Nitric Oxide Protocols 2nd Ed (Methods in Molecular Biology by Aviv Hassid

By Aviv Hassid

A suite of state of the art options for measuring nitric oxide and the enzyme that produces it in organic tissues and fluids. those simply reproducible equipment can be utilized to degree novel nitric oxide-related items similar to protein nitration and nitrosation, in addition to to precise nitric oxide synthase in easy examine and gene treatment utilizing viral vectors.

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From this, various NO-specific electrodes have been described that enable the direct and continuous measurement of NO in vitro and in vivo (12–24). We also developed a microcoaxial electrode for selective NO measurement with high spatial and temporal resolutions within a confined area of tissue (25). Because the working and reference electrodes are oriented coaxially and close together, the microcoaxial electrode can be used to great effect for NO measurements in tissues or in isolated cells. In this chapter, we describe in detail the procedure for making NO measurements with the microcoaxial electrode.

Treatment with anti-inflammatory corticosteroids that inhibit iNOS (33,34) reduces eNO in subjects with allergic asthma but not in normal subjects (35– 37). This suggests that eNO in healthy subjects is derived from cNOS, whereas in asthmatics, the additional amounts of eNO are generated by iNOS (12). Indeed, biopsies from asthmatic patients but not healthy subjects express iNOS in airway epithelial cells. Pro-inflammatory cytokines, known to be increased in the airways of asthmatics, have been shown to induce iNOS expression in human epithelial cells in vitro (38).

5C,D and Note 4). 2. 1. Urinary Nitrite and Nitrate Urinary nitrate reflects NO formed locally in organs or tissues located far from the urinary tract because gaseous NO is converted to nitrite and later to nitrate by local agents or other agents in systemic circulation, including oxyhemoglobins and ROS (14,16). Thus, increased urinary nitrate has been reported in atopic dermatitis and infective gastroenteritis (5,22,23). Furthermore, urinary nitrate has indicated increase of NO levels in systemic circulation, following cardiac and renal allograft rejections (7,24).

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