This thesis presents a highly innovative study of the ultrafast structural and vibrational dynamics of hydrated phospholipids, the basic constituents of cell membranes. As a novel approach to the water-phospholipid interface, the author studies phosphate vibrations using the most advanced methods of nonlinear vibrational spectroscopy, including femtosecond two-dimensional infrared spectroscopy. He shows for the first time that the structure of interfacial water undergoes very limited fluctuations on a 300 fs time scale and that the lifetimes of hydrogen bonds with the phospholipid are typically longer than 10 ps. Such properties originate from the steric hindrance of water fluctuations at the interface and the orienting action of strong electric fields from the phospholipid head group dipoles. In an extensive series of additional experiments, the vibrational lifetimes of the different vibrations and the processes of energy dissipation are elucidated in detail.

This volume includes chapters by experts around the world on many aspects of microtubule imaging in living and fixed cells; assays to study microtubule function in a wide array of model organisms and cultured cells; high resolution approaches to study of the cytoskeleton. The authors share their years of experience, outlining potential pitfalls and critical factors to consider in experimental design, experimental implementation and data interpretation.

Iincludes chapters by experts around the world on many aspects of microtubule imaging in living and fixed cells; assays to study microtubule function in a wide array of model organisms and cultured cells; high resolution approaches to study of the cytoskeleton. The authors share their years of experience, outlining potential pitfalls and critical factors to consider in experimental design, experimental implementation and data interpretation.

Guide to Techniques in Mouse Development, Part B, is an authoritative guide to different methods used in enzymology, focusing on investigating mouse development using technological advances. The text provides information regarding the principles of the methods in mouse development, and it offers readers reliable experimental protocols and recipes described comprehensively by leaders in the field of enzymology. The text is divided into three sections and organized into 25 chapters. Below are several concepts covered by the text: Lentivirus transgenesis o Germline modification using mouse stem cells Electroporation Applications of transposons in mouse genetics Functional genomics using transposon systems The use of DNA transposons in detecting cancer genes in mice Recombination, conditional mutagenesis and induction of tamoxifen Genetic fate mapping using recombinases Genetic screens mouse ES cells Gene trap mutagenesis Mouse mutagenesis Self- renewal and pluripotency Transgenic RNAi applications Gene knockdowns Tetracycline-controlled transcription Gene expression profiling of mouse embryos The book is a comprehensive guide for students and professionals in genetics, cytology and molecular biology, who will find this book invaluable for their learning and practice.

This volume comprehensively covers new technologies and methodologies that have appeared for the study of mouse development.

This volume is the first of a 2 part update of volume 225, "Guide to Techniques in Mouse Development," edited by P.M. Wassarman and M.L. DePamphilis and published in 1993.

Comprehensively covers new techniques for the cryopreservation of gametes and embryos, production of transgenic and null (knockout) animals (use of ES cells), generation of conditional/inducible mutant animals, use of gene-trap mutagenesis, analysis of allele-specific expression, use of new reporter constructs, humanizing of transgenic animals, transcript profiling of mouse development, imaging of mouse development, and rederivation of animals and use of mouse genomics.

With the aim of providing a deeper insight into possible mechanisms of biological self-organization, this thesis presents new approaches to describe the process of self-assembly and the impact of spatial organization on the function of membrane proteins, from a statistical physics point of view. It focuses on three important scenarios: the assembly of membrane proteins, the collective response of mechanosensitive channels and the function of the twin arginine translocation (Tat) system. Using methods from equilibrium and non-equilibrium statistical mechanics, general conclusions were drawn that demonstrate the importance of the protein-protein interactions. Namely, in the first part a general aggregation dynamics model is formulated, and used to show that fragmentation crucially affects the efficiency of the self-assembly process of proteins. In the second part, by mapping the membrane-mediated forces into a simplified many-body system, the dynamic and equilibrium behaviour of interacting mechanosensitive channels is derived, showing that protein agglomeration strongly impacts its desired function. The final part develops a model that incorporates both the agglomeration and transport function of the Tat system, thereby providing a comprehensive description of this self-organizing process.

This volume of Current Topics in Membranes focuses on Membrane Protein Crystallization, beginning with a review of past successes and general trends, then further discussing challenges of mebranes protein crystallization, cell free production of membrane proteins and novel lipids for membrane protein crystallization.
This publication also includes tools to enchance membrane protein crystallization, technique advancements, and crystallization strategies used for photosystem I and its complexes, establishing Membrane Protein Crystallization as a needed, practical reference for researchers.

