In the past few years there has been the increased recognition that the effects of oxidative stress are not limited to the damage of cellular constituents. There is now evidence that reactive oxygen species (ROS) can alter cell function by acting upon the intermediates, or second messengers, in signal transductions. Such effects on signaling mechanisms probably account for the role of oxidative stress in inflammation, aging, and cancer. This volume brings together internationally recognized researchers in both the major areas covered by the book, oxidative stress and signal transduction. The work is organized in three sections. The first deals with the immediate cellular responses to oxidative stress and the production of second messengers. The second details the connection between second messengers and the gene. The third part looks more closely at the level of the gene.

This detailed volume encompasses new technological developments that specifically address questions related to adenosine 3',5'-monophosphate (cAMP) compartmentalization, that probe relevant protein-protein interactions, that increase the spatial and temporal resolution of cAMP signal detection, and that can facilitate integration of the mounting complexity of the information that is becoming available on this signaling system. cAMP, the prototypical intracellular second messenger, regulates a large variety of cellular functions and biological processes, including gene transcription, cell metabolism, proliferation, development, as well as more specialized functions depending on the cell type, so the realization of its extremely complex spatial organization and local regulation is providing novel mechanistic insight into cell physiology and is producing a novel framework for the identification of disease mechanisms. Written in the highly successful Methods in Molecular Biology series format, chapters include introduction 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 authoritative, cAMP Signaling: Methods and Protocols serves as a vital resource for researchers working in this expanding, complex field.

Computational cell biology courses are increasingly obligatory for biology students around the world but of course also a must for mathematics and informatics students specializing in bioinformatics. This book, now in its second edition is geared towards both audiences. The author, Volkhard Helms, has, in addition to extensive teaching experience, a strong background in biology and informatics and knows exactly what the key points are in making the book accessible for students while still conveying in depth knowledge of the subject. Besides a greatly increased focus on RNA biology and epigenetics, new topics such as intracellular trafficking, particle tracking, and more cellular programs e.g. circadian rhythms, apoptosis and cell differentiation, have been included. In addition, the book features supplementary material on the web including slides for presentations.

This book is a broadly historical account of a remarkable and very exciting scientific story-the search for the number of human chromosomes. It covers the processes and people, culminating in the realization that discovering the number of human chromosomes brought as much benefit as unraveling the genetic code itself. With the exception of red blood cells, which have no nucleus and therefore no DNA, and sex cells, humans have 46 chromosomes in every single cell. Not only do chromosomes carry all of the genes that code our inheritance, they also carry them in a specific order. It is essential that the number and structure of chromosomes remains intact, in order to pass on the correct amount of DNA to succeeding generations and for the cells to survive. Knowing the number of human chromosomes has provided a vital diagnostic tool in the prenatal diagnosis of genetic disorders, and the search for this number and developing an understanding of what it means are the focus of this book.

Of all scientific instruments, probably none has had more applications in the life sciences than the light microscope. In Light Microscopy: Methods and Protocols, expert researchers explore the basics and the latest advances in microscope instrumentation, sample preparation, and imaging techniques, all of which have been producing fundamental insights into the functions of cells and tissues. Chapters cover a variety of bright field and fluorescence microscopy-based approaches that are central to the study of a range of biological questions, providing information on how to prepare cells and tissues for microscopic investigations, covering detailed staining procedures, and exploring methods to analyze images and interpret the results accurately. 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. Comprehensive and current, Light Microscopy: Methods and Protocols is an essential handbook for all researchers who are exploring the intriguing microscopic world of the cell.

Products from Cells - Cells as Products This book ist he "lasting" product, a resource ofup to date information in the scientific literature fort he field ofanimal cell tec hnology, as it was presented during a pleasant and s timulating mee ting that was held in Lugano Switzerland in April 1999. "Products" appeartwice int he title oft he conference. This clearly indicates the fact that the focus oft he papers presented during this meeting was really the application ofn ew technologies (novel reactors or novel vectors, for example for the preparation and/ort he more efficient generation ofproducts ) that could be used, mainly, int he medical field. Classical approaches forthe use ofa nimal cells, for example forthe p r oduction of virus vaccines for human and animal health, still remain an important technology and still have, surprisingly, quite significant potential for further development and improvement. How ever, it appears that major technological advances an d major growth from an economical point ofview are occurring in other areas. Most importantly, protein production on the basis of recombinant DNA molecules transferred into a nimal cells, appears to be an ever increasing field of interest and innovation, even though the first production scheme with this technology was approved more than 15 years ago.

