but are not limited, to the E. coli expression vector pUR278 (Ruther et al., EMBO J. 2:1791 (1983)), in which the antibody coding sequence may be ligated individually into the vector in frame with the lac Z coding region so that a fusion protein is produced; pIN vectors (Inouye & Inouye, Nucleic Acids Res.
13:3101-3109 (1985); Van Heeke & Schuster, J. Biol. Chem. 24:5503-5509 (1989)); and the like.
pGEX vectors may also be used to express foreign polypeptides as fusion proteins with glutathione S-transferase (GST). In general, such fusion proteins are soluble and can easily be purified from lysed cells by adsorption and binding to matrix glutathione-agarose beads followed by elution in the presence of free glutathione. The pGEX vectors are designed to include thrombin or factor Xa protease cleavage sites so that the cloned target gene product can be released from the GST moiety.
[0254] In an insect system, Autographa californica nuclear polyhedrosis virus (AcNPV) is used as a vector to express foreign genes. The virus grows in Spodoptera frugiperda cells. The antibody coding sequence may be cloned individually into non-essential regions (for example the polyhedrin gene) of the virus and placed under control of an AcNPV promoter. (for example the polyhedrin promoter).
[0255] In mammalian host cells, a number of viral-based expression systems may be utilized. In cases where an adenovirus is used as an expression vector, the antibody coding sequence of interest may be ligated to an adenovirus transcription/translation control complex, e.g., the late promoter and tripartite leader sequence. This chimeric gene may then be inserted in the adenovirus genome by in vitro or in vivo recombination.
Insertion in a non- essential region of the viral genome (e.g., region E1 or E3) will result in a recombinant virus that is viable and capable of expressing the antibody molecule in infected hosts. (e.g., see Logan & Shenk, Proc. Natl. Acad. Sci. USA 81:355-(1984)). Specific initiation signals may also be required for efficient translation of inserted antibody coding sequences. These signals include the ATG initiation codon and adjacent sequences. Furthermore, the initiation codon must be in phase with the reading frame of the desired coding sequence to ensure translation of the entire insert. These exogenous translational control signals and initiation codons can be of a variety of origins, both natural and synthetic. The efficiency of expression may be enhanced by the inclusion of appropriate transcription enhancer elements, transcription terminators, etc.
(see Bittner et al., Methods in Enzymol. 153:51-544 (1987)).
[0256] In addition, a host cell strain may be chosen which modulates the expression of the inserted sequences, or modifies and processes the gene product in the specific fashion desired. Such modifications (e.g., glycosylation) and processing (e.g., cleavage) of protein products may be important for the function of the protein.
Different host cells have characteristic and specific mechanisms for the post-translational processing and modification of proteins and gene products. Appropriate cell lines or host systems can be chosen to ensure the correct modification and processing of the foreign protein expressed.
To this end, eukaryotic host cells which possess the cellular machinery for proper processing of the primary transcript, glycosylation, and phosphorylation of the gene product may be used. Such mammalian host cells include but are not limited to CHO, VERY, BHK, Hela, COS, MDCK, 293, 3T3, WI38, and in particular, breast cancer cell lines such as, for example, BT483, Hs578T, HTB2, BT20 and T47D, and normal mammary gland cell line such as, for example, CRL7030 and Hs578Bst.
[0257] For long-term, high-yield production of recombinant proteins, stable expression is preferred. For example, cell lines, which stably express the antibody molecule may be engineered. Rather than using expression vectors which contain viral origins of replication, host cells can be transformed with DNA controlled by appropriate expression control elements (e.g., promoter, enhancer, sequences, transcription terminators, polyadenylation sites, etc.), and a selectable marker. Following the introduction of the foreign DNA, engineered cells may be allowed to grow for 1-2 days in an enriched media, and then are switched to a selective media. The selectable marker in the recombinant plasmid confers resistance to the selection and allows cells to stably integrate the plasmid into their chromosomes and grow to form foci which in turn can be cloned and expanded into cell lines. This method may advantageously be used , to engineer cell lines, which express the antibody molecule. Such engineered cell lines may be particularly useful in screening and evaluation of compounds that interact directly or indirectly with the antibody molecule.
[0258] A number of selection systems may be used, including but not limited to the herpes simplex virus thymidine kinase (Wigler et al., Cell 11:223 (1977)), hypoxanthine-guanine phosphoribosyltransferase (Szybalska & Szybalski, Proc. Natl. Acad.
Sci. USA
48:202 (1992)), and adenine phosphoribosyltransferase (Lowy et al., Cell 22:817 (1980)) genes can be employed in tk-, hgprt- or aprt- cells, respectively. Also, antimetabolite resistance can be used as the basis of selection for the following genes:
dhfr, which confers resistance to methotrexate (Wigler et al., Natl. Acad. Sci. USA 77:357 (1980);
O'Hare et al., Proc. Natl. Acad. Sci. USA 78:1527 (1981)); gpt, which confers resistance to mycophenolic acid (Mulligan & Berg, Proc. Natl. Acad. Sci. USA 78:2072 (1981));
neo, which confers resistance to the aminoglycoside G-418 Clinical Pharmacy 12:488-505; Wu and Wu, Biotherapy 3:87-95 (1991); Tolstoshev, Ann. Rev. Pharmacol.
Toxicol. 32:573-596 (1993); Mulligan, Science 260:926-932 (1993); and Morgan and Anderson, Ann. Rev. Biochem. 62:191-217 (1993); TIB TECH 11(5):155-215 (1993));
and hygro, which confers resistance to hygromycin (Santerre et al., Gene 30:147 (1984)).
Methods commonly known in the art of recombinant DNA technology may be routinely applied to select the desired recombinant clone, and such methods are described, for example, in Ausubel et al. (eds.), Current Protocols in Molecular Biology, John Wiley &
Sons, NY (1993); Kriegler, Gene Transfer and Expression, A Laboratory Manual, Stockton Press, NY (1990); and in Chapters 12 and 13, Dracopoli et al. (eds), Current Protocols in Human Genetics, John Wiley & Sons, NY (1994); Colberre-Garapin et al., J.
