Development of a Proteomic Assay for Menstrual Blood, Vaginal Fluid and Species Identification Author(S): Donald Siegel, Ph.D

Total Page:16

File Type:pdf, Size:1020Kb

Development of a Proteomic Assay for Menstrual Blood, Vaginal Fluid and Species Identification Author(S): Donald Siegel, Ph.D The author(s) shown below used Federal funding provided by the U.S. Department of Justice to prepare the following resource: Document Title: Development of a Proteomic Assay for Menstrual Blood, Vaginal Fluid and Species Identification Author(s): Donald Siegel, Ph.D. Document Number: 251932 Date Received: August 2018 Award Number: 2010-DN-BX-K192 This resource has not been published by the U.S. Department of Justice. This resource is being made publically available through the Office of Justice Programs’ National Criminal Justice Reference Service. Opinions or points of view expressed are those of the author(s) and do not necessarily reflect the official position or policies of the U.S. Department of Justice. Development of a Proteomic Assay for Menstrual Blood, Vaginal Fluid and Species Identification Final Draft Technical Report NIJ Grant 2010-DN-BX-K192 Principal Investigator: Donald Siegel, Ph.D. Principal Scientist Office of Chief Medical Examiner 421 East 26th Street New York, NY 10016 Tel: 212-323-1434 Fax: 212-323-1560 Email: [email protected] Web: www.nyc.gov/ocme This resource was prepared by the author(s) using Federal funds provided by the U.S. Department of Justice. Opinions or points of view expressed are those of the author(s) and do not necessarily reflect the official position or policies of the U.S. Department of Justice. Final Draft Technical Report NIJ Grant 2010-DN-BX-K192 Development of a Proteomic Assay for Menstrual Blood, Vaginal Fluid and Species Identification TABLE OF CONTENTS ABBREVIATIONS……………………………………………………………………………………………………………………..4 ABSTRACT .................................................................................................................................................................... 6 EXECUTIVE SUMMARY ............................................................................................................................................. 7 SYNOPSIS OF THE PROBLEM ........................................................................................................................................................... 7 PURPOSE ........................................................................................................................................................................................... 8 RESEARCH DESIGN ........................................................................................................................................................................... 8 Menstrual & Venous Blood Collection ........................................................................................................................... 8 Vaginal Fluid Collection ...................................................................................................................................................... 8 Sample Preparation and Processing .............................................................................................................................. 9 FINDINGS ....................................................................................................................................................................................... 12 Menstrual Blood .................................................................................................................................................................. 13 Markers Found in All Women but Also Detected in Other Body Fluids ................................................. 13 Markers Found Only in Menstrual Blood but Not Found in All Women ................................................ 14 Markers Enriched in Menstrual Blood but Detected in Other Body Fluids Not in All Women ..... 14 Vaginal Fluid ........................................................................................................................................................................ 15 Species Identification ........................................................................................................................................................ 16 CONCLUSIONS INCLUDING IMPLICATIONS FOR POLICY AND PRACTICE.................................................................................16 INTRODUCTION ....................................................................................................................................................... 18 STATEMENT OF THE PROBLEM ................................................................................................................................................... 18 Serology Assays ................................................................................................................................................................... 18 Menstrual Blood and Vaginal Fluid Testing ............................................................................................................ 19 LITERATURE REVIEW AND CITATIONS ...................................................................................................................................... 19 Current Serology Methods in Use ................................................................................................................................. 19 Chemical Assays .......................................................................................................................................................... 20 Enzymatic Assays ........................................................................................................................................................ 20 Microscopic Observations ........................................................................................................................................ 20 Immunochemical Assays .......................................................................................................................................... 20 Current Research ................................................................................................................................................................ 21 Messenger RNA Assays ............................................................................................................................................. 21 MicroRNA Assays ........................................................................................................................................................ 21 DNA Methylation Analysis for Forensic Tissue Identification ................................................................... 