A light-hearted look at the nature of academic science, intended for anyone interested in biology but particularly for biology students who want to find out what the future holds in store. The "Egg" of the title refers to the science of developmental biology, which is the speciality of the author, and which provides the material for many of the anecdotes. The "Ego" relates to the vanity of the scientists themselves. Academic scientists have to struggle to maintain their research funding. To do this they must persuade other scientists that they are very good, and that means working at a good institution, publishing papers in the most fashionable journals and giving lectures at the most prestigious meetings. Success often goes to those with the largest egos and it is their style of operation that is described in this book. The author is a well-known scientist who has worked at both universities and research institutes. He has published over 100 scientific papers and an influential book about embryonic development: "From Egg to Embryo".

The previous edition of Transmembrane Signaling Protocols was published in 1998. Since then the human genome has been completely sequenced and new methods have been developed for the use of microarrays and proteomics to analyze global changes in gene expression and protein profiles. These advances have increased our ability to understand transmembrane signaling processes in much greater detail. They have also simultaneously enhanced our ability to determine the role of a large number of newly identified molecules in signaling events. In addition, novel video microscopy methods have been developed to image transmembrane signaling events in live cells in real time. In view of these major advances, it is time to update the previous edition. Because of the success of that volume, we have chosen to keep the essential character of the book intact. Introductory chapters from experts have been included to provide overall perspective and an overview of recent advances in signal transduction pathways. The individual chapters now include comp- hensive detailed methods, studies in genetically tractable systems, fluorescence microscopy in live single cells, ex vivo analysis of primary cells from tra- genic mice, as well as genomic and proteomic approaches to the analysis of transmembrane signaling events. We would like to express our deep gratitude to the coauthors of this publi- tion. We hope that Transmembrane Signaling Protocols, Second Edition will serve as a valuable resource for future progress in the study of signal transd- tion pathways.

Adoptive Cell Transfer, Volume 371 in the International Review of Cell and Molecular Biology series highlights advances in the field, with this new volume presenting interesting chapters written by an international board of authors who expound on topics such as the Impact of tumor microenvironment on Adoptive Cell Transfer activity, Dendritic Cell Transfer, CAR-T Cell dysfunction and exhaustion, NK Cell-based cancer immunotherapy, Enabling CAR-T cells for solid tumors: rage against the suppressive tumor microenvironment, Improving Adoptive T-Cell therapy with cytokines administration, and What will (and should) be improved in Immunotherapy with CAR?

This volume of the Methods in Molecular Biology series is entirely devoted to the study of steroid receptor biology. Steroid hormone receptors represent a powerful system for the study of both the most fundamental molecular mec- nisms of gene regulation and control and the gross physiological responses of organisms to steroid hormones. Research in this field has brought forth advances in the treatment of cancer, endocrine disorders, and reproductive biology, and allowed elucidation of the fundamental biological mechanisms of gene expr- sion. In Steroid Receptor Methods: Protocols and Assays, the reader will find a collection of methods and protocols submitted by many fine steroid receptor researchers from throughout the world. These authors have been instructed to create a highly informative cross-section of the latest research techniques ava- able. The resulting work is timely, useful, and approachable for both the ex- rienced researcher and the novice to the field. Because the steroid receptor family is represented by a wonderfully diverse, yet strongly interrelated set of steroid receptor proteins, Steroid Receptor Methods contains protocols for the prod- tion and purification of a variety of receptor forms, including the progesterone, glucocorticoid, and androgen receptors. These procedures provide the raw ma- rial needed to conduct sophisticated biochemical analysis of receptor properties. Other techniques presented allow the reader to perform biochemical experiments on DNA binding characteristics, hormone binding assays, and protocols using combinatorial chemistry for drug discovery.

In the last few years, significant breakthroughs in transcription research expanded our appreciation for the complexity of molecular controls on gene expression in mammalian cells. In Transcription Factors: Methods and Protocols, experts in the field describe state-of-the-art approaches that investigators can use to probe critical mechanisms underlying transcription factor nuclear-cytoplasmic trafficking as well as to assess the functional impact of post-translational modifications on transcription factor function. The chapters are written by prominent scientists, many of whom developed these methods, and highlight protocols that focus on specific transcription factor family members with particular relevance to human disease. 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, Transcription Factors: Methods and Protocols compiles the latest techniques for elucidating controls on transcription factor intracellular localization and activity, and consequently is unlike any other methods-based text on transcriptional regulation today.

This volume compiles a broad range of step-by-step protocols, complementary to the ones published in the first edition of this book, to study various aspects of mitochondrial structure and function in different model organisms, both in vitro and in vivo. 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, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Mitochondria: Practical Protocols, Second Edition aims to be useful for beginners as well as for experienced researchers in the field.

