Molecular Biology in Cellular Pathology

by Crocker, John; Murray, Paul G.

• ISBN13: 9780470844755
• ISBN10: 0470844752
• Edition: 1st
• Format: Hardcover
• Copyright: 2003-06-02
• Publisher: WILEY

Summary

The latest edition of this highly successful text, covers the major advances in the methods used in cellular and molecular pathology. In recent years, knowledge of the molecular organization of the cell has led to the development of powerful new techniques that bring greater accuracy and objectives to the diagnosis, prognosis and management of many diseases and to the study of pathological states. This book describes the latest molecular techniques available for the analysis of diseases. In particular it includes new techniques using fluorescent dyes, DNA microarrays, protein chemistry, and mass spectrometry. It also incorporates information from the Human Genome Project, and the new disciplines of genomics and proteomics, where relevant to pathology. Color plates are a new feature of this edition, illustrating the advances in fluorescence labeling of cells.

Author Biography

John Crocker is the editor of Molecular Biology in Cellular Pathology, published by Wiley.

Paul G. Murray is the editor of Molecular Biology in Cellular Pathology, published by Wiley.

Table of Contents

Preface	xiii
Preface to Molecular Biology in Histopathology	xv
List of Contributors	xvii
1 Blotting Techniques: Methodology and Applications	1	(18)
Fiona Watson and C. Simon Herrington
1.1 Introduction	1	(1)
1.2 Blotting techniques	1	(14)
1.3 References	15	(4)
2 In-situ Hybridisation in Histopathology	19	(30)
Gerald Niedobitek and Hermann Herbst
2.1 Introduction	19	(1)
2.2 Experimental conditions	20	(3)
2.3 Probes and labels	23	(4)
2.4 Controls and pitfalls	27	(2)
2.5 Double-labelling	29	(2)
2.6 Increasing the sensitivity of ISH	31	(2)
2.7 What we do in our laboratories	33	(2)
2.8 Applications of ISH: examples	35	(4)
2.9 Perspective	39	(1)
2.10 References	40	(9)
3 DNA Flow Cytometry	49	(12)
M. G. Ormerod
3.1 Introduction	49	(1)
3.2 Definitions and terms	49	(1)
3.3 Dye used for DNA analysis	50	(2)
3.4 Sample preparation for DNA analysis	52	(1)
3.5 Analysis of the DNA histogram	53	(1)
3.6 Quality control	53	(2)
3.7 Computer analysis of the DNA histogram	55	(2)
3.8 Multiparametric measurement	57	(2)
3.9 Acknowledgements	59	(1)
3.10 References	59	(2)
4 Interphase Cytogenetics	61	(18)
Sara A. Dyer and Jonathan J. Waters
4.1 Introduction	61	(1)
4.2 Interphase cytogenetics	62	(5)
4.3 Applications	67	(9)
4.4 Conclusion	76	(1)
4.5 References	77	(2)
5 Oncogenes	79	(26)
Fiona Macdonald
5.1 Introduction	79	(1)
5.2 Identification of the oncogenes	79	(1)
5.3 Functions of the proto-oncogenes	80	(9)
5.4 Mechanism of oncogene activation	89	(2)
5.5 Oncogenes in colorectal cancer	91	(3)
5.6 Oncogenes in breast cancer	94	(1)
5.7 Oncogenes in lung cancer	95	(1)
5.8 Oncogenes in haematological malignancies	96	(3)
5.9 Other cancers	99	(1)
5.10 Conclusion	100	(1)
5.11 References	100	(5)
6 Molecular and Immunological Aspects of Cell Proliferation	105	(32)
Karl Baumforth and John Crocker
6.1 The cell cycle and its importance in clinical pathology	105	(3)
6.2 Molecular control of the cell cycle	108	(3)
6.3 Cell cycle control	111	(1)
6.4 The cell cycle and cancer	112	(3)
6.5 Immunocytochemical markers of proliferating cells	115	(18)
6.6 References	133	(2)
6.7 Further Reading	135	(2)
7 Interphase Nucleolar Organiser Regions in Tumour Pathology	137	(16)
Massimo Derenzini, Davide Treré, Marie-Françoise O'Donohue and Dominique Ploton
7.1 Introduction	137	(1)
7.2 The AgNORs	138	(4)
7.3 NOR silver-staining	142	(3)
7.4 Quantitative AgNOR analysis	145	(1)
7.5 AgNORs as a parameter of the level of cell proliferation	146	(1)
7.6 Application of the AgNOR technique to tumour pathology	147	(4)
7.7 What future for AgNORs in tumour pathology?	151	(1)
7.8 References	152	(1)
8 Apoptosis and Cell senescence	153	(40)
Lee B. Jordan and David J. Harrison
8.1 Introduction	153	(1)
8.2 Apoptosis	153	(21)
8.3 Cell senescence	174	(4)
8.4 Summary	178	(1)
8.5 References	179	(14)
9 The Polymerase Chain Reaction	193	(20)
