Nitric Oxide Protocols

Michael Titheradge provides both the experienced researcher and the newcomer with time-tested techniques to cover all aspects of NO synthase and metabolism. Topics range from the cloning and expression of the different isoforms of NO synthase to the role of NO in DNA damage and apoptosis and include indirect and direct measurements of NO production, measurement on NO synthase mRNA both in situ and in vitro, and Western blotting and immunohistochemical localization of different isoforms of NO synthase. A variety of alternative techniques are described to enable researchers to set up an assay using either the simplest equipment or the most expensive EPR machines.

The past two decades have seen an explosion in the number of research articles relating to both the physiological and pathological responses evoked by nitric oxide generation. Despite this, accurate quantification of nitric oxide in either in vitro or in vivo models remains challenging, due to the relatively unstable nature of the molecule. In Nitric Oxide: Methods and Protocols, experts in the field provide a detailed reference manual packed with step-by-step approaches to reproduce various complex protocols within your own laboratory. Divided into two parts, the volume first covers various disparate protocols relating to the direct detection and quantification of nitric oxide, including techniques which detail how to image real time in vivo generation of nitric oxide, quantify nitric oxide production in the rat brain, and the detect ultralow levels of nitric oxide in the pM range, and the book then continues with a section on techniques designed to either inhibit or enhance nitric oxide, with an aim to achieve therapeutic gain, featuring the inhibition of the nitric oxide synthase enzymes using viral, shRNA delivery systems to prevent cardiovascular dysfunction, peripheral neuropathy, and graft rejection. Written in the highly successful Methods in Molecular BiologyTM series format, chapters include introductions to their respective topics, lists of the necessary materials, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls. Authoritative and easy to use, Nitric Oxide: Methods and Protocols examines numerous complex methodologies detailed in each chapter and proves to be an invaluable resource for anyone with an interest in nitric oxide research.

This book presents WHO guidelines for the protection of public health from risks due to a number of chemicals commonly present in indoor air. The substances considered in this review, i.e. benzene, carbon monoxide, formaldehyde, naphthalene, nitrogen dioxide, polycyclic aromatic hydrocarbons (especially benzo[a]pyrene), radon, trichloroethylene and tetrachloroethylene, have indoor sources, are known in respect of their hazardousness to health and are often found indoors in concentrations of health concern. The guidelines are targeted at public health professionals involved in preventing health risks of environmental exposures, as well as specialists and authorities involved in the design and use of buildings, indoor materials and products. They provide a scientific basis for legally enforceable standards.

Carrying on the high standards of the much-praised first edition of Nitric Oxide Protocols, Aviv Hassid has brought together a panel of expert researchers and clinician scientists to describe in step-by-step detail the latest methodologies for the measurement of nitric oxide--and the enzyme that produces it-in biological tissues and fluids. The authors take advantage of the latest methodologies for the quantitation of biological fluids and tissues, including capillary electrophoresis, microcoaxial electrodes, in vivo measurement of nitric oxide in exhaled air, confocal microscopy, gas chromatography, in situ hybridization, and real-time polymerase chain reaction. Chapters on the measurement of the novel products of nitric oxide, such as nitrated proteins, S-nitrosylated proteins, and dioxygen-dependent NO metabolism, are also included. Additional chapters address the expression of nitric oxide synthase via the use of viral vectors in gene therapy for erectile dysfunction and cancer, as well as in retrovirus, adenovirus, or adenoassociated virus-mediated expression of nitric oxide synthase in vivo. The protocols follow the successful Methods in Molecular BiologyTM series format, each one offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. State-of-the-art and highly practical, Nitric Oxide Protocols, 2nd ed., offers investigators and clinican/scientists a gold-standard collection of readily reproducible analytical techniques for measuring levels of nitric oxide and determining its manifold functions and effects.

With numerous methods available for the quantification of nitric oxide (NO), this detailed book explores their advantages and disadvantages in order to provide a foundation for further research in plant nitric oxide. After providing a useful practical guide to choosing a technique for measuring NO from plant materials, the book continues with chapters on chemiluminescence, diaminofluorescence (DAF), EPR spectroscopy, a laser-based method, as well as many other topics. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory methods, and tips on troubleshooting and avoiding known pitfalls. Practical and authoritative, Plant Nitric Oxide: Methods and Protocols serves as a valuable guide to all researchers working and intending to work in the field of plant nitric oxide research.

Nitric oxide has a tantalizing role in health and disease: while many of its wide-ranging effects are well known, there remains much more to explore and to learn about the interactions of this fascinating molecule in physiological and pathophysiological processes. The volume reviews the myriad effects of nitric oxide as a chemical messenger in the central nervous system, peripheral nervous system, immune system and cardiovascular system. Furthermore, it provides a very practical introduction to the procedures and experimental protocols necessary to work with and study nitric oxide and its synthesizing enzyme, nitric oxide synthase, in the laboratory. In this respect the volume is unique, providing as it does a complete single-volume review of the role of nitric oxide in health and disease, and a very practical introduction to the methods and protocols involved in this intriguing and active area of biomedical research.

Contributors. -- Foreword. -- Preface. -- Getting Started. -- Assessing Available Information. -- Organizing and Preliminary Planning for Surgical Research -- Writing a Protocol: Animals, Humans, and Use of Biologic, Chemical, and Radiologic Agents. -- Grantsmanship. -- Informed Consent and the Protection of Human Research Subjects: Historical Perspectives and Guide to Current United States Regulations. -- Animal Care and Maintenance. -- Funding Strategies and Agencies: Academic-Industrial Relationships; Intellectual Property. -- Statistical Considerations. -- Use of Nonexperimental Studies to Evaluate Surgical Procedures and Other Interventions: The Challenge of Risk Adjustment. -- Measuring Surgical Outcomes. -- Design of Clinical Trials. -- Using Administrative Data for Clinical Research. -- Research in the Intensive Care Unit: Ethical and Methodological Issues. -- Research in the Operating Room. -- Effects of Age and Gender. -- Strategies, Principles, and Techniques Using Transgeni ...

The main objective of these updated global guidelines is to offer health-based air quality guideline levels, expressed as long-term or short-term concentrations for six key air pollutants: PM2.5, PM10, ozone, nitrogen dioxide, sulfur dioxide and carbon monoxide. In addition, the guidelines provide interim targets to guide reduction efforts of these pollutants, as well as good practice statements for the management of certain types of PM (i.e., black carbon/elemental carbon, ultrafine particles, particles originating from sand and duststorms). These guidelines are not legally binding standards; however, they provide WHO Member States with an evidence-informed tool, which they can use to inform legislation and policy. Ultimately, the goal of these guidelines is to help reduce levels of air pollutants in order to decrease the enormous health burden resulting from the exposure to air pollution worldwide.