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1. 1 Historical Aspects and Terminology Granulated metrial gland (GMG) cells are readily identified by their cytoplasmic granules and were observed a number of years before the term'metrial gland'was introduced. A series of papers by Duval in 1891 provided a comprehensive description and a critical review of earlier studies of the placenta of rodents, but it was not until 1902 that the first convincing illustrations of GMG cells appeared in the literature (Jenkinson 1902). Jenkinson described'maternal glycogen cells'in the pregnant mouse uterus and noted that they contained cytoplasmic granules which stained with a variety of dyes. From his detailed description of the appearance and distribution of these maternal glycogen cells it is clear that he had observed what are now called granulated metrial gland cells. In 1911 Ancel and Bouin used the phrase une glande myometriale endocrine to describe a structure appearing between the muscle layers of the uterus at the insertion site of the placenta in rabbits. They described one of the cell types present in the glande myometriale as having the characteristics of glandular cells and noted their content of safraninophilic cytoplasmic granules. A glande myometriale endo­ crine was also described in the pregnant rat uterus by Weill (1919). He reported that the cellules granuleuses contained acidophilic inclusions and despite the absence of any illustrations in his paper it is apparent that he also had observed GMG cells.

Both functions of the mammalian ovary, the endocrine and (synthesis and secretion of steroid hormones) and exocrine (production of ova), depend upon the presence and cyclic growth of follicles, as the depletion of primordial follicles from the ovary leads to cessation of these f-unctions or female reproduction in mammals, or to postmenopausal period in humans. Actually, various fertility and sterility problems at the ovarian level are related to follicles. Therefore, a thorough understanding of the biology of ovarian follicles in mammals is of fundamental interest to a wide variety of academic and scientific disciplines. Study of their structure, function, and control involves mor­ phology, including ultrastructure, cell biology, physiology, endocrinology, biochemis­ try, immunology, neurobiology and pharmacology. Zoologists take interest in comparative and evolutionary aspects of biology of ovarian follicles in many different groups of mammals. Agricultural scientists and wildlife biologists need a thorough knowledge of the biology of follicles to control more effectively fecundity in domestic animals and endangered species of mammals. Finally, clinical scientists, toxicologists and physicians want to know the normal and pathological features of ovarian follicles in women, especially in relation to health and the regulation of fertility. Having in view the great importance of studying various aspects of biology of ovarian follicles, numerous papers and reviews on several aspects of ovarian follicles are published annually in a wide variety of scientific journals.

Advances in Cell Biology has been initiated as a continuing, multi-volume series to report on the progress of a wide spectrum of problems of cell structure and cell function. Jn arranging these volumes individual contributors are asked not only to review the major new information, but especially to present the state of a given problem or area by discussing the current central issues, speculations, concepts, hypotheses, and technical problems. We intend, in addition, that these volumes will not be concerned with comprehensive reviews of the recent literature but will consist rather of presentations of an interpretive and integrative nature, based on selection of major research advances. It is our aim that these volumes should provide the means whereby cell biologists may keep themselves reasonably well informed about the current progress in research areas in cell biology in which they are not immediately or directly involved themselves. The articles, nevertheless, are expected to bring into focus the experimental objectives of the specialists in a given research area. D. M.P. L. G. E.M. vii Contents Contributors v Preface vii 1 1. The Regulation of DNA Synthesis in Eukaryotes James Douglas Watson 2. D·RNA Containing Ribonucleoprotein Particles and Messenger RNA Transport 47 G. P. Georgiev and 0. P. Samarina Recent Developments in the Synchronization of 3. Tetrahymena Cell Cycle 111 Eric Zeuthen 153 4. Repetitious DNA Christopher Bostock 5. Mitosis 225 R. Bruce Nicklas Specific Enzyme Production in Eukaryotic Cells 299 6.