There is no doubt that nowadays, biology benefits greatly from mathematics. In particular, cellular biology is, besides population dynamics, a field where tech niques of mathematical modeling are widely used. This is reflected by the large number of journal articles and congress proceedings published every year on the dynamics of complex cellular processes. This applies, among others, to metabolic control analysis, where the number of articles on theoretical fundamentals and experimental applications has increased for about 15 years. Surprisingly, mono graphs and textbooks dealing with the modeling of metabolic systems are still exceptionally rare. We think that now time is ripe to fill this gap. This monograph covers various aspects of the mathematical description of enzymatic systems, such as stoichiometric analysis, enzyme kinetics, dynamical simulation, metabolic control analysis, and evolutionary optimization. We believe that, at present, these are the main approaches by which metabolic systems can be analyzed in mathematical terms. Although stoichiometric analysis and enzyme kinetics are classical fields tracing back to the beginning of our century, there are intriguing recent developments such as detection of elementary biochemical syn thesis routes and rate laws for the situation of metabolic channeling, which we have considered worth being included. Evolutionary optimization of metabolic systems is a rather new field with promising prospects. Its goal is to elucidate the structure and functions of these systems from an evolutionary viewpoint."

This book provides an update of recent advances in the basic and clinical applications of cell-based therapies for myocardial repair and regeneration in ischemic heart disease (IHD) and heart failure (HF). The first sections of the book illuminate basic aspects of stem cells such as definitions, isolation criteria and characterization of embryonic and adult stem cells, as well as pluripotent stem cells and tissue specific progenitor and stem cells. In the following section, the text examines the role of critical regulators of stem cell differentiation in myocardial regeneration, that include circadian rhythyms, microRNAs, epigenetics, microvescicles, and exosomes. The text concludes with a review of the use of stem/progenitor cells in cardiac regeneration and discuss current controversies, unresolved issues, challenges, and future directions. Stem Cells and Cardiac Regeneration is addressed to a wide audience consisting of cardiologists, clinicians, and research scientists, who will learn new concepts in cardiovascular regeneration and repair with special focus on molecular mechanisms and therapeutic potential for cardiovascular regeneration.

Membrane fusion is an important molecular event which plays a pivotal role in many dynamic cellular processes, such as exocytosis, endocytosis, membrane biogenesis, fertilization, etc. While many reports on the physico-chem1cal process involved in membrane fusion have appeared in the literature and much information has accumulated, there has been no comprehensiv meeting of workers in the field. A recent symposium which took place at the Center for Tomorrow, State University of New York at Buffalo, New York, April 27-29, 1987, was the first meeting of its kind to specifically address the molecular mechanisms of membrane fusion. The Symposium consisted of oral, workshop and poster presentation sessions. The papers presented in the oral and workshop sessions are collected here and arranged approximately in the order presented at the Symposium. These papers are the most up-to-date and representative work at the for front of each aspect of membrane fusion. Although the readers may find some differences in interpretations regarding the molecular mechanisms of membrane fusion, there is an over all concensus that increased hydrophobicity and dehydration of the membrane surface are essential physico-chemical factors for membrane fusion to occur. We trust that these papers will contribute to your further understanding of the mechanisms of membrane fusion."

Defining and understanding cellular and molecular mechanisms that are relevant to women's health has become a critical area of scientific pursuit. Until recently, very little effort has been place on defining or understanding critical differences between women and men that may be critical to the overall health of the woman. In 1990, the National Institutes of Health recognized this gap in knowledge resulting in the creation of the Office of Research on Women's Health. One of the purposes of this office was to advance the understanding of health issues from the women's perspective from both a basic and clinical scientific perspective. From a scientific evolution of understanding, the existence of this office is new and thus there has not been enough time for new information to integrate itself in our current scientific thought process. This book will seek to capture and disseminate our current understanding of scientific advancements relevant to women's health and provide the information to a broad audience. The purpose of this work is to discuss recent advancements in basic science across three areas of concern for women's health. In addition, the book will provide "translational" chapters that attempt to place the basic science work in context within our current understanding of the human. Although it is well acknowledge that gender differences exist across organ function which translates into differences in whole body function, until recently little effort has been made to define basic mechanisms within various tissues within the woman. This work will focus on recent scientific findings that are relevant to women's health and to provide novel and relevant information to interested scientists and clinicians.

In the past few years, the scientific community has witnessed significant progress in the study of ion channels. Technological advancement in biophysics, molecular biology, and immunology has been greatly ac celerated, making it possible to conduct experiments which were deemed very difficult if not impossible in the past. For example, patch-clamp techniques can now be used to measure ionic currents generated by almost every type of cell, thereby allowing us to analyze whole-cell and single channel events. It is now possible to incorporate purified ion channel components into lipid bilayers to reconstitute an "excitable membrane." Gene cloning and monoclonal antibody techniques provide us with new approaches to the study of the molecular structure of ion channels. A variety of chemicals have now been found to interact with ion channels. One of the classical examples is represented by tetrodotoxin, a puffer fish poison, which was shown in the early 1960s to block the voltage-activated sodium channel in a highly specific and potent manner.