Mol. Biol. 150:1 (1981), which are incorporated by reference herein in their entireties.
[0259] The expression levels of an antibody molecule can be increased by vector amplification (for a review, see Bebbington and Hentschel, The use of vectors based on gene amplification for the expression of cloned genes in mammalian cells in DNA
cloning, Vol.3. (Academic Press, New York, 1987)). When a marker in the vector system expressing antibody is amplifiable, increase in the level of inhibitor present in culture of host cell will increase the number of copies of the marker gene.
Since the amplified region is associated with the antibody gene, production of the antibody will also increase (Grouse et al., Mol. Cell. Biol. 3:257 (1983)).
[0260] Vectors, which use glutamine synthase (GS) or DHFR as the selectable markers can be amplified in the presence of the drugs methionine sulphoximine or methotrexate, respectively. An advantage of glutamine synthase based vectors are the availabilty of cell lines (e.g., the murine myeloma cell line, NSO) which are glutamine synthase negative. Glutamine synthase expression systems can also function in glutamine synthase expressing cells (e.g., Chinese Hamster Ovary (CHO) cells) by providing additional inhibitor to prevent the functioning of the endogenous gene. A
glutamine synthase expression system and components thereof are detailed in PCT
publications:
W087/04462; W086/05807; W089/01036; W089/10404; and W091/06657, which are incorporated in their entireties by reference herein. Additionally, glutamine synthase expression vectors that may be used according to the present invention are commercially available from suplliers, including, for example Lonza Biologics, Inc.
(Portsmouth, NH).
Expression and production of monoclonal antibodies using a GS expression system in murine myeloma cells is described in Bebbington et al., Biotechnology 10:169(1992) and in Biblia and Robinson Biotechnol. Prog. 11:1 (1995) which are incorporated in their entirities by reference herein.
[0261] The host cell may be co-transfected with two expression vectors of the invention, the first vector encoding a heavy chain derived polypeptide and the second vector encoding a light chain derived polypeptide. The two vectors may contain identical selectable markers, which enable equal expression of heavy and light chain polypeptides.
Alternatively, a single vector may be used which encodes, and is capable of expressing, both heavy and light chain polypeptides. In such situations, the light chain should be placed before the heavy chain to avoid an excess of toxic free heavy chain (Proudfoot, Nature 322:52 (1986); Kohler, Proc. Natl. Acad. Sci. USA 77:2197 (1980)). The coding sequences for the heavy and light chains may comprise cDNA or genomic DNA.
[0262] Once an antibody molecule of the invention has been produced by an animal, chemically synthesized, or recombinantly expressed, it may be purified by any method known in the art for purification of an immunoglobulin molecule, for example, by chromatography (e.g., ion exchange, affinity, particularly by affinity for the specific antigen after Protein A, and sizing column chromatography), centrifugation, differential solubility, or by any other standard technique for the purification of proteins. In addition, the antibodies of the present invention or fragments thereof can be fused to heterologous polypeptide sequences described herein or otherwise known in the art, to facilitate purification.
[0263] The present invention encompasses antibodies recombinantly fused or chemically conjugated (including both covalently and non-covalently conjugations) to a polypeptide (or portion thereof, preferably at least 10, 20, 30, 40, 50, 60, 70, 80, 90 or 100 amino acids of the polypeptide) of the present invention to generate fusion proteins. The fusion does not necessarily need to be direct, but may occur through linker sequences.
The antibodies may be specific for antigens other than polypeptides (or portion thereof, preferably at least 10, 20, 30, 40, 50, 60, 70, 80, 90 or 100 amino acids of the polypeptide) of the present invention. For example, antibodies may be used to target the polypeptides of the present invention to particular cell types, either in vitro or in vivo, by fusing or conjugating the polypeptides of the present invention to antibodies specific for particular cell surface receptors. Antibodies fused or conjugated to the polypeptides of the present invention may also be used in in vitro immunoassays and purification methods using methods known in the art. See e.g., Harbor et al., supra, and PCT
publication WO
93/21232; EP 439,095; Naramura et al., linmunol. Lett. 39:91-99 (1994); U.S.
Patent 5,474,981; Gillies et al., PNAS 89:1428-1432 (1992); Fell et al., J. Immunol.
146:2446-2452 (1991), which are incorporated by reference in their entireties.
[0264] The present invention further includes compositions comprising the polypeptides of the present invention fused or conjugated to antibody domains other than the variable regions. For example, the polypeptides of the present invention may be fused or conjugated to an antibody Fc region, or portion thereof. The antibody portion fused to a polypeptide of the present invention may comprise the constant region, hinge region, CH1 domain, CH2 domain, and CH3 domain or any combination of whole domains or portions thereof. The polypeptides may also be fused or conjugated to the above antibody portions to form multimers. For example, Fc portions fused to the polypeptides of the present invention can form dimers through disulfide bonding between the Fc portions.
Higher multimeric forms can be made by fusing the polypeptides to portions of IgA and IgM. Methods for fusing or conjugating the polypeptides of the present invention to antibody portions are known in the art. See, e.g., U.S. Patent Nos. 5,336,603;
5,622,929;
5,359,046; 5,349,053; 5,447,851; 5,112,946; EP 307,434; EP 367,166; PCT.
publications WO 96/04388; WO 91/06570; Ashkenazi et al., Proc. Natl. Acad. Sci. USA
88:10535-10539 (1991); Zheng et al., J. Immunol. 154:5590-5600 (1995); and Vil et al., Proc. Natl.
Acad. Sci. USA 89:11337- 11341 (1992) (said references incorporated by reference in their entireties).