The beginning student of histology is frequently confronted by a paradox: diagrams in many books that illustrate human microanatomy in a simplified, cartoon-like manner are easy to understand, but are difficult to relate to actual tissue specimens or photographs. In turn, photographs often fail to show some important features of a given tissue, because no individual specimen can show all of the tissue's salient fea­ tures equally well. This atlas, filled with photo-realistic drawings, was prepared to help bridge the gap between the simplicity of diagrams and the more complex real­ ity of microstructure. All of the figures in this atlas were drawn from histological preparations used by students in my histology classes, at the level of light microscopy. Each drawing is not simply a depiction of an individual histological section, but is also a synthesis of the key structures and features seen in many preparations of similar tissues or organs. The illustrations are representative of the typical features of each tissue and organ. The atlas serves as a compendium of the basic morphological characteristics of human tissue which students should be able to recognize.

Recent studies have shown that cells from adipose tissue are capable of trafficking to tumors, thus enabling paracrine action of adipokines from within the tumor microenvironment. Increased tumor vascularization, immune system suppression and direct effects on malignant cell survival and proliferation have been investigated as mechanisms regulated by adipokines. The goal of this book is to discuss data pointing to the role of adipose tissue in cancer and to dissect individual mechanisms through which adipose tissue excess or restriction could influence cancer progression.

Motility is a fundamental property of living systems, from the cytoplasmic streaming of unicellular organisms to the most highly differentiated and de­ veloped contractile system of higher organisms, striated muscle. Although of motility have a long and in­ scientific investigations into the mechanisms teresting history, the knowledge of molecular processes, especially in the area of regulation of control of motility, has been developing at an ever more rapid pace with the utilization of multidisciplinary approaches from physiology, cell biology, genetics, biochemistry, pharmacology, and biophysics. In Volume 3, Cell and Muscle Motility continues the same philosophy as that of the preceding volumes. The essays are meant to focus on topics of current interest, to be critical rather than exhaustive, and to indicate the current trends of research efforts. The series is intended to foster an in­ terchange of concepts among various workers in a variety of disciplines and to serve as a reference for students and workers who wish to familiarize them­ selves with the most current progress in motility. Robert M. Dowben Jerry W. Shay Dallas vii Contents Chapter 1 Intermediate Filaments in Striated Muscle: A Review of Structural Studies in Embryonic and Adult Skelttal and Cardiac Muscle Maureen C. Price and Joseph W. Sanger 1. Introduction.................................................

Stem Cells: An Insider's Guide is an exciting new book that takes readers inside the world of stem cells guided by international stem cell expert, Dr. Paul Knoepfler. Stem cells are catalyzing a revolution in medicine. The book also tackles the exciting and hotly debated area of stem cell treatments that are capturing the public's imagination. In the future they may also transform how we age and reproduce. However, there are serious risks and ethical challenges, too. The author's goal with this insider's guide is to give readers the information needed to distinguish between the ubiquitous hype and legitimate hope found throughout the stem cell world. The book answers the most common questions that people have about stem cells. Can stem cells help my family with a serious medical problem such as Alzheimer's, Multiple Sclerosis, or Autism? Are such treatments safe? Can stem cells make me look younger or even literally stay physically young? These questions and many more are answered here.A number of ethical issues related to stem cells that spark debates are discussed, including risky treatments, cloning and embryonic stem cells. The author breaks new ground in a number of ways such as by suggesting reforms to the FDA, providing a new theory of aging based on stem cells, and including a revolutionary Stem Cell Patient Bill of Rights. More generally, the book is your guide to where the stem cell field will be in the near future as well as a thoughtful perspective on how stem cell therapies will ultimately change your life and our world.

In 1954, when I obtained my B. Sc. (Honours School) degree in Zoology, I was asked by my advisor, the late Professor Vishwa Nath, to make comparative cytological studies of oogenesis in birds for my M. Sc. (Honours School) thesis submitted to the Panjab University in 1956. Using the classical techniques of cytology, I studied the various aspects of follicle growth and atresia in five species of birds. Professor Vishwa Nath, who himself devoted most of his life to the study of the comparative cytology of gametogenesis in animals, was so much impressed by my findings on avian oogenesis that he asked me to continue my research work for the Ph. D. degree under his supervision. During my research work on avian oogenesis, I became very much interested in further comparative study of ovarian follicles in vertebrates and thus decided to make comparative, morphological and histochemical studies of ovarian follicles in reptiles, birds and mammals. Actually, very little information was available in this regard in 1957 when I started working for the Ph. D. degree. In subsequent years, I and my numerous dedicated and highly motivated M. Sc. and Ph. D. students con­ tinued to make comparative, morphological, histochemical, biochemical and physiological studies of ovarian follicles in all groups of vertebrates. In connection with the subject of this book, I would like to make special mention of my two brilliant students, namely Dr. S. K. Varma and Dr. R. K.

