PubMed · 9619243
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K R Dunster. 1998. Monitoring nitric oxide.. https://pubmed.ncbi.nlm.nih.gov/9619243/
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A biomimetic bilayer membrane immobilizing uricase (urate oxidase; EC 1.7.3.3) (UOx) and a redox agent of 1-methoxy-5-methylphenazinium (MMP) was fabricated on an Au electrode substrate with use of the Au substrate coated with a self-assembled monolayer of n-octanethiolate (OT/Au) and L-alpha-phosphatidylcholine beta-oleoyl-gamma-palmitoyl (PCOP). The preparation was carried out by successively immersing an Au electrode substrate in an ethanol solution of OT, an MMP aqueous solution, and a suspension of proteoliposome formed by PCOP containing UOx and MMP. The prepared electrode exhibited such fast steady amperometric responses to uric acid as to allow its determination within 20 s after injecting uric acid, indicating that UOx-catalyzed electrochemical oxidation of uric acid was accomplished with assistance of electron mediation by MMP between UOx and the Au substrate. An increase in the response currents with increasing concentration of uric acid was obtained in a concentration range of uric acid found in healthy human blood. Any interference in the current response that is caused by direct anodic oxidation of uric acid or ascorbic acid was not observed at the prepared sensor electrode because the densely packed bilayer effectively blocked the diffusion of these substrates toward the Au surface, making it possible to determine amperometrically uric acid at the electrode with high precision.
Nitric oxide (NO) is an important vasodilator with various activities in the cerebral vasculature. Although the response of NO levels to shear stress has been investigated in various models using systemic endothelium, no study has evaluated human cerebral endothelial cells (HCE). We determined the NO levels of HCE cultured in an artificial capillary system in response to changes in shear stress. With direct measurement by a porphyrinic microsensor, we found that NO levels increased immediately with a peak at 7 h after changes in shear stress, and by 24 h dropped to a constant elevated baseline. Shear stress-mediated increases in NO levels were confirmed by the measurement of citrulline, an indirect measure of NO. Furthermore, NO levels by HCE were shown to decrease with decreasing shear stress levels. This study presents a novel system to study NO production by microvascular HCE, and indicates a linear relationship between shear stress and NO levels. As cerebral vessels age and lose transmural compliance, shear stress-mediated production of NO may play a greater role in cerebrovascular function and dysfunction.
Trichothecene mycotoxins in animal feed and human food can cause fatalities in livestock and disease in humans. In addition, these toxins are suspected chemical warfare agents. Therefore, development of a simple and sensitive method for the screening of trichothecenes is important to prevent economic loss and health hazards. A simple and inexpensive method for the detection of type-B trichothecene mycotoxins has been developed in our laboratory. By hydrolyzing the toxin under basic conditions at 80 degrees C for 1 h it is possible to detect the toxin with simple electrochemical techniques. Deoxynivalenol (DON), commonly known as vomitoxin, was used as a representative compound for type-B trichothecenes in this detection scheme. The detection limit for DON using our procedures was determined to be 9.1 microM in solution, corresponding to 0.24 ppm in a 25-g grain sample if the final extraction volume is 2.2 mL. The linear dynamic detection range was determined to be from 0.32 ppm to greater than 32 ppm. In addition to standard solutions, this method was used on rice samples spiked with DON. It was demonstrated that there is no electrochemical interference from rice extract and that 1 ppm of DON in rice samples can be quantified. This method may be ideal for toxin screening in animal feeds or in runoff from sites that produce the compounds as chemical warfare agents. Since the active moiety in DON is common to virtually all type-B trichothecenes, our approach may be ideal for type-specific screening.