[0265] As discussed, supra, the polypeptides corresponding to a polypeptide, polypeptide fragment, or a variant of SEQ ID NO:Y may be fused or conjugated to the above antibody portions to increase the in vivo half life of the polypeptides or for use in immunoassays using methods known in the art. Further, the polypeptides corresponding to SEQ ID NO:Y may be fused or conjugated to the above antibody portions to facilitate purification. One reported example describes chimeric proteins consisting of the first two domains of the human CD4-polypeptide and various domains of the constant regions of the heavy or light chains of mammalian immunoglobulins. See EP 394,827;
Traunecker et al., Nature 331:84-86 (1988). The polypeptides of the present invention fused or conjugated to an antibody having disulfide- linked dimeric structures (due to the IgG) may also be more efficient in binding and neutralizing other molecules, than the monomeric secreted protein or protein fragment alone. See, for example, Fountoulakis et al., J. Biochem. 270:3958-3964 (1995). In many cases, the Fc part in a fusion protein is beneficial in therapy and diagnosis, and thus can result in, for example, improved pharmacokinetic properties. See, for example, EP A 232,262. Alternatively, deleting the Fc part after the fusion protein has been expressed, detected, and purified, would be desired. For example, the Fc portion may hinder therapy and diagnosis if the fusion protein is used as an antigen for immunizations. In drug discovery, for example, human proteins, such as hIL-5, have been fused with Fc portions for the purpose of high-throughput screening assays to identify antagonists of hIL-5. (See, Bennett et al., J.
Molecular Recognition 8:52-58 (1995); Johanson et al., J. Biol. Chem. 270:9459-(1995)).
[0266] Moreover, the antibodies or fragments thereof of the present invention can be fused to marker sequences, such as a peptide to facilitate purification. In preferred embodiments, the marker amino acid sequence is a hexa-histidine peptide, such as the tag provided in a pQE vector (QIAGEN, Inc., 9259 Eton Avenue, Chatsworth, CA, 91311), among others, many of which are commercially available. As described in Gentz et al., Proc. Natl. Acad. Sci. USA 86:821-824 (1989), for instance, hexa-histidine provides for convenient purification of the fusion protein. Other peptide tags useful for purification include, but are not limited to, the "HA" tag, which corresponds to an epitope derived from the influenza hemagglutinin protein (Wilson et al., Cell 37:767 (1984)) and the "flag" tag.
[0267] The present invention further encompasses antibodies or fragments thereof conjugated to a diagnostic or therapeutic agent. The antibodies can be used diagnostically to, for example, monitor the development or progression of a tumor as part of a clinical testing procedure to, e.g., determine the efficacy of a given treatment regimen. Detection can be facilitated by coupling the antibody to a detectable substance.
Examples of detectable substances include various enzymes, prosthetic groups, fluorescent materials, luminescent materials, bioluminescent materials, radioactive materials, positron emitting metals using various positron emission tomographies, and nonradioactive paramagnetic metal ions. The detectable substance may be coupled or conjugated either directly to the antibody (or fragment thereof) or indirectly, through an intermediate (such as, for example, a linker known in the art) using techniques known in the art. See, for example, U.S. Patent No. 4,741,900 for metal ions which can be conjugated to antibodies for use as diagnostics according to the present invention.
(0268] Further, an antibody or fragment thereof may be conjugated to a therapeutic moiety such as a cytotoxin, e.g., a cytostatic or cytocidal agent, a therapeutic agent or a radioactive metal ion, e.g., alpha-emitters such as, for example, 213Bi. A
cytotoxin or cytotoxic agent includes any agent that is detrimental to cells. Examples include paclitaxol, cytochalasin B, gramicidin D, ethidium bromide, emetine, mitomycin, etoposide, tenoposide, vincristine, vinblastine, colchicin, doxorubicin, daunorubicin, dihydroxy anthracin dione, mitoxantrone, mithramycin, actinomycin D, 1-dehydrotestosterone, glucocorticoids, procaine, tetracaine, lidocaine, propranolol, and puromycin and analogs or homologs thereof. Therapeutic agents include, but are not limited to, antimetabolites (e.g., methotrexate, 6-mercaptopurine, 6-thioguanine, cytarabine, 5-fluorouracil decarbazine), alkylating agents (e.g., mechlorethamine, thioepa chlorambucil, melphalan, carmustine (BSNU) and lomustine (CCNU), cyclothosphamide, busulfan, dibromomannitol, streptozotocin, mitomycin C, and cis-dichlorodiamine platinum (II) (DDP) cisplatin), anthracyclines (e.g., daunorubicin (formerly daunomycin) and doxorubicin), antibiotics (e.g., dactinomycin (formerly actinomycin), bleomycin, mithramycin, and anthramycin (AMC)), and anti-mitotic agents (e.g., vincristine and vinblastine).
[0269] The conjugates of the invention can be used for modifying a given biological response, the therapeutic agent or drug moiety is not to be construed as limited to classical chemical therapeutic agents. For example, the drug moiety may be a protein or polypeptide possessing a desired biological activity. Such proteins may include, for example, a toxin such as abrin, ricin A, pseudomonas exotoxin, or diphtheria toxin; a protein such as tumor necrosis factor, a-interferon, 13-interferon, nerve growth factor, platelet derived growth factor, tissue plasminogen activator, an apoptotic agent, e.g., TNF-alpha, TNF-beta, AIM I (See, International Publication No. WO 97/33899), AIM II
(See, International Publication No. WO 97/34911), Fas Ligand (Takahashi et al., Int.
Immunol., 6:1567-1574 (1994)), VEGI (See, International Publication No. WO

99/23105), a thrombotic agent or an anti- angiogenic agent, e.g., angiostatin or endostatin;
or, biological response modifiers such as, for example, lymphokines, interleukin-1 ("IL-1 "), interleukin-2 ("IL-2"), interleukin-6 ("IL-6"), granulocyte macrophage colony stimulating factor ("GM-CSF"), granulocyte colony stimulating factor ("G-CSF"), or other growth factors.
