Serum markers

本提案的目的是确定我们开发的用于未结合游离脂肪酸（FFAu）的简单血液检测是否可以早期准确诊断缺血性卒中。虽然使用t-PA的再灌注疗法是缺血性中风的有效治疗方法，但其采用缓慢，部分原因是它必须在缺血性中风的3小时内施用。脑卒中的快速准确诊断是有效实施溶栓再灌注治疗的主要障碍。所需要的是一种简单的测试，例如，在症状明显后的最早时间，以高灵敏度和特异性在少量血液中测定中风标志物的测试。荧光（ADIFAB）方法，我们有 所开发的方法仅需一个步骤，使用不到0.1毫升的血液，在几秒钟内完成，并且可以用手持设备完成。我们的初步结果表明，血浆FFAu水平可以检测到心肌缺血分钟内的侮辱和证据，从动物研究表明脑缺血相同。因此，我们将使用由UCSD卒中团队收集的血液标本来确定FFAu在卒中和拟卒中中的时间过程，FFAu水平是否可用于监测t-PA治疗的过程，以及FFAu水平是否可用于帮助患者进行治疗。使用该检测试剂盒对TIMI II试验标本进行检测的初步结果表明，呈现时（t-PA前）的FFAu水平 给药），预测死亡和严重出血事件的风险增加。因此，我们将确定在中风患者中是否存在类似的相关性，以及是否设计用于降低FFAu水平的治疗方法可以改善动物模型的结果。最后，我们将进行研究，以确定观察到的FFAu水平升高的起源，因为更好地了解这种现象的分子机制，可能会导致缺血性卒中的新的诊断和治疗策略。

The goal of this proposal is to determine if a simple blood-based assay we have developed for unbound free fatty acids (FFAu), allows for an early and accurate diagnosis of ischemic stroke. Although reperfusion therapy using t-PA is an effective treatment for ischemic stroke it has been adopted slowly, in part because it must be administered within 3 hours of an ischemic stroke. The rapid and accurate diagnosis of stroke is a major barrier to the effective implementation of thrombolytic reperfusion therapy. What is needed is a simple test, for example one that would assay for a stroke marker in a small quantity of blood, with high sensitivity and specificity at the earliest times after symptoms are apparent. The fluorescent (ADIFAB) method we have developed requires only a single step, uses less than 0.1 ml of blood, is complete in seconds, and can be done with a hand-held device. Our preliminary results indicate that plasma FFAu levels can detect myocardial ischemia within minutes of the insult and evidence from animal studies suggest the same for cerebral ischemia. We will therefore use blood specimens collected by the UCSD Stroke Team to determine, the time course of FFAu in stroke and stroke mimics, whether FFAu levels can be used to monitor the course of t-PA therapy, and if FFAu levels can be used to help group patients for treatment. Preliminary results using this assay for specimens from the TIMI II trial indicate that FFAu levels, at presentation (before t-PA administration), predict increased risk of death and severe hemorrhagic events. We will therefore determine if similar correlations exist in stroke code patients and if therapies designed to reduce FFAu levels improve outcome in animal models. Finally, we will undertake studies to determine the origin for the observed increased FFAu levels, because better understanding of the molecular mechanisms of this phenomenon, may lead to novel diagnostic and therapeutic strategies for ischemic stroke.