This book came about as a result of a review I had written earlier on fea tures of cellular changes occurring during anuran metamorphosis. Only a limited treatment of this subject was possible in such a circumscribed work and only specific examples of organic change were dealt with. Thus the sins of omission weighed heavily, for so much information could not be included to provide a more comprehensive and authenticated account of the elaborate, complex, and far-reaching changes that an aquatic larva undergoes to become a terrestrial froglet. A good deal of my working life has been spent investigating amphib ians, especially their larval developmental morphology during metamor phosis, first at the level of light microscopy and in later years by electronmicroscopy. Initially I was particularly concerned with morpho logical homologies of a variety of larval structures, such as the cranial and pharyngeal skeleton and the nerves and musculature, in order to learn more about amphibian phylogeny, for during my pre-and early postgrad uate years G. R. Beer and D. M. S. Watson inspired an undying interest in and respect for vertebrate comparative anatomy. However, it now seems to be that amphibian phylogenetic relationships are best dealt with by the paleontologists, so ably demonstrated by D. M. S. Watson and A. S. Romer and the contemporary enthusiasts in this field like A. L. Panchen, R. L. Carroll, E. Jarvik, and K. S. Thompson among a host of others, particularly in the USA."

This book contains 14 original review chapters each yielding new, exciting and intriguing data about the emerging understanding of nucleolar structure and function in normal, stressed and diseased cells. The goal of this work is to provide special insight into the nucleolus of the past, present and future, as well its regulation, translocation, and biomedical function. A multitude of topics are introduced and discussed in detail, including nucleologenesis, nucleolar architecture, nucleolar targeting, retention, anchoring, translocation, and the relationship between the nucleolus and cancer. This book also brings together work from several different species, from human to Drosophila to Dictyostelium and other eukaryotic microbes. The final chapter summarizes some of the issues brought up in the various chapters with a view to future research. This book supports the continued emergence of the nucleolus as a dynamic intranuclear region that oversees a vast diversity of events.

Given the vital importance of immune system research, the gathering of clear, consistent, and informative protocols involving the study of dendritic cells is paramount. Bringing the popular first edition fully up to date, Dendritic Cell Protocols, Second Edition presents protocols from experts in the field that cover the basics and more complex forays into the exploration of DC development and function, both in mice and humans. The first section of the volume involving humans explores topics such as the isolation of blood DC subtypes, primary skin Langerhans cells, and the generation of gene-manipulated human DCs with the inclusion of more clinically relevant methods as well, while the second section involving rodent models delves into DC and precursor generation in vitro, isolation ex vivo, disease models, as well as DC functions and properties. Written in the highly successful Methods in Molecular Biologya" series style, chapters include introductions to their respective subjects, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls.

Comprehensive and cutting-edge, Dendritic Cell Protocols, Second Edition aims to become a bench-side handbook for both beginners and experts in the field of DC research and a long-term reference for some of the most popular methods put forward by those who lead the field.

Biofilms affect the lives of all of us, growing as they do for example on our teeth (as plaque), on catheters and medical implants in our bodies, on our boats and ships, in food processing environments, and in drinking and industrial water treatment systems. They are highly complex biological communities whose detailed structure and functioning is only gradually being unravelled, with the development of increasingly sophisticated technology for their study. Biofilms almost always have a negative impact on human affairs (flocs in sewage treatment plants are a major exception) and a lot of research is being carried out to gain a better understanding of them, so that we will be in a better position to control them.
This volume, with contributions by international experts from widely diverse areas of this field, presents a state-of-the-art picture of where we are at present in terms of our knowledge of biofilms, the techniques being used to study them, and possible strategies for controlling their growth more successfully. It should provide a valuable reference source for information on biofilms and their control for many years to come.

Numerous animal species live in environments characterized by a seasonal reduction in the availability of water, which often but not always occurs when temperatures are highest. For many such animals, survival during the toughest season requires spending long periods of time in a rather inactive state known as aestivation. But aestivation is much more than remaining inactive. Successful aestivation requires the selection of a proper microhabitat, variable degrees of metabolic arrest and responsiveness to external stimuli, the ability to sense the proper time of year for emergence, the preservation of inactive tissue, and much more. So, aestivation involves a complex collection of behaviors, ecological associations and physiological adjustments that vary across species in their type, magnitude and course. This book seeks to explore the phenomenon of aestivation from different perspectives and levels of organization, ranging from microhabitat selection to genetic control of physiological adjustments. It brings together authors from across the world working on different systematic groups, approaches, and questions, but who are all ultimately working to better understand the complex issue of aestivation.