[0270] Antibodies may also be attached to solid supports, which are particularly useful for immunoassays or purification of the target antigen. Such solid supports include, but are not limited to, glass, cellulose, polyacrylamide, nylon, polystyrene, polyvinyl chloride or polypropylene.
[0271] Techniques for conjugating such therapeutic moiety to antibodies are well known. See, for example, Arnon et al., "Monoclonal Antibodies For Immunotargeting Of Drugs In Cancer Therapy", in Monoclonal Antibodies And Cancer Therapy, Reisfeld et al. (eds.), pp. 243-56 (Alan R. Liss, Inc. 1985); Hellstrom et al., "Antibodies For Drug Delivery", in Controlled Drug Delivery (2nd Ed.), Robinson et al. (eds.), pp.

(Marcel Dekker, Inc. 1987); Thorpe, "Antibody Carriers Of Cytotoxic Agents In Cancer Therapy: A Review", in Monoclonal Antibodies '84: Biological And Clinical Applications, Pinchers et al. (eds.), pp. 475-506 (1985); "Analysis, Results, And Future Prospective Of The Therapeutic Use Of Radiolabeled Antibody In Cancer Therapy", in Monoclonal Antibodies For Cancer Detection And Therapy, Baldwin et al. (eds:), pp.
303-16 (Academic Press 1985), and Thorpe et al., "The Preparation And Cytotoxic Properties Of Antibody-Toxin Conjugates", Immunol. Rev. 62:119-58 (1982).
[0272] Alternatively, an antibody can be conjugated to a second antibody to form an antibody heteroconjugate as described by Segal in U.S. Patent No. 4,676,980, which is incorporated herein by reference in its entirety.
[0273] An antibody, with or without a therapeutic moiety conjugated to it, administered alone or in combination with cytotoxic factors) and/or cytokine(s) can be used as a therapeutic.
Immunophenotyping (0274] The antibodies of the invention may be utilized for immunophenotyping of cell lines and biological samples. Translation products of the genes of the present invention may be useful as cell specific markers, or more specifically as cellular markers that are differentially expressed at various stages of differentiation and/or maturation of particular cell types. Monoclonal antibodies directed against a specific epitope, or combination of epitopes, will allow for the screening of cellular populations expressing the marker. Various techniques can be utilized using monoclonal antibodies to screen for cellular populations expressing the marker(s), and include magnetic separation using antibody-coated magnetic beads, "panning" with antibody attached to a solid matrix (i.e., plate), and flow cytometry (See, e.g., U.S. Patent 5,985,660; and Mornson et al., Cell, 96:737-49 (1999)).
[0275] These techniques allow for the screening of particular populations of cells, such as might be found with hematological malignancies (i.e. minimal residual disease (MRD) in acute leukemic patients) and "non-self' cells in transplantations to prevent Graft-versus-Host Disease (GVHD). Alternatively, these techniques allow for the screening of hematopoietic stem and progenitor cells capable of undergoing proliferation and/or differentiation, as might be found in human umbilical cord blood.
Assays For Antibody Binding [0276] The antibodies of the invention may be assayed for immunospecific binding -by any method known in the art. The immunoassays which can be used include but are not limited to competitive and non-competitive assay systems using techniques such as western blots, radioimmunoassays, ELISA (enzyme linked immunosorbent assay), "sandwich" immunoassays, immunoprecipitation assays, precipitin reactions, gel diffusion precipitin reactions, immunodiffusion assays, agglutination assays, complement-fixation assays, immunoradiometric assays, fluorescent immunoassays, and protein A immunoassays, to name but a few. Such assays are routine and well known in the art (see, e.g., Ausubel et al, eds, 1994, Current Protocols in Molecular Biology, Vol.
1, John Wiley & Sons, Inc., New York, which is incorporated by reference herein in its entirety). Exemplary immunoassays are described briefly below (but are not intended by way of limitation).
[0277] Immunoprecipitation protocols generally comprise lysing a population of cells in a lysis buffer such as RIPA buffer ( 1 % NP-40 or Triton X- 100, 1 % sodium deoxycholate, 0.1 % SDS, 0.15 M NaCI, 0.01 M sodium phosphate at pH 7.2, 1 Trasylol) supplemented with protein phosphatase and/or protease inhibitors (e.g., EDTA, PMSF, aprotinin, sodium vanadate), adding the antibody of interest to the cell lysate, incubating for a period of time (e.g., 1-4 hours) at 4° C, adding protein A and/or protein G
sepharose beads to the cell lysate, incubating for about an hour or more at 4° C, washing the beads in lysis buffer and resuspending the beads in SDS/sample buffer. The ability of the antibody of interest to immunoprecipitate a particular antigen can be assessed by, e.g., western blot analysis. One of skill in the art would be knowledgeable as to the parameters that can be modified to increase the binding of the antibody to an antigen and decrease the background (e.g., pre-clearing the cell lysate with sepharose beads). For further discussion regarding immunoprecipitation protocols see, e.g., Ausubel et al., eds., (1994), Current Protocols in Molecular Biology, Vol. 1, John Wiley & Sons, Inc., New York, section 10.16.1.
[0278] Western blot analysis generally comprises preparing protein samples, electrophoresis of the protein samples in a polyacrylamide gel (e.g., 8%- 20%
SDS-PAGE depending on the molecular weight of the antigen), transferring the protein sample from the polyacrylamide gel to a membrane such as nitrocellulose, PVDF or nylon, blocking the membrane in blocking solution (e.g., PBS with 3% BSA or non-fat milk), washing the membrane in washing buffer (e.g., PBS-Tween 20), blocking the membrane with primary antibody (the antibody of interest) diluted in blocking buffer, washing the membrane in washing buffer, blocking the membrane with a secondary antibody (which recognizes the primary antibody, e.g., an anti-human antibody) conjugated to an enzymatic substrate (e.g., horseradish peroxidase or alkaline phosphatase) or radioactive molecule (e.g., 32P or 125I) diluted in blocking buffer, washing the membrane in wash buffer, and detecting the presence of the antigen. One of skill in the art would be knowledgeable as to the parameters that can be modified to increase the signal detected and to reduce the background noise. For further discussion regarding western blot protocols see, e.g., Ausubel et al., eds., (1994), Current Protocols in Molecular Biology, Vol. l, John Wiley & Sons, Inc., New York, section 10.8.1.
