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ADVANCES IN CLINICAL CHEMISTRY
VOLUME 49
This page intentionally left blank
Advances in
CLINICAL
CHEMISTRY
Edited by

GREGORY S. MAKOWSKI
Department of Laboratory Medicine
University of Connecticut Health Center
Farmington, CT, USA

VOLUME 49

AMSTERDAM • BOSTON • HEIDELBERG • LONDON

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09 10 11 12 10 9 8 7 6 5 4 3 2 1
 CONTENTS

CONTRIBUTORS ................................................................................ ix

PREFACE . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xi

High-Sensitivity Troponin: A New Tool for Pathophysiological

Investigation and Clinical Practice
ALDO CLERICO, ALBERTO GIANNONI, CONCETTA PRONTERA,
AND STEFANIA GIOVANNINI

1. Abstract ... ................................................................................... 2

2. Background and Aim....................................................................... 2
3. Introduction: Troponin Framework Within Myocardial Cells and
Release Kinetics After Myocardial Damage ............................................. 4
4. Impact of the New Definition of Myocardial Infarction on Laboratory
Practice and Instrumentation: The Need for High-Sensitivity cTnI
and cTnT Methods ......................................................................... 5
5. The Impact of High-Sensitivity cTnI and cTnT Methods on Clinical Practice ...... 9
6. High-Sensitivity cTnI and cTnT Methods: A Powerful Tool for Monitoring
Physiological Renewal and Pathological Remodeling of the Myocardial Tissue? ... 19
7. Use of High-Sensitivity cTnI and cTnT Methods in a Multimarker
Approach for Early Screening: An Increase in Diagnostic and
Prognostic Efficiency? ...................................................................... 22
8. Conclusion ................................................................................... 23
References. ................................................................................... 24

Biochemical Biomarkers of Oxidative Collagen Damage

Y. HENROTIN, M. DEBERG, M. MATHY-HARTERT, AND G. DEBY-DUPONT

1. Abstract ... ................................................................................... 31

2. The Collagen Family: Structure and Biochemistry...................................... 32
3. Chemical Nature and Reactivity of Oxygen ............................................. 33
4. In Vivo Production of RNOS .............................................................. 35
5. RNOS Biological Activity.................................................................. 37
6. In Vivo Markers of Oxidant Stress .. ...................................................... 39
7. Oxidant-Induced Changes in Collagens .................................................. 40
8. Biomarkers of Collagen Oxidative Damage ............................................. 43
9. Critical Comments and Concluding Remarks ... ........................................ 48
References. ................................................................................... 49

v
vi CONTENTS

Biochemical Basis of Fabry Disease with Emphasis on

Mitochondrial Function and Protein Trafficking
A.M. DAS AND H.Y. NAIM

1. Abstract....................................................................................... 57
2. Introduction.................................................................................. 58
3. Inheritance ................................................................................... 58
4. Clinical Picture............................................................................... 59
5. Diagnosis ..................................................................................... 59
6. Treatment .................................................................................... 60
7. Biochemical Basis............................................................................ 62
References .................................................................................... 68

Urinary Biomarkers for the Detection of Renal Injury

MITCHELL H. ROSNER

1. Abstract....................................................................................... 73
2. Introduction.................................................................................. 74
3. Biomarker Development for Kidney Diseases ........................................... 75
4. Biomarkers for AKI......................................................................... 81
5. Biomarker Development and Implementation ........................................... 90
6. Summary ..................................................................................... 91
References .................................................................................... 92

Biomarkers of Bone and Mineral Metabolism Following

Bone Marrow Transplantation
KI HYUN BAEK AND MOO IL KANG

1. Abstract....................................................................................... 99
2. Introduction.................................................................................. 100
3. Clinical Features of BMT-Related Bone Loss ........................................... 100
4. Changes in Bone-Turnover Markers After BMT ........................................ 102
5. Calcium, Parathyroid Hormone (PTH), and Vitamin D.. .............................. 109
6. Sex Hormones ............................................................................... 110
7. RANKL and Osteoprotegerin ............................................................. 112
8. Cytokines and Growth Factors ............................................................ 114
9. Conclusion ................................................................................... 115
References .................................................................................... 116

Factor V Leiden and Activated Protein C Resistance

OLIVIER SEGERS AND ELISABETTA CASTOLDI

1. Abstract....................................................................................... 121
2. Introduction.................................................................................. 122
 CONTENTS vii

3. The Protein C System ...................................................................... 123

4. APC Resistance and FV Leiden........................................................... 130
5. Conclusions and Perspectives.............................................................. 143
References. ................................................................................... 144

Self-Assembled Tethered Bimolecular Lipid Membranes

EVA-KATHRIN SINNER, SANDRA RITZ, RENATE NAUMANN,
STEFAN SCHILLER, AND WOLFGANG KNOLL

1. Abstract ... ................................................................................... 159

2. Introduction ................................................................................. 160
3. Assembly of tBLMs from Telechelics and Reconstitution of Proteins................ 162
4. The Peptide-Tethered Lipid Bilayer Membrane (peptBLM) ........................... 165
5. Protein-Tethered Bilayer Lipid Membrane (protBLM) ................................ 170
6. Conclusions .................................................................................. 174
References. ................................................................................... 177

INDEX . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 181
COLOR PLATE SECTION AT THE END OF THE BOOK
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 CONTRIBUTORS

Numbers in parentheses indicate the pages on which the authors’ contributions begin.

