Animal cells are the preferred "cell factories" for the production of complex molecules and antibodies for use as prophylactics, therapeutics or diagnostics. Animal cells are required for the correct post-translational processing (including glycosylation) of biopharmaceutical protein products. They are used for the production of viral vectors for gene therapy. Major targets for this therapy include cancer, HIV, arthritis, cardiovascular and CNS diseases and cystic fibrosis. Animal cells are used as in vitro substrates in pharmacological and toxicological studies. This book is designed to serve as a comprehensive review of animal cell culture, covering the current status of both research and applications. For the student or R&D scientist or new researcher the protocols are central to the performance of cell culture work, yet a broad understanding is essential for translation of laboratory findings into the industrial production. Within the broad scope of the book, each topic is reviewed authoritatively by experts in the field to produce state-of-the-art collection of current research. A major reference volume on cell culture research and how it impacts on production of biopharmaceutical proteins worldwide, the book is essential reading for everyone working in cell culture and is a recommended volume for all biotechnology libraries.

The field of epigenetics has grown exponentially in the past decade, and a steady flow of exciting discoveries in this area has served to move it to the forefront of molecular biology. Although epigenetics may previously have been considered a peripheral science, recent advances have shown considerable progress in unraveling the many mysteries of nontraditional genetic processes. Given the fast pace of epigenetic discoveries and the groundbreaking nature of these developments, a thorough treatment of the methods in the area seems timely and appropriate and is the goal of Epigenetics Protocols. The scope of epigenetics is vast, and an exhaustive analysis of all of the techniques employed by investigators would be unrealistic. However, this (TM) volume of Methods in Molecular Biology covers three main areas that should be of greatest interest to epigenetics investigators: (1) techniques related to analysis of chromatin remodeling, such as histone acetylation and methylation; (2) methods in newly developed and especially promising areas of epigenetics such as telomere position effects, quantitative epigenetics, and ADP ribosylation; and (3) an updated analysis of techniques involving DNA methylation and its role in the modification, as well as the maintenance, of chromatin structure.

Over the past decade the importance of natural resources for sustainable agricultural development has been increasingly discussed at international forums and conferences. Aside from the sustainable management of soil, water, and air, it now seems to be accepted that the sustainable management of genetic resources is one of the four indispensable preconditions for a sustainable agriculture. The discussion on conservation of plant genetic resources for food and agriculture (PGRFA), however, has to reflect the costs of conservation as well. These have not yet been discussed intensively. The study analyzes the conservation costs of plant genetic resources; it also assesses the effectiveness of conservation and the efficiency of the different conservation instruments. It is based on extensive surveys in relevant countries. Following the detailed cost and impact analysis, the results show that the effectiveness of conservation strategies may be increased.

The growth of human population has increased the demand for improved yield and quality of crops and horticultural plants. However, plant productivity continues to be threatened by stresses such as heat, cold, drought, heavy metals, UV radiations, bacterial and fungal pathogens, and insect pests. Long noncoding RNAs are associated with various developmental pathways, regulatory systems, abiotic and biotic stress responses and signaling, and can provide an alternative strategy for stress management in plants. Long Noncoding RNAs in Plants: Roles in development and stress provides the most recent advances in LncRNAs, including identification, characterization, and their potential applications and uses. Introductory chapters include the basic features and brief history of development of lncRNAs studies in plants. The book then provides the knowledge about the lncRNAs in various important agricultural and horticultural crops such as cereals, legumes, fruits, vegetables, and fiber crop cotton, and their roles and applications in abiotic and biotic stress management.

The study of the prehistory of East Asia is developing very rapidly. In uncovering the story of the flows of human migration that constituted the peopling of East Asia there exists widespread debate about the nature of evidence and the tools for correlating results from different disciplines.

Drawing upon the latest evidence in genetics, linguistics and archaeology, this exciting new book examines the history of the peopling of East Asia, and investigates the ways in which we can detect migration, and its different markers in these fields of inquiry. Results from different academic disciplines are compared and reinterpreted in the light of evidence from others to attempt to try and generate consensus on methodology. Taking a broad geographical focus, the book also draws attention to the roles of minority peoples - hitherto underplayed in accounts of the region's prehistory - such as the Austronesian, Tai-Kadai and Altaic speakers, whose contribution to the regional culture is now becoming accepted.