[0279] ELISAs comprise preparing antigen, coating the well of a 96 well microtiter plate with the antigen, adding the antibody of interest conjugated to a detectable compound such as an enzymatic substrate (e.g., horseradish peroxidase or alkaline phosphatase) to the well and incubating for a period of time, and detecting the presence of the antigen. In ELISAs the antibody of interest does not have to be conjugated to a detectable compound; instead, a second antibody (which recognizes the antibody of interest) conjugated to a detectable compound may be added to the well.
Further, instead of coating the well with the antigen, the antibody may be coated to the well.
In this case, a second antibody conjugated to a detectable compound may be added following the addition of the antigen of interest to the coated well. One of skill in the art would be knowledgeable as to the parameters that can be modified to increase the signal detected as well as other variations of ELISAs known in the art. For further discussion regarding ELISAs see, e.g., Ausubel et al., eds, (1994), Current Protocols in Molecular Biology, Vol. 1, John Wiley & Sons, Inc., New York, section 11.2.1.
[0280] The binding affinity of an antibody to an antigen and the off rate of an antibody-antigen interaction can be determined by competitive binding assays.
One example of a competitive binding assay is a radioimmunoassay comprising the incubation of labeled antigen (e.g., 3H or 125I) with the antibody of interest in the presence of increasing amounts of unlabeled antigen, and the detection of the antibody bound to the labeled antigen. The affinity of the antibody of interest for a particular antigen and the binding off rates can be determined from the data by scatchard plot analysis.
Competition with a second antibody can also be determined using radioimmunoassays.
In this case, the antigen is incubated with antibody of interest conjugated to a labeled compound (e.g., 3H or 125I) in the presence of increasing amounts of an unlabeled second antibody.
[0281] Antibodies of the invention may be characterized using immunocytochemisty methods on cells (e.g., mammalian cells, such as CHO cells) transfected with a vector enabling the expression of a connective tissue antigen or with vector alone using techniques commonly known in the art. Antibodies that bind connective tissue antigen transfected cells, but not vector-only transfected cells, are connective tissue antigen specific.
Therapeutic Uses [0282] The present invention is further directed to antibody-based therapies which involve administering antibodies of the invention to an animal, preferably a mammal, and most preferably a human, patient for treating one or more of the disclosed, diseases, disorders, or conditions. Therapeutic compounds of the invention include, but are not limited to, antibodies of the invention (including fragments, analogs and derivatives thereof as described herein) and nucleic acids encoding antibodies of the invention (including fragments, analogs and derivatives thereof and anti-idiotypic antibodies as described herein). The antibodies of the invention can be used to treat, inhibit or prevent diseases, disorders or conditions associated with aberrant expression and/or activity of a polypeptide of the invention, including, but not limited to, any one or more of the diseases, disorders, or conditions described herein. The treatment and/or prevention of diseases, disorders, or conditions associated with aberrant expression and/or activity of a polypeptide of the invention includes, but is not limited to, alleviating symptoms associated with those diseases, disorders or conditions. Antibodies of the invention may be provided in pharmaceutically acceptable compositions as known in the art or as described herein.
[0283] In a specific and preferred embodiment, the present invention is directed to antibody-based therapies which involve administering antibodies of the invention to an animal, preferably a mammal, and most preferably a human, patient for treating one or more of the diseases, disorders, or conditions of connective tissues, including, but not limited to, rheumatoid arthritis, psoriatic arthritis, discoid lupus erythematosus, systemic lupus erythematosus, scleroderma, CREST syndrome, Sjogren's syndrome, polymyositis, dermatomyositis, mixed connective tissue disease, relapsing polychondritis, vasculitis, Henoch-Schonlein syndrome, erythema nodosum, polyarteritis nodosa, temporal (giant cell) arteritis, Takayasu's arteritis, Wegener's granulomatosis, Reiter's syndrome, Behcet's syndrome, ankylosing spondylitis, cellulitis, keloids, Ehler Danlos syndrome, Marfan syndrome, pseudoxantoma elasticum, osteogenese imperfecta, chondrodysplasias, epidermolysis bullosa, Alport syndrome, cubs laxa, genetic disorders affecting skeleton, skin and muscles; formation of excessive scar tissue; deposition of pathological amounts of connective tissue in body organs, including kidney, intestines and heart, and in liver by liver cirrhosis, in skin by scleroderma, in lung by lung fibrosis, in bone marrow by leukemia, in blood vessels by atherosclerosis, and in joints by rheumatic diseases.
Therapeutic compounds of the invention include, but are not limited to, antibodies of the invention (e.g., antibodies directed to the full length protein expressed on the cell surface of a mammalian cell; antibodies directed to an epitope of a connective tissue associated polypeptide of the invention (such as, a linear epitope (shown in Table 1A, column 6) or a conformational epitope), including fragments, analogs and derivatives thereof as described herein) and nucleic acids encoding antibodies of the invention (including fragments, analogs and derivatives thereof and anti-idiotypic antibodies as described herein). The antibodies of the invention can be used to treat, inhibit or prevent diseases, disorders or conditions associated with aberrant expression and/or activity of a polypeptide of the invention, including, but not limited to, any one or more of the diseases, disorders, or conditions of connective tissues described herein. The treatment and/or prevention of diseases, disorders, or conditions of connective tissues associated with aberrant expression and/or activity of a polypeptide of the invention includes, but is .
not limited to, alleviating symptoms associated with those diseases, disorders or conditions. Antibodies of the invention may be provided in pharmaceutically acceptable compositions as known in the art or as described herein.