KI HYUN BAEK (99), Department of Internal Medicine, The Catholic University

of Korea, College of Medicine, 137-701 Seoul, Korea

ELISABETTA CASTOLDI (121), Department of Biochemistry, Maastricht

University, 6200 MD Maastricht, The Netherlands

ALDO CLERICO (1), Scuola Superiore Sant’Anna, 56126 Pisa, Italy; and
Laboratory of Cardiovascular Endocrinology and Cell Biology, G. Monasterio
Foundation, CNR Regione Toscana, 56126 Pisa, Italy

A.M. DAS (57), Department of Pediatrics, Hannover Medical School,

Hannover, Germany

M. DEBERG (31), Bone and Cartilage Research Unit, Institute of Pathology,

University of Liège, CHU-Sart-Tilman, 4000 Liège, Belgium

G. DEBY-DUPONT (31), Center for Oxygen Research and Development,

Institute of Chemistry, University of Liège, 4000 Liège, Belgium

ALBERTO GIANNONI (1), Scuola Superiore Sant’Anna, 56126 Pisa, Italy; and
Laboratory of Cardiovascular Endocrinology and Cell Biology, G. Monasterio
Foundation, CNR Regione Toscana, 56126 Pisa, Italy

STEFANIA GIOVANNINI (1), Laboratory of Cardiovascular Endocrinology

and Cell Biology, G. Monasterio Foundation, CNR Regione Toscana, 56126
Pisa, Italy

Y. HENROTIN (31), Center for Oxygen Research and Development, Institute of

Chemistry, University of Liège, 4000 Liège, Belgium; and Bone and Cartilage
Research Unit, Institute of Pathology, University of Liège, CHU-Sart-Tilman,
4000 Liège, Belgium

ix
x CONTRIBUTORS

MOO IL KANG (99), Department of Internal Medicine, The Catholic University

of Korea, College of Medicine, 137-701 Seoul, Korea

WOLFGANG KNOLL (159), Austrian Institute of Technology, Vienna, Austria;

and Institute of Materials Research and Engineering, Singapore

M. MATHY-HARTERT (31), Bone and Cartilage Research Unit, Institute of

Pathology, University of Liège, CHU-Sart-Tilman, 4000 Liège, Belgium

H.Y. NAIM (57), Department of Physiological Chemistry, University of

Veterinary Medicine, Hannover, Germany

RENATE NAUMANN (159), Austrian Institute of Technology, Vienna, Austria;

and Max Planck Institute for Polymer Research, Mainz, Germany

CONCETTA PRONTERA (1), Laboratory of Cardiovascular Endocrinology

and Cell Biology, G. Monasterio Foundation, CNR Regione Toscana, 56126
Pisa, Italy

SANDRA RITZ (159), Max Planck Institute for Polymer Research, Mainz,
Germany

MITCHELL H. ROSNER (73), Division of Nephrology, University of Virginia

Health System, Box 800133, Charlottesville, Virginia 22908

STEFAN SCHILLER (159), Freiburg Institute for Advanced Studies, Freiburg,

Germany

OLIVIER SEGERS (121), Department of Biochemistry, Maastricht University,

6200 MD Maastricht, The Netherlands

EVA-KATHRIN SINNER (159), Institute of Materials Research and Engineering,

Singapore; and Max Planck Institute for Polymer Research, Mainz, Germany
 PREFACE

I am pleased to present volume forty-nine of Advances in Clinical

Chemistry series.
In this third volume for 2009, the lead chapter investigates the analytical
challenges of developing highly sensitive cardiac troponin immunoassays and
their role in advancing clinical diagnosis. Pathophysiologic mechanisms
of troponin release are discussed as well as their potential impact on monitor-
ing the heart in a number of chronic disease processes. The next chapter
explores eVect of oxidative stress on collagen, a key structural and functional
protein within the connective tissue family. The biochemistry of Fabry disease
is discussed in the following chapter with particular emphasis on the impact
of compromised energy metabolism and abnormal lipid composition.
The role of urinary markers in renal disease is presented next. This chapter
is noteworthy given the aging population worldwide and the significant cost
of chronic kidney disease in healthcare. Bone cell loss and osteoporosis follow-
ing bone marrow transplantation is a serious endocrine disorder that is eluci-
dated in the next chapter. The state of the art and new developments in factor V
Leiden and activated protein C resistance testing are also explored with
emphasis on the fundamental biochemistry of this coagulopathic condition.
Volume forty-nine is concluded with a revealing manuscript on the biophysical
and interfacial studies of lipid bilayers and the importance of lipid–protein
interactions within this unique system.
I extend my appreciation to each contributor of volume forty-nine and
thank colleagues who participated in the peer review process. I would also
like to thank my Elsevier editorial liaison, Gayathri Venkatasamy.
I hope the third and final volume of 2009 will be enjoyed. I warmly invite
comments and suggestions for future review articles for the Advances in
Clinical Chemistry readership.
In keeping with the tradition of the series, I would like to dedicate volume
forty-nine to my grandmother Stephanie and my grandfather Stephen.