A general review of lipid bilayer structure and dynamics is given, including such current topics as the hydration of lipid bilayers, the superstructural behaviour of bilayers at different states of hydration and external conditions, the role and behaviour of lipid bilayers on fusion and rupture and the interaction of lipid bilayers with small organic molecules and additives and of protein lipid bilayer interactions. In addition, recent research on lipid interaction with proteins and other molecules in monolayers is reviewed, and the use of highly aligned samples under biologically relevant conditions and the benefits derived from such preparations are addressed. Finally, the latest approach in simulation of impurities within a lipid bilayer is introduced. This book will be a comprehensive review of the current state of biologically relevant model membrane systems which will become an indispensible reference for the "working biophysicist."

Mitosis: Methods and Protocols provides state-of-the-art overviews on the most important approaches currently used in mitosis research spanning from the analysis of single molecules in isolation to their utilization within the complex environment of the cell. The volume is divided into four parts, each focused on methods pertaining to distinct aspects of mitosis research. Part I presents approaches for visualizing and analyzing the dynamic behaviors of the spindle apparatus, the microtubule based machine that drives chromosome segregation. Part II focuses more generally on methods for studying and manipulating the microtubule cytoskeleton in cells and complex cell free extracts. Part III provides state of the art biophysical and high resolution microscopy approaches for assessing complex interactions between microtubules and microtubule-associated proteins in isolation as well as microtubule structure in cells. Part IV provides methods for studying the effects of cell shape on cell division and methods for quantifying aneuploidy (aberrant chromosome number) which frequently results from mitotic defects and has been linked to human maladies ranging from birth defects to cancer. 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, Mitosis: Methods and Protocols seeks to provide diverse methods and new techniques to address new or old questions related to the mechanisms of mitosis.

Agrobacterium tumefaciens is a soil bacterium that for more than a century has been known as a pathogen causing the plant crown gall disease. Unlike many other pathogens, Agrobacterium has the ability to deliver DNA to plant cells and permanently alter the plant genome. The discovery of this unique feature 30 years ago has provided plant scientists with a powerful tool to genetically transform plants for both basic research purposes and for agric- tural development. Compared to physical transformation methods such as particle bomba- ment or electroporation, Agrobacterium-mediated DNA delivery has a number of advantages. One of the features is its propensity to generate single or a low copy number of integrated transgenes with defined ends. Integration of a single transgene copy into the plant genome is less likely to trigger "gene silencing" often associated with multiple gene insertions. When the first edition of Agrobacterium Protocols was published in 1995, only a handful of plants could be routinely transformed using Agrobacterium. Ag- bacterium-mediated transformation is now commonly used to introduce DNA into many plant species, including monocotyledon crop species that were previously considered non-hosts for Agrobacterium. Most remarkable are recent devel- ments indicating that Agrobacterium can also be used to deliver DNA to non-plant species including bacteria, fungi, and even mammalian cells.

This volume details computational techniques for analyses of a wide range of biological contexts, providing an overview of the most up-to-date techniques used in the field. Chapters guide the reader through available data resources and analysis methods and easy-to-follow protocols that allow the researcher to apply various computational tools to an array of different data types. 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 and computational protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Computational Cell Biology: Method and Protocols aims to ensure successful results in the further study of this vital field.

Modern cell biology has brought improvements in therapy for advanced malignant diseases through immunomodulation, hematopoietic stem cell transplantation, and other advanced techniques. Collected here are selected papers from the Fifth International Symposium of Keio University for Life Sciences and Medicine on Cell Therapy.
All chapters include innovative basic research for clinical application: immunotherapy, cancer vaccination, molecular biology of hematopoietic stem cells, stem cell processing, and gene therapy.
The book is divided into three parts: Immunotherapy for malignant diseases; Hematopoietic stem cell biology and clinical application; and International collaboration in hematopoietic stem cell transplantation. Included in the third part is information on bone marrow registries from around the world.
The book thus presents up-to-date information on biological and clinical aspects of treatment, with insight into the future of cell therapy.

Signaling networks are composed of numerous signaling pathways and each has its own intricate component parts. Signaling outputs are dynamic, extraordinarily complex and yet highly specific. In, Computational Modeling of Signaling Networks: Methods and Protocols, expert researchers in the field provide key techniques to study signaling networks. Focusing on Systems of ODEs, parameterization of signaling models, signaling pathways, mass-action kinetics and ODEs, and how to use modeling to plan experiments. 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, Computational Modeling of Signaling Networks: Methods and Protocols aids scientists in continuing study of how signaling networks behave in space and time to generate specific biological responses and how those responses impact biology and medicine.

