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In the past few years, the number of applications of tracers for in vivo biomedical studies has greatly increased. New analytical tools at the genetic and protein levels have spurred this growth, opening the door for a deeper understanding of metabolic events. This in turn promises to yield significant advances in the understanding and treatment of human disease. Now fully revised and expanded, Isotope Tracers in Metabolic Research, Second Edition is the established definitive text on stable and radioactive isotope tracers. In unique, multidisciplinary fashion, it presents comprehensive coverage of new methodological, mathematical, and theoretical approaches. This new Second Edition includes: All-new chapters on nuclear magnetic resonance, mass isotopomer analysis, and methods of protein metabolism analysis A completely updated categorized list of over 750 references Major advances in the development of mass isotopomer and positional isotopomer techniques, noninvasive isotope techniques for studying metabolic pathways, hyphenated techniques, and new tracer techniques The latest developments in quantification of DNA synthesis and mass spectrometry spurred by genome sequencing and proteomics New coverage of mathematical modeling Expanded coverage of microdialysis probes, laboratory procedures, and regulatory issues related to human studies In this complete guide to performing tracer studies, the authors systematically cover tracer selection, modeling considerations, sample derivitization, mass spectrometry analysis, and data interpretation. Problems and discussion questions highlight key points in each chapter. Isotope Tracers in Metabolic Research, Second Edition offers students and researchers a comprehensive, practical resource for utilizing the latest tracer methodologies.
An up-to-date, detailed, authoritative overview regarding all aspects of the theory and practice involved in utilizing isotopic tracers for the in vivo study of human metabolic function. Presents specific technical information on performing tracer studies including isotope infusion, analysis and interpretation of data.
The purpose of this monograph is to describe theoretical aspects of the interpretation of data obtained from experiments performed with labeled hormones. Quantitative endocrinologic studies involving the use of tracers include the determination of rates at which hormones are secreted by endocrine glands and are produced outside these glands by conversion of other secreted hor mones. Tracer experiments are also performed with the purpose of measuring rates of metabolic reactions. These measurements reveal the contribution of secreted hormones to the formation of circulating compounds and urinary metabolites. The estimation of rates of fetal and placental production and exchange of hormones characterizes a class of in vivo quantitative studies performed with isotopically labeled hormones (radioactive or not). In addi tion, tracers are used to measure permeability and rates of reaction in in vitro systems, and to study the uptake of hormones by tissues, both in vivo and in vitro. The stability of the steroid nucleus carrying the isotopic label and the large number of reversible metabolic reactions in which steroids are involved, both facilitated and motivated the development of a sophisticated theoretical treat ment of tracer experiments in the field of endocrinology. Although the prac tical examples used to illustrate the concepts and calculations presented in this monograph involve labeled hormones, the theory is presented in a general symbolic manner and is applicable to other fields of investigation.
This detailed volume focuses on recent technological, computational, and biostatistical advances in the field of high-throughput metabolomics. Chapters encompass methods, platforms, and analytical strategies for steady state measurements and metabolic flux analysis with stable isotope-labeled tracers, in biological matrices of clinical relevance and model organisms. Mass spectrometry-based or orthogonal methods are discussed, along with computational and statistical methods to address data sparsity in high-throughput metabolomics approaches. As a part of the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, High-Throughput Metabolomics: Methods and Protocols provides tools that can bring about the next generation of clinical biochemistry in a cost-effective, rigorous fashion, exponentially advancing our capacity to investigate nature while hastening the advent of personalized medicine.
Methods for studying metabolism. Determination of substrate kinetics. Constant isotope infusion technique. Determination of substrate kinetics. Bolus-injection technique. Determination of substrate oxidation. Bicarbonate kinetics and natural enrichment. Priming the pool. Nonsteady state kinetics. Selection of sites for infusing isotope andj drawing samples. Specific application. Glucose metabolism. Specific application. Urea. Specific application. Fat meltabolism. Amino acids. Whole-body protein turnover. Total body water. Clinical application of stable isotopes. Analytical methods. Instrumentation. Mass spectrometry. Selected ion monitoring. Preparation of samples. Chromatography. Determination of specific radioactivity. Collection of CO2. Isotope ratio mass spectrometry techniques. Gas chromatography-mass spectrometry methods. In vivo approaches. Isotopic tracers. Advantages of stable isotopes. Biological effects. Enzymatic effects. Definition of terms. Influence of uptake on calculation of rate of appearance. Influence of pool size and complexity. Net movement of tracer and tracee. Clearance. Stable versus radioisotopes. Single-pool analysis. Bicarbonate recovery. Natural abundance of 13C. Practical consideration of performing recovery studies. Determining NaH13 CO3 infusion rate. Determining VCO2. Theory of priming dose. Priming secondary pools. Steele equation. In vivo assessment of steele equation. Glucose. Effective volume of distribution. Urea. Arterial versus central venous infusion.