23 RATIONALE FOR RESEARCH ........................................................................................................................................................ 23 SECTION 1 – MALDI TOF/TOF ANALYSIS OF MENSTRUAL BLOOD ....................................................... 24 BODY FLUID COLLECTION METHODS.......................................................................................................................................... 24 BODY FLUID COLLECTION RESULTS............................................................................................................................................ 25 Length of Periods ................................................................................................................................................................ 25 Volunteer Demographics ................................................................................................................................................. 25 MENSTRUAL BLOOD SAMPLE PREPARATION METHODS ..........................................................................................................26 PROTEIN QUANTITATION RESULTS ............................................................................................................................................ 27 Varying Lengths of Menses ............................................................................................................................................. 27 Five Day Menses Only ........................................................................................................................................................ 31 REDUCTION OF PROTEIN DYNAMIC RANGE ............................................................................................................................... 34 SDS MICROFLUIDIC GEL ELECTROPHORESIS ............................................................................................................................39 PEPTIDE ISOBARIC LABELING FOR MULTIPLEXING AND QUANTIZATION ............................................................................40 How Isobaric Barcoding Works .................................................................................................................................... 41 NANO-HPLC ................................................................................................................................................................................... 42 MASS SPEC ANALYSIS ................................................................................................................................................................... 42 MENSTRUAL BLOOD / VENOUS BLOOD SAMPLE ANALYSIS .....................................................................................................44 2 This resource was prepared by the author(s) using Federal funds provided by the U.S. Department of Justice. Opinions or points of view expressed are those of the author(s) and do not necessarily reflect the official
Recommended publications
  • Quantitation of Relative Mobilities of Serum Transferrin in Quarter Horses and Thoroughbreds Using Crossed Immunoelectrophoresis
    QUANTITATION OF RELATIVE MOBILITIES OF SERUM TRANSFERRIN IN QUARTER HORSES AND THOROUGHBREDS USING CROSSED IMMUNOELECTROPHORESIS BY RUTH GREER KAGEN II Bachelor of Science in Biology East Central Oklahoma State University Ada. Oklahoma 1982 Submitted to the Faculty of the Graduate College of the Oklahoma State University in partial fulfillment of requirements for the degree of MASTERS OF SCIENCE December. 1985 4 ' ;_ '" SERUM TRANSFERRIN IN QUARTER HORSES AND THOROUGHBREDS USING CROSSED IMMUNOELECTROPHORESIS Thesis Approved ii 1236431 ~ ACKNOWLEDGEMENTS The author wishes to express sincere gratitude to Dr. T. E. Staley for serving as the principle advisor and for providing the facilities and financial support necessary to carry out this work. I am most in­ debted to Dr. Staley for his continued faith in my work. his constant support and patient advice. and for his attentive and meticulous instruc­ tion as well as superb organization--all without which this study would not have been possible. I also wish to thank the members of the committee: Dr. C. L. Ownby and Dr. L. G. Stratton. I wish to extend my sincere thanks to Dr. C. L. Ownby for her continued interest in my career and her invaluable advice. both which greatly assisted me in the endeavors to carry out this work. I am very grateful for all the time which Dr. Ownby has afforded me and for her objective and sincere counsel. I am most appreciative of Dr. L. G. Stratton's interest in this project. Through his generous support and concern. we were provided with Pxcellent facilities in which to carry out this work and we were also provided with access to the animals needed for this project.
    [Show full text]
  • Human Placenta Exosomes: Biogenesis, Isolation, Composition, and Prospects for Use in Diagnostics
    International Journal of Molecular Sciences Review Human Placenta Exosomes: Biogenesis, Isolation, Composition, and Prospects for Use in Diagnostics Evgeniya E. Burkova 1,* , Sergey E. Sedykh 1,2 and Georgy A. Nevinsky 1,2 1 SB RAS Institute of Chemical Biology and Fundamental Medicine, 630090 Novosibirsk, Russia; [email protected] (S.E.S.); [email protected] (G.A.N.) 2 Department of Natural Sciences, Novosibirsk State University, 630090 Novosibirsk, Russia * Correspondence: [email protected]; Tel.: +7-(383)-363-51-27 Abstract: Exosomes are 40–100 nm nanovesicles participating in intercellular communication and transferring various bioactive proteins, mRNAs, miRNAs, and lipids. During pregnancy, the placenta releases exosomes into the maternal circulation. Placental exosomes are detected in the maternal blood even in the first trimester of pregnancy and their numbers increase significantly by the end of pregnancy. Exosomes are necessary for the normal functioning of the placenta and fetal development. Effects of exosomes on target cells depend not only on their concentration but also on their intrinsic components. The biochemical composition of the placental exosomes may cause various complications of pregnancy. Some studies relate the changes in the composition of nanovesicles to placental dysfunction. Isolation of placental exosomes from the blood of pregnant women and the study of protein, lipid, and nucleic composition can lead to the development of methods for early diagnosis of pregnancy pathologies. This review describes the biogenesis of exosomes, methods Citation: Burkova, E.E.; Sedykh, S.E.; of their isolation, analyzes their biochemical composition, and considers the prospects for using Nevinsky, G.A. Human Placenta exosomes to diagnose pregnancy pathologies.