This volume includes comprehensive descriptions of miRNA biogenesis and their role in the development and progression of various human diseases. The first few chapters of MicroRNA Profiling: Methods and Protocols discuss the effects of over-expressing and repressing of a target miRNA and their effects on cell viability and proliferation. The next few chapters explore the protocols for total RNA isolation from cells and cell-derived product including formalin fixed paraffin embedded tissue and plant tissue. The last few chapters discuss isolation and characterization of exosomes from medium conditioned by cell lines, serum, and plasma specimens. This book also includes discussions of several software tools, such as miRandola, PicTar, DIANA, and miRWalk. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Comprehensive and cutting-edge, MicroRNA Profiling: Methods and Protocols is a valuable resource for anyone interested in the field of Micro RNAs.

Soil salinity is a key abiotic-stress and poses serious threats to crop yields and quality of produce. Owing to the underlying complexity, conventional breeding programs have met with limited success. Even genetic engineering approaches, via transferring/overexpressing a single 'direct action gene' per event did not yield optimal results. Nevertheless, the biotechnological advents in last decade coupled with the availability of genomic sequences of major crops and model plants have opened new vistas for understanding salinity-responses and improving salinity tolerance in important glycophytic crops. Our goal is to summarize these findings for those who wish to understand and target the molecular mechanisms for producing salt-tolerant and high-yielding crops. Through this 2-volume book series, we critically assess the potential venues for imparting salt stress tolerance to major crops in the post-genomic era. Accordingly, perspectives on improving crop salinity tolerance by targeting the sensory, ion-transport and signaling mechanisms were presented in Volume 1. Volume 2 now focuses on the potency of post-genomic era tools that include RNAi, genomic intervention, genome editing and systems biology approaches for producing salt tolerant crops.

This new volume reviews current progress on different approaches of in vivo reprogramming technology. Leaders in the field discuss how in vivo cell lineage reprogramming can be used for tissue repair and regeneration in different organs, including brain, spinal cord, pancreas, liver and heart. Recent studies on in vivo cell reprogramming towards pluripotency are reviewed; examples are given to show its potential in regenerative medicine. In each chapter, the regenerative potential of different in vivo reprogramming approaches is discussed in detail. More specifically, how different tissue failures or damages can be treated with this technology is explained. Examples from various animal models are given and the regenerative potential of in vivo reprogramming is compared to that of cell transplantation studies. The last chapter discusses current challenges of these preclinical studies and gives suggestions in order to improve the current strategies. Future directions are indicated for the transition of in vivo reprogramming technology to clinical settings. This is among the first books in the literature which specifically focuses on the in vivo reprogramming technology in regenerative medicine and these chapters collectively cover one of the most important and exciting topics of regenerative medicine.

This book serves as an introduction to targeted genome editing, beginning with the background of this rapidly developing field and methods for generation of engineered nucleases. Applications of genome editing tools are then described in detail, in iPS cells and diverse organisms such as mice, rats, marine invertebrates, fish, frogs, and plants. Tools that are mentioned include zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and CRISPR/Cas9, all of which have received much attention in recent years as breakthrough technologies. Genome editing with engineered nucleases allows us to precisely change the target genome of living cells and is a powerful way to control functional genes. It is feasible in almost all organisms ranging from bacteria to plants and animals, as well as in cultured cells such as ES and iPS cells. Various genome modifications have proven successful, including gene knockout and knock-in experiments with targeting vectors and chromosomal editing. Genome editing technologies hold great promise for the future, for example in biomedical research, clinical medicine, and generation of crops and livestock with desirable traits. A wide range of readers will find this book interesting, and with its focus on applications in a variety of organisms and cells, the book will be valuable for life scientists in all fields.

Computers have revolutionized the analysis of sequencing data. It is unlikely that any sequencing projects have been performed in the last few years without the aid of computers. Recently their role has taken a further major step forward. Computers have become smaller and more powerful and the software has become simpler to use as it has grown in sophistication. This book reflects that change since the majority of packages described here are designed to be used on desktop computers. Computer software is now available that can run gels, collect data, and assess its accuracy. It can assemble, align, or compare multiple fragments, perform restriction analyses, identify coding regions and specific motifs, and even design the primers needed to extend the sequencing. Much of this soft ware may now be used on relatively inexpensive computers. It is now possible to progress from isolate d DNA to database submission without writing a single base down. To reflect this progression, the chapters in our Sequence Data Analysis Guidebook are arranged, not by software package, but by fimction. The early chapters deal with examining the data produced by modem automated sequenc ers, assessing its quality, and removing extraneous data. The following chap ters describe the process of aligning multiple sequences in order to assemble overlapping fragments into sequence contigs to compare similar sequences from different sources. Subsequent chapters describe procedures for compar ing the newly derived sequence to the massive amounts of information in the sequence databases."