The generation of genetically modified mice is absolutely crucial to gene function studies today, primarily because mice are genetically similar to man and because gene function studies in mice are in the context of a whole organism, making them particularly useful. In Transgenic Mouse Methods and Protocols, Second Edition, expert research explore current advances in the field through detailed laboratory protocols. Chapters provide a general introduction outlining how to deal with mice and how to generate transgenic mouse models, explore the generation of conditional and induced knockout and transgenic mice, and offer alternative routes to studying gene function in mice. 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 state of the art, Transgenic Mouse Methods and Protocols, second Edition is the ideal guide for all researchers interested in the latest information about the production and analysis of transgenic and knockout mice.

This volume provides protocols for the generation of various biomaterials for tissue engineering and regenerative medicine applications. The chapters in this book include a look at a range of biomaterials including hydrogels and other matrices (natural, synthetic, self-healing) for various applications including drug and gene delivery, surface modification and functionalization of biomaterials. In addition, techniques described include those for controlling biomaterial geometry, such as three-dimensional printing and electrospinning. 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. Cutting-edge and thorough, Biomaterials for Tissue Engineering: Methods and Protocols is a valuable resource for scientists and engineers interested in this vital field of study.

Biotechnology has emerged as one of the key environmentally safe technologies for the future which enables use of biomass to develop novel smart materials and to replace oil derived products. Fungi are the most efficient producers of the enzymes needed for this purpose and in addition they produce a plethora of secondary metabolites, among which novel antibiotics can be found. Industrial application and exploitation of the metabolic capacities of fungi requires highly productive and robust gene expression systems, which can be achieved by selection of appropriate species and strain improvement. In this book we aim to summarize homologous and heterologous gene expression systems of fungi for production of enzymes and secondary metabolites. A broad overview on requirements, challenges and successful applications shall serve as a basis for further development of fungi as biotechnological workhorses in research and industry.

This two-volume work surveys the entire range of general aspects of chromosome research on plants. This first volume is divided into two sections. Section A consists of 11 chapters covering the entire range of general aspects of chromosome research in plants (including a chapter on genetic engineering in crop improvement). Section B is devoted to cytogenetics of cereals and millets (wheat, rye, barley, triticale, oats, maize, rice, pearl millet, and minor millets). More than one chapter is devoted to the same crop to give a detailed treatment of chromosome research (including molecular biology) in these crops.

The second volume deals with cytogenetics of plant materials including legumes, vegetable and oil crops, sugar crops, forage crops, fibre crops, medicinal crops and ornamentals. This work will be useful both as a reference work and a teaching aid to satisfy a wide range of workers. Every chapter has been written by an expert who has been involved in chromosome research on a particular plant material for many years.

Synthetic biology is becoming one of the most dynamic new fields of biology, with the potential to revolutionize the way we do biotechnology today. By applying the toolbox of engineering disciplines to biology, a whole set of potential applications become possible ranging very widely across scientific and engineering disciplines. Some of the potential benefits of synthetic biology, such as the development of low-cost drugs or the production of chemicals and energy by engineered bacteria are enormous. There are, however, also potential and perceived risks due to deliberate or accidental damage. Also, ethical issues of synthetic biology just start being explored, with hardly any ethicists specifically focusing on the area of synthetic biology. This book will be the first of its kind focusing particularly on the safety, security and ethical concerns and other relevant societal aspects of this new emerging field. The foreseen impact of this book will be to stimulate a debate on these societal issues at an early stage. Past experiences, especially in the field of GM-crops and stem cells, have shown the importance of an early societal debate. The community and informed stakeholders recognize this need, but up to now discussions are fragmentary. This book will be the first comprehensive overview on relevant societal issues of synthetic biology, setting the scene for further important discussions within the scientific community and with civil society.

Enzymes and whole cells are able to catalyze the most complex chemical processes under the most benign experimental and environmental conditions. In this way, enzymes and cells could be excellent catalysts for a much more sustainable chemical industry. However, enzymes and cells also have some limitations for nonbiological applications: fine chemistry, food chemistry, analysis, therapeutics, and so on. Enzymes and cells may be unstable, difficult to handle under nonconventional conditions, poorly selective toward synthetic substrates, and so forth. From this point of view, the transformation-from the laboratory to industry-of chemical processes catalyzed by enzymes and cells may be one of the most complex and exciting goals in biotechnology. For many industrial applications, enzymes and cells have to be immobilized, via very simple and cost-effective protocols, in order to be re-used over very long periods of time. From this point of view, immobilization, simplicity, and stabilization have to be strongly related concepts. Over the last 30 years, a number of protocols for the immobilization of cells and enzymes have been reported in scientific literature. However, only very few protocols are simple and useful enough to greatly improve the functional properties of enzymes and cells, activity, stability, selectivity, and related properties.