This concise introductory textbook uses carefully chosen examples from clinical and experimental observations to provide an insight into the principles underlying the immune system. As a result, it encourages readers to ask critical questions in order to further advance our understanding of this unique organ. Both authors are experienced lecturers and highly regarded researchers.The book is professionally illustrated in four color throughout with beautiful artwork which by itself distinguish the title from any comparable title.Website: www.wiley-vch.de/home/immunology

The Atlas provides a complete overview of all major structures of the mouse brain that can be identified in Golgi preparations. The most important feature is its three-dimensional integrity since all structures and nerve tracts can be followed from one section to the next one with uninterrupted continuity. The Golgi Atlas presents a series of camera lucida drawings of the entire telencephalon and upper brain stem of the young postnatal mouse in 24 transverse, 11 sagittal and 15 horizontal planes. The drawings were prepared from selected brains stained in toto with the Golgi method, that have been serially sectioned in the three orthogonal planes. The text includes an introduction of the material and methods used for the construction of this Atlas and a survey with a complete bibliography on the previous studies made with the Golgi method in Rodents. In this account, a number of issues concerning particular anatomical details are considered in relation to the interpretations obtained by other students. Reference is made to some relevant reviews and key articles.

Worldwide, numerous textbooks and publications have dealt with research on muscle fibres carried out under different points of view. In addition, comprehensive works such as Myology (Engel and Franzini-Armstrong 1994), Disorders of Voluntary Muscle (Walton et al. 1994), and Skeletal Muscle (Schmalbruch 1985) as a volume ofthe work Handbook of Microscopic Anatomy, have been published. Moreover, proceedings from myology symposiums give us access to the present state of the art in muscle research. The book The Dynamic State of Muscle Fibres (Pette 1990a) summarizes the contributions to the symposium of the same name, which was held in Constance in 1989. Considering these outstanding works one has to ask the question: Why do we need the present book? The first reason is that results from ongoing research expand scientific knowledge continuously. When dealing with muscle research one soon realizes that muscle tissue is a fascinating subject, whose secrets have not yet been revealed completely. The application of new techniques in muscle fibre research enables and provokes us to go deeper into the nature of muscle tissue. The results are findings that add a new dimension to what is already known. For instance, the detailed metabolic charac­ terization of muscle fibre types in the context of an intact histological section has been performed only recently using cytophotometrical quantification of enzyme activities. The second reason for this book is of a more pragmatic nature.

Advances in Stem Cell Research discusses recent advances in stem cell science, including therapeutic applications. This volume covers such topics as biomanufacturing iPS cells for therapeutic applications, techniques for controlling stem cell fate decisions, as well as current basic research in such areas as germ line stem cells, genomics and proteomics in stem cell research. It is a useful book for biology and clinical scientists, especially young investigators and stem cell biology students who are newly entering the world of stem cells research. The editors hope that the new knowledge and research outlined in this book will help contribute to new therapies for a wide variety of diseases that presently afflict humanity.

The Centrosome: Cell and Molecular Mechanisms of Functions and Dysfunctions in Disease includes chapters on classic and modern aspects of centrosome research to cover topics of current interest that have not been covered in depth in most books on the market so far. It extends on previous topics and includes new exciting aspects of centrosome research focused on primary cilia and their dysfunctions that are implicated in numerous diseases. Each chapter will be written by experts in their fields who will contribute their unique expertise in specific research fields and include cell and molecular details that are important for the specific subtopics. The book will be comprehensive, concise and will include reviews of key topics in the field. Cutting edge new information will be balanced with background information that will be readily understandable for the newcomer and the experienced centrosome researcher alike.

Leading international experts contribute to this selection of reviews covering the merging fields of growth factors, differentiation factors, and cytokines. These appear to play fundamental roles in a wide variety of physiological and pathophysiological processes that include the regulation of growth in normal and malignant cells, embryogenesis, the immune response, wound healing, inflammation, and atherogenesis. The major aspects of recent research and development are discussed, providing an important update in this pioneering field.

