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.

将使用麻风杆菌接种物进行研究， 从nu/nu小鼠的感染足垫新鲜收获， 丰富的活生物体，探索最佳的生物物理和 营养参数的代谢维持 麻风杆菌使用下列敏感组合 代谢测定：a）细胞内腺苷的定量 三磷酸（ATP），B）将14 C-棕榈酸酯掺入PGL-1， c）测量从14 C-1释放的14 CO2的速率。 棕榈酸盐 综合使用这些净能量指标 状态，合成代谢和分解代谢活动将克服 使用单一代谢测定固有的局限性。 初步研究表明，酸性pH值的最佳值（5.5-6.5）和 气体环境连续保持要求 氧气浓度低于环境（2.5-10%） 维持和体外药物敏感性试验， 条件明显上级封闭系统， 中性pH值。这种方法应导致逐步 开发一种能够长期支持 维持和随后的真正增长的M。麻风病人 少 总的来说，成功仍然可以使一个 在体外培养基中对M.麻风病可能是 通过代谢活动的变化进行评估。