GREGORY S. MAKOWSKI

xi
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 ADVANCES IN CLINICAL CHEMISTRY, VOL. 49

HIGH-SENSITIVITY TROPONIN: A NEW TOOL

FOR PATHOPHYSIOLOGICAL INVESTIGATION
AND CLINICAL PRACTICE

Aldo Clerico,*,†,1 Alberto Giannoni,*,†

Concetta Prontera,* and Stefania Giovannini*

*Laboratory of Cardiovascular Endocrinology and

Cell Biology, G. Monasterio Foundation, CNR Regione
Toscana, 56126 Pisa, Italy
†
Scuola Superiore Sant’Anna, 56126 Pisa, Italy

1. Abstract . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
2. Background and Aim. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
3. Introduction: Troponin Framework Within Myocardial Cells and Release
Kinetics After Myocardial Damage. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
4. Impact of the New Definition of Myocardial Infarction on Laboratory
Practice and Instrumentation: The Need for High-Sensitivity
cTnI and cTnT Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
4.1. Quality Specifications for cTnI and cTnT Immunoassays. . . . . . . . . . . . . . . . . . . 5
4.2. The Development of High-Sensitivity Immunoassays
for cTnI and cTnT Measurement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
4.3. Definition of Highly (Ultra) Sensitive Immunoassay for cTnI and cTnT . . . . 8
5. The Impact of High-Sensitivity cTnI and cTnT Methods on Clinical Practice . . . . 9
5.1. The Problem of Reliable Definition and Accurate Estimation of the 99th
Percentile Upper Reference Limit: Can Reference Values Be AVected by any
Characteristics of the Reference Population? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
5.2. More Myocardial Infarctions or More False Positive Results?. . . . . . . . . . . . . . 14
5.3. Clinical Relevance of Serially Measured Troponin Circulating Levels. . . . . . . 15
5.4. High-Sensitivity Troponin Methods in Patients with HF:
A Better Stratification of Cardiovascular Risk . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
5.5. Early Detection of Myocardial Injury in Patients with Extracardiac Diseases
or Assuming Potentially Cardiotoxic Drugs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
6. High-Sensitivity cTnI and cTnT Methods: A Powerful
Tool for Monitoring Physiological Renewal and Pathological Remodeling
of the Myocardial Tissue? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19

1
Corresponding author: Aldo Clerico, e-mail: [email protected]

0065-2423/09 $35.00 Copyright 2009, Elsevier Inc.

DOI: 10.1016/S0065-2423(09)49001-2 All rights reserved.
2 CLERICO ET AL.

7. Use of High-Sensitivity cTnI and cTnT Methods in a Multimarker Approach for

Early Screening: An Increase in Diagnostic and Prognostic EViciency?. . . . . . . . . . . 22
8. Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24

1. Abstract

At the dawn of the new century, the advent of more specific myocardial
tissue markers, such as cardiac troponin I (cTnI) and T (cTnT), has led to a
new definition of acute myocardial infarction (AMI) by international guide-
lines. If we accept the concept that AMI is the portion of acutely necrotic
myocardial tissue (irrespective of size), some patients previously diagnosed
with severe angina may be currently considered to present minimal (even
microscopic) quantities of myocardial necrosis. Although increased cTnI or
cTnT values always indicate myocardial tissue damage, a positive test is not
able to identify the mechanism responsible for that cardiac damage (which
could be not due to ischemia). New cTnI and cTnT immunoassays with
increased analytical sensitivity may increase ‘‘false positive’’ results in
patients with cardiovascular disease, especially those with advanced age,
heart failure (HF), severe comorbidities (such as chronic renal insuViciency),
or assuming potential cardiotoxic drugs. Hence, it may be not clear for most
patients and physicians whether high-sensitivity cTnI and cTnT methods will
lead to more clarity or confusion. The aim of this review is to update the
present knowledge in the field of cTnI and cTnT with particular attention on
the impact of immunoassays with increased analytical sensitivity on both
laboratory and clinical practice.

2. Background and Aim

About 25 years ago, few diagnostic tests were available for clinical practice
and considered useful in the assessment of cardiac necrosis, such as those
measuring the total enzymatic activity of creatine kinase (CK) and lactate
dehydrogenase. Unfortunately, those methods were characterized by low
sensitivity and specificity for cardiac damage. Some immunoassay methods
for structural proteins and cardiac isoenzymes, such as CK-MB isoenzyme
and myoglobin, were then developed (Fig. 1). These markers showed an in-
creased sensitivity, but only a relative specificity for cardiac disease, because
these proteins are also present in the skeletal tissue.
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