    [Show full text]
  • Wilson Disease
    Wilson Disease What is Wilson disease? Wilson disease is a hereditary disease in which excessive amounts of copper accumulate in the body, mainly in the liver. The disease affects approximately one in every 30,000 Canadians. Small amounts of copper are essential to good health. One of the liver’s jobs is to maintain the balance of copper in the body. The liver is also the main organ to store copper. In Wilson disease, when its storage capacity is full, copper is released into the blood stream. It then accumulates in various organs such as the brain and the cornea of the eye. This copper overload damages these organs. Left untreated, Wilson disease can be fatal. What causes the disease? Wilson disease is inherited. In order to have the disease, a person must get two defective genes, one from each parent. The liver begins to retain copper at birth and it may take years before symptoms manifest themselves. Having only one defective gene does not lead to Wilson disease. What are the symptoms of Wilson disease? Wilson disease is sometimes difficult to diagnose. Affected individuals may have no symptoms for years. When symptoms develop, they can be subtle. Sometimes symptoms of Wilson disease resemble hepatitis. Alternatively, some individuals have an enlarged liver and spleen and liver test abnormalities. Copper accumulation in the brain can present itself in two ways: (1) as physical symptoms such as slurred speech, failing voice, drooling, tremors or difficulty in swallowing or (2) as psychiatric disorders such as depression, manic behaviour or suicidal impulses. Very rarely, Wilson disease may cause the liver to fail.
    [Show full text]
  • Characteristic Changes in Decidual Gene Expression Signature in Spontaneous Term Parturition
    Journal of Pathology and Translational Medicine 2017; 51: 264-283 ▒ ORIGINAL ARTICLE ▒ https://doi.org/10.4132/jptm.2016.12.20 Characteristic Changes in Decidual Gene Expression Signature in Spontaneous Term Parturition Haidy El-Azzamy1,* · Andrea Balogh1,2,* Background: The decidua has been implicated in the “terminal pathway” of human term parturi- Roberto Romero1,3,4,5 · Yi Xu1 tion, which is characterized by the activation of pro-inflammatory pathways in gestational tissues. Christopher LaJeunesse1 · Olesya Plazyo1 However, the transcriptomic changes in the decidua leading to terminal pathway activation have Zhonghui Xu1 · Theodore G. Price1 not been systematically explored. This study aimed to compare the decidual expression of devel- Zhong Dong1 · Adi L. Tarca1,6 opmental signaling and inflammation-related genes before and after spontaneous term labor in Zoltan Papp7 · Sonia S. Hassan1,6 order to reveal their involvement in this process. Methods: Chorioamniotic membranes were 1,6 Tinnakorn Chaiworapongsa obtained from normal pregnant women who delivered at term with spontaneous labor (TIL, n = 14) 1,8,9 Chong Jai Kim or without labor (TNL, n = 15). Decidual cells were isolated from snap-frozen chorioamniotic mem- Nardhy Gomez-Lopez1,6 branes with laser microdissection. The expression of 46 genes involved in decidual development, Nandor Gabor Than1,6,7,10,11 sex steroid and prostaglandin signaling, as well as pro- and anti-inflammatory pathways, was ana- lyzed using high-throughput quantitative real-time polymerase chain reaction (qRT-PCR). Chorio- 1Perinatology Research Branch, NICHD/NIH/DHHS, amniotic membrane sections were immunostained and then semi-quantified for five proteins, and Bethesda, MD, and Detroit, MI, USA; 2Department of Immunology, Eotvos Lorand University, Budapest, immunoassays for three chemokines were performed on maternal plasma samples.
