Recent stem cell research has revealed that miRNA and RNAi-mediated gene regulation is one of the vital determinates controlling the state of cell differentiation, with the small RNAs serving as key elements involved in regulatory network control of pluripotent cell fate determination. In RNAi and microRNA-Mediated Gene Regulation in Stem Cells: Methods, Protocols, and Applications, expert authors from laboratories across the globe contribute an accessible compendium of up-to-date, proven methods focused on the study of the titular topic. Divided into three sections, the book first gives a brief introduction to RNAi and miRNAs in stem cells, with a focus on the current status of research and future perspectives, then it continues with detailed methods and protocols for RNAi screening, transfection, and the knockdown of specific genes and pathways in several animal species, including humans and mice, concluding with a section on recently developed methods for identification of miRNAs, including a general protocol for preparation and analysis of miRNA libraries for deep sequencing, knock down of a specific gene using miRNA-based shRNA, and miRNA expression analysis using qRT-PCR. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes highlighting tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, RNAi and microRNA-Mediated Gene Regulation in Stem Cells: Methods, Protocols, and Applications serves as a valuable resource for scientists and aspiring graduate students interested in the intersection of RNAi, miRNA, and stem cell molecular biology and the exciting areas of medicine, including regenerative medicine, aging, cancer, and neurological disorders, that can be advanced through this expanding area of research.

Cultured cells have combined accessibility and the ability to expand a homogeneous cell population from a relatively limited source, thus opening up a wealth of possibilities for researchers. In Mouse Cell Culture: Methods and Protocols, expert researchers provide a number of methods for the culture of a wide range of specific cells and tissues isolated from the key genetic model of the fetal or adult mouse. Including protocols for the explant of fetal tissues and stem cells that allow developmental processes to be followed ex vivo as well as protocols for the culture of isolated cell types that allow for the study of relatively homogeneous cell populations, this volume brings together a selection of the most current methods in order to make them available in one convenient source. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Practical and authoritative, Mouse Cell Culture: Methods and Protocols serves as an immediately applicable springboard for the development of new tissue culture methods in order to advance the study and treatment of human disorders.

Muller glial cells ensheath all retinal neurons in vertebrate retinae. There are a multitude of functional interactions between neurons and Muller cells, including delivery of the light stimuli to the photoreceptor cells in the inverted vertebrate retina, a 'metabolic symbiosis' with the neurons, and the processing of visual information. Muller cells are also responsible for the maintenance of the homeostasis of the retinal extracellular milieu (ions, water, neuro-transmitter molecules, and pH). In vascularized retinae, Muller cells may also be involved in the control of angiogenesis, and the regulation of retinal blood flow. Virtually every disease of the retina is associated with a reactive Muller cell gliosis which, on the one hand, supports the survival of retinal neurons but, on the other hand, may accelerate the progress of neuronal degeneration:

Muller cells protect neurons via a release of neurotrophic factors. However, gliotic Muller cells display a dysregulation of various neuron-supportive functions. This contributes to a disturbance of retinal glutamate metabolism and ion homeostasis, and causes the development of retinal edema and neuronal cell death. Moreover, there are diseases evoking a primary Muller cell insufficiency, such as hepatic retinopathy and certain forms of glaucoma. Any impairment of supportive functions of Muller cells, primary or secondary, must cause and/or aggravate a dysfunction and loss of neurons, by increasing the susceptibility of neurons to stressful stimuli in the diseased retina.

Muller cells may be used in the future for novel therapeutic strategies to protect neurons against apoptosis (i.e. somatic gene therapy), or to differentiate retinal neurons from Muller/stem cells. Meanwhile, a proper understanding of the gliotic responses of Muller cells in the diseased retina, and of their protective vs. detrimental effects, is essential for the development of efficient therapeutic strategies that use and stimulate the neuron-supportive/-protective - and prevent the destructive - mechanisms of gliosis.

