This volume collects a series of protocols describing the kinds of infrastructures, training, and standard operating procedures currently available to actualize the potential of stem cells for regenerative therapies. Stem Cells and Good Manufacturing Practices: Methods, Protocols, and Regulations pulls together key GMP techniques from laboratories around the world. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Inclusive and authoritative, Stem Cells and Good Manufacturing Practices: Methods, Protocols, and Regulations will be an invaluable resource to both basic and clinical practitioners in stem cell biology.

David Kuter and a host of leading international researchers summarize in one volume all the knowledge of thrombopoietins (TPO) available today. The distinguished experts review the history of the search to discover TPO, describe the molecular and biological characteristics of this new molecule, and present the results of the preclinical animal experiments that will guide clinical use of this new hormone. Along the way they provide the most recent and comprehensive guide to the biology of megakaryocytes and platelets.

Progress in Cell Cycle Research is a new annual series designed to be the source for up-to-date research on this rapidly expanding field. Review articles by international experts examine various aspects of cell division regulation from fundamental perspectives to potential medical applications. Researchers as well as advanced undergraduate and graduate students in cell biology, biochemistry, and molecular biology will benefit from this series.

One of the mysteries of mammalian reproduction is the physiologic process that determines the length of gestation. The proper timing of birth ensures that the young individual is sufficiently developed to survive and adapt in the extrauterine environment, and that the mother is capable to provide nutrition and protection to the newborn. This volume summarizes new knowledge obtained by many researchers seeking to unravel the compile mechanisms that contribute to the maintenance and termination of pregnancy. The most important common goal of these efforts is to reduce the incidence of preterm birth, which is the leading cause of perinatal morbidity and mortality in numerous countries.
Separate chapters are devoted to the best-studied animal models of parturition. In sheep, the fetus is in control of the timing of its own birth, while in avian species, oviposition is evidently determined by the female laying the feritlized egg. In humans and non-human primates, the roles of the fetus and the mother are more balanced, and involve a complicated and poorly understood interplay between the mother, the fetus, and the placenta. Some major aspects of these interactions, such as trophoblast function, myometrial contractility, and the endocrine-paracrine systems, are discussed in further chapters.

There are probably few people who do not dream of the good old times, when do ing science often meant fascination, excitement, even adventure. In our time, do ing science involves often technology and, perhaps, even business. But there are still niches where curiosity and fascination have their place. The subject of this book, technological as its title may sound, is one of the fortunate examples. It will report on lasers generating the coldest places in the Universe, and on table top laser microtools which can produce a heat "inferno" as it prevails in the interior of the Sun, or simulate, for specific plant cells, microgravity of the space around our plan et Earth. There will be some real surprises for the reader. The applications range from basic studies of the driving forces of cell division (and thus life) via genetic modification of cells (for example, for plant breeding) to medical applications such as blood cell analysis and finally in vitro fertilization. What are these instruments: laser microbeams and optical tweezers? Both are lasers coupled with a fluorescence microscope. The laser microbeam uses a pulsed ultraviolet laser. Light is focused, as well as possible, in space and time, in order to obtain extremely high light intensities - high enough to generate, for a very short instant, extremely hot spots which can be used to cut, fuse or perforate biological material."

This is the first in a series of volumes concerning the properties of the eukaryotic nucleus. Contributions from several of the most active laboratories are brought together to present a focused overview of a selected aspect of nuclear structure and function.

For many years, the authors have investigated the adaptive role of heat shock proteins (HSPs) in different animals, including the representatives of homothermic and poikilothermic organisms that inhabit regions with contrasting thermal conditions. This book will summarize the data accumulated in the course of these studies and describe the general molecular mechanisms underlying the adaptation of various organisms to aggressive environments. We also concentrate on different evolutionary trends characteristic for HSP systems in the course of adaptation to fluctuating environmental conditions. In addition, we describe the peculiarities in the regulatory regions of heat shock genes necessary for fine tuning of these systems providing the adaptation to adverse conditions. Special emphasis is given to the role of mobile elements in the evolution and functioning of various groups of HSP genes. The book combines the results of field studies and laboratory analysis of stress genes systems.

