Chromosomal Alterations (eBook)

Both editors have a year long experience in cytogenetic research and published many papers in peer reviewed journals on different topics of cytogenetics.

Preface 5
Contents 7
List of Contributors 19
1 Human Chromosomes: Structural and Functional Aspects 25
Abstract 25
1.1 History of Chromosome Research 26
1.2 Composition and Compartmentalization of Human Chromosomes 28
1.3 The Human Karyotype and Clinical Cytogenetics 29
1.4 Cell Cycle and Chromosome Cycle 33
1.5 Shaping the Metaphase Chromosome 34
1.6 Cohesion and Condensins 35
1.7 DNA Repair 37
1.8 Mendelian Disorders Affecting Chromosome Integrity 38
References 41
2 DNA Lesions Induced by Ionizing Radiation 45
Abstract 45
2.1 Introduction 45
2.2 Energy Deposition by Ionizing Radiation 47
2.3 Direct and Indirect Effect 48
2.4 Cerenkov Radiation 49
2.5 Hole and Electron Transfer Through DNA 49
2.6 Formation of Base Radicals 49
2.7 Base Damage 51
2.8 Single-Strand Breaks 54
2.9 Double Lesions 55
2.10 DNA–DNA Cross-Links 58
2.11 DNA–Protein Cross-Links 58
2.12 Clustered Lesions Beyond Double Lesions 58
2.13 Oxygen Effect and Chemical Repair 58
2.14 DNA Damage by UV-C and Ionizing Radiation – a Comparison 59
References 60
3 Effects of Ultraviolet Radiation on DNA 63
Abstract 63
3.1 Introduction 63
3.2 Primary Photoproducts in DNA 66
3.3 Action Spectra 68
3.4 Reversal and Repair of Primary Photoproducts 70
3.5 Cytological Effects 71
3.6 Conclusions 75
References 75
4 Double Strand Break Repair Mechanisms in Mammalian Cells 78
Abstract 78
4.1 Introduction 78
4.2 NHEJ Reaction 79
4.3 DSB Repair by HRR 82
4.4 Impaired DSB Repair and Chromosomal Aberrations 84
4.5 Fragile Sites Are Associated with DNA DSBs 85
4.6 Conclusions 86
References 86
5 Backup Pathways of Nonhomologous End Joining May Have a Dominant Role in the Formation of Chromosome Aberrations 89
Abstract 89
5.1 Introduction 89
5.2 Pathways of DSB Repair 91
5.3 Homology-Independent End Joining and Chromosome Aberration Formation 99
5.4 DSBs, NHEJ and Theories of Chromosome Aberration Formation 100
References 105
6 Targeted and Non-Targeted Induction of Chromosomal Rearrangements After Exposure to Ionizing Radiation 108
Abstract 108
6.1 Introduction 108
6.2 Radiation-Induced Chromosomal Instability 109
6.3 The Death-Inducing Effect 111
6.4 Chromosomal Instability Induced by DIE Medium from Unstable Clones 114
6.5 A Role for Non-Targeted Bystander Effects in Radiation- Induced Chromosomal Instability 114
6.6 Implications of Non-Targeted Effects for Cytogenetic Biodosimetry 115
6.7 Conclusions 116
References 116
7 DNA Methylation Damage: Formation, Repair and Biological Consequences 120
Abstract 120
7.1 Methylating Agents as Environmental Carcinogens 120
7.2 Anticancer Drugs 121
7.3 DNA Lesions, Sites of Attack 122
7.4 Repair of DNA Methylation Damage 125
7.5 Mechanism of Mutagenicity and Carcinogenicity 129
7.6 Mechanism of Cytotoxicity of Methylating Agents by Apoptosis 130
7.7 Mechanism of Formation of SCEs and Aberrations 132
References 136
8 Adducts, Sister-Chromatid Exchanges and Mutations Following Benzo[ a] pyrene Exposure: a Review of Quantitative Data Followed by Some Considerations Regarding Risk 143
Abstract 143
8.1 Introduction 143
8.2 Methodological Approach 144
8.3 Results 145
8.4 Discussion 152
References 158
9 Analysis of DNA Double-Strand Breaks by Means of .-H2AX Foci 164
Abstract 164
9.1 Introduction 164
9.2 Histone H2AX 165
9.3 Repair and Checkpoint Factors Are Recruited into y-H2AX Foci 168
9.4 Early DSB Rejoining Can Proceed Without Foci Formation 169
9.5 Foci May Be Present Long After DSB Are Rejoined 169
9.6 Practical Aspects of .-H2AX Estimation 170
9.7 Concluding Remarks 176
References 177