* Discusses cancer cell biology in relation to Genome stability and Cell cycle regulation Unique assembly of experts in these fields who wrote a comprehensive and deep up-to-date overview Discusses models for the understanding of DNA damage-dependent signal transduction and regulation in human cells Since the establishment of the DNA structure researchers have been highly interested in the molecular basis of the inheritance of genes and of genetic disorders. Scientific investigations of the last two decades have shown that, in addition to oncogenic viruses and signalling pathways alterations, genomic instability is important in the development of cancer. This view is supported by the findings that aneuploidy, which results from chromosome instability, is one of the hallmarks of cancer cells. Chromosomal instability also underpins our fundamental principles of understanding tumourigenesis: It thought that cancer arises from the sequential acquisition of genetic alterations in specific genes. In this hypothesis, these rare genetic events represent rate-limiting bottlenecks' in the clonal evolution of a cancer, and pre-cancerous cells can evolve into neoplastic cells through the acquisition of somatic mutations. This book is written by international leading scientists in the field of genome stability. Chapters are devoted to genome stability and anti-cancer drug targets, histone modifications, chromatin factors, DNA repair, apoptosis and many other key areas of research. The chapters give insights into the newest development of the genome stability and human diseases and bring the current understanding of the mechanisms leading to chromosome instability and their potential for clinical impact to the reader.

The Biogenesis of Cellular Organelles represents a comprehensive summary of recent advances in the study of the biogenesis and functional dynamics of the major organelles operating in the eukaryotic cell. This book begins by placing the study of organelle biogenesis in a historical perspective by describing past scientific strategies, theories, and findings and relating these foundations to current investigations. Reviews of protein and lipid mediators important for organelle biogenesis are then presented, and are followed by summaries focused on the endoplasmic reticulum, Golgi, lysosome, nucleus, mitochondria, and peroxisome. All chapters are written by experts in their fields and, though concentrated on particular topics, are integrated under the general themes of organelle structure, function, dynamics, and biogenesis. An understanding of these concepts is important for all researchers and students interested in general cell biology and particularly to those with interests in organelle function.

This book lays out numerous simple techniques for growing and carrying out experiments with many varieties of neurons. Subjects include peripheral and central neurons from vertebrate and invertebrate sources, as well as neuron-like cell lines. It also explains recent advances in our ability to introduce exogenous proteins and genes to neurons in culture. Procedures for successful protein infiltration, biolistic transfection, electroporation, and viral transgenic methods in neurons are also presented.
* Contains culture methodology for more than a dozen types of CNS and PNS neurons
* Includes most recent and reliable techniques from expert practitioners for specific experimental applications
* Addresses the latest strategies for transfecting neurons

Published since 1959, International Review of Neurobiology is a well-known series appealing to neuroscientists, clinicians, psychologists, physiologists, and pharmacologists. Led by an internationally renowned editorial board, this important serial publishes both eclectic volumes made up of timely reviews and thematic volumes that focus on recent progress in a specific area of neurobiology research.
This volume is a collection of chapters covering recent advances in the field of neurobiology. Chapters address anesthetic binding sites on the nicotinic acetylcholine receptors, NMDA receptor signal regulation, alcohol self-administration in rodents, and dopamine receptor mutations in mice.
Published since 1959, International Review of Neurobiology is a well-known series appealing to neuroscientists, clinicians, psychologists, physiologists, and pharmacologists. Led by an internationally renowned editorial board, this important serial publishes both eclectic volumes made up of timely reviews and thematic volumes that focus on recent progress in a specific area of neurobiology research.
This volume is a collection of chapters covering recent advances in the field of neurobiology. Chapters address anesthetic binding sites on the nicotinic acetylcholine receptors, NMDA receptor signal regulation, alcohol self-administration in rodents, and dopamine receptor mutations in mice.

Scientific interest in regulatory T cells has revived during the last decade. Initially described in the early seventies as suppressor T cells, the concept of suppressor/regulatory T cells went through turbulent times during the eighties when molecular analysis failed to identify putative suppressor genes. The constructive and elegant cellular experiments on regulatory T cells during the nineties, initiated by Shimon Sakaguchi and co-workers, however have brought these cells back into the limelight. Nowadays, regulatory T cells are regarded as essential components of the immune system, and several different subsets of regulatory T cells have been described. Considerable regulatory function has been attributed to the CD4+CD25+ T cell subset. These cells act by suppressing adaptive and possibly also innate immune responses thereby maintaining or restoring the balance between immunity and tolerance. The suppressive effects of CD4+CD25+ regulatory T cells are cell-contact dependent but a role for soluble factors, particularly in vivo, has been suggested as well.
The aim of this book is to bring together recent developments and viewpoints in the field of CD4+CD25+ regulatory T cells and to discuss the potential use of regulatory T cells in immunotherapy of inflammatory diseases. By linking data on regulatory T cells from experimental models with recent findings from the clinic, this topical book will be of interest to immunologists and other biomedical researchers as well as clinicians that are interested in regulation and manipulation of the immune response during (chronic) inflammatory disease.