The migration of stem cells has been found to be critical during early development for the organization of the embryonic body as well as during adult life with tissue homeostasis and regeneration of organ function. Due to the low frequency of these cells in vivo, problems in identifying and prospectively purifying tissue specific stem cells near homogeneity, and, most importantly, a lack of adequate technologies and protocols to study stem cell migration in vivo, this vital research has been quite difficult until recently. In "Stem Cell Migration: Methods and Protocols," experts in the field compile and highlight the standard and novel techniques that allow the studying of the migration of stem cells in one succinct manual. Including protocols on germ, neuronal, and hematopoietic stem cells, during development and adulthood with a clear emphasis on in vivo technologies, the volume also extends its coverage to in vitro approaches toward several developmentally-conserved signaling pathways. 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.

Practical and convenient, "Stem Cell Migration: Methods and Protocols" provides key, state-of the art information on experimental techniques for studying stem cell migration both at a cellular and molecular level in development, regeneration, and disease.

The incentive for putting together Volume 4 of this series was to review the wealth of new information that has become available in prokaryotic organisms in protein export and membrane biogenesis. Just in the last several years, protein translocation has now been efficiently reconstituted using defined components and the mechanism by which proteins are moved across membrane bilayers is now being examined at a higher resolution. In addition, because of a new technical breakthrough using osmolytes, it is now possible to reconstitute a number of channel proteins, ATPase, receptors, and transporters. In many cases, it is possible to successfully predict the membrane topology of these types of proteins using both "hydrophobicity analysis" and the "positive inside" rule.
In this volume, two chapters focus on protein translocation across membranes ("Biochemical Analyses of Components Comprising the Protein Translocation Machinery of E. Coli; Protein Translocation Genetics"), while several others on how proteins assemble into the ineer membrane of E. Coli ("Membrane Protein Assembly; Membrane Insertion of Small Proteins: Evolutionary and Functional Aspects; Pigment-Protein Complex Assembly in Rhodobacter sphaeroides and Rhodobacter Capsulatus"). Other sections review recent progress on transporters ("Identification and Reconstitution of Anion Exchange Mechanisms in Bacteria; Helic Packing in the C-Terminal Half of Lactose Permease") and signal transduction ("Mechanism of Transmembrane Signaling in Osmoregulation") as well as the assembly of prints into the outer membrane ("Export and Assembly of Outer Membrane Proteins in E. coli"). Although the emphasis of the book is on proteins, the role of phospholipids in controlling various cell surface processes is reviewed ("Role of Phospholipids in coli Cell Function"). I should point out the reason for the rapid progress in bacteria research is because of the possibility to apply biochemistry and genetics in this organism.

This book addresses the most recent advances in the transport of proteins across a variety of biological membranes. In addressing this topic, this volume includes several new twists not previously addressed in the literature. In the last few years, the study of protein translocation has been revolutionized by the availability of structural information on many of the components and complexes involved in the process. Unlike earlier books written on protein translocation, this volume considers these advances. In addition, several chapters discuss facets of protein translocation from a systems biology perspective, considered by many to be the next paradigm for biological study. Readers of this book will come away with a deeper understanding of the problems facing researchers of protein translocation and see how the most modern biological techniques and approaches are being recruited to answer those questions. The chapters are also written such that problems awaiting future investigation are clearly presented.

This detailed volume provides thorough protocols describing how to use genetics to study mouse and zebrafish erythropoiesis in whole animal models and for genetically manipulating cultured mouse and human erythroid cells. Protocols include strategies to analyze circulating red blood cell parameters, as well as progenitor cell status, stage of differentiation, and enucleation. Methods for the study of erythroid cell gene regulation using chromatin immunoprecipitation and chromatin conformation capture are detailed. Furthermore, protocols are provided to examine stress erythropoiesis, erythroblastic islands, and erythroblast structure. The collection also includes reviews on the available mouse models of erythropoiesis, as well as on future considerations for good manufacturing practice in order to translate the manufacture of erythrocytes to the clinic. 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, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Erythropoiesis: Methods and Protocols provides a definitive source for molecular and cellular biology protocols to study erythropoiesis and to move this vital field forward.