Epithelia are one of the commonest tissue types in the animal kingdom. Chapters from leading scientists in the major international research laboratories use examples from different systems to illustrate the form and function of epithelia. An important theme is the way in which epithelial cells differentiate to specialized tissue - reversal of this process occurs when cells become tumorigenic.

The fungus fusarium is a major plant pathogen that causes disease in nearly every agriculturally important plant. In addition, some strains produce mycotoxins that can cause serious illness in humans and livestock. The enormous economic importance of and health hazards posed by fusarium have fuelled research into its biochemistry, genetics, genomics, proteomics and metabolomics by scientists worldwide. In this book, an international group of researchers critically review the most important research on the genomics and molecular and cellular biology of fusarium.

The plant' is often the most neglected part of plant-based medicine. Throughout time, humans have searched, collected, and effectively used plants for healing. Currently, the medicinal plant-based business is flourishing at a dramatic pace and at the expense of an already declining population of plant species, many of which are on the verge of extinction. In spite of this history and popularity, the mystery of what transforms a plant into a medicinal plant persists, and there are chronic problems with ensuring the safety and efficacy of medicinal plant products. Therefore, there is a real need for a full characterization of medicinal plant species and for the development and application of novel technologies for the production of plant-based medicines. This book highlights some of the recent advances and new approaches to the development of technologies for plant-based medicines and is intended to stimulate new discussions among researchers, regulatory authorities, and pharmaceutical organizations, leading to significant advancements in the field.

Radiation Oncology and Radiotherapy, Part A, Volume 172 in the Methods in Cell Biology series, highlights advances in the field, with this new volume presenting interesting chapters on timely topics, including DNA damage quantification by the COMET assay, Immunofluorescence microscopy-assisted quantification of ATM and ATR activation in irradiated cells, Immunoblotting-based characterization of the DNA damage response, Assessment of lipid peroxidation in irradiated cells, A simple method to assess clonogenic survival of irradiated cancer cells, Quantification of beta-galactosidase activity as a marker of radiation-driven cellular senescence, Cytofluorometric assessment of cell cycle progression in irradiated cells, and more. Other sections cover Assessment of transcription inhibition as a characteristic of immunogenic cell death, Assessment of eIF2a phosphorylation during immunogenic cell death, Quantification of cytosolic DNA species by immunofluorescence and automated image analysis, Flow cytometry-assisted quantification of CALR exposure during immunogenic cell death, Interference of immunogenic anticancer therapy by artificially controlled calreticulin secretion from tumor cells, along with many additional topics of interest.

Despite many technological challenges faced by the xenotransplantation field, many major advances have been made in the last two decades. The field seeks to overcome the limitations and difficulties in organ procurement, which also apply to human cells and tissues, and facilitate the development of new therapies based on cell and engineered-tissue. Xenogeneic cells are simpler than solid organs and seem to pose less hurdles to attain long-term graft survival. In, Xenotransplantation: Methods and Protocols expert researchers study characterizations of xenogeneic interactions at the cellular and molecular levels and describe the use of relevant small-animal and pig-to-primate models. Related ethical and legal considerations are also covered. Written in the highly successful Methods in Molecular Biology (TM) series format, the chapters include the kind of detailed description and implementation advice that is crucial for getting optimal results in the laboratory. Thorough and intuitive, Xenotransplantation: Methods and Protocols aids scientists in continuing to study xenotransplantation and its multiple aspects.

In situ hybridization has developed as a means of localizing specific DNA and RNA sequences within tissues. The great strength of this approach is the ability to relate the distribution of specific nucleic acids with cell structures and the protein products of the target gene by means of immunohistochemistry. Complementary DNA, RNA or oligonucleotide probes, suitably labelled, are hybridized to specific DNA or RNA targets within tissues. The spatial information thus obtained has contributed greatly to our understanding of such diverse areas of research as gene mapping, viral infection, cytogenetics, protein synthesis, prenatal diagnosis and tissue grafting. This book is not intended as another recipe book, although it does describe theoretical and practical aspects of the technology. Rather, the authors critically describe the contribution made by in situ hybridization to specific areas of medical research.