1. Introduction. 2. Fundamentals of Tracer Kinetics. 3. The Noncompartmental Model of Multipool Systems. 4. The Compartmental Model. 5. Identifiability of the Tracer Model. 6. Using the Tracer Model to Estimate Kinetic Parameters. 7. Compartmental Versus Noncompartmental Kinetic Parameters. 8. Parameter Estimation: Some Fundamentals of Regression Analysis. 9. Parameter Estimation in Noncompartmental Models. 10. Parameter Estimation in Compartmental Models. 11. Precursor-Product Models. Appendices. Index.
This special volume "Tools and Applications of Biochemical Engineering Science" is dedicated to Professor Wolf-Dieter Deckwer on the occasion of his 60th bir- day. It was a great pleasure for me to act together with Professor Karl Schtigerl as volume editor and to present here a collection of 11 outstanding review articles written mainly by former students, associates, colleagues and friends of Wolf- Dieter Deckwer. The title of this special volume well reflects the research interests and sci- tific pursuit of Wolf-Dieter Deckwer during his more than 20 years’ work in the area of biochemical engineering, particularly during the last 15 years when he was the head of the Biochemical Engineering Division of GBF (German Nat- nal Research Center for Biotechnology). He has decisively pushed the devel- ment not only of "software tools" ranging from analytical means and mathe- tical models for monitoring and understanding cellular processes to gene expression systems for designing microorganisms, but also of "hardware tools" such as computer control systems, bioreaction and separation devices for eff- tively producing a variety of bioproducts on semi-production scale. New developments in some of these important tools in biochemical engineering are reviewed in articles included in this volume. Wolf-Dieter Deckwer was among the leading biochemical engineers who timely pointed out the necessity of applying these tools in an integrated manner for bioprocess development. By establishing "Integrated Bioprocess Development" as one of the GBF main - search topics as early as 1990 he also actively promoted this idea.
This book aims to aid the selection of the most appropriate methods for use in early phase (1 and 2) clinical studies of new drugs for diabetes, obesity, non-alcoholic fatty liver disease (NAFLD) and related cardiometabolic disorders. Clinical research methods to assess the pharmacokinetics and pharmacodynamics of new diabetes drugs, e.g. the euglycemic clamp technique, have become well-established in proof-of-mechanism studies. However, selection of the most appropriate techniques is by no means straightforward. Moreover, the application of such methods must conform to the regulatory requirements for new drugs. This book discusses the need for new pharmacotherapies for diabetes, obesity and NAFLD and the molecular targets of drugs currently in development. Emerging technologies including functional imaging, circulating biomarkers and omics are considered together with practical and ethical issues pertaining to early phase clinical trials in subjects with cardiometabolic disorders. Translational Research Methods in Diabetes, Obesity, and Non-Alcoholic Fatty Liver Disease is of interest to biomedical scientists, pharmacologists, academics involved in metabolic research and clinicians practicing in these specialties.
This book covers the new Omics area, Metallomics. As Metallomics is intrinsically a transdisciplinary area, this book is authored by experts in the field on such diverse topics as Environmental, Nuclear, and Human Metallomics. Within these topics metals play important role, as being part of biomolecules, controlling different biochemical process, being signaling agents, being catalyst of biochemical reactions, among others. This volume demonstrates the importance of more investigation about metals and their interactions with biomolecules. As the knowledge in this field is growing and growing daily, then new challenges concerning studies involving Metallomics is appearing, such as comparative metallomics, speciation metallomics, real-time metallomics, new predictions of metals in biomolecules, metalloprotein databank expansion, interactions between metalloprotein-metalloprotein, among others.
Radioactive isotopes and enriched stable isotopes are used widely in medicine, agriculture, industry, and science, where their application allows us to perform many tasks more accurately, more simply, less expensively, and more quickly than would otherwise be possible. Indeed, in many casesâ€"for example, biological tracersâ€"there is no alternative. In a stellar example of "technology transfer" that began before the term was popular, the Department of Energy (DOE) and its predecessors has supported the development and application of isotopes and their transfer to the private sector. The DOE is now at an important crossroads: Isotope production has suffered as support for DOE's laboratories has declined. In response to a DOE request, this book is an intensive examination of isotope production and availability, including the education and training of those who will be needed to sustain the flow of radioactive and stable materials from their sources to the laboratories and medical care facilities in which they are used. Chapters include an examination of enriched stable isotopes; reactor and accelerator-produced radionuclides; partnerships among industries, national laboratories, and universities; and national isotope policy.