In the last decade, a number of powerful kernel-based learning methods have been proposed in the machine learning community: support vector machines (SVMs), kernel fisher discriminant (KFD) analysis, kernel PCA/ICA, kernel mutual information, kernel k-means, and kernel ARMA. Successful applications of these algorithms have been reported in many fields, such as medicine, bioengineering, communications, audio and image processing, and computational biology and bioinformatics. ""Kernel Methods in Bioengineering, Signal and Image Processing"" covers real-world applications, such as computational biology, text categorization, time series prediction, interpolation, system identification, speech recognition, image de-noising, image coding, classification, and segmentation. ""Kernel Methods in Bioengineering, Signal and Image Processing"" encompasses the vast field of kernel methods from a multidisciplinary approach by presenting chapters dedicated to adaptation and use of kernel methods in the selected areas of bioengineering, signal processing and communications, and image processing.

Protein hydrolysates, otherwise commonly known as peptones or peptides, are used in a wide variety of products in fermentation and biotechnology industries. The term "peptone" was first introduced in 1880 by Nagelli for growing bacterial cultures. However, later it was discovered that peptones derived from the partial digestion of proteins would furnish organic nitrogen in readily available form. Ever since, p- tones, which are commonly known as protein hydrolysates, have been used not only for growth of microbial cultures, but also as nitrogen source in commercial fermen- tions using animal cells and recombinant microorganisms for the production of value added products such as therapeutic proteins, hormones, vaccines, etc. Today, the characterization, screening and manufacturing of protein hyd- lysates has become more sophisticated, with the introduction of reliable analytical instrumentation, high throughput screening techniques coupled with statistical design approaches, novel enzymes and efficient downstream processing equipment. This has enabled the introduction of custom-built products for specialized appli- tions in diverse fields of fermentation and biotechnology, such as the following. 1. Protein hydrolysates are used as much more than a simple nitrogen source. For example, the productivities of several therapeutic drugs made by animal cells and recombinant microorganisms have been markedly increased by use of p- tein hydrolysates. This is extremely important when capacities are limited. 2. Protein hydrolysates are employed in the manufacturing of vaccines by ferm- tation processes and also used as vaccine stabilizers.

This volume arranged into three sections describes biochemical, in vitro, and in vivo protocols on Semaphorins. Chapters focus on approaches that would allow the novice to study Semaphorins and employ robust assays to characterize mechanisms of action. 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 cutting-edge, Semaphorin Signaling: Methods and Protocols aims to ensure successful results in the further study of this vital field.

sector. This ensured eventual transfer of the technology demonstrated at the wo- shops and Technical Meetings to marketable devices. BIOSET provided assistance for researchers from European laboratories to meet to exchange ideas, use equ- ment, and establish a basis for new joint projects. The secretariat of the Concerted Action BIOSET supported the Technical Meetings. There were three Technical Meetings held, two in Berlin in 1997 and 1998, and the third in Barcelona, in April 2000. The goal of these technical meetings was to join different research and industrial teams to evaluate the performance of their biosensor technology in field conditions with common and standardized surface and waste waters. As a result of these field experiments, the additional information that biosensors can offer to environmental monitoring was also evaluated. Thus, these three Technical Meetings were useful accompanying measures and practical additions to the currently organized yearly workshops. The concerted action BIOSET was f- lowed by the SENSPOL network. The 1st SENSPOL Workshop was held on the 9-11 May 2001 on Sensing Technologies for Contaminated Sites and Groundwater at the University of Alcala. There was one special Workshop on "Genotoxicity Biosensing (TECHNOTOX)" supported by the European Commission DG XII D-1 and BIOSET in the year 2000. The TECHNOTOX meeting at the Flemish Institute for Technological Research (VITO) in Mol was organized by Phillippe Corbisier (VITO), Peter-D. Hansen (TU Berlin) and Damia Barcelo (CSIC Barcelona).

