COURSE ON 1H / 13C / 2H METHODS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 此活动有两个组件可供用户选择。这些活动是对每周教学的补充，并在研究小组会议上讨论13C核磁共振和新陈代谢的各个方面。 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核磁共振实验的有经验的研究人员提供这门课程，他们希望在自己的实验室中介绍基于氘和碳-13的实验。所涵盖的材料包括使用D2O和13C丙酸盐来测量体内的通量。向受试者提供D2O和通过葡萄糖的2H核磁共振测量氢交换提供了糖异生作用(来自PEP或甘油)和糖原分解的相对贡献。给出胶囊中的[U-13C]专一性，并通过13C核磁共振分析葡萄糖的C2碳，给出了PEP的糖异生的相对速率和柠檬酸循环的通量。当两种方法的测量结果结合在一起时，葡萄糖的产量可以用柠檬酸循环周转率来表示。与葡萄糖周转率的测量(通过以恒定速度注入的葡萄糖在血液中的稀释)相结合，上述各种相对测量(各种来源的葡萄糖产量、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方方面面，从设计方案到数据分析和解释。学生将观察处理准备、处理样品、收集核磁共振数据、执行光谱分析以及应用代谢模型完成的程序。提供了在不同阶段所需软件的使用说明。该课程尽可能贴近个人需求。 其次，用户被一对一地教授如何解释人类受试者的1H核磁共振谱。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，拟合和将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. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. 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.