[0284] A summary of the ways in which the antibodies of the present invention may be used therapeutically includes binding polynucleotides or polypeptides of the present invention locally or systemically in the body or by direct cytotoxicity of the antibody, e.g.
as mediated by complement (CDC) or by effector cells (ADCC). Some of these approaches are described in more detail below. Armed with the teachings provided herein, one of ordinary skill in the art will know how to use the antibodies of the present invention for diagnostic, monitoring or therapeutic purposes without undue experimentation.
[0285] The antibodies of this invention may be advantageously utilized in combination with other monoclonal or chimeric antibodies, or with lymphokines or hematopoietic growth -factors (such as, e.g., IL-2, IL-3 and IL,-7), for example, which serve to increase the number or activity of effector cells which interact with the antibodies.
[0286] The antibodies of the invention may be administered alone or in combination with other types of treatments (e.g., radiation therapy, chemotherapy, hormonal therapy, immunotherapy and anti-tumor agents). Generally, administration of products of a species origin or species reactivity (in the case of antibodies) that is the same species as that of the patient is preferred. Thus, in a preferred embodiment, human antibodies, fragments derivatives, analogs, or nucleic acids, are administered to a human patient for therapy or prophylaxis.

[0287] It is preferred to use high affinity and/or potent in vivo inhibiting and/or neutralizing antibodies against polypeptides or polynucleotides of the present invention, fragments or regions thereof, for both immunoassays directed to and therapy of disorders related to polynucleotides or polypeptides, including fragments thereof, of the present invention. Such antibodies, fragments, or regions, will preferably have an affinity for polynucleotides or polypeptides of the invention, including fragments thereof.
Preferred binding affinities include those with a dissociation constant or Kd less than 5 X 10-Z M, 10-Z M, 5 X 10-3 M, 10-3 M, 5 X 10~ M, 10-4 M, 5 X 10-5 M, 10-5 M, 5 X 10-6 M, 10-~ M, S X 10-' M, 10-' M, S X 10-$ M, 10-g M, 5 X 10-9 M, 10-9 M, 5 X 10-' °
M, 10-' ° M, 5 X
10-" M, 10-" M, 5 X 10-' 2 M, 10-' 2 M, 5 X 10-' 3 M, 10-' 3 M, 5 X 10-' 4 M, 10-' 4 M, 5 X
10-' S M, and 10-' S M.
Gene Therapy [0288] In a specific embodiment, nucleic acids comprising sequences encoding antibodies or functional derivatives thereof, are administered to treat, inhibit or prevent a disease or disorder associated with aberrant expression and/or activity of a polypeptide of the invention, by way of gene therapy. Gene therapy refers to therapy performed by the administration to a subject of an expressed or expressible nucleic acid. In this embodiment of the invention, the nucleic acids produce their encoded protein that mediates a therapeutic effect.
[0289] Any of the methods for gene therapy available in the art can be used according to the present invention. Exemplary methods are described below.
[0290] For general reviews of the methods of gene therapy, see Goldspiel et al., Clinical Pharmacy 12:488-505 (1993); Wu and Wu, Biotherapy 3:87-95 (1991);
Tolstoshev, Ann. Rev. Pharmacol. Toxicol. 32:573-596 (1993); Mulligan, Science 260:926-932 (1993); and Morgan and Anderson, Ann. Rev. Biochem. 62:191-217 (1993); May, TIBTECH 11(5):155-215 (1993). Methods commonly known in the art of recombinant DNA technology which can be used are described in Ausubel et al.
(eds.), Current Protocols in Molecular Biology, John Wiley & Sons, NY (1993); and Kriegler, Gene Transfer and Expression, A Laboratory Manual, Stockton Press, NY (1990).
[0291] In a preferred embodiment, the compound comprises nucleic acid sequences encoding an antibody, said nucleic acid sequences being part of expression vectors that express the antibody or fragments or chimeric proteins or heavy or light chains thereof in a suitable host. In particular, such nucleic acid sequences have promoters operably linked to the antibody coding region, said promoter being inducible or constitutive, and, optionally, tissue-specific. In another particular embodiment, nucleic acid molecules are used in which the antibody coding sequences and any other desired sequences are flanked by regions that promote homologous recombination at a desired site in the genome, thus providing for intrachromosomal expression of the antibody encoding nucleic acids (Koller and Smithies, Proc. Natl. Acad. Sci. USA 86:8932-8935 (1989); Zijlstra et al., Nature 342:435-438 (1989). In specific embodiments, the expressed antibody molecule is a single chain antibody; alternatively, the nucleic acid sequences include sequences encoding both the heavy and light chains, or fragments thereof, of the antibody.
[0292] Delivery of the nucleic acids into a patient may be either direct, in which case the patient is directly exposed to the nucleic acid or nucleic acid- carrying vectors, or indirect, in which case, cells are first transformed with the nucleic acids in vitro, then transplanted into the patient. These two approaches are known, respectively, as in vivo or ex vivo gene therapy.
[0293] In a specific embodiment, the nucleic acid sequences are directly administered in vivo, where it is expressed to produce the encoded product. This can be accomplished by any of numerous methods known in the art, e.g., by constructing them as part of an appropriate nucleic acid expression vector and administering it so that they become intracellular, e.g., by infection using defective or attenuated retrovirals or other viral vectors (see U.S. Patent No. 4,980,286), or by direct injection of naked DNA;
or by use of microparticle bombardment (e.g., a gene gun; Biolistic, Dupont), or coating with lipids or cell-surface receptors or transfecting agents, encapsulation in liposomes, microparticles, or microcapsules, or by administering them in linkage to a peptide which is known to enter the nucleus, by administering it in linkage to a ligand subject to receptor-mediated endocytosis (see, e.g., Wu and Wu, J. Biol. Chem. 262:4429-( 1987)) (which can be used to target cell types specifically expressing the receptors), etc.