Timothy Diss
9.1 Introduction	193	(1)
9.2 Principles	194	(3)
9.3 Analysis of products	197	(2)
9.4 RT-PCR	199	(1)
9.5 Quantitative PCR	200	(1)
9.6 DNA and RNA extraction	200	(1)
9.7 Correlation of the PCR with morphology	201	(1)
9.8 Problems	202	(1)
9.9 Applications	202	(1)
9.10 Diagnostic applications	203	(6)
9.11 Infectious diseases	209	(1)
9.12 Identity	209	(1)
9.13 The future	210	(1)
9.14 References	210	(2)
9.15 Online information	212	(1)
10 Laser Capture Microdissection: Techniques and Applications in the Molecular Analysis of the Cancer Cell	213	(20)
Amanda Dutton, Victor Lopes and Paul G. Murray
10.1 Introduction	213	(1)
10.2 The principle of LCM	214	(2)
10.3 Technical considerations	216	(1)
10.4 Advantages and disadvantages of LCM	217	(5)
10.5 Applications of LCM	222	(7)
10.6 Future perspectives	229	(1)
10.7 Acknowledgements	229	(1)
10.8 References	229	(4)
11 The In-situ Polymerase Chain Reaction	233	(18)
John J. O'Leary, Cara Martin and Orla Sheils
11.1 Introduction	233	(1)
11.2 Overview of the methodology	234	(1)
11.3 In-cell PCR technologies	235	(3)
11.4 In-cell amplification of DNA	238	(4)
11.5 Detection of amplicons	242	(1)
11.6 Reaction, tissue and detection controls for use with in-cell DNA PCR assays	243	(1)
11.7 In-cell RNA amplification	244	(2)
11.8 Problems encountered with in-cell PCR amplification	246	(1)
11.9 Amplicon diffusion and back diffusion	247	(1)
11.10 Future work with in-cell PCR-based assays	247	(2)
11.11 References	249	(2)
12 TagMan® Technology and Real-Time Polymerase Chain Reaction	251	(18)
John J. O'Leary, Orla Sheils, Cara Martin, and Aoife Crowley
12.1 Introduction	251	(1)
12.2 Probe technologies	252	(2)
12.3 TagMan® probe and chemistry (first generation)	254	(2)
12.4 Second generation TagMan® probes	256	(2)
12.5 Hybridisation	258	(1)
12.6 TagMan® PCR conditions	259	(1)
12.7 Standards for quantitative PCR	260	(1)
12.8 Interpretation of results	261	(1)
12.9 End-point detection	262	(1)
12.10 Real-time detection	263	(1)
12.11 Relative quantitation	263	(1)
12.12 Reference genes	264	(1)
12.13 Specific TagMan® PCR applications	265	(3)
12.14 References	268	(1)
13 Gene Expression Analysis Using Microarrays	269	(18)
Sophie E. Wildsmith and Fiona J. Spence
13.1 Introduction	269	(1)
13.2 Microanay experiments	269	(4)
13.3 Data analysis	273	(11)
13.4 Recent examples of microarray applications	284	(1)
13.5 Conclusions	284	(1)
13.6 Acknowledgements	284	(1)
13.7 References	284	(2)
13.8 Further Reading	286	(1)
13.9 Useful websites	286	(1)
14 Comparative Genomic Hybridisation in Pathology	287	(20)
Marjan M. Weiss, Mario A.J.A. Herensen, Antoine Snijders, Horst Buerger, Werner Boecker, Ernst J. Kuipers, Paul J. van Diest and Gerrit A. Meijer
14.1 Introduction	287	(2)
14.2 Technique	289	(3)
14.3 Data analysis	292	(1)
14.4 Applications	293	(6)
14.5 Clinical applications	299	(1)
14.6 Screening for chromosomal abnormalities in fetal and neonatal genomes	299	(1)
14.7 Future perspectives	300	(1)
14.8 Acknowledgements	301	(1)
14.9 References	301	(6)
15 DNA Sequencing and the Human Genome Project	307	(22)
Philip Bennett
15.1 Introduction	307	(1)
15.2 DNA sequencing: the basics	308	(10)
15.3 Applications of DNA sequencing	318	(2)
15.4 The Human Genome Project	320	(7)
15.5 References	327	(1)
15.6 Further Reading	327	(1)
15.7 Useful websites	328	(1)
16 Monoclonal Antibodies: The Generation and Application of 'Tools of the Trade' Within Biomedical Science	329	(22)
Paul N. Nelson, S. Jane Astley and Philip Warren
16.1 Introduction	329	(2)
16.2 Antibodies and antigens	331	(1)
16.3 Polyclonal antibodies	332	(1)
16.4 Monoclonal antibody development	333	(5)
16.5 Monoclonal antibody variants	338	(3)
16.6 Monoclonal antibody applications	341	(4)
16.7 Therapy	345	(1)
16.8 Specific applications	346	(1)
16.9 Conclusions	347	(1)
16.10 Acknowledgements	347	(1)
16.11 References	347	(4)
17 Proteomics	351	(20)
Kathryn Lilley, Azam Razzaq and Michael J. Deery
17.1 Introduction	351	(1)
17.2 Definitions and applications	352	(1)
17.3 Stages in proteome analysis	352	(16)
17.4 Future directions	368	(1)
17.5 References	368	(3)
Index	371