Signal Transduction now in paperback, is a text reference on cellular signalling processes. Starting with the basics, it explains how cells respond to external cues (hormones, cytokines, neurotransmitters, adhesion molecules, extracellular matrix, etc), and shows how these inputs are integrated and co-ordinated. The first half of the book provides the conceptual framework, explaining the formation and action of second messengers, particulary cyclic nucleotides and calcium, and the mediation of signal pathways by GTP-binding proteins. The remaining chapters deal with the formation of complex signalling cascades employed by cytokines and adhesion molecules, starting at the membrane and ending in the nucleus, there to regulate gene transcription. In this context, growth is an important potential outcome and this has relevance to the cellular transformations that underlie cancer. The book ends with a description at the molecular level of how signalling proteins interact with their environment and with each other through their structural domains. Each main topic is introduced with a historical essay, detailing the sources key observations and experiments that set the scence for recent and current work.
* Coherent, precise text providing insight in depth to a subject that is central to cell biology and fundamental to many areas of biomedicine
* Conceptual colour artwork assists with the comprehension of key topics
* Extensive referencing provides an invaluable link to the core and historical literature
* Margin notes highlighting milestones in the evolution of our understanding of signalling mechanisms

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.

Gene therapy offers considerable potential for the treatment of various incurable diseases of the nervous system. This volume describes a number of different viral vectors developed for achieving high efficiency gene delivery to the brain. Vectors described include those based on adenovirus, adeno-associated virus, Herpes Simplex Virus, lentivirus, and other retroviruses. It also discusses the potential application of such viruses in treating brain tumors, Parkinson's disease, and other diseases of the nervous system.
* Provides up-to-date account of gene therapy approaches for incurable neurological disorders
* Describes a range of gene delivery methods based on different viruses

The acclaimed International Review of Cytology series presents current advances and reviews in cell biology, both plant and animal. Aricles address structure and control of gene expression, nucleocytoplasmic interactions, control of cell development and differentiation, and cell transformation and growth. Contributors to this volume include Yosef Gruenbaum, Sergey Razin, Johanna M. van der Wouden, J. M. Mitchison, Ora A. Weisz, and
Anne Regnier-Vigourous.
The acclaimed International Review of Cytology series presents current advances and reviews in cell biology, both plant and animal. Aricles address structure and control of gene expression, nucleocytoplasmic interactions, control of cell development and differentiation, and cell transformation and growth. Contributors to this volume include Yosef Gruenbaum, Sergey Razin, Johanna M. van der Wouden, J. M. Mitchison, Ora A. Weisz, and
Anne Regnier-Vigourous.

International Review of Cytology presents current advances and comprehensive reviews in cell biology--both plant and animal. Articles address structure and control of gene expression, nucleocytoplasmic interactions, control of cell development and differentiation, and cell transformation and growth. Authored by some of the foremost scientists in the field, each volume provides up-to-date information and directions for future research.
Key Features
* Each volume provides up-to-date information and directions for future research
* Authored by some of the foremost international scientists in the field
* Fully illustrated in color and black and white

Mitochondria are complex organelles, possessing a double-membrane and even their own genome, the mtDNA. They play a pivotal role in cellular metabolism, respiration, and production of ATP essential for the normal function of all human organ systems. It is not surprising, therefore, that genetic defects of mitochondrial functions cause a wide spectrum of human diseases. This book provides the first modern and truly comprehensive coverage of the biochemistry, genetics, and pathology of mitochondria in different organisms. It particularly focuses on the recent advances in our understanding of basic mitochondrial research to the consequences of dysfunction at the molecular level.

The 13 contributions written by leading researchers in the field include topics such as: mitochondrial genome evolution and mtDNA stability, mitochondrial biogenesis and protein quality control, mitochondrial morphology, assembly and function of the mitochondrial energy generation apparatus and mitochondrial metabolic pathways. These are particularly oriented to link in these various mitochondrial pathways to the clinical consequences of their dysfunctions.