Nearly 10 years have elapsed since I finished writing the first edition of Intro­ duction to Molecular Embryology. During this period, molecular embryology has made great strides forward, but without undergoing a major revolution; there­ fore, the general philosophy and outline of the book have remained almost un­ changed. However, all the chapters had to be almost completely rewritten in or­ der to introduce new facts and to eliminate findings which have lost interest or have been disproved. There was a major gap in the first edition of this book: very little was said about mammalian eggs despite their obvious interest for mankind. Research on mammalian eggs and embryos is so active today that this important topic deserves a full chapter in a book concerned with molecular embryology. Therefore, I am very thankful to my colleague Dr. Henri Alexandre, who has written a chapter on mammalian embryology (Chap. 9) and has prepared all the illustrations for this book.

This monograph has been written in the hope that it will prove of value to medical students and clinicians, to Honours undergradu­ ates in appropriate branches of the natural sciences, and to repro­ ductive biologists in general. It would be pleasing if the text also caught the attention of veterinary undergraduates, since there is much information bearing on reproduction in domestic animals. First and foremost, however, the intended audience is a medical one, for scientific studies of human reproduction have been cata­ lyzed by the intense interest in procedures of fertilization in vitro. Some would judge that this very activity has narrowed our view of physiological events occurring within the Fallopian tubes. The pre­ sent work may therefore serve as a useful counterbalance to the overwhelming series of publications on procedures of in vitro fer­ tilization, and offer opportunities to those in the clinical field for extending their knowledge of the scientific background to much of the current work.

The mouse is a perfect model organism to study mammalian, and thus indirectly also human, embryology. Most scientific achievements that have had an important impact on the understanding of basic mechanisms governing embryo development in humans, originated from mouse embryology. Stem cell research, which now offers the promise of regenerative medicine, began with the isolation and culture of mouse embryonic stem cells by Martin Evans (who received the Nobel Prize in medicine in 2007 for this achievement) and Matthew Kaufman. This book provides an overview of mouse development, spanning from oocytes before fertilization to the state-of-the-art description of embryonic and adult stem cells. The chapters, written by the leading specialists in the field, deal with the most recent discoveries in this extremely fast-developing area of research.

This book provides a detailed account of the physico-chemical properties and biological functions of the outer membrane vesicles (OMVs) of different pathogenic and non-pathogenic Gram-negative bacteria. It also includes an authentic record of the first systematic study that discovered the mechanism of OMV formation by a pathogen, Vibrio cholerae, and proposed that the process represented a novel secretory activity of bacteria. Furthermore, the authors present clinical and laboratory data on the use of OMVs as immunogens, as effective and licensed vaccines against Neisseria meningitidis serogroup B infections and on the development of more effective vaccines against other human and animal pathogens including Vibrio cholerae. This volume thus bears witness to the emerging revolution in the field of vaccines against pathogens and closes with a discussion of open questions and future research on OMVs.

This book will describe the nuclear encoded genes and their expressed proteins of mitochondrial oxidative phosphorylation. Most of these genes occur in eukaryotic cells, but not in bacteria or archaea. The main function of mitochondria, the synthesis of ATP, is performed at subunits of proton pumps (complexes I, III, IV and V), which are encoded on mitochondrial DNA. The nuclear encoded subunits have mostly a regulatory function. However, the specific physiological functions of the nuclear encoded subunits of complexes I, III, IV, and V are mostly unknown. New data indicates that they are essential for life of higher organisms, which is characterized by an adult life without cell division (postmeiotic stage) in most tissues, after the juvenile growth. For complex IV (cytochrome c oxidase) some of these subunits occur in tissue-specific (subunits IV, VIa, VIb, VIIa, VIII), developmental-specific (subunits IV, VIa, and VIIa) as well as species-specific isoforms. Defective genes of some subunits were shown to induce mitochondrial diseases. Mitochondrial genes and human diseases will also be covered.