The current explosive progress in molecular biological research can be definitively traced to the development of molecular cloning technology. The ability to insert specific gene sequences into cloning vectors and their subse quent expansion is the cornerstone of modem molecular biology. A direct practical outcome of molecular cloning technology is its application to ex press specific recombinant genes. Currently, recombinant gene products are used in a wide spectrum of applications, including gene therapy, production of bioactive pharmaceuticals, synthesis of novel biopolymers, in agriculture and animal husbandry, and so on. A fundamental requirement for successful recombinant gene expression is the design of the cloning vector and the choice of the host organism for expression. Recombinant Gene Expression Protocols grows out of the need for a laboratory manual that provides the reader the background and rationale, as well as the practical protocols for the preparation of "expression constructs" and their introduction into appropriate host cells and/or organisms. The chap ters in this book are grouped by their expression hosts, including E. coli, yeast, mammalian cells, nonmammalian eukaryotes such as plants, Xenopus, and insects, as well as in transgenic organisms. In-depth information is presented on the important characteristics of expression cloning vectors and the various methods for efficiently introducing expression constructs into target cells and/ or organisms. Throughout Recombinant Gene Expression Protocols, the authors have consistently striven for a balanced presentation of both background informa tion and actual laboratory details.

''Interesting with many useful ideas and references. It covers a broad range and it is a good introduction to this field.'' ---Analyst

Since the initial establishment of Robert Koch's postulates in the nineteenth century, microbial protein toxins have been recognized as a major factor of bacterial and fungal virulence. An increasing number of proteins produced and secreted by various bacteria, yeasts and plants are extremely toxic and most of them developed remarkably "intelligent" strategies to enter, to penetrate and to finally kill a eukaryotic target cell by modifying or blocking essential cellular components.

This book describes the strategies employed by protein toxins to render their pro- and eukaryotic producers a selective growth advantage over competitors. In providing an up-to-date overview on the mode of protein toxin actions, it accommodates biomedically and biologically relevant toxin model systems. As a result, it significantly broadens our perspective on biochemical architecture and molecular ploy behind the lethal principles of pro- and eukaryotic toxins.

Research into and interest in the role of stromal cells in immunology has exploded over the past 15 years. The conventional view that placed non-hematopoietic stromal cells as passive, structural, and supportive entities has now been replaced with an appreciation that these cells have active, dynamic roles during immune responses, and thus impact on the pathophysiology of multiple immune-mediated diseases. This book serves to provide solid grounding in the fundamentals of stromal immunology, focusing on the biological aspects of their function in addition to highlighting key areas for the development of the field in the future. The book is also a unique source of information on emerging concepts that place stromal cells from outside lymphoid organs as major contributors to the biology of diverse conditions, such as rheumatoid arthritis, chronic parasitic infection, inflammatory bowel disease, and cancer.

Koval provides an interdisciplinary forum for the diverse studies involved in the stress biology of eukaryotic cells. Readers gain access to the most recent information available for eukaryotic systems ranging from plants to humans. For the student, this format introduces a source of potentially unifying concepts and hypotheses. Scientists will find a unique opportunity to conveniently examine the similarities among inducible responses initiated by a variety of agents.

The receptor-associated JAK protein kinases and their substrates, the STAT transcriptional activators, transmit signals following cytokine and growth factor binding to receptors expressed on the cell surface, to result in specific transcriptional and cellular responses. Over the last two decades, the field has progressed from identification of the individual components through to an understanding of the activation and deactivation mechanisms, and the complex structural detail of the proteins involved. We now know that these pathways are important in many biological processes, including growth and development, hematopoiesis, and the innate and adaptive immune response. JAK-STAT Signalling: Methods and Protocols provides detailed methodologies for examining many aspects of the pathway. Divided into four sections, topics include JAK and STAT specific approaches, the negative regulators of the pathway (SOCS proteins), and the production and crystallization of JAK and STAT proteins, among others. Written in the 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 notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, JAK-STAT Signalling: Methods and Protocols will be of use not only to those working in the area but also to new investigators who are led to delve into the complexities of JAK and STAT responses.