This book focuses on the intersection between cell cycle regulation and embryo development. Specific modifications of the canonical cell cycle occur throughout the whole period of development and are adapted to fulfil functions coded by the developmental program. Deciphering these adaptations is essential to comprehending how living organisms develop. The aim of this book is to review the best-known modifications and adaptations of the cell cycle during development. The first chapters cover the general problems of how the cell cycle evolves, while consecutive chapters guide readers through the plethora of such phenomena. The book closes with a description of specific changes in the cell cycle of neurons in the senescent human brain. Taken together, the chapters present a panorama of species - from worms to humans - and of developmental stages - from unfertilized oocyte to aged adult.

Integrins play pivotal roles not only across a wide range of physiological processes including tissue morphogenesis, immune responses, wound healing, and regulation of cell growth and differentiation, but also in numerous pathological phenomena such as autoimmunity, thrombosis, and cancer metastasis/progression. Therefore, investigations on integrins often demand multi-disciplinary approaches, making researchers long for a handy collection of comprehensive and practical protocols that detail experimental methods for studying integrin and related cell adhesion molecule functionality. "Integrin and Cell Adhesion Molecules: Methods and Protocols "aims to provide readers not only with basic protocols in studying integrin functions, but also with summaries on those state-of-the-art technologies that have been utilized for understanding integrin functionality at the cellular, molecular, structural, and organismal levels. Divided into six convenient sections, this detailed volume covers basic protocols for the study of integrin and related cell adhesion molecule functionality "in vitro," illustrates structural biology approaches for studying integrins and related cell adhesion molecules, focuses on emerging imaging technologies for investigating cell migration, presents strategies to elucidate signaling through cell adhesion molecules, includes experimental techniques to investigate integrin functions at organismal levels in a physiological context, and showcases the most promising methods and technologies for the development of novel therapeutics and diagnostics. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls.

Both experts and non-experts in the scientific community who wish to study cell adhesion molecules and diagnostics will find "Integrin and Cell Adhesion Molecules: Methods and Protocols" authoritative, easily accessible, and vastly informative.

Now in two volumes, this completely updated and expanded edition of Embryonic Stem Cells: Methods and Protocols provides a diverse collection of readily reproducible cellular and molecular protocols for the manipulation of nonhuman embryonic stem cells. Volume two, Embryonic Stem Cell Protocols: Differentiation Models, Second Edition, covers state-of-the-art methods for deriving many types of differentiating cells from ES cells. The first volume, Embryonic Stem Cell Protocols: Isolation and Characterization, Second Edition, provides a diverse collection of readily reproducible cellular and molecular protocols for the isolation, maintenance, and characterization of embryonic stem cells. Together, the two volumes illuminate for both novices and experts our current understanding of the biology of embryonic stem cells and their utility in normal tissue homeostasis and regenerative medicine applications.

In "Animal Models for Stem Cell Therapy: Methods and Protocols, "expert researchers in the field detail disease models of hepatic, cardiovascular, neurological diseases, connective and contractile tissue. Chapters focus on a wide range of diseases and application of different kinds of stem cells and reprogrammed tissue cells (iPS).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 key tips on troubleshooting and avoiding known pitfalls.

Authoritative and practical, "Animal Models for Stem Cell Therapy: Methods and Protocols, "covers interest of basic scientists and clinicians to assess the biological as well as the therapeutic potential of stem cell therapy."

The book focuses on the aqueous interface of biomolecules, a vital yet overlooked area of biophysical research. Most biological phenomena cannot be fully understood at the molecular level without considering interfacial behavior. The author presents conceptual advances in molecular biophysics that herald the advent of a new discipline, epistructural biology, centered on the interactions of water and bio molecular structures across the interface. The author introduces powerful theoretical and computational resources in order to address fundamental topics such as protein folding, the physico-chemical basis of enzyme catalysis and protein associations. On the basis of this information, a multi-disciplinary approach is used to engineer therapeutic drugs and to allow substantive advances in targeted molecular medicine. This book will be of interest to scientists, students and practitioners in the fields of chemistry, biophysics and biomedical engineering.