This volume covers data describing the role of free radicals and antioxidants that deal with clinical and pre-clinical trials, as well as basic research in the area of women's health. There is increasing evidence that oxidative stress is a causative, or at least a supporting factor in female pathology and infertility. During advancing gestational age, oxidative stress biomakers rise. Oxidative stress plays a regulatory role in transcription, signal transduction, gene expression and membrane trafficking. A search on Pub Med shows 449 papers have been published to date related to women's health disorders and use of antioxidants in a variety of disease that are prevalent in women, such as hypertension and cardiovascular disease, osteoporosis, obesity and restless leg syndrome.

Cilia: From Mechanisms to Disease Part B, Volume 176 of Methods in Cell Biology series, highlights new advances in the field, with this new volume presenting interesting chapters. Each chapter is written by an international board of authors.

This volume provides a comprehensive list of protocols for molecular biologists, biochemists and geneticists. Chapters cover protocols that further the study into protein complexes that modify chromatin either by adding or removing post-translational modifications, or by exchanging histone variants within the nucleosome. 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. Authoritative and cutting-edge, Histones: Methods and Protocols aims to ensure successful results in the further study of this vital field.

This book focuses on issues in clinical practice and research that are of general interest. The articles primarily focus on understanding the pathogenic mechanisms of diseases, their prevention, and therapy. The topics addressed include cardiovascular regulation with regard to blood pressure and heart rate variability, and to coupling blood pressure changes with subarachnoid fluid oscillations. In addition, the book discusses recent advances in the diagnostics of and targeted molecular therapy for renal and pancreatic malignancies, growth disorders, vitamin D and calcium homeostasis in children in the context of neonatal urolithiasis, and neurosurgical interventions in multifarious age-related diseases of the vertebrae. Neuropsychological aspects of patients' quality of life and of shaping medical staff's attitude toward patients are also addressed. The respective articles are intended to build a bridge between basic and clinical research. Further, the book enhances the current body of knowledge on diagnostics and patient treatment and offers valuable new perspectives on practical clinical issues. As such, it offers a unique resource for clinicians, family physicians, medical scholars, and professionals engaged in patient management.

Mechanics plays a central role in determining form and function in biology. This holds at the cellular, molecular and tissue scales. At the cellular scale, mechanics in?uences cell adhesion, cytoskeletal dynamics and the traction that the cell can generate on a given substrate. All of these in turn - fect the cellular functions of migration, mitosis, phagocytosis, endocytosis and stem cell differentiation among others. Indeed, if cells do not develop the appropriate stresses, they are unviable and die. These aspects of cell mechanics are frequently used by mainstream biologists, as traditional mechanicians may be surprised to learn. There is a growing view that many functions of the cell are mechanical in nature even though chemical signals play crucial roles in the processes. Free energy barriers control transitions between different conformations of vir- ally every macromolecule including DNA, RNA, the adhesion protein integrin, the motor protein myosin, and the proteins vinculin and talin that link the cytoskeleton to focal adhesions. The strain energy can be a signi?cant component of the total free energy barrier. For binding to take place, the macromolecules need to be in conf- mational states that expose chemical groups without steric hinderance. The kinetics of chemical reactions are therefore strongly in?uenced by the conformational strain energy.

The technique of in situ hybridization is now vital to molecular biologists and their understanding of the pathophysiology of cellular functions. This practical guide covers all aspects of in situ hybridization, describing: practical procedures and protocols; the scientific background; areas of application; and the limitations of the technology. This edition has been completely rewritten to take into acocunt the many advances in the seven years since its initial publication.