Past Human Migrations in East Asia presents a full picture of the latest research on the peopling of East Asia, and will be of interest to scholars of all disciplines working on the reconstruction of the peopling of East and North East Asia.

This volume is designed to provide a framework for studying the public policy implications of a broad range of biomedical technologies. Each chapter focuses on the policy issues and political activities surrounding a single technology. Contributors address such issues as new reproductive technologies, animal experimentation, contraceptive drugs, genetic markers and technology and the aging society.

2. IMPORTANCE OF NITROGEN METABOLISM 2. 1. Range of naturally occurring nitrogenous components in forest trees 2. 2. Gene expression and mapping 2. 3. Metabolic changes in organized and unorganized systems 2. 4. Nitrogen and nutrition 2. 5. Aspects of intermediary nitrogen metabolism 3. NITROGEN METABOLISM IN GROWTH AND DEVELOPMENT 3. 1. Precultural factors 3. 2. Callus formation 3. 3. Cell suspensions 3. 3. 1. Conifers 3. 3. 2. Acer 3. 4. Morphogenesis 3. 4. 1. Nitrogen metabolism of natural embryos 3. 4. 2. Somatic embryogenesis 3. 4. 2. 1. Sweetgum (Liquidambar styraciflua) 3. 4. 2. 2. Douglar-fir and loblolly pine 3. 4. 3. Organogenesis 4. OUTLOOK 11. CARBOHYDRATE UTILIZATION AND METABOLISM - T. A. Thorpe 325 1. INTRODUCTION 2. NUTRITIONAL ASPECTS 3. CARBOHYDRATE UPTAKE 4. CARBOHYDRATE METABOLISM 4. 1. Sucrose degradation 4. 2. Metabolism of other carbon sources 4. 3. Hexose mobilization and metabolism 4. 3. 1. Cell cycle studies 4. 3. 2. Growth studies 4. 3. 3. Organized development 4. 4. Cell wall biogenesis 4. 4. 1. Primary cell walls 4. 4. 2. Cell wall turnover 4. 4. 3. Secondary cell walls 4. 5. Carbon skeleton utilization 5. OSMOTIC ROLE 6. CONCLUDING THOUGHTS 369 12. THE USE OF IN VITRO TECHNIQUES FOR GENETIC MODIFICATIO~FOREST TREES - E. G. Kirby 1. INTRODUCTION 2. IN VITRO SELECTION 2. 1. Natural variation 2. 2. Induction of variation 2. 3. Selection techniques 2. 4. Plant regeneration 2 . * 5. Applications x 3. SOMATIC HYBRIDIZATION 3. 1.

In the field of molecular evolution, inferences about past evolutionary events are made using molecular data from currently living species. With the availability of genomic data from multiple related species, molecular evolution has become one of the most active and fastest growing fields of study in genomics and bioinformatics.

Most studies in molecular evolution rely heavily on statistical procedures based on stochastic process modelling and advanced computational methods including high-dimensional numerical optimization and Markov Chain Monte Carlo. This book provides an overview of the statistical theory and methods used in studies of molecular evolution. It includes an introductory section suitable for readers that are new to the field, a section discussing practical methods for data analysis, and more specialized sections discussing specific models and addressing statistical issues relating to estimation and model choice. The chapters are written by the leaders of field and they will take the reader from basic introductory material to the state-of-the-art statistical methods.

This book is suitable for statisticians seeking to learn more about applications in molecular evolution and molecular evolutionary biologists with an interest in learning more about the theory behind the statistical methods applied in the field. The chapters of the book assume no advanced mathematical skills beyond basic calculus, although familiarity with basic probability theory will help the reader. Most relevant statistical concepts are introduced in the book in the context of their application in molecular evolution, and the book should be accessible for most biology graduate students with an interest in quantitative methods and theory.

Rasmus Nielsen received his Ph.D. form the University of California at Berkeley in 1998 and after a postdoc at Harvard University, he assumed a faculty position in Statistical Genomics at Cornell University. He is currently an Ole Romer Fellow at the University of Copenhagen and holds a Sloan Research Fellowship. His is an associate editor of the Journal of Molecular Evolution and has published more than fifty original papers in peer-reviewed journals on the topic of this book.