Apoptosis is a fascinating concept for the basic scientist. This is not only because of the multifaceted variety of proposed and discovered mechanisms, but because apoptosis represents a fundamental pathway for cell renewal. The study of apoptosis has resulted in an array of discoveries on signal transduc­ tion and downstream effects that have facilitated and advanced many fields in biology, including research on cancer and other diseases. Thus, the apoptotic process can be viewed as the largest effort of the scientific community to understand how cells work and tissues assemble or remodel. The most direct consequence of this accumulated knowledge is a greater understanding of disease and pathological mechanisms. The end result of these efforts will be significant contributions to health and the adoption of new, never anticipated, therapeutic approaches. This book represents the summation of considerable effort from a significant group of contributors from all over the world as well as from its editors. In this fashion, many viewpoints have been collected and SUbjected to thorough academic discussion. The concepts contained in this medically important volume will stimulate and renew the ideas of scientists and indeed, will generate additional work to advance biological knowledge even further. The emphasis of this volume cements what has been established, adds what has not been explored fully, and creates a fertile ground for further hypotheses that will lead to a more complete understanding of the apoptotic process.

Progression of chronic diseases in general and chronic kidney disease in particular has been traditionally viewed in the light of various contributors to development of glomerulosclerosis and tubulointerstitial scarring culminating in renal fibrosis. Indeed, this dogma prevailed for decades underscoring experimental attempts to halt fibrotic processes. Breakthrough investigations of the past few years on stem/progenitor cell involvement in organ regeneration caused a conceptual shift in tackling the mechanisms of nephrosclerosis. It has become clear that the rate of progression of chronic kidney disease is the net sum of the opposing trends: degenerative fibrotic processes and regenerative repair mechanisms. The latter part of this equation has been by and large ignored for years and only recently attracted investigative attention. This book revisits the problem of kidney disease by focusing on regenerative mechanisms in renal repair and on the ways these regenerative processes can become subverted by an intrinsic disease process eventuating in its progression. Cutting-edge investigations are summarized by the most experienced international team of experts. Presents a comprehensive, translational source for all aspects of renal stem cells, tissue regeneration, and stem cell therapies for renal diseases in one reference work. This will ultimately result in time savings for academic, medical and pharma researchers Experts in the renal stem cell system in kidney repair and regeneration take readers from the bench research to new therapeutic approaches, providing a common language for nephrology researchers, fellows and other stem cell researchers. This enables the discussion of development of stem cells and their use in the repair and regeneration of the kidney

Research into the field of stem cell biology has developed exponentially over recent years, and is beginning to offer significant promise for unravelling the molecular basis of a multitude of disease states. Importantly, in addition to offering the opportunity to delve deeply into the mechanisms that drive disease aetiology the research is realistically opening the doors for development of targeted and personalized therapeutic applications that many considered, until recently, to be nothing more that a far fetched dream. This volume provides a timely glimpse into the methods that have been developed to instigate, and the mechanisms that have been identified to drive, the process of nuclear reprogramming, chronicling how the field has developed over the last 50-60 years. Since the early 1950s a small number of notable experiments have provided significant impetus to the field, primarily the demonstration of reprogramming ability, first by the complex cytoplasmic milieu that constitutes the amphibian egg, then that of the mammalian egg, and finally that of the mammalian embryonic stem cell. Most recently, the demonstration that a limited pool of defined molecules is capable of reprogramming a multitude of cell types has provided massive impetus and facilitated transition towards realistic therapeutic application. We have therefore reproduced some of the key articles that elegantly document these dramatic stages of development of the field in an inclusive appendix to the book, for the benefit of readers keen to investigate the history of how the field of stem cell biology has evolved. Owing to the ever broadening nature of this field, and the incredible rate at which it is evolving, the main content of this volume focuses on areas that have shown significant movement in recent years, are most likely to translate into personalized therapeutic application, and thus provide greatest potential for significant impact on human health inthe not too distant future. We recognize that research into many other disease states and cell types are all equally worthy of discussion. We would therefore like to acknowledge those researchers involved whose work we have not been able to include in this volume. Nuclear Reprogramming and Stem Cells will serve as a valuable resource for all researchers in the field of stem cell biology, including those just setting out on their career path as well as those already established in the field.

