BIOSENSOR TEST ANESTHETIC EFFECTS ON CARDIAC MEMBRANES

这项拟议的研究将测试使用电容的可行性。 基于交叉指电极阵列的生物传感器 挥发性麻醉药与心肌膜蛋白的相互作用 在钙稳态中发挥作用，被认为是钙平衡的关键 挥发性麻醉药会抑制心脏的收缩能力。生物传感器可以 被定义为生物识别系统耦合的系统 带有换能器，以提供直接的电子输出 可用于测量或控制目的。如果能证明 这种生物传感器具有足够的灵敏度，其输出可以 与特定的双分子过程有关，那么这项技术就具有 测试药理活性及其相互作用的巨大潜力 对膜受体进行ct的药物。 心肌的电压依赖性钙通道和钙/镁-三磷酸腺苷酶 将被重组成膜状结构，位于 平面电容式生物传感器。系统对电压的电容响应 激动剂、拮抗剂、氟烷的应用 这些药物的组合将被测量。膜组件 用同样的技术制备的样品将用傅里叶变换进行研究 利用衰减全反射的红外光谱 (ATR)从表面膜获得光谱数据的技术。 含有膜蛋白的生物膜以及形成的组件 将通过FTIR观察单独来自磷脂的成分，以确定 药物的结合部位及其对蛋白质或脂肪的影响 结构。FTIR获得的结构信息将与 来自生物传感器的电响应来确定这种类型的 生物传感器有助于机械研究和/或快速监测 药物和麻醉剂的相互作用。

The proposed study will test the feasibility of using a capacitive biosensor based on an interdigitated electrode array to monitor the interaction of volatile anesthetics with cardiac membrane proteins that play a role in calcium homeostasis and are thought to be sties at which the volatile anesthetics act to depress cardiac contractility. A biosensor can be defined as a system in which a biological recognition system is coupled with a transducing device to give an electrical output that is directly usable for either measurement or control purposes. If it can be shown that such biosensors are sufficiently sensitive and that the outputs can be related to specific bimolecular processes, then this technology has enormous potential for testing pharmacologic activity and the interaction of agents that ct on membrane receptors. The voltage-dependent Ca2+ channel and Ca2+/Mg2+-ATPase from cardiac muscle will be reconstituted into membrane-like structures at the surface of a planar capacitive biosensor. The capacitance response of the system to the application of agonists, antagonists, halothane, and to various combinations of these agents will be measured. Membrane assemblies prepared by the same technique will be studied by Fourier transform infrared (FTIR) spectroscopy through use of attenuated total reflectance (ATR) techniques for obtaining spectral data from surface films. Biomembranes incorporating membrane proteins as well as assemblies formed from phospholipids alone will be observed by FTIR in order to identify the sites at which the agents bind and their effects on protein or lipid structure. Structural information obtained by FTIR will be correlated with electrical response from the biosensor to determine whether this type of biosensor can contribute to mechanistic studies and/or rapid monitoring of drug and anesthetic interactions.