Plant microtubules are key elements of cell growth, division and morphogenesis. In addition to their role in plant development and architecture, they have emerged as regulatory elements of signalling and important targets of evolution.

Since the publication of the first edition of Plant Microtubules in 2000, our understanding of microtubules and their manifold functions have advanced substantially. Consisting of the following three parts, this book highlights the morphogenetic potential of plant microtubules from three general viewpoints: Microtubules and Morphogenesis: control of cell axis during division and expansion, cross-talk with actin filaments, mechanical properties of the cell wall. Microtubules and Environment: the role of microtubules during the sensing or response of environmental factors such as pathogens or abiotic stresses. Microtubules and Evolution: complexity and specialization of plant microtubules in the context of plant evolution.

The book is an invaluable source of information for researchers as well as for graduate and advanced students.

Kinetochores orchestrate the faithful transmission of chromosomes from one generation to the next. Kinetochores were first depicted over 100 years ago, but kinetochore research has progressed by leaps and bounds since the first description of their constituent DNA and proteins in the 1980s. "The Kinetochore: from Molecular Discoveries to Cancer Therapy" presents a thorough up-to-date analysis of kinetochore and centromere composition, formation, regulation, and activity, both in mitosis and meiosis, in humans and "model" eukaryotic species, and at natural and mutant neocentromeres. Recently initiated translational research on kinetochores is also discussed as kinetochores are being mined as a very rich target for the next generations of anti-cancer drugs.

RNA technologies are the driving forces of modern medicine and biotechnology. They combine the fields of biochemistry, chemistry, molecular biology, cell biology, physics, nanotechnology and bioinformatics. The combination of these topics is set to revolutionize the medicine of tomorrow. After more than 15 years of extensive research in the field of RNA technologies, the first therapeutics are ready to reach the first patients. Thus we are witnessing the birth of a very exciting time in the development of molecular medicine, which will be based on the methods of RNA technologies. This volume is the first of a series. It covers various aspects of RNA interference and microRNAs, although antisense RNA applications, hammerhead ribozyme structure and function as well as non-coding RNAs are also discussed. The authors are internationally highly respected experts in the field of RNA technologies.

Single Cell Diagnostics: Methods and Protocols applies modern
molecular diagnostic techniques to the analysis of single cells, small
numbers of cells, or cell extracts. Emphasis is placed on non-invasive
analysis of single cell metabolites and the direct analysis of RNA and
DNA from single cells, with a focus on polymerase chain reaction and
fluorescence in situ hybridization. The authors, all leading
scientists in the field, provide current and innovative methods for
extracting useful, accurate, and reliable data from single-cell
samples. The stepwise, technically precise protocols presented in this
volume thoroughly describe single-cell assays, from material collection
and isolation of cells to data interpretation. Cell-types analyzed
include embryonic blastomeres, fibroblasts, and lymphocytes. Among the
techniques described are real-time quantitative PCR, isothermal whole
genome amplification, comparative genomic hybridization, real-time
genetic expression analysis, and the production of RNA and cDNA
libraries. This handbook will be essential for researchers and
clinicians, and practitioners providing care for couples seeking
treatment for infertility or preimplantation genetic diagnosis.

This volume of the acclaimed Methods in Cell Biology series provides specific examples of applications of confocal microscopy to cell biological problems. It is an essential guide for students and scientists in cell biology, neuroscience, and many other areas of biological and biomedical research, as well as research directors and technical staff of microscopy and imaging facilities.
An integrated and up-to-date coverage on the many various techniques and uses of the confocal microscope (CM).
Key Features
* Includes detailed protocols accessible to new users
* Details how to set up and run a "Confocal Microscope Core Facility"
* Contains over 170 figures

In this volume of "Cell and Molecular Responses to Stress" articles provide up-to-date information on key areas of signal sensing (sensing of pain, heat, cold, light, infrared radiation), molecules involved in the intracellular transmission of these signals, metabolic responses to stress including changes in gene expression and production of specialized proteins that aid cell responses to factors including interrupted blood supply (ischemia), oxygen limitation (hypoxia/anoxia), freezing and dehydration, amino acid limitation, radiation and processing drugs. There are chapters which also provide insights into new technologies (such as cDNA arrays), analysis of metabolic control theory (a key method for analysing stress effects on cells), and examine how enzymes evolve in the face of stress.