When new fellows join my lab, I give them some reading materials so that they can orient themselves in their assignment in a new eld. When fellows leave my lab, some after writing their dissertations, I prefer to give them a book as a symbolic present. I was longing for a book that contained something on more or less eve- thing about the islets. At the same time, I wished it contained information as recent as possible. There are a few such books in the market but they are pretty outdated. I started picking islets myself from October 1990, when I joined the Rolf Luft Center, Karolinska Institutet. Over the years my fascination for islet research remained high. Since last year, I felt a stronger urge to do more for these mysterious and hidden mini-organs that are directly or indirectly involved in the pathogenesis of all forms of diabetes that affects?250 million people in the world. After I launched the Islet (landesbioscience. com/journals/islets) and founded the Islet Society (isletso- ety. org), there was a momentum that could be utilized to create something equally meaningful i. e. this book. The idea cracked in September 2008. Starting September 19, 2008, I contacted an estimated 90% of the authors who published anything on the islets during 2007–2008 and who could be traced from the internet.

The writing of this book is based on: 1. earlier experience writing textbooks for biology students with a university-level background in biology and biochemistry with many (>200)?gures in these books (in Hungarian), 2. and on the necessity to present university lectures as power point presentations to catch the interest of s- dents. The author realizes that young readers who were grown up in an information society are relactant to read too much unless they have to take exams. Even then they prefer books which contain illustrations for a better understanding. In collaboration with colleagues, referees and members of the publishing staff an extensive set of photographs and illustrations were collected to provide a graphic follow-up to the text. Further aids for the student, instructor and the curious reader are provided by summaries, extensive sets of readings and references for each chapter, a glossary of the terms, list of abbreviations and a DVD with a red-blue eyeglass to visualize three dimensional chromatin structures. The reader could ask: Why another book on apoptosis? The answer to the question is related to the de?nition of the process. The term apoptosis has been introduced to describe typical morphological changes leading to controlled se- destruction of cells. The?rst demonstrated biochemical feature of this type of cell death was internucleosomal fragmentation, which was occasionally preceded by the generation of large DNA fragments.

Stem cells appear to be fundamental cellular units associated with the origin of multicellular organisms and have evolved to function in safeguarding the cellular homeostasis in organ t- sues. The characteristics of stem cells that distinguish them from other cells have been the fascinating subjects of stem cell research. The important properties of stem cells, such as ma- tenance of quiescence, self-renewal capacity, and differentiation potential, have propelled this exciting?eld and presently form a common theme of research in developmental biology and medicine. The derivation of pluripotent embryonic stem cells, the prospective identi?cation of multipotent adult stem cells, and, more recently, the induced pluripotent stem cells (popularly called iPS) are important milestones in the arena of stem cell biology. Complex networks of transcription factors, different signaling molecules, and the interaction of genetic and epi- netic events constantly modulate stem cell behavior to evoke programming and reprogramming processes in normal tissue homeostasis during development. In any given cellular scenario, the regulatory networks can pose considerable complexity and yet exert an orderly control of stem cell differentiation during normal development. An aberration in these?nely tuned processes during development usually results in a spectrum of diseases such as cancers and neurological disorders. Thisunderscorestheimminentneedforamorecompleteunderstandingofmolecular mechanisms underlying the regulatory circuitries required for stem cell maintenance. Overthepast3–5years,adiversegroupofbenchandphysicianscientistshaveprospectively enhanced our knowledge of stem cell biology. These studies are unveiling many unrecognized or previously unknown fundamentals ofdevelopmental biology.

Chloroplasts are vital for life as we know it. At the leaf cell level, it is common knowledge that a chloroplast interacts with its surroundings – but this knowledge is often limited to the benefits of oxygenic photosynthesis and that chloroplasts provide reduced carbon, nitrogen and sulphur. This book presents the intricate interplay between chloroplasts and their immediate and more distant environments. The topic is explored in chapters covering aspects of evolution, the chloroplast/cytoplasm barrier, transport, division, motility and bidirectional signalling. Taken together, the contributed chapters provide an exciting insight into the complexity of how chloroplast functions are related to cellular and plant-level functions. The recent rapid advances in the presented research areas, largely made possible by the development of molecular techniques and genetic screens of an increasing number of plant model systems, make this interaction a topical issue.