    [Show full text]
  • K392-100 Total Iron-Binding Capacity (TIBC) and Serum Iron Assay Kit (Colorimetric)
    FOR RESEARCH USE ONLY! Total Iron-Binding Capacity (TIBC) and Serum Iron Assay Kit (Colorimetric) rev 08/19 (Catalog # K392-100; 100 assays; Store at -20°C) I. Introduction: BioVision’s TIBC and Serum Iron Assay Kit measures both Total iron-binding capacity (TIBC) and Serum iron. Those values indicate the requisite iron for transferrin saturation and Serum Iron respectively. In humans, Transferrin is a blood protein that binds and transports iron throughout the body. Iron bound to transferrin and not bound are reflected in the following: 1) Total Iron Binding Capacity, 2) Unbound Iron, 3) Transferrin Saturation Bound Iron, and 4) Free Iron. Those measurements can be used for to detect and monito transferrin saturation and also iron-deficiency anemia and chronic inflammatory diseases. Part A: TIBC Part B: Serum Iron 1 1 2 2 3 3 4 II. Application: Determination of TIBC, Unbound Iron, Transferrin Saturation, Serum Iron III. Sample Type: Serum or plasma. Serum-off-the clot is preferable to normal serum. IV. Kit Contents: Components K392-100 Cap Code Part Number TIBC Assay Buffer 25 ml WM K392-100-1 Iron Solution 100 µl Blue K392-100-2 TIBC Detector 2 x 1.5 ml Brown K392-100-3 TIBC Developer 5 ml NM K392-100-4 Iron Standard (100 mM) 100 µl Yellow K392-100-5 V. User Supplied Reagents and Equipment: • 96-well plate clear plate with flat bottom • Microplate reader capable of absorbance reading VI. Storage Conditions and Reagent Preparation: Store kit at -20°C, protected from light. Briefly centrifuge small vials prior to opening.
    [Show full text]
  • Understanding Your Blood Test Lab Results
    Understanding Your Blood Test Lab Results A comprehensive "Health Panel" has been designed specifically to screen for general abnormalities in the blood. This panel includes: General Chemistry Screen or (SMAC), Complete Blood Count or (CBC), and Lipid examination. A 12 hour fast from all food and drink (water is allowed) is required to facilitate accurate results for some of the tests in this panel. Below, is a breakdown of all the components and a brief explanation of each test. Abnormal results do not necessarily indicate the presence of disease. However, it is very important that these results are interpreted by your doctor so that he/she can accurately interpret the findings in conjunction with your medical history and order any follow-up testing if needed. The Bernards Township Health Department and the testing laboratory cannot interpret these results for you. You must speak to your doctor! 262 South Finley Avenue Basking Ridge, NJ 07920 www.bernardshealth.org Phone: 908-204-2520 Fax: 908-204-3075 1 Chemistry Screen Components Albumin: A major protein of the blood, albumin plays an important role in maintaining the osmotic pressure spleen or water in the blood vessels. It is made in the liver and is an indicator of liver disease and nutritional status. A/G Ratio: A calculated ratio of the levels of Albumin and Globulin, 2 serum proteins. Low A/G ratios can be associated with certain liver diseases, kidney disease, myeloma and other disorders. ALT: Also know as SGPT, ALT is an enzyme produced by the liver and is useful in detecting liver disorders.
    [Show full text]
  • Ocular Copper Deposition Associated with Monoclonal Gammopathy of Undetermined Significance: Case Report
    RELATOS DE CASOS Ocular copper deposition associated with monoclonal gammopathy of undetermined significance: case report Depósito ocular de cobre associado a gamopatia monoclonal de significância indeterminada: relato de caso Patrick F. Tzelikis1 ABSTRACT Peter R. Laibson2 Marco P. Ribeiro3 To report a case of ocular copper deposition in both eyes at the level of 4 Christopher J. Rapuano Descemet’s membrane associated with a monoclonal gammopathy of 5 Kristin M. Hammersmith undetermined significance (MGUS). A 49-year-old white woman had Elisabeth J. Cohen6 golden-brown metallic dust-like deposits on Descemet’s membrane of both eyes. A systemic examination revealed an elevated serum copper, normal serum ceruloplasmin and a normal level of total protein. Serum protein electrophoresis demonstrated a single peak (M-spike) in the gamma region (M-protein in serum = 11 g/l). Flow cytometric analysis of the marrow aspirate identified a monoclonal plasma cell population that represents approximately 2% of the total marrow cells consistent with the diagnosis of monoclonal gammopathy of undetermined significance. Copper deposits at the level of Descemet’s membrane may be a finding in a patient with monoclonal gammopathy of undetermined significance. Keywords: Copper/analysis; Blood protein electrophoresis; Descemet’s membrane; Gama-globulins/diagnosis; Paraproteinemias/diagnosis INTRODUCTION Bilateral deposition of copper on Descemet’s membrane occurs in a variety of conditions associated with hypercupremia, including multiple (1-3) (4) From the Cornea Service, Wills Eye Hospital, Jefferson myeloma , pulmonary carcinoma , and also benign monoclonal gamma- Medical College of Thomas Jefferson University, nopathies(5-6). We describe the third case in the literature of deposition of Philadelphia, Pennsylvania, U.S.A.