Available for the first time with Macmillan's new online learning tool, Achieve, Molecular Cell Biology remains the most authoritative and cutting-edge resource available for the cell biology course. The author team, consisting of world-class researchers and teachers, incorporates medically relevant examples where appropriate to help illustrate the connections between cell biology and health and human disease. Emphasis on experimental techniques that drive advances in biomedical sciences and introduce students to cutting edge research teach students the skills they need for their careers. Achieve, Macmillan's new online learning platform, supports educators and students throughout the full range of instruction, including assets suitable for pre-class preparation, in-class active learning, and post-class study and assessment. Featuring new digital resources to engage students and help them to master cell biology concepts, the pairing of a powerful new platform with outstanding biology content provides an unrivalled learning experience.

Phase transitions occur throughout nature. The most familiar example is the one that occurs in water the abrupt, discontinuous transition from a liquid to a gas or a solid, induced by a subtle environmental change. Practically magical, the ever-so-slight shift of temperature or pressure can induce an astonishing transition from one entity to another entity that bears little resemblance to the first.

So "convenient" a feature is seen throughout the domains of physics and chemistry, and one is therefore led to wonder whether it might also be common to biology. Indeed, many of the most fundamental cellular processes are arguably attributable to radical structural shifts triggered by subtle changes that cross a critical threshold. These processes include transport, motion, signaling, division, and other fundamental aspects of cellular function.

Largely on the basis of this radical concept, a symposium was organized in Poitiers, France, to bring together people who have additional evidence for the role of phase transitions in biology, and this book is a compendium of some of the more far-reaching of those presentations, as well as several others that seemed to the editors to be compelling.

The book should be suitable for anyone interested in the nature of biological function, particularly those who tire of lumbering along well trodden pathways of pursuit, and are eager to hear something fresh. The book is replete with fresh interpretations of familiar phenomena, and should serve as an excellent gateway to deeper understanding."

This volume describes the state-of-knowledge in the study of the relationships between mechanical loading states in tissues and common pathophysiologies related to increase in mass of adipose tissues and/or hyperglycemia which eventually lead to obesity, diabetes, insulin resistance, hyperlipidemia, metabolic inflammations, certain types of cancer and other related diseases. There appears to be an interaction between the loading states in tissues and cells and these chronic conditions, as well as with factors such as age, gender and genetics of the individual. Bioengineering has made key contributions to this research field in providing technologies for cell biomechanics experimentation, microscopy and image processing, tissue engineering and multi-scale, multi-physics computational modeling. Topics at the frontier of this field of study include: the continuous monitoring of cell growth, proliferation and differentiation in response to mechanical factors such as stiffness of the extracellular matrix (ECM) and mechanical loads transferred through the ECM; mechanically-activated signaling pathways and molecular mechanisms; effects of different loading regimes and mechanical environments on differentiation fates of mesenchymal stem cells (MSCs) into myogenic and osteogenic versus adipogenic lineages; the interactions between nutrition and mechanotransduction; cell morphology, focal adhesion patterns and cytoskeletal remodeling changes in adipogenesis; activation of receptors related to diabetes by mechanical forces; brown and white adipose plasticity and its regulation by mechanical factors.

This book highlights recent advances in the field of biomaterials design and the state of the art in biomaterials applications for biomedicine. Addressing key aspects of biomaterials, the book explores technological advances at multi-scale levels (macro, micro, and nano), which are used in applications related to cell and tissue regeneration. The book also discusses the future scope of bio-integrated systems. The contents are supplemented by illustrated examples, and schematics of molecular and cellular interactions with biomaterials/scaffolds are included to promote a better understanding of the complex biological mechanisms involved in material-to-biomolecule interactions. The book also covers factors that govern cell growth, differentiation, and regeneration in connection with the treatment and recovery of native biological systems. Tissue engineering, drug screening and delivery, and electrolyte complexes for biomedical applications are also covered in detail. This book offers a comprehensive reference guide for multi-disciplinary communities working in the area of biomaterials, and will benefit researchers and graduate students alike.