In another embodiment, nucleic acid-ligand complexes can be formed in which the ligand comprises a fusogenic viral peptide to disrupt endosomes, allowing the nucleic acid to avoid lysosomal degradation. In yet another embodiment, the nucleic acid can be targeted in vivo for cell specific uptake and expression, by targeting a specific receptor (see, e.g., PCT Publications WO 92/06180; WO 92/22635; W092/20316; W093/14188, WO 93/20221). Alternatively, the nucleic acid can be introduced intracellularly and incorporated within host cell DNA for expression, by homologous recombination (Koller and Smithies, Proc. Natl. Acad. Sci. USA 86:8932-8935 (1989); Zijlstra et al., Nature 342:435-438 (1989)).
[0294] In a specific embodiment, viral vectors that contains nucleic acid sequences encoding an antibody of the invention are used. For example, a retroviral vector can be used (see Miller et al., Meth. Enzymol. 217:581-599 (1993)). These retroviral vectors contain the components necessary for the correct packaging of the viral genome and integration into the host cell DNA. The nucleic acid sequences encoding the antibody to be used in gene therapy are cloned into one or more vectors, which facilitates delivery of the gene into a patient. More detail about retroviral vectors can be found in Boesen et al., Biotherapy 6:291-302 (1994), which describes the use of a retroviral vector to deliver the mdrl gene to hematopoietic stem cells in order to make the stem cells more resistant to chemotherapy. Other references illustrating the use of retroviral vectors in gene therapy are: Clowes et al., J. Clin. Invest. 93:644-651 (1994); Kiem et al., Blood 83:1467-1473 (1994); Salmons and Gunzberg, Human Gene Therapy 4:129-141 (1993); and Grossman and Wilson, Curr. Opin. in Genetics and Devel. 3:110-114 (1993).
[0295] Adenoviruses are other viral vectors that can be used in gene therapy.
Adenoviruses are especially attractive vehicles for delivering genes to respiratory epithelia. Adenoviruses naturally infect respiratory epithelia where they cause a mild disease. Other targets for adenovirus-based delivery systems are liver, the central nervous system, endothelial cells, and muscle. Adenoviruses have the advantage of being capable of infecting non-dividing cells. Kozarsky and Wilson, Current Opinion in Genetics and Development 3:499-503 (1993) present a review of adenovirus-based gene therapy. Bout et al., Human Gene Therapy 5:3-10 (1994) demonstrated the use of adenovirus vectors to transfer genes to the respiratory epithelia of rhesus monkeys. Other instances of the use of adenoviruses in gene therapy can be found in Rosenfeld et al., Science 252:431-434 (1991); Rosenfeld et al., Cell 68:143- 155 (1992); Mastrangeli et al., J.
Clin. Invest.
91:225-234 (1993); PCT Publication W094/12649; and Wang, et al., Gene Therapy 2:775-783 (1995). In a preferred embodiment, adenovirus vectors are used.

[0296] Adeno-associated virus (AAV) has also been proposed for use in gene therapy (Walsh et al., Proc. Soc. Exp. Biol. Med. 204:289-300 (1993); U.S. Patent No.
5,436,146).
[0297] Another approach to gene therapy involves transferring a gene to cells in tissue culture by such methods as electroporation, lipofection, calcium phosphate mediated transfection, or viral infection. Usually, the method of transfer includes the transfer of a selectable marker to the cells. The cells are then placed under selection to isolate those cells that have taken up and are expressing the transferred gene. Those cells are then delivered to a patient.
[0298] In this embodiment, the nucleic acid is introduced into a cell prior to administration in vivo of the resulting recombinant cell. Such introduction can be carried out by any method known in the art, including but not limited to transfection, electroporation, microinjection, infection with a viral or bacteriophage vector containing the nucleic acid sequences, cell fusion, chromosome-mediated gene transfer, microcell-mediated gene transfer, spheroplast fusion, etc. Numerous techniques are known in the art for the introduction of foreign genes into cells (see, e.g., Loeffler and Behr, Meth.
Enzymol. 217:599-618 (1993); Cohen et al., Meth. Enzymol. 217:618-644 (1993);
Cline, Pharmac. Ther. 29:69-92m (1985) and may be used in accordance with the present invention, provided that the necessary developmental and physiological functions of the recipient cells are not disrupted. The technique should provide for the stable transfer of the nucleic acid to the cell, so that the nucleic acid is expressible by the cell and preferably heritable and expressible by its cell progeny.
[0299] The resulting recombinant cells can be delivered to a patient by various methods known in the art. Recombinant blood cells (e.g., hematopoietic stem or progenitor cells) are preferably administered intravenously. The amount of cells envisioned for use depends on the desired effect, patient state, etc., and can be determined by one skilled in the art.
[0300] Cells into which a nucleic acid can be introduced for purposes of gene therapy encompass any desired, available cell type, and include but are not limited to epithelial cells, endothelial cells, keratinocytes, fibroblasts, muscle cells, hepatocytes; blood cells such as T lymphocytes, B lymphocytes, monocytes, macrophages, neutrophils, eosinophils, megakaryocytes, granulocytes; various stem or progenitor cells, in particular DEMANDE OU BREVET VOLUMINEUX
LA PRESENTE PARTIE DE CETTE DEMANDE OU CE BREVET COMPREND
PLUS D'UN TOME.

