INTEGRATION OF FTIR AND A CAPTIVE BUBBLE SURFACTOMETER

目前的方法，无论是从仪器灵敏度还是设计，都不能有效地研究肺表面活性物质蛋白的结构和/或功能。确定NRDS和ARDS的作用机制对于开发NRDS和ARDS的治疗方法至关重要。这将通过将灵敏的红外光谱技术(ATR-FTIR)与独特的自带气泡表面测量仪(CBS)相结合来实现，从而产生一种能够准确地模拟生理条件的制剂的脂类/蛋白质相互作用和表面活性蛋白质结构的方法。拟议研究的目标是优化CBS/ATR-FTIR样品室中FTIR光谱采集的参数，优化CBS组件测量表面张力的参数和仪器，并集成CBS/ATR-FTIR采样附件的所有组件，以实现完整的计算机化仪器控制和数据采集。这种配置将允许同时测量1)单层表面压力，2)蛋白质结构(作为表面压力的函数)，3)蛋白质和脂肪取向(与表面压力相比)，以及4)表面活性蛋白质和其他蛋白质(例如，膜受体、抗菌肽)在脂质单层界面上的蛋白质/脂肪相互作用。通过CBS/ATR-FTIR的同时结构/功能测量，将有可能描绘出每种表面活性物质蛋白在健康和疾病肺组织中肺表面活性物质生物学功能中的作用。在不久的将来，对更复杂的表面活性物质蛋白质/脂肪混合物的实验可能有助于改进和开发针对婴儿、儿童和成人表面活性物质缺乏性疾病的治疗方法。

Current methods, whether from instrumental sensitivity or design, have not been capable of effectively studying the structure and/or function of pulmonary surfactant proteins. It is crucial for the development of treatments for NRDS and ARDS that the mechanisms by which these proteins function be determined. This will be accomplished by combining a sensitive infrared spectroscopic technique (ATR-FTIR) with a unique captive bubble surfactometer (CBS) to yield a method that will be capable of defining lipid/protein interactions and surfactant protein structure of preparations that accurately mimic physiological conditions. The goals of the proposed research are to optimize parameters for FTIR spectral acquisition in the CBS/ATR-FTIR sample chamber, to optimize parameters and instrumentation for surface tension measurements by the CBS component, and to integrate all components of the CBS/ATR-FTIR sampling accessory for complete computerized instrumental control and data acquisition. This configuration will permit simultaneous measurement of 1) monolayer surface pressure, 2) protein structure (as a function of surface pressure), 3) protein and lipid orientation (vs. surface pressure), and 4) protein/lipid interactions of surfactant proteins and other proteins (e.g., membrane receptors, antimicrobial peptides) at a lipid monolayer interface. Simultaneous structure/function measurements by CBS/ATR-FTIR will make it possible to delineate the role of each surfactant protein in the biological function of pulmonary surfactant in healthy and diseased lungs. In the near future, experiments with more complex surfactant protein/lipid mixtures may aid in the improvement and development of treatments for surfactant deficient diseases in infants, children and adults.