From the reviews:

..".Overall this is a very useful book in an area of increasing importance." Journal of the Royal Statistical Society

"I find Statistical Methods in Molecular Evolution very interesting and useful. It delves into problems that were considered very difficult just several years ago...the book is likely to stimulate the interest of statisticians that are unaware of this exciting field of applications. It is my hope that it will also help the 'wet lab' molecular evolutionist to better understand mathematical and statistical methods." Marek Kimmel for the Journal of the American Statistical Association, September 2006

"Who should read this book? We suggest that anyone who deals with molecular data (who does not?) and anyone who asks evolutionary questions (who should not?) ought to consult the relevant chapters in this book." Dan Graur and Dror Berel for Biometrics, September 2006

"Coalescence theory facilitates the merger of population genetics theory with phylogenetic approaches, but still, there are mostly two camps: phylogeneticists and population geneticists. Only a few people are moving freely between them. Rasmus Nielsen is certainly one of these researchers, and his work so far has merged many population genetic and phylogenetic aspects of biological research under the umbrella of molecular evolution. Although Nielsen did not contribute a chapter to his book, his work permeates all its chapters. This book gives an overview of his interests and current achievements in molecular evolution. In short, this book should be on your bookshelf." Peter Beerli for Evolution, 60(2), 2006"

The determination of protein function has been a major goal of molecular biology since the founding of the discipline. However, as we learn more about gene function, we discover that the context within which a gene is expressed controls the specific function of that gene. It has become critical to establish the background in which gene function is determined and to perform experiments in multiple applicable backgrounds.In "Gene Function Analysis, Second Edition," a number of computational and experimental techniques are presented for identifying not only the function of an individual gene, but also the partners that work with that gene. The theme of data integration runs strongly through the computational techniques, with many focusing on gathering data from different sources and different biomolecular types. Experimental techniques have evolved to determine function in specific tissues and at specific times during development. 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.

Authoritative and easily accessible, "Gene Function Analysis, Second Edition" seeks to serve both professionals and novices with a growing understanding of the complexity of gene function."

General inspection of a role performed in the cell by RNAs allows us to distinguish three major groups of transcripts: I. protein-coding mRNAs, II. non-coding housekeeping and III. regulatory RNAs.

The housekeeping RNAs include RNA classes that are generally, constitutively expressed and whose presence is required for normal function and viability of the cells. On the other hand, a group of regulatory RNAs includes RNA species that are expressed at certain stages of organism development or cell differentiation or as a response to external stimuli and can affect expression of other genes on the levels of transcription or translation.

Non-coding RNA transcripts form a heterogeneous class of RNAs that can not be characterized by a single specific function. Initially, the term non-coding RNA (ncRNA) was used primarily to describe polyadenylated and a capped eukaryotic RNAs transcribed by RNA polymerase II, but lacking long open reading frames. Now, this definition can be extended to cover all RNA transcripts that do not show protein-coding capacity and is sometimes used to describe any RNA that does not encode protein, including introns.

This book is an in-depth look at the function of Non-Coding RNAs and their relationship to Molecular Biology and Molecular Biology.

This volume provides up-to-date and novel techniques for various screening technologies currently used in metagenomics and related areas. Starting with DNA/RNA isolation from environmental samples, the book continues by delving into areas such as current methods used to isolate DNA and construct metagenomic libraries, establishment of metagenome libraries in non-E. coli hosts, and topics like function-driven mining of metagenomic DNA, screening and analyzing protocols for a wide array of different genes encoding enzymes, bacterial viruses and much more. 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 and readily reproducible protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Metagenomics: Methods and Protocols, Third Edition provides a comprehensive collection of up-to-date metagenome protocols and tools for the recovery of many major types of biocatalysts and allows for the easy setup of these screens in microbiology laboratories.

Genetically Modified Plants, Second Edition, provides an updated roadmap and science-based methodology for assessing the safety of genetic modification technologies, as well as risk assessment approaches from regulators across different agroecosystems. This new edition also includes expanded coverage of technologies used in plant improvement, such as RNA-dependent DNA methylation, reverse breeding, agroinfiltration, and gene-editing technologies such as CRISPR and TALENS. This book is an essential resource for anyone interested in crop improvement, including students and researchers, practitioners in regulatory agencies, and policymakers involved in plant biotechnology risk assessment.

