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This volume focuses on protein analysis, including a wide range of the use of mass spectrometry and other protein methods within neurobiological disciplines. Chapters cover topics such as cerebrospinal fluid (CSF) processing and biobanking; label-free quantitative proteomics; SWATH; top-down proteomics; and experimental strategies based on other –omics applied to CSF metabolome, lipidome, and microRNAome. Written in the highly successful Methods in Molecular Biology series format, 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. Cutting-edge and thorough, Cerebrospinal Fluid (CSF) Proteomics: Methods and Protocols is a valuable resource for graduate students and post-doctoral fellows interested in learning more about CSF proteotyping. It is also useful to established researchers seeking further insight into this growing field.
The neuronal ceroid lipofuscinoses are an extremely rare group of inherited neurodegenerative diseases that primarily affect children. Core symptoms of these conditions typically include epilepsy, cognitive decline and visual failure. These diseases are so rare that professionals who come into contact with them need a consultative reference work that enables them to become expert, or identify who to contact for more details. Fully updated and revised, this second edition continues to be the definitive volume on this devastating group of disorders. Written by an international collection of authorities in the field, it provides invaluable advice on their diagnosis, patient care, and new treatments that are available. This new edition of the definitive reference text on the neuronal ceroid lipofuscinoses will prove useful for clinicians, family physicians, research scientists, diagnostic laboratories, families affected by the disease as well as by workers in industry planning translational research.
Cerebrospinal Fluid in Neurologic Disorders, Volume 146 provides a brief overview on the current use of CSF in clinical routine, the physiology of CSF, and its usefulness and potential as a biomarker. The second part addresses the main purpose of the volume, describing CSF from a research perspective in context with the most important diagnostic entities in neurology. The book's authors provide insight into the current understanding of CSF changes in these various conditions and what it tells us about the nature of neurological diseases. Furthermore, methodological aspects are discussed, as are shortcomings that need to be addressed. Finally, the book provides an outlook for potential directions that can be explored to improve the various aspects of CSF research with the ultimate goal of being incorporated in clinical practice. Provides a brief overview on the current use of CSF in clinical routine, the physiology of CSF, and its usefulness and potential as a biomarker Addresses relevant research in context with the most important diagnostic entities in neurology Edited by leading authors in CSF research from around the globe, presenting the broadest, most expert coverage available
The cerebrospinal fluid (CSF) is an invaluable diagnostic tool in clinical neurology, not only in the evaluation of inflammatory, degenerative, and malignant diseases of the nervous system, but also in the diagnosis of all forms of cerebral and subarachnoidal bleedings. The CSF can be easily obtained by lumbar puncture and a set of basic analyses can be conducted using relatively simple laboratory methods. By combining different CSF parameters, a wide range of diagnostic entities can be identified. However, properly interpreting the test results requires a high level of expertise and cannot be achieved by just reporting on individual analytic values. This book covers essential aspects of cerebrospinal fluid analysis and its use in the diagnosis of common neurological diseases. The first part addresses preclinical aspects such as the history of CSF, as well as the anatomical, physiological, and biological background of this valuable fluid. In addition, CSF collection, its preanalytical and methodological implications, and the increasing number of disease-specific markers in CSF are discussed in detail. Lastly, CSF analyses are put into context with clinical syndromes, demonstrating their diagnostic value in neurological clinical practice. Cerebrospinal Fluid in Clinical Neurology helps readers understand the preanalytical and analytical aspects of CSF diagnostics and offers a valuable reference guide for interpreting CSF results during the clinical work-up for neurological patients.
This volume covers the latest methods used in clinical neurochemistry laboratories for both clinical practice and research. Chapters in this book discuss topics such as techniques for cerebrospinal fluid (CSF) collection, pre-analytical processing, and basic CSF analysis; an examination of biomarkers including ELISA and automated immunochemical assays for amyloid and tau markers for Alzheimer’s disease; the analysis of neurofilaments by digital ELISA; and an example of successful novel immunoassay development. In the Neuromethods series style, chapters include the kind of detail and key advice from the specialists needed to get successful results in your laboratory. Cutting-edge and thorough, Cerebrospinal Fluid Biomarkers is a valuable resource for clinicians and researchers to use in CSF labs and CSF courses.
