Core--Eicosanoid Analysis

描述：(申请人描述)二十烷类化合物分析核心的具体目标是为本计划项目资助中的研究人员进行复杂生物液和组织中二十烷类化合物的分析。该计划项目拨款中提出的四个项目涉及环氧合酶和二十烷类化合物在大肠肿瘤中的作用的研究，并将需要对前列腺素(PG)进行量化。二十烷基类化合物将通过毛细管色谱(GC)/电子捕获负离子化学电离(NICI)质谱仪(MS)方法在细胞培养物以及动物和人类组织中进行定量，该方法使用由Core Leader开发的目前正在使用的稳定同位素稀释技术。参与该计划项目赠款的研究人员使用这些分析方法已经进行了一些基本的观察，这些观察有助于描述二十烷类化合物在与胃肠道恶性肿瘤相关的病理生理过程中的作用。此外，Core还将开发一种GC/NICI MS分析方法，以测定从环氧合酶生成的双环内过氧物中间体PGH2中提取的化合物12-羟基-十七碳三烯酸(12-HHT)。这个测试将被用于项目4(Marnett博士)，以进一步探索12-HHT在环氧合酶过度表达情况下的作用。因此，二十烷类化合物核心的总体目标是为二十烷类化合物的定量分析提供技术支持和实施高度敏感的方法。

DESCRIPTION: (Applicant's Description) The specific goals of the Eicosanoid Analysis Core are to carry out the analyses of eicosanoids in complex biological fluids and tissues for investigators in this Program Project Grant. The four projects proposed in this Program Project Grant involve research examining the role of the cyclooxygenase and eicosanoids in colorectal neoplasia and will require quantification of prostaglandins (PGs). Eicosanoids will be quantified in cellular incubations and animal and human tissues by methodology employing capillary gas chromatography (GC)/electron capture negative ion chemical ionization (NICI) mass spectrometry (MS) using stable isotope dilution techniques that were developed by the Core Leader and which are currently in use. Use of these assays by investigators involved in this Program Project Grant has already allowed a number of fundamental observations to be made that have helped to delineate the role of eicosanoids in pathophysiological processes related to gastrointestinal malignancy. In addition, the Core will also develop a GC/NICI MS assay to measure the compound 12- hydroxy-heptadecatrienoic acid (12-HHT) derived from the cyclooxygenase-generated bicycloendoperoxide intermediate PGH2. This assay will be utilized in Project 4 (Dr. Marnett) to further explore the role of 12-HHT in situations of cyclooxygenase overexpression. The overall objectives of the Eicosanoid Core, therefore, are to provide technical support and implementation of highly sensitive methodology for the quantitative analysis of eicosanoids.