Signal transduction comprises the intracellular biochemical signals which induce the appropriate cell response to an external stimulus. The players in signal transduction are diverse, from small molecules as first messengers, to proteins, receptors, transcription factors, among many others. The different signaling pathways and the crosstalk between them originates the unique signaling profile of every cell type in the human body. The cell signaling specificity depends on several aspects including protein composition, subcellular localization and complexes and gene promoters. This textbook provides a comprehensive overview of the specific signaling pathways on a variety of human tissues. This information can be of great value for health science researchers, professionals and students to understand key pathways for tissue-specific functions in the plethora of signals, signals receptors, transducers and effectors. Chapter 3 and 15 are available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.

This volume reviews the most important recent findings of the studies on pathogenic E. coli providing a timely overview of the field. The topics covered include epidemiology of the disease in humans and animals and the biological mechanisms that shaped the pathogenic types of E. coli; shiga toxins; subtilase cytotoxin; cell cycle modulating toxins; the heat stable and heat labile exterotoxins; and much more.

"International Review of Cell and Molecular Biology "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. Impact factor for 2008: 4.935.

* Authored by some of the foremost scientists in the field * Provides up-to-date information and directions for future research * Valuable reference material for advanced undergraduates, graduate students and professional scientists

Bone marrow stem cells are the most transplanted cells worldwide. These cells are used as a replacement therapy for patients suffering from a diverse number of hematopoietic diseases and immunodeficiencies. However, the use of bone marrow cells in regenerative medicine has so far remained without much success. In the new era of pluripotent stem cells, great opportunities for establishing new therapies have opened up. The discovery of human embryonic stem cells and that of induced pluripotent (iPS) stem cells has made it possible to derive any desired tissues for regenerative medicine as iPS cell derived cells are only limited by the lack of established protocols that can be applied in humans. There is no doubt that stem cells present a new and innovative platform for establishing novel cell based therapies. The challenge is to establish new protocols that allow the successful differentiation of these cells into lineage committed cells. Embryonic Stem Cell Immunobiology: Methods and Protocols covers a variety of relevant topics, such as hematopoietic stem cells derived from ES cells, the interaction of these cells with natural killer cells or with cytotoxic T cells, and specific protocols for the derivation of hematopoietic cells and neuronal cells, to name a few. Written in the highly successful Methods in Molecular Biology 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 on troubleshooting and avoiding known pitfalls.

Authoritative and accessible, Embryonic Stem Cell Immunobiology: Methods and Protocols serves as an ideal guide to experts and non-experts interested in different aspects of stem cells.

The present edited book is an attempt to update the state of art of the knowledge on metabolism of ROS and antioxidants and their relationship in plant adaptation to abiotic stresses involving physiological, biochemical and molecular processes. The chapters are much focused on the current climate issues and how ROS metabolism can manipulate with antioxidant system to accelerate detoxification mechanism. It will enhance the mechanistic understanding on ROS and antioxidants system and will pave the path for agricultural scientists in developing tolerant crops to achieve sustainability under the changing environmental conditions. The increase in abiotic stress factors has become a major threat to sustainability of crop production. This situation has led to think ways which can help to come out with potential measures; for which it is necessary to understand the influence of abiotic stress factors on crops performance and the mechanisms by which these factors impact plants. It has now become evident that abiotic stress impacts negatively on plant growth and development at every stage of plant's life. Plants adapt to the changing environment with the adjustment at physiological, biochemical and molecular levels. The possible mechanisms involved in the negative effects of abiotic stress factors are excess production of reactive oxygen species (ROS). They alter physiological and molecular mechanisms leading to poor performance of plants. Plants however, are able to cope with these adverse effects by inducing antioxidant systems as the priority. Nevertheless, the dual role of ROS has now been ascertained which provides an evidence for regulation of plant metabolism positively on a concentration-dependent manner. Under conditions of high ROS production, the antioxidant system plays a major role in diminishing the effects of ROS. Thus, ROS production and antioxidant system are interwoven with abiotic stress conditions. The antioxidants have the capacity to hold the stability in metabolism in order to avoid disruption due to environmental disturbances.

This timely volume explores the impact of autophagy in various human diseases, emphasizing the cell biological aspects and focusing on therapeutic approaches to these diseases. The chapters cover autophagy and its potential applications on diseases ranging from obesity, osteoarthritis, pulmonary fibrosis, and inflammation, through ALS, Parkinson's, retinal degeneration, breast cancer, alcoholic liver disease and more. The final chapters round out the book with a discussion of autophagy in drug discovery and 'bench to bedside'. Chapters are contributed by leading authorities and describe the general concepts of autophagy in health and disease, marrying cell biology and pharmacology and covering: studies derived from preclinical experiments, manufacturing considerations,and regulatory requirements pertaining to drug discovery and manufacturing and production. This volume will be useful for basic scientists as well as already practicing clinicians and advanced graduate students.