Programmed cell death (PCD) is a genetically encoded, active process which results in the death of individual cells, tissues, or whole organs. PCD plays an essential role in plant development and defense, and occurs throughout a plant's lifecycle from the death of the embryonic suspensor to leaf and floral organ senescence. In plant biology, PCD is a relatively new research area, however, as its fundamental importance is further recognized, publications in the area are beginning to increase significantly. The field currently has few foundational reference books and there is a critical need for books that summarizes recent findings in this important area. This book contains chapters written by several of the world's leading researchers in PCD. This book will be invaluable for PhD or graduate students, or for scientists and researchers entering the field. Established researchers will also find this timely work useful as an up-to-date overview of this fascinating research area.

The discovery of adult neurogenesis caused a paradigm shift in the neurosciences. For more than 100 years, it was believed that adult neurons do not regenerate. Joseph Altman and Fernando Nottebohm found proof to the contrary and changed the course of history. Their research, included here, provides the foundations of the field. Today, adult neurogenesis is a rapidly expanding discipline applicable to the study of brain development and diseases, learning and memory, aging, and neuropsychiatric disorders. With multiple authors, the 27 chapters of this book contain the latest work in two volumes. The first presents the basic biology of adult neurogenesis in non-mammalian vertebrates and in the mammalian hippocampus and olfactory bulb, and the second discusses clinical implications and delves into adult neurogenesis and brain injury as well as neurodegenerative and neuropsychiatric pathologies. With details of the anatomy, physiology, and molecular biology of the two neurogenic brain regions, this book provides indispensable knowledge for many areas of neuroscience and for experimental and clinical applications of adult neurogenesis to brain therapy.

Plant protoplasts have proved to be an excellent tool for in vitro manipulation, somatic hybridization, DNA uptake and genetic trans- formation, and for the induction of somac1onal variation. These studies reflect the far-reaching impact of protoplast research in agriculture and forest biotechnology. Taking these aspects into consideration, the series of books on Plant Protoplasts and Genetic Engineering provides a survey of the literature, focusing on recent information and the state of the art in protoplast manipulation and genetic transformation. This book, Plant Protoplasts and Genetic Engineering VII, like the previous six volumes published in 1989, 1993, 1994, and 1995, is unique in its approach. It comprises 27 chapters dealing with the regeneration of plants from protoplasts, and genetic transformation in various species of Agrostis, Allium, Anthriscus, Asparagus, Avena, Boehmeria, Carthamus, Coffea, Funaria, Geranium, Ginkgo, Gladiolus, Helianthus, Hordeum, Lilium, Lithospermum, Mentha, Panax, Papaver, Passiflora, Petunia, Physcomi- trella, Pinus, Poa, Populus, Rubus, Saintpaulia, and Swertia. This book may be of special interest to advanced students, teachers, and research scientists in the field of plant tissue culture, molecular biology, genetic engineering, plant breeding, and general biotechnology. New Delhi, June 1996 Professor y. P. S. BAJAJ Series Editor Contents Section I Regeneration of Plants from Protoplasts 1. 1 Regeneration of Plantlets from Protoplasts of Allium cepa (Onion) E. E. HANSEN, J. F. HUBSTENBERGER, and G. C. PHILLIPS (With 3 Figures) 1 Introduction ...3 2 Protoplast Isolation ...4 3 Protoplast Culture ...8 4 Regeneration of Plantlets ...9 5 Summary...

This volume assembles protocols for chromosome engineering and genome editing in two recently developed approaches for manipulating chromosomal and genomic DNA in plants. The first approach is a "plant chromosome vector" system, which allows the introduction of desired genes or DNA into target sites on the chromosome vector, particularly by sequence-specific recombination. The second approach is "genome-editing," which makes it possible to introduce mutations into any of the genes of DNA that we wish to change. In addition, this book also covers other related techniques used to accelerate progress in plant chromosome and genome engineering. 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, Chromosome and Genomic Engineering in Plants: Methods and Protocols provides a comprehensive source of protocols and other necessary information to anyone interested in this field of study.