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.

he biological sciences are dominated by the idea that cells are the functionally autonomous, physically separated, discrete units of life. TThis concept was propounded in the 19th century by discoveries of the cellular structuring of both plants and animals. Moreover, the ap parent autonomy of unicellular eukaryotes, as well as the cellular basis of the mammalian brain (an organ whose anatomy for a long while defied attempts to validate the idea of the cellular nature of its neurons), seemed to provide the final conclusive evidence for the completeness of *cell theory', a theory which has persisted in an almost dogmatic form up to the present day. However, it is very obvious that there are numerous observations which indicate that it is not the cells which serve as the basic units of biological life but that this property falls to some other, subcellular assemblage. To deal with this intricate problem concerning the fundamental unit of living matter, we proposed the so-called Cell Body concept which, in fact, devel ops an exceedingly original idea proposed by Julius Sachs at the end of the 19th century. In the case of eukaryotic cells, DNA-enriched nuclei are intimately associated with a microtubular cytoskeleton. In this configuration-as a Cell Body-these two items comprise the fundamental functional and struc tural unit of eukaryotic living matter. The Cell Body seems to be inherent to all cells in all organisms.

Recent work has revealed that stabilizing G-quadruplexes in telomeric DNA inhibits telomerase activity, providing impetus for the development of G-quartet-interacting drugs, while G-quartet-containing oligonucleotides have been recognized as a potent class of aptamers effective against STAT3 and other transcription factors implicated in oncogenesis, proving these guanine-quartets to be a vital and rich area for future study. In "G-Quadruplex DNA: Methods and Protocols", experts in the field present a collection of detailed techniques for studying G-quartet formation, dynamics, and molecular recognition. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters include brief introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, "G-Quadruplex DNA: Methods and Protocols "promises to be a useful resource for those familiar with G-quartets as well as an easy entry point for those researchers from diverse fields who are just developing an interest in the exciting implications of G-quadruplex DNA.

Bidirectional traffic of macromolecules across the nuclear envelope is an active and essential transport process in all eukaryotic cells. Work on various model systems has led to a tremendous increase in our understanding of nuclear transport in recent years.This volume summarizes our current knowledge of protein and RNA transport into and out of the nucleus. It contains nine up-to-date reviews which cover various aspects of nucleocytoplasmic transport, including the structure and function of the nuclear pore complex, the role of soluble transport factors in protein and RNA transport, and the regulation of protein transport through the nuclear pore.

Investigation of the mechanisms of cellular response to different mechanical stimuli, as well as mechano-electrical feedback (MEF) in the intact heart is one of the main topics in fundamental and clinical cardiology. The present volume of "Mechanosensitivity in Cells and Tissues: Mechanosensitivity of the Heart" c- bines excellent reviews written by worldwide leaders in this ?eld. The 3rd volume is a great addition to this excellent series of books edited by Andre Kamkin and Irina Kiseleva. This volume successfully combines reviews, aimed at academic, physiology and clinical cardiology communities, devoted to mechanosensitivity of the normal and diseased heart at the ion channel, cell, tissue and organ levels. Kamkin and Kiseleva have made signi?cant contributions to the investigation of mechanosentive ion channels in cardiomyocytes and ?broblasts. Their ba- ground, in addition to extensive collaborations helped them to ?nd and consolidate valuable research ?ndings from prominent specialists in the ?eld of cardiac mechanosensitivity. In the last decade, interest in the role of MEF in the heart has increased sign- cantly. MEF within cardiac tissue is a complex phenomenon in which electroph- iological changes are triggered by myocardial stretch. This phenomenon has been studiedintheclinicalcommunityforoveracenturyandmayhavebothpro-rhythmic and arrhythmogenic consequences. While signi?cant advances have been made in understanding of the effects of mechanical forces on cardiac cells, many questions remain regarding the mechanisms whereby mechanical forces are transduced into changes which alter the behavior of various cardiac cells.

Sequencing projects have revealed the presence of at least several hundred receptor kinases in a typical plant genome. Receptor kinases are therefore the largest family of primary signal transducers in plants, and their abundance suggests an immense signaling network that we have only just begun to uncover. Recent research findings indicate that individual receptor kinases fulfill important roles in growth and development, in the recognition of pathogens and symbionts or, in a few examples, in both growth and defense. This volume will focus on the roles of receptor kinases, their signaling pathways, and the ways in which these important signaling proteins are regulated.