The identification of normal and breast cancer stem cells has offered a new vision of this heterogeneous disease and new hopes for its prognosis and treatment. This volume provides an overview of recent developments in mammary stem cell research and discusses the many varieties of approaches used by researchers to investigate the properties and functions of mammary stem cells. The beginning chapters provide readers with an introduction to mammary stem cells, and the processes used to characterize stem cells and isolate them via fluorescent activated cell sorting. The next few chapters discuss DNA and mRNA sequencing, proteomic techniques to help profile cells, lentiviral cell transduction for gene expression, and in vivo lineage tracing. The final few chapters are dedicated to following stem cells from their initial niche to the new microenvironment at their metastasis site, and to studying these cells using physical and mathematical approaches. Written in the highly successful Methods in Molecular Biology series format, the chapters include the kind of detailed description and implementation advice that is crucial for getting optimal results in the laboratory. Authoritative and cutting-edge, Mammary Stem Cells: Methods and Protocols aims to help members of the scientific community explore the behavior of stem cells and how to work with them in order to guide the design of new and complimentary strategies to be applied in the clinic with the ultimate end goal of fighting breast cancer.

A paradigm-shifting book from an acclaimed Harvard Medical School scientist and one of Time’s most influential people.

It’s a seemingly undeniable truth that aging is inevitable. But what if everything we’ve been taught to believe about aging is wrong? What if we could choose our lifespan?

In this groundbreaking book, Dr. David Sinclair, leading world authority on genetics and longevity, reveals a bold new theory for why we age. As he writes: “Aging is a disease, and that disease is treatable.”

This eye-opening and provocative work takes us to the frontlines of research that is pushing the boundaries on our perceived scientific limitations, revealing incredible breakthroughs—many from Dr. David Sinclair’s own lab at Harvard—that demonstrate how we can slow down, or even reverse, aging. The key is activating newly discovered vitality genes, the descendants of an ancient genetic survival circuit that is both the cause of aging and the key to reversing it. Recent experiments in genetic reprogramming suggest that in the near future we may not just be able to feel younger, but actually become younger.

Through a page-turning narrative, Dr. Sinclair invites you into the process of scientific discovery and reveals the emerging technologies and simple lifestyle changes—such as intermittent fasting, cold exposure, exercising with the right intensity, and eating less meat—that have been shown to help us live younger and healthier for longer. At once a roadmap for taking charge of our own health destiny and a bold new vision for the future of humankind, Lifespan will forever change the way we think about why we age and what we can do about it.

Why do modern agricultural techniques, which are environmentally damaging, continue to be used? This book seeks the answer to that question, by looking at the evolution of agricultural research in its cultural context. The theoretical framework is supported by historical case studies concerning hybrid maize in the United States, and the Green Revolution in Mexico. A chapter is also devoted to biotechnology, and its implications for the disturbing trend towards genetic uniformity.

Biotic stresses cause yield loss of 31-42% in crops in addition to 6-20% during post-harvest stage. Understanding interaction of crop plants to the biotic stresses caused by insects, bacteria, fungi, viruses, and oomycetes, etc. is important to develop resistant crop varieties. Knowledge on the advanced genetic and genomic crop improvement strategies including molecular breeding, transgenics, genomic-assisted breeding and the recently emerging genome editing for developing resistant varieties in technical crops is imperative for addressing FHEE (food, health, energy and environment) security. Whole genome sequencing of these crops followed by genotyping-by-sequencing have facilitated precise information about the genes conferring resistance useful for gene discovery, allele mining and shuttle breeding which in turn opened up the scope for 'designing' crop genomes with resistance to biotic stresses. The 15 chapters dedicated to 13 technical crops and 2 technical crop groups in this volume will deliberate on different types of biotic stress agents and their effects on and interaction with crop plants; will enumerate on the available genetic diversity with regard to biotic stress resistance among available cultivars; illuminate on the potential gene pools for utilization in interspecific gene transfer; will brief on the classical genetics of stress resistance and traditional breeding for transferring them to their cultivated counterparts; will enunciate the success stories of genetic engineering for developing biotic stress resistant varieties; will discuss on molecular mapping of genes and QTLs underlying biotic stress resistance and their marker-assisted introgression into elite varieties; will enunciate on different emerging genomics-aided techniques including genomic selection, allele mining, gene discovery and gene pyramiding for developing resistant crop varieties with higher quantity and quality; and will also elaborate some case studies on genome editing focusing on specific genes for generating disease and insect resistant crops.