Bioethanol has been recognized as a potential alternative to petroleum-derived transportation fuels. Even if cellulosic biomass is less expensive than corn and sugarcane, the higher costs for its conversion make the near-term price of cellulosic ethanol higher than that of corn ethanol and even more than that of sugarcane ethanol. Conventional process for bioethanol production from lignocellulose includes a chemical/physical pre-treatment of lignocellulose for lignin removal, mostly based on auto hydrolysis and acid hydrolysis, followed by saccharification of the free accessible cellulose portions of the biomass. The highest yields of fermentable sugars from cellulose portion are achieved by means of enzymatic hydrolysis, currently carried out using a mix of cellulases from the fungus Trichoderma reesei. Reduction of (hemi)cellulases production costs is strongly required to increase competitiveness of second generation bioethanol production. The final step is the fermentation of sugars obtained from saccharification, typically performed by the yeast Saccharomyces cerevisiae. The current process is optimized for 6-carbon sugars fermentation, since most of yeasts cannot ferment 5-carbon sugars. Thus, research is aimed at exploring new engineered yeasts abilities to co-ferment 5- and 6-carbon sugars. Among the main routes to advance cellulosic ethanol, consolidate bio-processing, namely direct conversion of biomass into ethanol by a genetically modified microbes, holds tremendous potential to reduce ethanol production costs. Finally, the use of all the components of lignocellulose to produce a large spectra of biobased products is another challenge for further improving competitiveness of second generation bioethanol production, developing a biorefinery.

Interest in the study of kinship, a key area of anthropological enquiry, has recently reemerged. Dubbed 'the new kinship', this interest was stimulated by the 'new genetics' and revived interest in kinship and family patterns. This volume investigates the impact of biotechnology on contemporary understandings of kinship, of family and 'belonging' in a variety of European settings and reveals similarities and differences in how kinship is conceived. What constitutes kinship for different publics? How significant are biogenetic links? What does family resemblance tell us? Why is genetically modified food an issue? Are 'genes' and 'blood' interchangeable? It has been argued that the recent prominence of genetic science and genetic technologies has resulted in a 'geneticization' of social life; the ethnographic examples presented here do show shifts occurring in notions of 'nature' and of what is 'natural'. But, they also illustrate the complexity of contemporary kinship thinking in Europe and the continued interconnectedness of biological and sociological understandings of relatedness and the relationship between nature and nurture.

There is hardly an area of research developing so quickly and raising so many promises as stem cell research. Adult, embryonic and recently available induced pluripotent stem cells not only foster our understanding of differentiation of endo-, ecto- and mesodermal lineages to all organs of the body, but foremost nourish the hope that cells grown in culture can be used for regeneration of diseased organs such as the heart damaged by myocardial infarction. This book focuses on perspectives of stem cells for regenerative therapy of cardiovascular diseases. Based on the EC consortium INELPY, it reviews the field and disseminates major outcomes of this project. Thus it introduces the reader to this fascinating area of research and incorporates very recent findings interesting to the expert, spanning the field from bench to bedside. The compilation of contributions is unique as there is yet no similar comprehensive overview combining stem cell research with preclinical and clinical evaluation as well as engineering of tissue patches for transplantation. As such it will be an invaluable source of information for all researchers in the stem cell and tissue regeneration field including bioengineers as well as for all clinicians interested in regenerative therapies, especially for ischemic cardiomyopathies.

This timely volume explores the use of CRISPR-Cas9 for genome editing, presenting cutting-edge techniques and their applications in treatment of disease. The chapters describe latest methods such as use of targetable nucleases, investigation of the non-coding genome, mouse genome editing, increasing of knock-in efficiency in mouse zygotes, and generation of reporter stem cells; the text contextualizes these methods in treatment of cardiovascular disease, diabetes mellitus, retinitis pigmentosa, and others. The final chapters round out the book with a discussion of controversies and future directions. Genome Editing is an essential, of-the-moment contribution to this rapidly growing field. Drawing from a wealth of international perspectives, it presents novel techniques and applications for the engineering of the human genome. This book is essential reading for all clinicians and researchers in stem cells, regenerative medicine, genomics, biochemical and biomedical engineering- especially those interested in learning more about genome editing and applying it in a targeted, specific way.

This 2nd edition begins with an overview of NMR development and applications in biological systems. It describes recent developments in instrument hardware and methodology. Chapters highlight the scope and limitation of NMR methods. While detailed math and quantum mechanics dealing with NMR theory have been addressed in several well-known NMR volumes, chapter two of this volume illustrates the fundamental principles and concepts of NMR spectroscopy in a more descriptive manner. Topics such as instrument setup, data acquisition, and data processing using a variety of offline software are discussed. Chapters further discuss several routine stategies for preparing samples, especially for macromolecules and complexes. The target market for such a volume includes researchers in the field of biochemistry, chemistry, structural biology and biophysics.

