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E-raamat: Cyclooxygenases: Methods and Protocols

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Since the discovery of the pharmacological and toxicological importance of inhibiting the cyclooxygenase (COX) enzymes by non-steroidal anti-inflammatory drugs (NSAIDs), much research has gone into the development of methods to study the biological functions of COX-1 and COX-2.  In Cyclooxygenases: Methods and Protocols, experts and pioneers in the field present the most up-to-date in vitro and in vivo techniques routinely used in COX research.  The volume delves into essential topics such as the purification, cloning, and expression of COX enzymes as well as in vitro assays aimed at determining the inhibitory potency of drugs on COX-1 and COX-2 activities, with chapters describing protocols used for the extraction and measurement of the prostanoids. This volume also describes in vivo disease models used to study the roles of COX-1 and COX-2 in gastrointestinal injury, inflammation, and pain.  As a book in the highly successful Methods in Molecular Biology series, the protocols chapters include brief introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, Cyclooxygenases: Methods and Protocols serves as an indispensable tool for all scientists seeking the treatment of inflammation, pain, fever, and other harmful conditions.
Preface v
Contributors ix
PART I IN VITRO PROTOCOLS FOR STUDYING EXPRESSION AND ACTIVITY OF CYCLOOXYGENASE ENZYMES AND CHARACTERISATION OF THEIR INHIBITION PATTERNS
1 In Vitro Cyclo-oxygenase Expression and Activity Protocols: Introduction to Part I
3(8)
Roderick J. Flower
2 Techniques Used to Study Regulation of Cyclooxygenase-2 Promoter Sites
11(10)
Hiroyasu Inoue
Rieko Nakata
3 Purification of Recombinant Human COX-1 and COX-2
21(10)
James K. Gierse
4 Cloning and Expression of Cyclooxygenase-1 and Cyclooxygenase-2
31(14)
Nicholas R. Staten
Beverly A. Reitz
5 Expression of Cyclooxygenase Isoforms in the Baculovirus Expression System
45(10)
John C. Hunter
Natalie M. Myres
Daniel L. Simmons
6 Peroxidase Active Site Activity Assay
55(12)
Kelsey C. Duggan
Joel Musee
Lawrence J. Marnett
7 Study of Inhibitors of the PGH Synthases for which Potency Is Regulated by the Redox State of the Enzymes
67(24)
Olivier Boutaud
John A. Oates
8 Different Methods for Testing Potential Cyclooxygenase-1 and Cyclooxygenase-2 Inhibitors
91(26)
Stefan Laufer
Sabine Luik
9 In Vitro Cyclooxygenase Activity Assay in Tissue Homogenates
117(4)
Samir S. Ayoub
10 Techniques Used to Characterize the Binding of Cyclooxygenase Inhibitors to the Cyclooxygenase Active Site
121(10)
William F. Hood
11 In Vitro Assays for Cyclooxygenase Activity and Inhibitor Characterization
131(16)
Mark C. Walker
James K. Gierse
PART II EXTRACTION AND MEASUREMENT OF PROSTANOIDS AND ISOPROSTANES
12 Extraction and Measurement of Prostanoids and Isoprostanes: Introduction to Part II
147(6)
Paola Patrignani
13 Prostanoid Extraction and Measurement
153(12)
Lorenzo Polenzani
Samir S. Ayoub
14 Measurement of 8-Iso-Prostaglandin F2α in Biological Fluids as a Measure of Lipid Peroxidation
165(16)
Stefania Tacconelli
Marta L. Capone
Paola Patrignani
PART III IN VIVO MODELS TO STUDY INVOLVEMENT OF CYCLOOXYGENASE PRODUCTS IN HEALTH AND DISEASE
15 In Vivo Models to Study Cyclooxygenase Products in Health and Disease: Introduction to Part III
181(8)
Derek W. Gilroy
Melanie Stables
Justine Newson
16 Protocols to Assess the Gastrointestinal Side Effects Resulting from Inhibition of Cyclo-Oxygenase Isoforms
189(12)
Brendan J.R. Whittle
17 Cyclooxygenase Enzymes and Their Products in the Carrageenan-Induced Pleurisy in Rats
201(6)
Adrian R. Moore
Samir S. Ayoub
Michael P. Seed
18 Iloprost-Induced Nociception: Determination of the Site of Anti-nociceptive Action of Cyclooxygenase Inhibitors and the Involvement of Cyclooxygenase Products in Central Mechanisms of Nociception
207(10)
Samir S. Ayoub
Retina M. Botting
Index 217