Mitochondrial Function In Vivo Evaluated by NADH Fluorescence (eBook)

About the Author Avraham Mayevsky, Ph.D. is a Professor Emeritus in theFaculty of Life Sciences and the Brain Research Center at Bar Ilan University, Israel.He has published  more than two hundred papers in the field of mitochondrial function and tissue physiology in vivo under pathophysiological conditions.

Dedication 6
Preface 8
Acknowledgments 10
Contents 12
1: Introduction 16
1.1 The Mitochondrion 16
1.2 Collaboration of Avraham Mayevsky (A.M.) with Britton Chance (B.C.) 18
References 20
2: Tissue Energy Metabolism and Mitochondrial Function 22
2.1 Tissue Energy Metabolism 22
2.2 Evaluation of Mitochondrial Function In Vivo 23
References 27
3: Spectroscopic Monitoring of NADH: Historical Overview 30
3.1 Introduction 30
3.1.1 Historical Introduction: Written by Prof. Britton Chance in 2006 (The next three Paragraphs) 30
3.2 Monitoring of NADH UV Absorbance 31
3.3 Monitoring NADH Fluorescence 33
3.4 Fluorescence Emission Spectra of NADH 35
3.4.1 NADH Spectra in Solution 35
3.4.2 NADH Spectra in Isolated Mitochondria 36
3.4.3 Intact Cells 37
3.4.4 Tissue Slices and Blood-Free Perfused Organs 38
3.4.5 Organs In Vivo 40
3.5 Comparison Between Fluorescence Monitoring and Biochemical Analysis of the Pyridine Nucleotides 41
3.6 Intracellular Origin of NADH Fluorescence Signal 42
References 46
4: Technological Aspects of NADH Monitoring 57
4.1 Introduction 57
4.2 Old Types of NADH Fluorometers 57
4.3 Monitoring NADH Fluorescence and Reflectance 58
4.4 Fiber-Optic Fluorometer–Reflectometer 58
4.4.1 The “MitoViewer” 60
4.5 Factors Affecting NADH Fluorescence and Reflectance Signals 62
4.5.1 Movement Artifacts 63
4.5.2 Intracellular and Extracellular Space Events 63
4.5.3 Vascular Events 65
4.5.3.1 Blood Oxygenation 65
4.5.3.2 Blood Volume Changes 65
4.6 Principles of NADH Artifact Correction 66
4.7 Calibration of NADH in Solution 67
4.7.1 Aims of the Study 68
4.7.2 Basic Experimental Approach 68
4.7.3 Experimental Methodology 69
4.7.4 Choice of Concentrations of NADH Solutions 69
4.7.5 Materials and Methods 69
4.7.6 Results 70
4.7.7 Conclusions 70
4.8 Calibration of the Monitored Signals 71
4.9 Preparation of Animals for Monitoring 71
4.9.1 Surgical Procedures 72
4.9.1.1 Monitoring the Brain 72
4.9.1.2 Monitoring the Spinal Cord 74
4.9.1.3 Monitoring of Heart Muscle In Situ 75
4.9.1.4 Monitoring of Visceral Organs 75
4.9.1.5 Experimental Protocols 75
Testing the Safety of Anticancer Drugs 76
Testing the Efficacy of Anticancer Drugs 77
Acute Model 77
Chronic Model 77
References 77
5: Monitoring of NADH Together with Other Tissue Physiological Parameters 82
5.1 Introduction 82
5.2 Brain Energy Metabolism 82
5.3 Methods 85
5.3.1 NADH Monitoring 85
5.3.2 Microcirculatory Blood Flow 85
5.3.3 Oxygen Electrodes 85
5.3.4 Ion-Selective Electrodes and DC Potential 85
5.3.5 Reference Electrode 86
5.3.6 Electrocorticography (ECoG) 86
5.3.7 Temperature Measurements 86
5.3.8 Data Collection and Analysis 86
5.3.9 Animal Preparation for Monitoring 86
5.4 Results and Discussion 86
5.4.1 Fiber-Optic-Based Fluorometer and EEG 86
5.4.2 Addition of K+ Monitoring 87
5.4.3 NADH and pO2 Measurements 87
5.4.4 The First Multiparametric Monitoring System 89
5.4.5 An Upgraded Multiparametric Monitoring System 91