"International Review of Cell and Molecular Biology "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. Impact factor for 2008: 4.935.

* Authored by some of the foremost scientists in the field * Provides up-to-date information and directions for future research * Valuable reference material for advanced undergraduates, graduate students and professional scientists

Volume 6 of Biomembranes covers transmembrane receptors and channels. A particularly important role for the membrane is that of passing messages between a cell and its environment. Part I of this volume covers receptors for hormones and growth factors. Here, as in so many other areas of cell biology, the application of the methods of molecular biology have led to the recognition of a number of families of receptors. Typically, such receptors contain an extracellular ligand binding domain, a transmembrane domain, and an intracellular catalytic domain whose activation, as a result of ligand binding, leads to generation of second messengers within the cell and stimulation of a range of cytosolic enzymes. An alternative signaling strategy, exploited in particular in the nervous system, is to use ion channels to allow controlled movement of monovalent (Na+, K+) or divalent (Ca2+) cations in or out of the cell, resulting in changes in membrane potential or alterations in the intracellular concentration of Ca2+. Part II of this volume is concerned with these ion channels and with other, often simpler, ion channel systems whose study can throw light on channel mechanism.

This book discusses the paradigm-shifting phenomenon of intrinsically disordered proteins (IDPs) and hybrid proteins containing ordered domains and functional IDP regions (IDPRs). The properties of IDPs and IDPRs are highly complementary to those deriving from the presence of a unique and well-defined three-dimensional fold. Ignored for a long time in high-resolution studies of proteins, intrinsic protein disorder is now recognized as one of the key features for a large variety of cellular functions, where structural flexibility presents a functional advantage in terms of binding plasticity and promiscuity and this volume explores this exciting new research. Recent progress in the field has radically changed our perspective to study IDPs through NMR: increasingly complex IDPs can now be characterized, a wide range of observables can be determined reporting on the structural and dynamic properties, computational methods to describe the structure and dynamics are in continuous development and IDPs can be studied in environments as complex as whole cells. This volume communicates the new exciting possibilities offered by NMR and presents open questions to foster further developments. Intrinsically Disordered Proteins Studied by NMR Spectroscopy provides a snapshot to researchers entering the field as well as providing a current overview for more experienced scientists in related areas.

"International Review of Cell and Molecular Biology "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. Impact factor for 2008: 4.935.

* Authored by some of the foremost scientists in the field * Provides up-to-date information and directions for future research * Valuable reference material for advanced undergraduates, graduate students and professional scientists

This manual offers detailed protocols for fluorescence in situ hybridization (FISH) and comparative genomic hybridization approaches, which have been successfully used to study various aspects of genomic behavior and alterations. Methods using different probe and cell types, tissues and organisms, such as mammalians, fish, amphibians (including lampbrush-chromosomes), insects, plants and microorganisms are described in 57 chapters. In addition to multicolor FISH procedures and special applications such as the characterization of marker chromosomes, breakpoints, cryptic aberrations, nuclear architectures and epigenetic changes, as well as comparative genomic hybridization studies, this 2nd edition describes how FISH can be combined with other techniques. The latter include immunostaining, electron microscopy, single cell electrophoresis and microdissection. This well-received application guide provides essential protocols for beginning FISHers and FISH experts alike working in the fields of human genetics, microbiology, animal and plant sciences.

The current scenario of increasing sensitivity towards the sustainable agriculture has given a large space to extensively utilize natural resources that are environmental friendly and are a good replacement of chemicals in agriculture. Application of organic additives in the sustainable disease management can provide new insight in sustenance of plant productivity along with improved host stress tolerance. In the present book we have focussed upon a range of organic strategies to control plant pathogens of wide spectrum in addition to maintaining robust plant health. A detailed account on the application of organic additives has been discussed, irrespective of their origin and nature. In addition, the methods of utilising these organic supplements in the management of plant diseases and promotion of plant yield in more economic way have also been presented with reference to developing, underdeveloped and developed countries. The book has included the works of eminent scholars from across the world thus flashing light on the key literature related to application of organic matters including phytoextracts, chopped leaves, composted organic manures and liquid manures in eco-friendly agriculture. The mechanisms underlying the effectiveness of these organic amendments in promoting plant health has also been presented and discussed in understandable ways.