Dictyostelium discoideum Protocols presents a comprehensive collection of cutting-edge molecular biological protocols for studying D. discoideum, with emphasis on its utility in the study of fundamental cellular processes including signal transduction, chemotaxis, cell motility, cytokinesis, phagocytosis, and aspects of development such as cell sorting, pattern formation, and cell type differentiation. Written by experts in the field, this important guide provides easy access to current methods and techniques for the study of D. discoideum. For the newcomer to the field, it offers a complete and inclusive introduction to this important model organism. The book is divided into four main parts, which include an introduction to the organism that provides integral community resources and genome-wide approaches; basic methods and available molecular genetic techniques for study of the organism; imaging and localization methods; and a discussion of D. discoideum, with emphasis on its unique advantages as a model system.
This in-depth guide provides a complete introduction to and thorough handbook for the study of D. discoideum, including a discussion of the implications of the recently-completed Dictyostelium genome project in strengthening the position of D. discoideum as a model organism for studying fundamental cell processes and aspects of development. Researchers at all levels of experience will benefit from the presentations throughout of the most current, useful and innovative techniques for the study of D. discoideum from leading Dictyostelium scientists.

The fermentation of sugar by cell-free yeast extracts was demonstrated more than a century ago by E. Buchner (Nobel Prize 1907). Buchner's observations put an end to previous animistic theories regarding cellular life. It became clear that metabolism and all cellular functions should be accessible to explication in chemical terms. Equally important for an understanding of living systems was the concept, explained in physical terms, that all living things could be cons- ered as energy converters [E. Schroedinger (Nobel Prize 1933)] which generate complexity at the expense of an increase in entropy in their environment. Bioenergetics was established as an essential branch of the biochemical sciences by the investigations into the chemistry of photosynthesis in i- lated plant organelles [O. Warburg (Nobel Prize 1931)] and by the discovery that mitochondria were the morphological equivalent that catalyzed cellular respiration. The ?eld of bioenergetics also encompasses a large variety of ad- tional processes such as the molecular mechanisms of muscle contraction, the structure and driving mechanisms of microbial ?agellar motors, the energetics of solute transport, the extrusion of macromolecules across membranes, the transformation of quanta of light into visual information and the maintenance of complex synaptic communications. There are many other examples which, in most cases, may perform secondary energy transformations, utilizing - ergy stored either in the cellular ATP pool or in electrochemical membrane potentials.

Researched and written by a group of highly respected professionals in the fields of biochemistry, microbiology, and molecular genetics, this resource offers a comprehensive treatment on the role of metal ions in regulating genes. In addition to looking at the toxicity effects of metal, this text explores the role of metal ions in normal metabolisms, examining both prokaryotes and eukaryotes. Metal Ions in Gene Regulation should prove an essential reference for all microbiologists, biochemists, bioinorganic chemists, and molecular biologists, especially those interested in gene regulation.

The fluid-mosaic model of membrane structure formulated by Singer and Nicolson in the early 1970s has proven to be a durable concept in terms of the principles governing the organization of the constituent lipids and proteins. During the past 30 or so years a great deal of information has accumulated on the composition of various cell membranes and how this is related to the dif ferent functions that membranes perform. Nevertheless, the task of explaining particular functions at the molecular level has been hampered by lack of struc tural detail at the atomic level. The reason for this is primarily the difficulty of crystallizing membrane proteins which require strategies that differ from those used to crystallize soluble proteins. The unique exception is bacteriorhodopsin of the purple membrane of Halobacterium halobium which is interpolated into a membrane that is neither fluid nor in a mosaic configuration. To date only 50 or so membrane proteins have been characterised to atomic resolution by diffraction methods, in contrast to the vast data accumulated on soluble proteins. Another factor that has been difficult to explain is the reason why the lipid compliment of membranes is often extremely complex. Many hundreds of different molecular species of lipid can be identified in some membranes. Remarkably, the particular composition of each membrane appears to be main tained within relatively narrow limits and its identity distinguished from other morphologically-distinct membranes.

Over time, it has become clear that changes in stem cells do occur during aging, not only in their number but also in their relationship to their microenvironment and their functionality as reflected in changes to their metabolome. Stem Cells and Aging: Methods and Protocols brings together chapters from expert contributors with protocols critical for exploring the biology of stem cell aging, all of which is key for understanding these age-related stem cell changes at a basic biology level and at the level of their impacts for regenerative medicine. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and tips on troubleshooting and avoiding known pitfalls. Concise and easy to use, Stem Cells and Aging: Methods and Protocols serves as an ideal reference to guide investigators toward further valuable answers to the problems of our aging population.

