ANALYSIS OF MICRODISSECTED CATARACTOUS HUMAN LENSES

长期目标是了解原因和 导致人类白内障形成的机制。这个 具体目标是：1.继续我们长期的临床生物化学 与年龄和药物相关的人类白内障的调查。 将对患者进行全面的术前评估 手术，患者的白内障进展将会是 临床随访，并将使用裂隙灯记录 程序、偏振后照射法和Scheimpflug 摄影。在白内障手术后，晶状体将被 用生化方法进行显微解剖和评估。2.研究 新鲜切取的人晶状体上皮细胞的生化特性 在白内障手术后。我们将特别注意以下事项 谷胱甘肽还原酶和其他黄素酶。由于黄素酶 取决于饮食中的核黄素，饮食中核黄素的地位将 监测谷胱甘肽在白内障患者中的应用 这些患者的红细胞中的还原酶活性。其他 还将研究关键酶。3.研究 甲状腺激素(T3和甲状腺素)、β-受体阻滞剂(如 心得安)和类固醇(如泼尼松)对酶活性的影响 在人类晶状体上皮细胞中。牛、兔晶状体器官培养， 老鼠也将被用来研究激素的影响 还有毒品。4.以已知多肽为基础合成短肽 各种晶状体蛋白和其他特定晶状体的序列 蛋白质。这些多肽将被用于产生抗体 这些多肽，然后将用来探测 患有白内障的人类晶状体。5.研究晶状体的稳定性 晶体蛋白。人晶状体晶体蛋白的热稳定性 AGE的功能将用圆二色谱(CD)来研究， 吸收光谱和荧光光谱。6.继续并继续 晶状体质膜主要结构和功能的研究 内源性多肽(HIP-26)，并试图将其结晶为 X射线衍射研究。MIP-26有可能具有 将探索cGMP的结合部位。7.澄清 新药的理化性质及免疫定位 发现了晶状体纤维特有的固有膜蛋白MP-17。 这将包括提纯蛋白质的特性 CD法和原位法研究其二级和三级结构 本地化。8.检查白内障和透明晶状体 遗传性视网膜变性(RCS)大鼠。白内障 形成是次要现象，这些老鼠是极好的 氧化侮辱晶状体的动物模型，可能导致 白内障。

The long term objectives are to understand the causes and mechanisms which lead to the formation of cataract in humans. The specific aims are: 1. To continue our long term clinicobiochemical investigation of aging-related and drug-related human cataracts. A comprehensive presurgical evaluation of the patient will be performed, and the progression of cataract in the patient will be followed clinically, and will be documented using slit-lamp procedures, polarized retroillumination, and Scheimpflug photography. Following cataract surgery, the lens will be microdissected and evaluated biochemically. 2. To study the biochemical properties of freshly excised human lens epithelium following cataract surgery. Special attention will be given to glutathione reductase and other flavoenzymes. Since flavoenzymes depend on dietary riboflavin, the status of dietary riboflavin will be examined in patients with cataract by monitoring glutathione reductase activity in the red blood cells of these patients. Other key enzymes will also be studied. 3. To study the effects of thyroid hormones (T3 and thyroxine), beta-blockers (such as propranolol) and steroids (such as prednisone) on enzyme activity in human lens epithelium. Lens organ culture of bovine, rabbits, and rats will also be used to study the effects of the hormones and drugs. 4. To synthesize short peptides based on the known sequence of the various crystallins and other lens-specific proteins. These peptides will be used to produce antibodies specific to these peptides, and then will be used to probe cataractous human lens. 5. To study the stability of lens crystallins. The thermal stability of human lens crystallin as a function of age will be studied using circular dichroism (CD), absorption, and fluorescence spectroscopy. 6. To continue and study the structure and function of the lens plasma membrane major intrinsic polypeptide (HIP-26), and to try to crystallize it for X-ray diffraction studies. The possibility that MIP-26 has a binding site for cGMP will be explored. 7. To elucidate the physico-chemic properties and to immunolocalize the newly discovered lens fiber-specific intrinsic membrane protein MP-17. This will include purification of the protein characterization of its secondary and tertiary structure using CD and in situ localization. 8. To examine the cataractous and clear lenses of rats with inherited retinal degeneration (RCS). The cataract formation is a secondary phenomenon, these rats are an excellent animal model for oxidative insults to the lens which may cause cataract.