This fully updated edition of the best-selling three-part "Methods in Enzymology "series, "Guide to Yeast Genetics and Molecular Cell Biology "is specifically designed to meet the needs of graduate students, postdoctoral students, and researchers by providing all the up-to-date methods necessary to study genes in yeast. Procedures are included that enable newcomers to set up a yeast laboratory and to master basic manipulations. Relevant background and reference information given for procedures can be used as a guide to developing protocols in a number of disciplines. This volume serves as an essential reference for any beginning or experienced researcher in the field.
Provides up-to-date methods necessary to study genes in yeast.
Includes proceedures that enable newcomers to set up a yeast laboratory and to master basic manipulations.
This volume serves as an essential reference for any beginning or experienced researcher in the field.

This fully updated edition of the best-selling three-part "Methods in Enzymology "series, "Guide to Yeast Genetics and Molecular Cell Biology "is specifically designed to meet the needs of graduate students, postdoctoral students, and researchers by providing all the up-to-date methods necessary to study genes in yeast. Procedures are included that enable newcomers to set up a yeast laboratory and to master basic manipulations. This volume serves as an essential reference for any beginning or experienced researcher in the field.
Provides up-to-date methods necessary to study genes in yeast.
Includes proceedures that enable newcomers to set up a yeast laboratory and to master basic manipulations.
This volume serves as an essential reference for any beginning or experienced researcher in the field.

This volume presents an assortment of traditional and emerging experimental procedures relevant to Schwann cell research. The chapters are divided into four parts. Part I contains protocols for in vitro culture, purification, and characterization of primary Schwann cells from diverse species and stages of nerve development. It also contains protocols to create cancer cell lines and engineered Schwann cells from unconventional sources via chemical conversion, induced differentiation or genetic intervention. Parts II and III outline a wide range of methodologies used to study Schwann cells within in vitro and in vivo systems relevant to the analysis of peripheral nerve development, cancer, axon degeneration/regeneration, and myelination. Last but not least, part IV outlines protocols for Schwann cell production, collection, labeling and transplantation in the injured peripheral nerve and spinal cord of experimental animals and human subjects. Authoritative and practical, Schwann Cells: Methods and Protocols aims to aid both experienced and new investigators to make progress in their research endeavors involving Schwann cells.

This is the first book to provide a broad framework for obtaining an in depth understanding of the state-of-the-art knowledge on abnormalities of non-coding RNAs found to be associated with colorectal cancer pathogenesis. Readers will discover possible mechanisms underlying the substantial roles played by non-coding RNAs in molecular hallmarks of colorectal cancer. This work further provides the comprehensive overview and novel insights into using of non-coding RNAs as colorectal cancer biomarkers enabling early detection of the disease, prognostic stratification of the patients and prediction of therapeutic response. The reader is introduced to the overview of modern non-coding RNAs-based therapeutic strategies, and summary of their preclinical testing performed in colorectal cancer. The work is written for researchers who want to explore current state of the knowledge in this interesting field of molecular oncology.

This volume offers an analysis of the scale and nature of the immunological issues facing regenerative medicine, drawing on the expertise of laboratories around the world who have taken up the challenge of applying their expertise in immunology to the vagaries of stem cell biology. In Part I, we explore the extent to which the principles of allograft rejection, learned over several decades from our experiences of whole organ transplantation, apply within the unique context of cell replacement therapy. Part II discusses various innovative ways of addressing the issues of immunogenicity, while, in Part III, we focus exclusively on the induction of immunological tolerance through a variety of novel approaches. It is our hope that this systematic analysis of the current state of the field will galvanise efforts to solve an issue which has so far remained intractable.

Ataxia-telangiectasia (A-T) is a rare and severe genetic disorder affecting children. A-T is a multisystem disease characterized by progressive neurodegeneration, immunodeficiency and cancer predisposition. This detailed volume explores the ever expanding field of research into the ATM (ataxia-telangiectasia, mutated) gene and the role played by ATM kinase in DNA damage signaling and diverse cellular processes. What follows is a handy desktop reference for both seasoned A-T researchers and postgraduate students, as it demonstrates the breadth of recent developments in A-T studies. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Both classic and cutting-edge techniques are described, including ATM gene mutation detection, assays for radiosensitivity and radioresistant DNA synthesis, new methods to measure ATM kinase activity by imaging microscopy and high content screening as well as proteomics, phosphoproteomics and bioinformatics approaches to decipher ATM-dependent signalling pathways. Additional methods include generation of patient-specific stem cells and approaches to study ATM functions in the nervous system. Comprehensive and practical, ATM Kinase: Methods and Protocols aims to ignite and attract the interest of colleagues from diverse fields to A-T research in an effort to bring their expertise and fresh ideas to resolve many A-T puzzles still waiting to be pieced together and to alleviate the suffering of A-T children and their families.