    [Show full text]
  • 904 Genetics of Human Complement Component C4
    [Frontiers in Bioscience 6, d904-913, August 1, 2001] GENETICS OF HUMAN COMPLEMENT COMPONENT C4 AND EVOLUTION THE CENTRAL MHC O. Patricia Martinez,1 Natalie Longman-Jacobsen,1 Richard Davies,1 Erwin K. Chung,2 Yan Yang,2 Silvana Gaudieri,1 Roger L. Dawkins,1 and C. Yung Yu2 1 Centre for Molecular Immunology and Instrumentation, University of Western Australia, PO Box 5100, Canning Vale WA 6155, Australia, 2 Children’s Research Institute, Columbus, Ohio, and Department of Pediatrics, Department of Molecular Virology, Immunology and Medical Genetics, The Ohio State University, 700 Children’s Drive, Columbus Ohio 43205 TABLE OF CONTENTS 1. Abstract 2. The Genetic Diversity and the Nomenclature of Human C4A and C4B 3. Evolution of the MHC-Complement Proteins and the Central MHC 4. Paralogy Mapping Could Help Identify Candidate Genes for MHC – Associated Diseases: Prospects for the Central MHC 5. Acknowledgments 6. References 1. ABSTRACT 2. THE GENETIC DIVERSITY AND THE NOMENCLATURE OF HUMAN C4A AND C4B The two classes of human complement The human complement component C4 is one of component C4 proteins C4A and C4B manifest differential the most complex and polymorphic molecules. The chemical reactivities and binding affinities towards target activated product of C4, C4b, is a non-catalytic subunit C3 surfaces and complement receptor CR1. There are multiple, and C5 convertase in the classical and lectin pathways polymorphic allotypes of C4A and C4B proteins. A (reviewed in refs. 1, 2). Downstream of C4, the complex multiplication pattern of C4A and C4B genes with complement pathway includes the generation of variations in gene size, gene dosage and flanking genes anaphylatoxins C3a and C5a, and the assembly of the exists in the population.
    [Show full text]
  • The Structure and Function of Large Biological Molecules 5
    The Structure and Function of Large Biological Molecules 5 Figure 5.1 Why is the structure of a protein important for its function? KEY CONCEPTS The Molecules of Life Given the rich complexity of life on Earth, it might surprise you that the most 5.1 Macromolecules are polymers, built from monomers important large molecules found in all living things—from bacteria to elephants— can be sorted into just four main classes: carbohydrates, lipids, proteins, and nucleic 5.2 Carbohydrates serve as fuel acids. On the molecular scale, members of three of these classes—carbohydrates, and building material proteins, and nucleic acids—are huge and are therefore called macromolecules. 5.3 Lipids are a diverse group of For example, a protein may consist of thousands of atoms that form a molecular hydrophobic molecules colossus with a mass well over 100,000 daltons. Considering the size and complexity 5.4 Proteins include a diversity of of macromolecules, it is noteworthy that biochemists have determined the detailed structures, resulting in a wide structure of so many of them. The image in Figure 5.1 is a molecular model of a range of functions protein called alcohol dehydrogenase, which breaks down alcohol in the body. 5.5 Nucleic acids store, transmit, The architecture of a large biological molecule plays an essential role in its and help express hereditary function. Like water and simple organic molecules, large biological molecules information exhibit unique emergent properties arising from the orderly arrangement of their 5.6 Genomics and proteomics have atoms. In this chapter, we’ll first consider how macromolecules are built.