10 Comet Assay 180
Abstract 180
10.1 Introduction 180
10.2 Method 181
10.3 Factors That Might Affect the Outcome of Comet Analyses 185
10.4 Advantages/Disadvantages 187
10.5 Applications 188
10.6 Final Remark 191
References 191
11 Origin of Chromosome Aberrations: Mechanisms 196
Abstract 196
11.1 Introduction 196
11.2 Primary DNA Damage 198
11.3 Models of Chromosome Aberration Formation 200
11.4 Models of Chromatid Aberration Formation 206
11.5 Conclusions 213
References 213
12 Chromatin Structure and the Formation of Chromosomal Alterations 219
Abstract 219
12.1 Introduction 219
12.2 Modelling of Virtual Radiation Events 220
12.3 Modelling Results 221
12.4 Conclusions 225
References 226
13 Role of Chromatin Structure and Activity in the Induction of Chromosomal Aberrations 228
Abstract 228
13.1 Introduction 228
13.2 Methodological Aspects 230
13.3 Distribution of Chromosome Breakpoints in CHO Cells 232
13.4 Chromosome Damage in Human Cell Lines with Multiple X Chromosomes 236
13.5 Conclusion 238
References 238
14 A Biomedical Perspective of Telomere Structure and Function 242
Abstract 242
14.1 Introduction 242
14.2 The Complex Molecular Structure of the Terminal Chromosome Region 243
14.3 Cryptic Subtelomeric Aberrations: Their Importance in Human Health 245
14.4 Light Microscope Methods for the Study of Telomere Structure 246
14.5 Microdensitometric Analysis of T-banded Human and CHO Chromosomes 248
14.6 Final Remarks 251
References 252
15 Cytokinesis-Block Micronucleus Assay: a Comprehensive “ Cytome” Approach for Measuring Chromosomal Instability, Mitotic Dysfunction and Cell Death Simultaneously in One Assay 258
Abstract 258
15.1 Introduction 258
15.2 Nucleoplasmic Bridges 261
15.3 Nuclear Buds 263
15.4 Micronuclei, NPBs and Nuclear Buds Induced by Folic Acid Deficiency 265
15.5 Breakage–Fusion–Bridge Cycles Explain Micronuclei, NPBs and Nuclear Buds Induced by Folic Acid Deficiency 266
15.6 Micronucleus Formation Caused by Hypomethylation of Heterochromatin and Silencing of Cell Cycle Checkpoint Genes 268
15.7 Conclusion 269
References 270
16 In Vivo Rodent Micronucleus Assay 273
Abstract 273
16.1 Introduction 273
16.2 Historical Overview 275
16.3 The Rodent Micronucleus Assay for Evaluation of Chromosomal Aberration Induction of Agents 276
16.4 The Micronucleus Assay Using Tissues Other Than Bone Marrow 279
16.5 Automation of the Micronucleus Assay 281
16.6 Multiple End Point Assays 282
References 283
17 Sister-Chromatid Exchanges 287
Abstract 287
17.1 Introduction 287
17.2 The Influence of BrdU on the Spontaneous SCE Level 289
17.3 BrdU As a Source of DNA Damage Leading to SCE Formation 291
17.4 The Influence of BrdU on the SCE Level Induced by Mutagenic Compounds 292
17.5 The Problem of “False” and “True” SCE 293
17.6 The Problem of Scoring SCEs in the Second Posttreatment Mitoses 295
17.7 Conclusions 296
References 296
18 Fluorescence in Situ Hybridisation 300
Abstract 300
18.1 Introduction 300
18.2 The Different Classes of DNA Probes 301
18.3 Chromosome Painting Probes 302
18.4 FISH Probes Composed of Repetitive DNA Sequences 305
18.5 Locus-Specific Probes 306
18.6 Comparative Genomic Hybridisation and Array Comparative Genomic Hybridisation 307
18.7 Probe Labelling 308
18.8 The Hybridisation Procedure 309
18.9 Applications 310
18.10 Conclusion 312
References 312
19 The CAB and S& S Systems, and an Approach Towards the Classification of Complex Chromosome Exchanges
Abstract 315
19.1 Introduction 315
19.2 Configuration Versus Pattern 316
19.3 The CAB System 318
19.4 The S& S System
19.5 mFISH Painting 320
19.6 The S& S Descriptor
19.7 Conclusions 327
References 327
20 Aberration Patterns and Cell Cycle Progression Following Exposure of Lymphocytes to the Alkylating Agent Trenimon 329