The analysis of the protein content of the Cerebrospinal Fluid (CSF) is a central tool in the diagnosis of a number of diseases, including Multiple Sclerosis and Encephalitis. Proteins of the Cerebrospinal Fluid is a complete revision of the first edition published 16 years ago and reflects the advances in the field in that period. Containing the most comprehensive reference list available today with over 1300 references cross-indexed according to topics, it also includes many classical works that cannot be easily found through the public literature database (like PubMed). This book covers new areas of research such as the use of the analysis of protein content in the CSF to guide the prognosis of patients with severe head injury and/or stroke (currently being tested using technology which can be applied at the bedside to yield results within 15 minutes). With many new or completely revised illustrations, this book serves as not only a reference on the topic, but also a lab manual for bench work with recipes, and a clinical reference on individual diseases. Introduces the Eastern blotting, the technique considered the "gold standard" by the US Food & Drug Administration (FDA) for the demonstration of IgG by immunofixation following isoelectric focusing Provides numerous tables for ease-of-use Extensively cross-referenced with over 1300 references
Protein modifications and changes made to them, as well as the quantities of expressed proteins, can define the various functional stages of the cell. Accordingly, perturbations can lead to various diseases and disorders. As a result, it has become paramount to be able to detect and monitor post-translational modifications and to measure the abundance of proteins within the cell with extreme sensitivity. While protein identification is an almost routine requirement nowadays, reliable techniques for quantifying unmodified proteins (including those that escape detection under standard conditions, such as protein isoforms and membrane proteins) is not routine. Quantitative Methods in Proteomics gives a detailed survey of topics and methods on the principles underlying modern protein analysis, from statistical issues when planning proteomics experiments, to gel-based and mass spectrometry-based applications. The quantification of post-translational modifications is also addressed, followed by the “hot” topics of software and data analysis, as well as various overview chapters which provide a comprehensive overview of existing methods in quantitative proteomics. Written in the successful Methods in Molecular BiologyTM series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Quantitative Methods in Proteomics serves as a comprehensive and competent overview of the important and still growing field of quantitative proteomics.
The treatment of both cancer pain and non-cancer chronic pain is still suboptimal. The overall aim of this PhD thesis was to conduct translational pain research at the interface between clinical pain medicine and the field of human proteomics, using the practice of intrathecal analgesia at our institution as a starting point. Hence, the cerebrospinal fluid (CSF) is at the centre of the present dissertation, both as a target for infusing analgesics (Papers I and II – clinical and pharmacological aspects) and as an important biofluid for human biomarker studies (Papers III and IV – proteomic aspects). In Paper I, 28 cases of intrathecal analgesia in cancer patients were prospectively followed. Movement-evoked breakthrough pain remained a major clinical problem throughout the study month despite otherwise successful intrathecal analgesia (defined as good control of spontaneous resting pain paralleled by a marked decrease of concomitant systemic opioid doses). This study therefore illustrates the importance of considering not only spontaneous resting pain but also movement-evoked breakthrough pain. In Paper II, an expert-based algorithm for trialing the intrathecal analgesic ziconotide by bolus injections was evaluated in an open-label study of 23 patients with chronic neuropathic pain. We found few responders (13%) according to the strict criteria of the algorithm, but ziconotide bolus injection trialing seems feasible. The predictive power of ziconotide bolus trialing remains unclear, and the pharmacological profile of ziconotide (with very slow tissue penetration due to high hydrophilicity) calls the rationale for ziconotide bolus trialing into question. In Paper III, we found low levels of beta-endorphin in the CSF of chronic neuropathic pain patients (n=15) compared to healthy controls (n=19). We speculate that this might indicate dysfunctional top-down control of nociception. Substance P levels in the CSF did not differ by univariate statistics. InPaper IV, the CSF proteome of 11 patients with chronic neuropathic pain and 11 healthy controls was exploratively studied, combining gel-based proteomics with multivariate data analysis. After eliminating four proteins associated with age, 32 proteins were found to highly discriminate between groups. Among these, the seven proteins having the highest discriminatory power between patients and controls were: one isoform of angiotensinogen, two isoforms of alpha-1-antitrypsin, three isoforms of haptoglobin, and one isoform of pigment epithelium-derived factor. In conclusion, this PhD thesis demonstrates the fruitfulness of studying the CSF, both as a target for infusing analgesics and as a potential mirror of the neurobiological processes involved in pathological pain conditions. The thesis points to the need for more research into the mechanisms of different pain conditions, in order to hopefully achieve the vision of mechanism-based pain diagnoses.
Cerebrospinal fluid (CSF) is a clear colourless fluid and the major part of the extracellular fluid of the central nervous system. The CSF fills the brain ventricles and central spinal canal and forms a thin layer around the brain and spinal cord in the subarachnoid space and cisterns. In this book, the authors present current research in the study of the functions, composition and disorders relating to the cerebrospinal fluid. Topics include the proteomics of cerebral microdialysate; neuronal cytoskeleton components in cerebrospinal fluid in selected neurological diseases; composition of normal and pathological cerebrospinal fluid in neuropathology; cerebrospinal fluid flow dynamics; and cerebrospinal fluid biomarkers for Alzheimer's disease.