ANALYSIS OF LIGHT EMITTED DURING OXIDATIVE STRESS

描述：（改编自申请人的摘要）氧化应激是一种 共同点在氧化应激期间， 活性氧物质（ROS）启动和/或传播链式反应， 导致大多数生物分子的氧化，导致细胞损伤， 死亡因此，重要的是要有非侵入性技术， 氧化损伤的程度在真实的时间。然而，没有一个可用的 方法可以做到这一点而不干扰该过程。我们建议 在蛋白质和脂质氧化过程中发出的超弱光， 与ROS的稳态浓度成比例，可能是监测 氧化应激的动力学，而不干扰 反应或必须等待氧化产物的积累。光 发射是由产生的电子激发物质的衰变引起的 在蛋白质和脂质氧化过程中。为了放大这些信号， 重要的是要知道哪些是主要的发射波长。初步 研究表明，大多数发射的光在500和500之间的范围内。 700 nm，但现有的光子计数器不够灵敏，无法使用 进行更详细的光谱分析这项技术的灵敏度是 使用具有更高量子效率的二极管阵列（CCD） 效率，但缺乏关于峰值发射波长的信息 限制了它们对单光子计数的应用。本研究的目的是 重新设计我们的光子计数器， 在包括游离氨基酸，蛋白质， 脂质体、脂质/蛋白质混合物，最后是完整的细胞。 将使用具有大表面积和宽光谱范围的光电管作为 检测器拟议的变化将增加电流灵敏度约 一个数量级。本研究中获得的详细光谱分析 也将为今后高科技产业的发展提供重要信息。 敏感的CCD仪器，以监测氧化应激在各种 场景本申请中提出的大部分研究将在 在PI的指导下由本科生进行。

DESCRIPTION: (Adapted from the applicant's abstract) Oxidative stress is a common denominator in many pathological scenarios. During oxidative stress, reactive oxygen species (ROS) start and/or propagate chain reactions which result in the oxidation of most biological molecules leading to cell damage and death. Thus, it is important to have non-invasive techniques that will estimate the magnitude of an oxidative insult in real time. Yet, none of the available methods can do this without interfering with the process. We propose that the ultraweak light emitted during the oxidation of proteins and lipids, which is proportional to the steady state concentration of ROS, may be a tool to monitor the kinetics of oxidative stress without interfering with the course of the reaction or having to wait for the accumulation of oxidation products. Light emission results from the decay of electronically excited species produced during protein and lipid oxidation. In order to amplify these signals, it is important to know which are the primary emission wavelengths. Preliminary studies indicate that most of the emitted light is in the range between 500 and 700 nm, but the available photon counters are not sensitive enough to be used for more detailed spectral analysis. The sensitivity of this technique would be increased substantially using diode arrays (CCDs) with higher quantum efficiency, but the lack of information regarding peak emission wavelengths limits their application to single-photon counting. The aim of this study is to re-design our photon counter to characterize the spectra of light emitted during the oxidation of model systems including free amino acids, proteins, liposomes, lipid/protein mixtures and, finally, intact cells. A phototube with large surface area and broad spectral range will be used as a detector. The proposed changes will increase the current sensitivity by about one order of magnitude. The detailed spectral analysis obtained in this study will also provide important information for future development of highly sensitive CCD-based instrumentation to monitor oxidative stress in a variety of scenarios. Most of the studies proposed in this application will be carried out by undergraduate students under the direction of the PI.