COURSE ON 1H / 13C / 2H METHODS

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 此活动有两个组件供用户选择。 这些活动是除了每周的教学和讨论在各个方面的13 C NMR和代谢的研究小组会议。 首先，应要求提供为期3-4天的实践课程。我们为已经熟悉进行NMR实验的经验丰富的研究人员提供本课程，他们希望介绍基于在自己的实验室中使用氘和碳-13的实验。 所涵盖的材料包括使用D2 O和13 C丙酸盐测量体内通量。 向受试者提供D2 O和通过葡萄糖的2 H NMR测量氘交换提供了糖原生成（来自PEP或甘油）和糖原分解的相对贡献。 在胶囊中给予[U-13 C]丙酸盐，并通过13 C NMR分析葡萄糖的C2碳，得到PEP的异构化的相对速率和通过柠檬酸循环的通量。 当两种方法的测量结果结合起来时，葡萄糖的产生可以用柠檬酸循环周转率来表示。 将其与葡萄糖周转率的测量（通过以恒定速率输注的葡萄糖在血液中的稀释）相结合，上述各种相对测量（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。学生有机会参与这些测量的各个方面，从设计协议到数据分析和解释。 学生将观察处理制备，处理样品，收集NMR数据，进行光谱分析和应用代谢模型完成的程序。 提供了各个阶段所需软件的使用说明。 该课程尽可能符合个人需求。 其次，用户被教导，一对一，如何解释1H NMR光谱从人类受试者。 这包括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，拟合和原始数据转换为生理变量。

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.