Although much is known with respect to blood cell formation and function, many new concepts in the areas of the regulation of hematopoietic stem cell commitment and the subsequent survival, proliferation, and differentiation of progenitors have been elucidated in the last five years. Our understanding of the microenvironment where stem cells reside and commit to distinct blood types (the niche) has grown significantly in recent years. Furthermore, blood cells have been used as the key model system to study microRNA function and the role of microRNAs in the transformation of normal cells into cancer cells. The current volume Molecular Basis of Hematopoiesis, edited by Amittha Wickrema & Barbara Kee, provides the most recent developments in the area in addition to a chapter on the utilization of basic science knowledge for the treatment of blood diseases. Each chapter in this book has been written and edited by faculty in major academic and research institutions around the world, who are pushing the frontiers of research in this important area.

This book presents the latest findings in mechanosensitivity of the nervous system. The nervous system stands out from a number of tissues because besides reacting to the mechanical stress it is transmitting its own response to other organs and tissues, which are located downstream of its signaling pathway. For this reason any type of mechanical stimulation of the nervous system, which is capable of triggering a physiological response, has high scientific and practical significance, since it allows its use beyond a particular experimental model anywhere where it is contributing to a particular pathological condition. This book is a unique collection of reviews outlining the current knowledge and the future developments in this rapidly growing field. Currently, investigations of the effects of mechanical stress on the nervous system are focused on several issues. The majority of studies investigate the effects of mechanical stimulation on mechanosensitive channels, as its primary target and interactive agent, and aim on description of downstream intracellular signaling pathways together with addressing general issues of biomechanics of the nervous system. Knowledge of biomechanics, and mechanisms, which underlie it on organism, organ, tissue and cellular level, is necessary for understanding of the normal functioning of living organisms and allows to predict changes which arise due to alterations of their environment, and possibly will allow to develop new methods of artificial intervention. The book brings up the problem closer to the experts in related medical and biological sciences as well as practicing doctors besides just presenting the latest achievements in the field.

The use of directed cell models to study the manner in which two or more cells interact has revolutionized our understanding of protein-protein interactions in a complex milieu and has provided an invaluable platform for the identification of new molecules. In Cell-Cell Interactions: Methods and Protocols, highly skilled experts in the development and utilization of cell-cell interaction assays describe in detail their best methods-based approaches, both in vitro and in vivo, for uncovering cellular interactions at the molecular level. The protocols cover a diverse set of cell-cell interaction models in both normal and pathological states, are readily adaptable to nearly any cell type and organ system, and include primary data and outcome analysis. In addition, the protocols follow the successful Methods in Molecular Biology series format, each offering step-by-step laboratory instructions, an introduction outlining the principles behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. Insightful to beginners and inspiring to seasoned professionals, Cell-Cell Interactions: Methods and Protocols provides the cutting-edge and versatile laboratory tools needed to gain a basic understanding of cell-cell interactions.

Although nuclear reprogramming-the cellular process underlying cloning and the artificial generation of stem cells from somatic cells-may be considered a'black box'whose interior events are either completely unknown or at least imperfectly characterized, tantalizing clues about its operation are beginning to emerge. In Nuclear Reprogramming: Methods and Protocols, hands-on laboratory scientists describe readily reproducible methods for actually performing nuclear reprogramming by nuclear transfer in several different species, by fusion induced both chemically and electrically, and by the in vivo treatment of cells with cell extracts. Isolation of an adult stem cell type is also described. In addition, several methods of monitoring nuclear reprogramming are presented, including the use of transgenic markers, activation of telomerase as an ES-specific marker, light and electron microscopic observation of structural changes in the nucleus, and the verification of surface marker expression and differentiation potential of stem cells. Biochemical methods are provided for the examination of chromatin protein modifications, nucleosomal footprinting, transcription factor binding, and the study of DNA methylation changes both at the specific locus level and at the level of the whole nucleus. The protocols follow the successful Methods in Molecular Biology series format, each offering step-by-step laboratory instructions, an introduction outlining the principles behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. Wide-ranging and highly practical, Nuclear Reprogramming: Methods and Protocols significantly expands the repertoire of methods available to perform, study, and understand nuclear programming, not only for cloning, but also for reprogramming somatic cells to stem cells