This volume explores base editors (BEs), an invaluable CRISPR-based genome editing tool with a wide variety of versatile applications. Beginning with an overview of BEs, their diverse variants, and computational tools, the book continues with experimental applications of BEs for disease modeling in mammalian cells and generating mutagenic mice, therapeutic base editing strategies, which covers delivery methods of BE-encoded DNA plasmids, mRNAs, or ribonucleoproteins through viruses or non-viral lipid nanoparticles, and lastly, the use of BEs in plants and bacteria. 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 and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Base Editors: Methods and Protocols serves as an ideal guide for researchers looking to use base editors to continue their studies in an array of fields.

This volume covers a wide range of various fields of research, with the common thread being Next Generation Sequencing (NGS) related methods and applications, as well as analysis and interpretation of the data obtained. Chapters guide readers through the highly dynamic processes of translational and transcriptional profiling of a cell, method to detect copy number alterations (CNAs), targeted sequencing applications, method called "Hi-Plex" to characterize known polymorphic loci, single-cell of DNA or RNA, identify and characterize rare circulating CD4 T cells, and computational pipeline for RNAseq analysis. 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. Authoritative and practical, Next Generation Sequencing: Methods and Protocols aims to be useful and informative for further study into this vital field.

Developmental biology is seemingly well understood, with development widely accepted as being a series of programmed changes through which an egg turns into an adult organism, or a seed matures into a plant. However, the picture is much more complex than that: is it all genetically controlled or does environment have an influence? Is the final adult stage the target of development and everything else just a build-up to that point? Are developmental strategies the same in plants as in animals? How do we consider development in single-celled organisms? In this concise, engaging volume, Alessandro Minelli, a leading developmental biologist, addresses these key questions. Using familiar examples and easy-to-follow arguments, he offers fresh alternatives to a number of preconceptions and stereotypes, awakening the reader to the disparity of developmental phenomena across all main branches of the tree of life.

Stem cells have the ability to differentiate into all types of cells within the body, thus have great therapeutic potential for regenerative medicine to treat complicated disorders, like Parkinson s disease and spinal cord injury. There will also be many applications in drug development. However, several roadblocks, such as safety issues and low efficiency of pluripotent stem cell (PSC) line derivation need to be resolved before their clinical application. This thesis focuses on these two areas, so as to find methods to overcome the limitation. It covers deriving embryonic stem cells (ESCs) from several different species and reports an efficient system to generate induced pluripotent stem cells (iPSCs), and the first iPSC mice in the world. The results in this thesis confirm that somatic cells can be fully reprogrammed with the four Yamanaka factors. In addition, we have found that the Dlk1-Dio3 region can be a potential molecular marker to distinguish the fully reprogrammed iPSCs from partially reprogrammed ones. All of these results will help improve the safety of PSCs in the clinical applications and increase the current low induction efficiency of their production."

A better "casting" could not be conceived. The authors of this book are gold smiths on the subject. I have followed their work since their "entry" into cyto genetics and I have a high esteem for them. I consider it an honour to be asked to write the preface of their opus. Paul Popescu, Directeur de Recherche at INRA, has also played a promi nent part in the development of animal cytogenetics, especially in domestic animals. He is able to tell you the cost of a translocation in a pig breeding farm or a cow population: a fortune! P. Popescu has played a great part in gene mapping of these species using "in situ DNA hybridisation". His contributions are recognised world-wide. His laboratory receives many visitors every year and it serves as a reference for domestic animal cytogenetics. Helene Hayes, Charge de Recherche at INRA, has collaborated with P. POPESCU in the elaboration of the "at hand" techniques and in many other discoveries which are listed in her bibliography. She showed the fascinating correspondence between bovine and human chromosomes and the com pared gene maps of domestic bovidae.