生物化学技术

The nucleus tractus solitarius (NTS) of the brainstem is an important site for the regulation of sympathetic nerve activity. In the brain, nitric oxide (NO) has been shown to reduce blood pressure by inhibiting sympathetic nerve activity. However, most studies were performed in an acute state of ...

The generation of retrovectors (retroviral vectors) with tumor specificity is a promising and effective approach to targeted gene therapy of cancer. Carcinoembryonic antigen (CEA), overexpressed by various tumor cells, provides a specific tool for tumor-specific targeting by r ...

Erectile dysfunction (ED) is defined as the consistent inability to obtain or maintain an erection for satisfactory sexual intercourse. In the last decade, significant advancements in the pathophysiology of male ED have occurred. Basic science research on erectile physiology has be ...

Accumulating evidence shows that S-nitrosothiols, formed by the addition of nitric oxide (NO) to a cysteine thiol, S-nitrosylation, are involved in basal cellular regulation. It has been proposed that SNO formation/removal may be disrupted in a variety of pathophysiological conditi ...

Biologic nitration of protein tyrosine (to form 3-nitrotyrosine) is a recently described phenomenon that is associated with many diseases. We have devised a proteomic methodology to identify these modified proteins. This utilizes protein fractionation by two-dimensional pol ...

Nitric oxide (NO) serves critical signaling, energetic, and toxic functions throughout the biosphere. NO steady-state levels and functions are controlled in part by NO metabolism or degradation. Dioxygen-dependent NO dioxygenases (EC 1.14.12.17) and dioxygen-independent NO re ...

The endothelial isoform of nitric oxide synthase (eNOS) plays a major role in vascular biology and emerging evidence suggests it may serve diverse biological functions in nonvascular cells. To be able to study the eNOS at the molecular level, including regulation of mRNA levels, we developed r ...

Functional genomic increasingly requires the cell-specific localization of gene expression. The histological analysis of mRNA expression is performed by in situ hybridization (ISH), whereas proteins are detected by immunohistochemistry. ISH allows the rapid detection of a ...

We describe a fluorescence assay for nitric oxide synthase activity based on a new indicator, 4,5-diaminofluorescein (DAF-2). The method offers the advantage of being safer and more convenient than the citrulline radioassay in common use. The rapid and irreversible binding of DAF-2 to oxid ...

Measurement of the nitric oxide (NO) metabolites nitrite and nitrate in biological matrices is a reliable method to assess NO synthase (NOS) activity. Unlike the “L-citrulline assay,” the “nitrite/nitrate assay” is generally applicable and permits acquisition of maximum informati ...

The instability and low concentration of nitric oxide (NO) in specimens make it difficult to be detected directly. In this chapter, a method for imaging nitric oxide using laser scanning confocal microscopy (LSCM) is presented. A cultured hippocampal neuron is dyed with DAF-2 and observed und ...

Assessment of the value of exhaled NO (eNO) is an attractive tool for studying pulmonary disease, considering its wide advantages (i.e., fast analysis, noninvasive sampling, ability to measure large numbers of subjects , and inexpensive in use). Increased concentrations of eNO have been ob ...

Nitric oxide (NO) is a vital molecule contributing to numerous physiological phenomena in various biological systems. To investigate the physiological role of NO, a range of NO-specific electrodes allowing direct and continuous NO measurement have been developed for in vitro and in vivo ...

Nitrite and nitrate represent relatively stable nitric oxide (NO) end products in several biological fluids. The amounts of nitrite and/or nitrate in urine have been shown to reflect local and/or systemic involvement of NO in pathological conditions, including urinary tract infecti ...

Nitrite and nitrate are widely used reporters of endogenous nitric oxide (NO) and nitric oxide synthase (NOS) activity, which are crucial for a broad spectrum of physiological and pathophysiological pathways. Because of the great variety in spatial expression and activity of NOS in animal ...

A simple and sensitive method is presented to measure the unstable molecule nitric oxide (NO) by reconversion of nitrate/nitrite to NO. Nitrate and nitrite are the stable degradation products of NO that accumulate in supernatants of biological samples that release nitric oxide. First, ni ...

Reactive oxidants are thought to protect the host by destroying invading microbes (1). However, owing to their indiscriminate nature, reactive oxidants might also damage the host if generated inappropriately. Indeed, they have been implicated in a wide array of pathologies (2–4).

Synaptosomal protein oxidation, most often indexed by increased protein carbonyl content (1,2), is one biomarker of oxidative stress in brain (2). Protein carbonyls are detected by UV-Vis spectrometric, fluorescence, or immunochemical methods (2), and are increased in a variety of oxi ...

The thiobarbituric acid (TBA) assay was developed to quantitatively determine lipid peroxidation for aldehydic compounds in biological matrices. Kohn and Liversedge introduced this methodology in 1944 (1,2). Since its introduction, the TBA assay has generated widespread inte ...

Unsaturated aldehydes such as acrolein and hydroxyalkenals are produced in vivo through lipid peroxidation chain reactions under conditions of oxidative stress or carcinogenic insult, and are causally involved in the pathogenesis of numerous diseases (1,2). The ability of cytot ...