Following in the footsteps of its much-acclaimed first edition, Plant Cell Culture Protocols, Second Edition has been expanded and revised to include the most up-to-date and frequently used techniques for plant cell and tissue culture. Readily reproducible and extensively annotated, the methods range from general methodologies, such as culture induction, growth and viability evaluation, and contamination control, to such highly specialized techniques as chloroplast transformation involving the laborious process of protoplast isolation and culture. Most of the protocols are currently used in the research programs of the authors or represent important parts of business projects aimed at the generation of improved plant materials. The protocols follow the successful Methods in Molecular Biology series format, each offering step-by-step laboratory instructions, an introduction outlining the principles behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. Two new appendices have also been added to explain the principles for formulating culture media and the composition of the eight most commonly used media formulations, as well as to list more than 100 very useful internet sites. Comprehensive and highly practical, Plant Cell Culture Protocols, Second Edition offers today's plant biotechnologists indispensable state-of-the-art techniques for carrying out plant research, for developing new transgenic plants, and for today's many and diverse commercial applications.

Despite political and ethical controversies surrounding the study of human embryonic stem (hES) cells, interest remains high in understanding the regulatory mechanisms of stem cell self-renewal, their differentiation along various lineages, and their potential use in regenerative medicine. In Human Embryonic Stem Cell Protocols, internationally respected researchers describe in detail their most useful techniques for the molecular and cellular manipulation of these intriguing cells. This diverse collection of readily reproducible methods has been optimized for the derivation, characterization, and differentiation of hES cells, with special attention given to regenerative medicine applications. The protocols follow the successful Methods in Molecular Biology series format, each offering step-by-step laboratory instructions, an introduction outlining the principles behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. A companion CD provides color versions of all illustrations in the book. Comprehensive and cutting-edge, Human Embryonic Stem Cell Protocols offers both novice and expert researchers powerful tools essential to understanding the maintenance and differentiation of human embryonic stem cells, as well as their applications in regenerative medicine today.

The book provides an updated overview of molecular analysis of human tissues, and the impact this analysis has on diagnosis and prognosis of human diseases. Special emphasis is placed on human cancer and the future directions of the field. Methods of handling clinical tissue samples, including the impact of handling on subsequent molecular analysis, are also discussed. In addition, detailed protocols for molecular analysis of DNA, RNA and protein, with special emphasis on molecular analysis of highly complex human tissue samples containing mixtures of cell populations, are provided.

Rational pharmacotherapy of the inner ear will become an important issue in treatment of inner ear diseases today and in the future. The contributions of some of the experts in the field are gathered in the present volume dedicated to all aspects of the pharmacology and the possible local and systemic treatment of inner ear diseases. Major topics include new aspects of human inner ear anatomy, neurotransmission, ion channels, vascular system, immunology, excitotoxicity, modulation of cell cycle, regeneration and differentiation, as well as new aspects of basic mechanisms at the cellular and molecular levels. A major feature of this volume is that it brings together both basic researchers and physicians to evaluate new experimental therapies and discuss future clinical strategies in the treatment of cochleo-vestibular disorders. Therefore, it will prove interesting to both scientists and clinicians in sensory physiology and auditory systems.

In Natural Killer Cell Protocols: Cellular and Molecular Methods, Kerry S. Campbell and Marco Colonna have assembled a comprehensive collection of readily reproducible methods designed to study natural killer (NK) cells from the broadest variety of viewpoints. These include not only classic techniques, but also new approaches to standard methods, newly evolved techniques that have become valuable for specific applications, and unique models for manipulating and studying NK cells. Among the advanced methods covered are those for in vitro transendothelial migration, in vivo detection of cells migrating into tumors, immunofluorescence staining of intracellular cytokines, and in vitro NK cell development. Valuable techniques for specific applications include vaccinia virus protein expression, soluble KIR-Fc fusions for HLA class I binding assays, calcium mobilization in cell conjugates, and identification of heterodimeric receptor complexes using cDNA library expression cloning. No less important are accounts of such classic methods as hybrid resistance, ADCC, viral defense, target cell cytotoxicity assays, cloning and culturing, tumor immunotherapy, and generation of HLA class I transfected target cells. Natural Killer Cell Protocols: Cellular and Molecular Methods offers immunologists, cancer researchers, virologists, and cell biologists today's most comprehensive collection of both established and cutting-edge techniques, methods that will contribute significantly to advancing our understanding of this fascinating and critically important class of cells.