Septic shock remains a serious medical condition with high mortality. Despite many advances in intensive care medicine and antibiotic devel- ment, this has not changed appreciably in the last 20 years. Frustratingly, over the same period of time, enormous advances have been made in understa- ing the underlying pathogenic mechanisms of this condition. This has resulted in the development of several novel therapies for septic shock, which, despite excellent theoretical grounds for their efficacy, have failed in altering mort- ity attributable to sepsis. The reasons for these failures are multiple, but it is clear that further research is required aimed at increasing our understanding of the basic pat- physiological processes that occur following infection. Research into septic shock draws upon a number of different disciplines, ranging from molecular and cellular biology to physiological measurements on whole animals. Septic Shock Methods and Protocols is an attempt to draw together into one volume a number of protocols that are of use in the investigation of the mechanisms of septic shock. I have divided the book into five sections. The first deals with endotoxin, the lipopolysaccharide component of the Gram-negative cell m- brane that can mimic many of the features of septic shock. Gram-positive organisms are found increasingly as causes of septic shock, and several - pects of toxins produced from these bacteria are considered in the second s- tion.

Klappentext

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.

Zusammenfassung
Septic shock remains a serious medical condition with high mortality. Septic Shock Methods and Protocols is an attempt to draw together into one volume a number of protocols that are of use in the investigation of the mechanisms of septic shock.

Inhalt
Endotoxin.- Assay of Endotoxin by Limulus Amebocyte Lysate.- Preparation of Endotoxin from Pathogenic Gram-Negative Bacteria.- Assay of Anti-Endotoxin Antibodies.- Isolation of the Bactericidal/Permeability-Increasing Protein from Polymorphonuclear Leukocytes by Reversible Binding to Target Bacteria.- Purification of Lipopolysaccharide Binding Protein.- Other Bacterial Products.- Purification of Streptococcal Pyrogenic Exotoxin A.- Assay for Superantigens.- Cytokines.- Bioassay for Tumor Necrosis Factors-? and ?.- Assay of Soluble Tumor Necrosis Factor Receptors.- Whole-Blood Assays for Cytokine Production.- Nitric Oxide and Other Reactive Nitrogen Intermediates.- Nitric Oxide Synthase Inhibitors.- In Situ Detection of Nitric Oxide.- Immunochemical Detection of Nitric Oxide Synthase in Human Tissue.- Anti-Nitrotyrosine Antibodies for Immunohistochemistry.- Detection of Peroxynitrite in Biological Fluids.- Cell Culture Techniques.- Myocardial Cells in Culture to Study Effects of Cytokines.- Culture of Primary Human Bronchial Epithelial Cells.- Primary Culture of Human Proximal Renal Tubular Epithelial Cells.