~~ TTENANT LES PAGES 213 A 340 NOTE : Pour les tomes additionels, veuillez contacter 1e Bureau canadien des brevets JUMBO APPLICATIONS/PATENTS
THIS SECTION OF THE APPLICATION/PATENT CONTAINS MORE THAN ONE
VOLUME

NOTE: For additional volumes, please contact the Canadian Patent Office NOM DU FICHIER / FILE NAME
NOTE POUR LE TOME / VOLUME NOTE:

Claims

What Is Claimed Is:

An isolated nucleic acid molecule comprising a polynucleotide having a nucleotide sequence at least 95% identical to a sequence selected from the group consisting of:
(a) a polynucleotide fragment of SEQ ID NO:X or a polynucleotide fragment of the cDNA sequence contained in Clone ID NO:Z, which is hybridizable to SEQ ID
NO:X;
(b) a polynucleotide encoding a polypeptide fragment of SEQ m NO:Y or a polypeptide fragment encoded by the cDNA sequence contained in cDNA Clone >D
NO:Z, which is hybridizable to SEQ ID NO:X;
(c) a polynucleotide encoding a polypeptide fragment of a polypeptide encoded by SEQ m NO:X or a polypeptide fragment encoded by the cDNA sequence contained in cDNA Clone ID NO:Z, which is hybridizable to SEQ ID NO:X;
(d) a polynucleotide encoding a polypeptide domain of SEQ ID NO:Y or a polypeptide domain encoded by the cDNA sequence contained in cDNA Clone ID
NO:Z, which is hybridizable to SEQ ID NO:X;
(e) a polynucleotide encoding a polypeptide epitope of SEQ ID NO:Y or a polypeptide epitope encoded by the cDNA sequence contained in cDNA Clone ID
NO:Z, which is hybridizable to SEQ ID NO:X;
(f) a polynucleotide encoding a polypeptide of SEQ ID NO:Y or the cDNA
sequence contained in cDNA Clone ID NO:Z, which is hybridizable to SEQ ID NO:X, having biological activity;
(g) a polynucleotide which is a variant of SEQ ID NO:X;
(h) a polynucleotide which is an allelic variant of SEQ ID NO:X;
(i) a polynucleotide which encodes a species homologue of the SEQ ID NO:Y;
(j) a polynucleotide capable of hybridizing under stringent conditions to any one of the polynucleotides specified in (a)-(i), wherein said polynucleotide does not hybridize under stringent conditions to a nucleic acid molecule having a nucleotide sequence of only A
residues or of only T residues.
2. The isolated nucleic acid molecule of claim 1, wherein the polynucleotide fragment comprises a nucleotide sequence encoding a protein.
3. The isolated nucleic acid molecule of claim 1, wherein the polynucleotide fragment comprises a nucleotide sequence encoding the sequence identified as SEQ ID NO:Y
or the polypeptide encoded by the cDNA sequence contained in cDNA Clone ID
NO:Z, which is hybridizable to SEQ ID NO:X.
4. The isolated nucleic acid molecule of claim 1, wherein the polynucleotide fragment comprises the entire nucleotide sequence of SEQ ID NO:X or the cDNA
sequence contained in cDNA Clone ID NO:Z, which is hybridizable to SEQ ID NO:X.
5. The isolated nucleic acid molecule of claim 2, wherein the nucleotide sequence comprises sequential nucleotide deletions from either the C-terminus or the N-terminus.
6. The isolated nucleic acid molecule of claim 3, wherein the nucleotide sequence comprises sequential nucleotide deletions from either the C-terminus or the N-terminus.
7. A recombinant vector comprising the isolated nucleic acid molecule of claim 1.
8. A method of making a recombinant host cell comprising the isolated nucleic acid molecule of claim 1.
9. A recombinant host cell produced by the method of claim 8.
10. The recombinant host cell of claim 9 comprising vector sequences.
11. An isolated polypeptide comprising an amino acid sequence at least 90%
identical to a sequence selected from the group consisting of-.
(a) a polypeptide fragment of SEQ ID NO:Y or the encoded sequence contained in cDNA Clone ID NO:Z;
(b) a polypeptide fragment of SEQ >D NO:Y or the encoded sequence contained in cDNA Clone >D NO:Z, having biological activity;

(c) a polypeptide domain of SEQ ID NO:Y or the encoded sequence contained in cDNA Clone ID NO:Z;
(d) a polypeptide epitope of SEQ ID NO:Y or the encoded sequence contained in cDNA Clone ID NO:Z;
(e) a full length protein of SEQ ID NO:Y or the encoded sequence contained in cDNA
Clone ID NO:Z;
(f) a variant of SEQ ID NO:Y;
(g) an allelic variant of SEQ ID NO:Y; or (h) a species homologue of the SEQ ID NO:Y.
12. The isolated polypeptide of claim 11, wherein the full length protein comprises sequential amino acid deletions from either the C-terminus or the N-terminus.
13. An isolated antibody that binds specifically to the isolated polypeptide of claim 11.
14. A recombinant host cell that expresses the isolated polypeptide of claim 11.
15. A method of making an isolated polypeptide comprising:
(a) culturing the recombinant host cell of claim 14 under conditions such that said polypeptide is expressed; and (b) recovering said polypeptide.
16. The polypeptide produced by claim 15.
17. A method for preventing, treating, or ameliorating a medical condition, comprising administering to a mammalian subject a therapeutically effective amount of the polynucleotide of claim 1.
18. A method of diagnosing a pathological condition or a susceptibility to a pathological condition in a subject comprising:

(a) determining the presence or absence of a mutation in the polynucleotide of claim 1; and (b) diagnosing a pathological condition or a susceptibility to a pathological condition based on the presence or absence of said mutation.
19. A method of diagnosing a pathological condition or a susceptibility to a pathological condition in a subject comprising:
(a) determining the presence or amount of expression of the polypeptide of claim 11 in a biological sample; and (b) diagnosing a pathological condition or a susceptibility to a pathological condition based on the presence or amount of expression of the polypeptide.
20. A method for identifying a binding partner to the polypeptide of claim 11 comprising:
(a) contacting the polypeptide of claim 11 with a binding partner; and (b) determining whether the binding partner effects an activity of the polypeptide.
21. The gene corresponding to the cDNA sequence of SEQ ID NO:Y.
22. A method of identifying an activity in a biological assay, wherein the method comprises:
(a) expressing SEQ ID NO:X in a cell;
(b) isolating the supernatant;
(c) detecting an activity in a biological assay; and identifying the protein in the supernatant having the activity.
23. The product produced by the method of claim 20.
24. A method for preventing, treating, or ameliorating a medical condition, comprising administering to a mammalian subject a therapeutically effective amount of the polypeptide of claim 11.