    [Show full text]
  • (12) United States Patent (10) Patent No.: US 6,342,350 B1 Tanzi Et Al
    USOO6342350B1 (12) United States Patent (10) Patent No.: US 6,342,350 B1 Tanzi et al. (45) Date of Patent: Jan. 29, 2002 (54) ALPHA-2-MACROGLOBULIN DIAGNOSTIC with Chinese late onset Alzheimer's Disease,” Neuroscience TEST Letters 269:173–177 (1999). Dodel, R.C., et al., “C-2 Macroglobulin and the Risk of (75) Inventors: Rudolph E. Tanzi, Hull; Bradley T. Alzheimer's Disease,” Neurology 54:438-442 (2000). Hyman, Swampscott; George W. Dow, D.J., et al., “C-2 Macroglobulin Polymorphism and Rebeck, Somerville; Deborah L. Alzheimer Disease risk in the UK,” Nature Genetics Blacker, Newton, all of MA (US) 22:16–17 (May 1999). (73) Assignee: The General Hospital Corporation, Du, Y., et al., “c-Macroglobulin Attenuates B-Amyloid Boston, MA (US) Peptide 1-40 Fibril Formation and Associated Neurotoxicity of Cultured Fetal Rat Cortical Neurons,” J. of Neurochem. (*) Notice: Subject to any disclaimer, the term of this 70: 1182–1188 (1998). patent is extended or adjusted under 35 Gauderman, W.J., et al., “Family-Based Association Stud U.S.C. 154(b) by 0 days. ies.” Monogr: Natl. Canc. Inst. 26:31-37 (1999). Hampe, J., et al., “Genes for Polygenic Disorders: Consid (21) Appl. No.: 09/148,503 erations for Study Design in the Complex Trait of Inflam (22) Filed: Sep. 4, 1998 matory Bowel Disease,” Hum. Hered 50:91-101 (Mar.-Apr. 2000). Related U.S. Application Data Horvath, S., and Laird, N.M., “A Discordant-Sibship Test (60) Provisional application No. 60/093,297, filed on Jul. 17, for Disequilibrium and Linkage: No Need for Parental 1998, and provisional application No.
    [Show full text]
  • 2021 Undergraduate Research Abstract Booklet
    1 | P a g e Table of Contents FOREWORD ................................................................................................................................................... 4 Abidemi Awojuyigbe ..................................................................................................................................... 5 Aijalon Shantavia........................................................................................................................................... 6 Aminata Diagne ............................................................................................................................................. 8 The Exploration of BRAF Gene .................................................................................................................... 10 Araceli Estrada Martinez ............................................................................................................................. 10 Ashlee Young ............................................................................................................................................... 11 Ayanna D. Montegut ................................................................................................................................... 12 Brandon Bernäl ........................................................................................................................................... 13 Caleb Riggins ..............................................................................................................................................
    [Show full text]
  • Macrophage Tumoricidal Activity Induced by Human C-Reactive Protein1
    (CANCER RESEARCH 46, 5077-5083, October 1986] Macrophage Tumoricidal Activity Induced By Human C-Reactive Protein1 Kamyar Zahedi and Richard F. Mortensen Department of Microbiology, The Ohio State University, Columbus, Ohio 43210 ABSTRACT second binding site for certain polycations (15). The binding and structural properties of CRP have been conserved through Purified C-reactive protein (CRP), the prototypical acute phase reac- out vertebrate evolution, although it is not an APR in some tant of humans, activated inflammatory mouse macrophages to a tumor- species (7). CRP is opsonic and facilitates phagocytosis by both icidal state. The activation by CRP was not due to small amounts of macrophages (16, 17) and neutrophils (18); it is also a potent contaminating lipopolysaccharide. CRP at 10 fig/ml induced significant tumoricidal capacity in resident macrophages; the mouse macrophage activator of the classical complement cascade (19, 20) but cell lines PUS 1.8, RAW 264.7, and J774; as well as elicited macrophages restricts alternative pathway activation (21). CRP also binds to from two lipopolysaccharide nonresponder strains, C3H/HeJ and a subset of the Fc receptor bearing lymphocyte populations and C57BL/10SC. Macrophages obtained from bone marrow-derived mono- affects several in vitro T-cell and B-cell lymphocyte activities cytes grown in vitro and exúdate macrophages depleted of T-cells were (22, 23). Although these activities are associated with host also readily activated by Mg/ml amounts of CRP. Removal of CRP from defense, a clearly defined role for CRP in preimmune host culture medium using anti-CRP antibodies or phosphorylcholine-agarose resistance remains unknown.
    [Show full text]