Septic Shock: Methods and Protocols

Front Cover
Thomas J. Evans
Springer Science & Business Media, 2000 - Medical - 208 pages
In Septic Shock Methods and Protocols, an interdisciplinary panel of expert investigators describe proven methods for investigating the often deadly mechanisms of septic shock. Drawing on insights from a wide variety of fields-ranging from molecular and cellular biology to the physiological measurement of whole animals-these experienced laboratorians detail the most important techniques for studying sepsis involving endotoxins, toxins derived from gram-positive organisms, cytokines, nitric oxide and other reactive nitrogen intermediates, and the use of primary cell cultures. They also treat a number of significant techniques that are not well described in the literature, including the Limulus endotoxin assay, the bioassay of TNF, the purification of bacterial endotoxin and superantigens, and the culture of primary human cells. Multidisciplinary and state-of-the-art, Septic Shock Methods and Protocols offers both experimental and clinical investigators a gold-standard collection of key methods for illuminating the basic pathophysiological processes that occur following infection. Its readily reproducible methods will allow both newcomers and those with more experience to apply today's powerful techniques successfully in their work.
 

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Contents

ENDOTOXIN
3
Preparation of Endotoxin from Pathogenic GramNegative Bacteria
13
Assay of AntiEndotoxin Antibodies
27
Isolation of the BactericidalPermeabilityIncreasing Protein
37
Purification of Lipopolysaccharide Binding Protein
45
Purification of Streptococcal Pyrogenic Exotoxin
59
Assay for Superantigens
67
Bioassay for Tumor Necrosis Factorsα and
83
In Situ Detection of Nitric Oxide
133
Immunochemical Detection of Nitric Oxide Synthase
145
vii
149
AntiNitrotyrosine Antibodies for Immunohistochemistry
159
Detection of Peroxynitrite in Biological Fluids
171
Myocardial Cells in Culture to Study Effects of Cytokines
181
Culture of Primary Human Bronchial Epithelial Cells
189
Primary Culture of Human Proximal Renal Tubular Epithelial Cells
197

Assay of Soluble Tumor Necrosis Factor Receptors
91
WholeBlood Assays for Cytokine Production
101
NITRIC OXIDE AND OTHER REACTIVE NITROGEN INTERMEDIATES
113

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