Abstract 329
20.1 Introduction 329
20.2 Methods 330
20.3 Results 333
References 337
21 Cytogenetic Analysis and Occupational Health 339
Abstract 339
21.1 Introduction 339
21.2 History 340
21.3 Standard Method 341
21.4 Chromosomal Aberrations and Maximum Allowable Concentrations 341
21.5 Cytogenetic Analysis and Public Health Practice 342
21.6 Examples of Occupational Exposures to Carcinogens 345
21.7 Conclusions 350
References 351
22 Biological Dosimetry 355
Abstract 355
22.1 Introduction 355
22.2 Blood Sampling and Culture Conditions 356
22.3 Background Frequency of Asymmetrical and Symmetrical Chromosome Exchanges 357
22.4 Dose–Response Relationships 359
22.5 In Vitro and in Vivo Radiation-Induced Aberration Frequencies 361
22.6 Dicentric Chromosomes or Symmetrical Translocations for Dose Estimation 361
22.7 Summary 362
References 363
23 Statistical Methods for Biological Dosimetry 365
Abstract 365
23.1 Introduction 365
23.2 Statistical Approach 366
23.3 Examples of Dose Estimation 376
23.4 Summary 382
References 383
24 Retrospective Biological Dosimetry by FISH 385
Abstract 385
24.1 Introduction 385
24.2 Method 386
24.3 Which Chromosomes To Paint? 386
24.4 What To Score 387
24.5 Control Levels 388
24.6 Persistence of Translocations 389
24.7 Calibration 390
24.8 Sensitivity 391
24.9 Conclusion 392
References 392
25 Chromosomal Aberrations in Astronauts 395
Abstract 395
25.1 Introduction 395
25.2 Measured Yields of Exchanges in the Peripheral Blood Lymphocytes of Astronauts 397
25.3 Biologically Based Risk Estimates 399
25.4 Cytogenetic Signatures of Space Radiation Exposure 401
25.5 Persistence of Space Radiation Induced Cytogenetic Damage 402
25.6 Methodology Considerations and Suggestions for Standardized Protocols 405
25.7 Conclusions 409
References 409
26 Internet Information System and Literature Database on Biomedical Effects of Electromagnetic Fields: Cancer and Cytogenetic 411
Abstract 411
26.1 Introduction 411
26.2 Classification of Publications 412
26.3 The EMF-Portal: Internet Information System on the Effects of Electromagnetic Fields 415
26.4 Discussion 422
References 423
27 Interpreting Studies on Magnetic Fields and the Risk of Childhood Leukaemia: an Ongoing Challenge 424
Abstract 424
27.1 Introduction 424
27.2 Leukaemia in Children 425
27.3 Exposure to Magnetic Fields and the Risk of Childhood Leukaemia 431
27.4 Conclusions 435
References 437
28 Cytogenetic and Carcinogenic Effects of Exposure to Radiofrequency Radiation 440
Abstract 440
28.1 Introduction 440
28.2 Challenges When Conducting RFR Research 442
28.3 Cytogenetic Effects of RFR 444
28.4 DNA Strand Breaks 458
28.5 Epigenetic Effects of RFR 465
28.6 Cancer Studies in Animals 467
28.7 Summary 477
References 478
29 Radiofrequency Radiation and Replication Studies 483
Abstract 483
29.1 Introduction 483
29.2 DNA Strand Breaks 484
29.3 Micronucleus Test 485
29.4 Combination Studies – RFR and Other Mutagens 486
29.5 Discussion 488
References 489
30 Chromosome Analysis in Cancer Patients: Applications and Limitations 491
Abstract 491
30.1 Introduction 491
30.2 Methods 492
30.3 Hematologic Cancers 493
30.4 Solid Tumors 499
30.5 Molecular Cytogenetics 500
30.6 Future for Cytogenetics in Cancer Investigations 504
References 504
31 Chromosomal Aberration in Peripheral Blood Lymphocytes of Healthy Subjects and Risk of Cancer 506
Abstract 506
31.1 Biological Rationale 506
31.2 The Epidemiologic Approach 507
31.3 Literature Review of Cohort Studies Linking CA Frequency to the Risk of Cancer 508
31.4 Pooled Analysis of Published Data 510
31.5 Preliminary Results and Open Issues 513
31.6 Conclusion 514
References 514
Index 516