Every cell has developed mechanisms to respond to changes in its environment and to adapt its growth and metabolism to unfavorable conditions. The unicellular eukaryote yeast has long proven as a particularly useful model system for the analysis of cellular stress responses, and the completion of the yeast genome sequence has only added to its powerThis volume comprehensively reviews both the basic features of the yeast genral stress response and the specific adapations to different stress types (nutrient depletion, osmotic and heat shock as well as salt and oxidative stress). It includes the latest findings in the field and discusses the implications for the analysis of stress response mechanisms in higher eukaryotes as well.

The Fifth International Lymphokine Workshop was convened in Clearwater Beach, Florida, January 11-15, 1987. The theme chosen for the meeting was 'The Molecular Basis of Lymphokine Action," which reflected the opinion of the organizers as to how far the field had moved since the first Lymphokine Workshop only eleven years ago. As was evident at the last Lymphokine Workshop held in 1985, the contribution of molecular biology, particularly in the cloning of lymphokine genes, continues to play an important role in clarifying the structure of lymphokines, providing recombinant (read "pure") proteins for biological studies, and suggesting directions for studies of the molecular basis of lymphokine activity. The most recent lymphokines to yield to molecular cloning meth odology were the B-cell growth and differentiation factors, in partic ular BSF-1 or, as it is sometimes termed, interleukin 4. One of the surprises from this research is the broad spectrum of activities that can be attributed to this molecule, aside from its effects on B-cells, thus perhaps justifying its being called an interleukin. The interleukin 2 symposium demonstrated that even in a well-established research area, controversy and excitement can continue, when evidence was presented by several investigators indicating the presence of a second "converter" protein that changes the affinity of the now classical Tac antigen from a low to a high affinity IL-2 receptor."

Current biological research demands the extensive use of sophisticated mathematical methods and computer-aided analysis of experiments and data. This highly interdisciplinary volume focuses on structural, dynamical and functional aspects of cellular systems and presents corresponding experiments and mathematical models. The book may serve as an introduction for biologists, mathematicians and physicists to key questions in cellular systems which can be studied with mathematical models. Recent model approaches are presented with applications in cellular metabolism, intra- and intercellular signaling, cellular mechanics, network dynamics and pattern formation. In addition, applied issues such as tumor cell growth, dynamics of the immune system and biotechnology are included.

This volume explores pericytes' roles under distinct pathological conditions, ranging from tumors, ALS, Alzheimer's disease, Multiple Sclerosis, stroke, diabetes, atherosclerosis, muscular dystrophies and more. Together with its companion volumes Pericyte Biology in Different Organs and Pericyte Biology - Novel Concepts, Pericyte Biology in Disease presents a comprehensive update on the latest information and most novel functions attributed to pericytes. To those researchers newer to this area, it will be useful to have the background information on these cells' unique history. It will be invaluable for both advanced cell biology students as well as researchers in cell biology, stem cell biology and clinicians involved with these specific diseases.

Free radicals and oxidative damage in biology and medicine: An introduction.- Oxidative metabolism in skeletal muscle.- Strategies to assess oxidative stress.- The course of exercise-induced skeletal muscle fibre injury.- Free radical mechanisms in exercise-related muscle damage.- The effects of exercise, ageing and caloric restriction on protein oxidation and DNA damage in skeletal muscle.- Antioxidant enzyme response to exercise and training in the skeletal muscle.- Glutathione: A key role in skeletal muscle metabolism.- Vitamin E and its effect on skeletal muscle.- Differential susceptibility of skeletal muscle proteins to free radical-induced oxidative damage in vitro.- Oxidative stress and Ca2+ transport in skeletal and cardiac sarcoplasmic reticulum.- Oxidative stress in skeletal muscle atrophy induced by immobilization.- Effect of growth hormone on oxidative stress in immobilized muscles of old animals.- The diaphragm and oxidative stress.- Oxidative damage after ischemia/reperfusion in skeletal muscle.- Oxidative damage in rat skeletal muscle after excessive L-tryptophan and atherogenic diets.- Oxidative stress and muscle wasting of cachexia.- Free radicals and antioxidants in the pathogenesis of alcoholic myopathy.- Drug-induced muscle damage.- Free radicals and diseases of animal muscle.- Therapeutic trials of antioxidants in muscle diseases.

