COURSE ON 1H / 13C / 2H METHODS

1H / 13C / 2H 方法课程

• 批准号: 8363921
• 负责人: MARK JEFFREY
• 金额: $ 0.8万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8363921
• 关键词: Blood Glucose; Carbon; Citric Acid Cycle; Computer software; Data; Data Analyses; Data Files; Deuterium; Deuterium Exchange Measurement; Educational process of instructing; Funding; Gluconeogenesis; Glucose; Glycerol; Grant; Group Meetings; Individual; Instruction; Laboratories; Measurement; Measures; Metabolic; Metabolism; Methods; Modeling; National Center for Research Resources; Physiological; Preparation; Principal Investigator; Procedures; Process; Propionates; Protocols documentation; Relative (related person); Research; Research Infrastructure; Research Personnel; Resources; Sampling; Source; Staging; Students; United States National Institutes of Health; base; capsule; cost; design; experience; glucose production; glycogenolysis; human subject; in vivo; research study

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. This activity has two components to be selected by the user. These activities are in addition to weekly for teaching and discussion in various aspects of 13C NMR and metabolism in research group meetings. First, a hands-on course conducted over 3-4 days is made available on request. We offer this course for experienced investigators already familiar with conducting NMR experiments who would like an introduction to experiments based on the use of deuterium and carbon-13 in their own laboratories. The material covered includes the use of D2O and 13C propionate for measuring fluxes in vivo. The provision of D2O to subjects and measurement of deuterium exchange via 2H NMR of glucose provides the relative contributions of gluconeogenesis (from PEP or glycerol) and glycogenolysis. Giving [U-13C]proprionate in capsules and analysis of the C2 carbon of glucose via 13C NMR gives the relative rate of gluconeogenesis from PEP and flux through the citric acid cycle. When measurements from both approaches are combined, glucose production can be expressed in terms of citric acid cycle turnover. Combining this with measurement of glucose turnover (via the dilution in blood of glucose infused at a constant rate), the various relative measurements mentioned above (glucose production from various sources, citric acid cycle activity) can be expressed as absolute fluxes. The student has the opportunity to be involved in all aspects these measurements, from designing protocols to data analysis and interpretation. The student will observe the procedures for handling the preparation, processing samples, collecting NMR data, performing spectral analysis and finishing with applying metabolic models. Instruction in the use of software required at various stages is provided. The course is tailored to individual needs as closely as possible. Second, users are taught, one-on-one, how to interpret 1H NMR spectra from human subjects. This involves showing them how to open spectral data files, convert to format readable by ACD, transformation, fitting and conversion of raw data to physiological variables.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 此活动具有用户选择的两个组件。 这些活动是在研究小组会议中的13C NMR和代谢的各个方面的每周进行教学和讨论的补充。 首先，应要求进行3-4天的动手课程。我们为经验丰富的调查人员提供了这项课程，他们想根据自己的实验室中使用氘和碳13来介绍实验的NMR实验。 涵盖的材料包括使用D2O和13C丙酸酯在体内测量通量。 通过2H NMR的葡萄糖提供了D2O对受试者和氘交换的测量，这提供了糖异生（来自PEP或甘油）和糖基解体的相对贡献。 在胶囊中给出[U-13C] - 通过13C NMR对葡萄糖的C2碳进行分析，从而通过柠檬酸循环从PEP和通量中提供了糖异生的相对速率。 当将两种方法的测量结果合并时，葡萄糖的产生可以用柠檬酸周期周转率表示。 将其与葡萄糖更新的测量结合（通过以恒定速率注入的葡萄糖的血液稀释），上面提到的各种相对测量值（来自各种来源的葡萄糖产生，柠檬酸循环活性）可以表示为绝对通量。从设计协议到数据分析和解释，学生有机会参与各个方面。 学生将观察处理制备，处理样品，收集NMR数据，进行光谱分析并使用代谢模型完成的程序。 提供了在各个阶段使用所需软件的说明。 该课程应尽可能地针对个人需求量身定制。 其次，教授用户，一对一，如何解释人类受试者的1H NMR光谱。 这涉及向他们展示如何打开光谱数据文件，转换为ACD可读的格式，转换，拟合和转换原始数据为生理变量。