5.4.6 Addition of Hb Saturation to the MPA 91
Hemoglobin Spectrophotometry 91
5.4.7 A New Model of the MPA 93
5.4.8 Multiparametric Monitoring of Neurosurgical Patients 93
5.4.9 Use of MPA Inside an NMR Magnet 93
5.4.10 Propagation of CSD Wave 95
5.4.11 Addition of ICP Probe to the MPA 96
5.4.12 Use of MPA in Traumatic Brain Injury 97
5.4.13 Use of MPA in Monitoring the Beating Heart 97
5.4.14 Use of MPA in Monitoring the Kidney 98
References 100
6: Multisite Monitoring of NADH 102
6.1 Introduction 102
6.2 Multisite Monitoring of NADH in the Same Organ 102
6.2.1 NADH Monitoring of Two Sites in the Brain 102
6.2.2 NADH Monitoring of Two Sites in the Same Heart 104
6.2.3 NADH Monitoring of Four Sites in the Same Brain 107
6.3 NADH Monitoring of Four Different Organs in the Same Animal 109
6.4 Multisite Monitoring of NADH and DC Potential 112
6.5 Monitoring of NADH and Tissue Blood Flow in More Than One Organ 113
6.6 Multisite Monitoring of NADH, CBF, and DC Potential in the Brain 114
6.7 Two-Dimensional Mapping of NADH Fluorescence 115
References 121
7: Responses of NADH to Physiological and Pathophysiological Conditions 123
7.1 Introduction 123
7.2 Perturbation of Oxygen Supply In Vivo 123
7.2.1 Introduction 123
7.2.2 Anoxia and Hypoxia 124
7.2.3 Ischemia (Decreased Blood Flow) 132
7.2.4 Hyperoxia (Normobaric and Hyperbaric Increase in FiO2) 136
7.2.5 Changes in Inspired CO2 and CO 144
7.2.6 Oscillations of NADH Fluorescence 146
7.3 Responses to Energy Consumption Changes 150
7.3.1 Introduction 150
7.3.2 Direct Cortical Stimulation 150
7.3.3 Brain Activation by Epileptic Activity 151
7.3.4 Responses to Cortical Spreading Depression 154
7.3.5 Activation of the Brain Under Restricted Oxygen Supply 162
7.3.6 Activation of Body Organs 169
7.4 Effects of Pharmacological Agents 170
7.5 Effects of Other Pathophysiological Conditions 181
7.5.1 Hemorrhage 181
7.5.2 Effects of Animal Age 185
7.5.3 Effects of Hypothermia 188
7.5.4 Effects of Elevated ICP and Head Injury 189
7.5.5 Sepsis and Septic Shock 192
7.5.6 Monitoring of NADH During Organ Transplantation 194
References 195
8: Monitoring of Various Organs in Different Animal Models 217
8.1 Introduction 217
8.2 Monitoring the Brain 217
8.2.1 Studies of Large Animal Brains (Pigs, Dogs, and Monkeys) 217
8.2.2 Monitoring the Brain in Cats 220
8.3 Monitoring the Heart 226
8.4 Monitoring of Skeletal Muscle 229
8.5 Liver Monitoring 229
8.6 Monitoring the Kidney 232
8.7 NADH Monitoring in the Gastrointestinal Tract 232
8.8 Monitoring of Other Organs 232
References 236
9: Monitoring of NADH in Human Brain and Body Organs 253
9.1 Introduction 253
9.2 History of NADH Monitoring in Patients 253
9.2.1 Monitoring the Human Brain 253
9.2.2 Monitoring the Heart and Skeletal Muscle 254
9.2.3 Monitoring of Visceral Organs 255
9.2.4 Monitoring of Cancer Cells and Tissues 256
9.3 Monitoring of Patients in Clinical Practice 256
9.3.1 Methods and Results 258
9.3.1.1 The Multiparametric Monitoring System 258
9.3.1.2 The “Tissue Spectroscope” 262
9.3.1.3 The CritiView 264
References 269
10: Discussion and Conclusions 272
10.1 From Isolated Mitochondria to Clinical Monitoring of NADH 272
10.2 From the Single-Parameter to Multiparameter Monitoring Approach 276
10.3 Tissue Vitality Index 277
10.4 Future Perspectives 281
References 281
About the Author 283
Index 284