METABOLIC MAINTENANCE & GROWTH OF MYCOBACTERIUM LEPRAE

代谢维持

• 批准号: 3454648
• 负责人: Scott G. Franzblau
• 金额: $ 4.6万
• 依托单位: NATIONAL HANSEN'S DISEASE PROGRAM
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-06-01 至 1993-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3454648
• 关键词: Mycobacterium leprae; acid base balance; adenosine triphosphate; athymic mouse; bacterial antigens; bioassay; cellular respiration; glycolipids; growth media; laboratory mouse; leprosy; microorganism classification; microorganism culture; microorganism growth; microorganism metabolism; microscopy; oxidation; oxygen tension; palmitates; radiotracer; scintillation counter

Studies will be carried out with inocula of Mycobacterium leprae, freshly harvested, from the infected foot pads of nu/nu mice and enriched for viable organisms, to explore optimal biophysical and nutritional parameters for the metabolic maintenance of the leprosy bacillus using the following combination of sensitive metabolic assays: a) quantitation of intracellular adenosine triphosphate (ATP), b) incorporation of 14C-palmitate into PGL-1, c) measurement of the rate of 14CO2 released from 14C- palmitate. The combined use of these indicators of net energy status, anabolic and catabolic activity will overcome the limitations inherent in using a single metabolic assay. Preliminary studies indicate an acidic pH optimum (5.5-6.5) and a requirement for continuous maintenance of a gaseous environment with an oxygen concentration below ambient (2.5-10%) Metabolic maintenance and in vitro drug susceptibility testing under these conditions are markedly superior to closed systems employing neutral pH. This approach should lead to the step-wise development of a medium which will support long-term maintenance and subsequently genuine growth of M. leprae. Less than total success should still allow significant improvement in an in vitro medium in which drug activity against M. leprae could be assessed via changes in metabolic activity.

研究将与麻风分枝杆菌接种进行研究， 刚收获的Nu/Nu小鼠的脚垫， 富含可行的生物，探索最佳的生物物理和 用于代谢维持的营养参数 麻风病芽孢杆菌使用以下敏感的组合 代谢测定：a）定量细胞内腺苷 三磷酸盐（ATP），b）将14c-钙米酸盐掺入PGL-1， c）测量从14C-释放的14CO2的速率 棕榈酸酯。 这些净能量指标的联合使用 状态，合成代谢和分解代谢活动将克服 使用单一代谢测定法固有的局限性。 初步研究表明酸性pH最佳（5.5-6.5）和A 持续维护气态环境的要求 氧气浓度低于环境（2.5-10％）代谢 维护和体外药物敏感性测试 条件明显优于采用封闭的系统 中性pH。 这种方法应导致逐步 开发将支持长期支持的媒介 维护和随后的麻风树先生的真实生长。 较少的 超过全部成功应该可以显着改善 在体外培养基中，可能是对麻风策的药物活性 通过代谢活性的变化评估。