This detailed volume provides a comprehensive resource covering the existing and state-of-the-art tools in the field of profiling chromatin accessibility and its dynamics. Beginning with a section on bulk-cell methods for profiling chromatin accessibility and nucleosome positioning that rely on enzymatic cleavage of accessible DNA and produce information about relative accessibility, the book continues with methods that use single-molecule and enzymatic approaches to solving the problem of mapping absolute occupancy/accessibility, emerging tools for mapping DNA accessibility and nucleosome positioning in single cells, imaging-based methods for visualizing accessible chromatin in its nuclear context, as well as computational methods for the processing and analysis of chromatin accessibility datasets. Written for 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 and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and up-to-date, Chromatin Accessibility: Methods and Protocols serves as an extensive and useful reference for researchers studying different facets of chromatin accessibility in a wide variety of biological contexts. Chapter 6 is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.

In the recent years, a significant amount of research has emerged connecting the link between alcohol and cancer. The field has rapidly advanced, especially since the complex connection between alcohol and cancer has several unique sub areas that are being investigated. This proceedings volume will contain chapters based upon the presentation of the 2nd International Conference on Alcohol and Cancer in Colorado, 2013. The various topics explore the affects of alcohol on: liver and breast cancer; cell signaling and cancer; stem cells; biomarkers and metabolomics; aerodigestive cancers; cancer and the immune system and more.

The potato (Solanum tuberosum L. ) tuber is a major food source in many countries of the world, and subsequently potato has been the target of a good deal of effort directed at engineering disease and herbicide tolerance, and improvements in various crop characteristics. Consequently investigations into the regulation of gene expression in tubers is relevant to these endeavours, as tubers are the main target organ for modification of gene expression. We have been interested in the regulation of genes in tubers for these reasons. Morphologically tubers are modified stems, which have enlarged radially by limited cell division and substantial expansion. At the molecular level, tuber development is characterised by a massive increase in starch deposition and the synthesis of a limited number of abundant proteins. These include proteinase inhibitors and a 40kd group of proteins called patatin, which are acyl hydrolases. Together these proteins account for over 50% of tuber proteins (reviewed by Bevan, 1991). The synthesis of these proteins has parallels to the synthesis of other somatic storage proteins, especially the VSP proteins of soybean. In both potato and soybean, removal of the sink for these proteins (tubers and pods, respectively) causes deposition in other tissues (Staswick, 1990). It is hypothesised that transcriptional control of the genes encoding these proteins is regulated in part by source-sink relationships of metabolites or other factors. In the case of VSPs, both amino acid levels and jasmonic acid play a major regulatory role (Staswick et aI.

This introductory guide provides novice researchers and lab students with a thorough step-by-step approach to standard animal cell culture techniques. Coverage includes lab safety and best practices, sterility management, preparation, ethical considerations, and troubleshooting for common pain points. This is an up-to-date, indispensable handbook for early-career researchers and students, as well as established scientists in biotechnology, cell and developmental biology, pharmaceutical toxicology, cytogenetics, and more.

This detailed volume provides a comprehensive overview of state-of-the-art metabolomics methods based on mass spectrometry (MS), and their application in food, nutrition, and biomedical research. The chapters assembled herein cover hot topics related to sample preparation, chromatographic and electrophoretic separation, MS-based analysis, as well as data processing and analysis. Written for 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 and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Mass Spectrometry for Metabolomics serves as a timely guide for chemists, biochemists, biologists, nutritionists, clinicians, and other experts working in the growing and exciting field of metabolomics.

In recent years, a new understanding of cell-penetrating peptides has emerged, helping researchers to expand beyond a number of long-held dogmas. In Cell-Penetrating Peptides: Methods and Protocols, expert researchers explore the latest information on cell-penetrating peptides (CPPs), providing insight into the most important and contemporary areas of CPP research. Chapters address the historical background of CPP studies, provide an overview of the growing field of research, investigate methods for testing CPP mechanisms, present a summary of methods that attempt to use properties of CPPs to study biochemical intracellular mechanisms of interaction and signal transduction, and include new ideas for turning CPP-based strategies into drugs. Composed in the highly successful Methods in Molecular Biology(TM) series format, each chapter contains a brief introduction, step-by-step methods, a list of necessary materials, and a Notes section which shares tips on troubleshooting and avoiding known pitfalls. Innovative and current, Cell-Penetrating Peptides: Methods and Protocols is an essential guide that allows researchers to study intracellular mechanisms in new ways, and to promote the future development of novel drugs.

All the information necessary to set up and run a tissue culture facility is provided in this introductory book.; ; Includes an overview of all the basic tissue culture techniques and describes in detail both the theoretical background and the practical a

This book describes these exciting new developments, and presents experimental and computational findings that altogether describe the frontier of knowledge in cellular and biomolecular mechanics, and the biological implications, in health and disease. The book is written for bioengineers with interest in cellular mechanics, for biophysicists, biochemists, medical researchers and all other professionals with interest in how cells produce and respond to mechanical loads.