Advanced biotechnologies enable breeders to produce a whole generation of new crops for specialist needs (designer crops), including raw materials for the energy, chemical and pharmaceutical industries. This book provides concerns useful to promote an increase of the productivity of crops by using functional genomics (to understand the regulation of plant metabolism at molecular, cellular and whole plants), and the improvement of photosynthetic efficiency (to design new plants with enhanced raw materials percent and recovery). Fundamental thematics have been addressed: metabolic engineering, plant breeding tools, renewable biomass for energy generation, fibres and composites, biopharmaceuticals. The gained know how is relevant to identify bottlenecks in the major production chains and to propose actions for moving these issues forward.

Until recently, knowledge of ruminant gut microbiology was primarily obtained using classical culture based techniques, which probably only account for 10 to 20% of the rumen microbial population. New gene-based technologies can now be employed to examine microbial diversity through the use of small sub-unit ribosomal DNA analysis (e.g. 16S rDNA) and to understand the function of complex microbial ecosystems in the rumen through metagenomic analysis. These technologies have the potential to revolutionize the understanding of rumen function and will overcome the limitations of classical based techniques, including isolation and taxonomic identification of strains important to efficient rumen function and better understanding of the roles of microorganisms in relation to achieving high productivity and decreasing environmental pollutants. This book presents a comprehensive up-to-date account of the methodologies and protocols for conventional and modern molecular techniques that are currently in use for studying the gut microbial ecology of ruminants. Each chapter has been contributed by experts in the field and methods have been presented in a recipe-like format designed for direct practical use in the laboratory and also to provide insight into the most appropriate techniques, their applications and the type of information that could be expected. The techniques and procedures described are also relevant and adaptable to other gastrointestinal ecosystems and the microbiology of anaerobic environments in general. This manual will a ~demystifya (TM) the methods in molecular microbial ecology for readers who are novice in the field but are excited by the prospects of the technology. Itwould also be invaluable for the experienced workers striving for giving new dimension to their research a" expanding the work in other fields and initiating cross-cutting activities.

Dynamic Single-Use Bioreactors Used in Modern Liter- and m3- Scale Biotechnological Processes: Engineering Characteristics and Scaling Up, by Christian Loffelholz, Stephan C. Kaiser, Matthias Kraume, Regine Eibl, Dieter Eibl. Orbitally Shaken Single-Use Bioreactors, by Wolf Klockner, Sylvia Diederichs, Jochen Buchs.

Therapeutic Human Cells: Manufacture for Cell Therapy/Regenerative Medicine by Christian van den Bos, Robert Keefe, Carmen Schirmaier, Michael McCaman.

Fast Single-Use VLP Vaccine Productions Based on Insect Cells and the Baculovirus Expression Vector System: Influenza as Case Study by Regine Eibl, Nina Steiger, Sabine Wellnitz, Tiago Vicente, Corinne John, Dieter Eibl.

Microbial High Cell Density Fermentations in a Stirred Single-Use Bioreactor by Thomas Dreher, Bart Walcarius, Ute Husemann, Franziska Klingenberg, Christian Zahnow, Thorsten Adams, Davy de Wilde, Peter Casteels, Gerhard Greller.

Quorus Bioreactor: A New Perfusion-Based Technology for Microbial Cultivation by Sheena J. Fraser, Christian Endres.

Cultivation of Marine Microorganisms in Single-Use Systems by Friederike Hillig, Maciej Pilarek, Stefan Junne, Peter Neubauer.

Flexible Biomanufacturing Processes that Address the Needs of the Future by Bernhard Diel, Christian Manzke, Thorsten Peuker.

An Approach to Quality and Security of Supply for Single-Use Bioreactors by Magali Barbaroux, Susanne Gerighausen, Heiko Hackel.

A Risk Analysis for Production Processes with Disposable Bioreactors by Tobias Merseburger, Ina Pahl, Daniel Muller, Markus Tanner."

Altogether, the biochemical, technical and economic limitations on existing proka- otic and eukaryotic expression systems and the growing clinical demand for complex therapeutic proteins have created substantial interest in developing new expression systems for the production of therapeutic proteins. To that end, plants have emerged in the past decade as a suitable alternative to the current production systems, and today their potential for production of high quality, much safer and biologically active complex recombinant pharmaceutical proteins is largely documented. The chapters in this volume, contributed by leaders in the field, sum up the state-- the-art methods for using a variety of different plants as expression hosts for phar- ceutical proteins. Several production platforms are presented, ranging from seed- and leaf-based production in stable transgenic plant lines, to plant cell bioreactors, to viral or Agrobacterium-mediated transient expr ession systems. Currently, antibodies and their derived fragments represent the largest and most important group of biote- nological products in clinical trials. This explains why the potential of most prod- tion platforms is illustrated here principally for antibodies or antibody fragments with acknowledged potential for immunotherapy in humans. In addition, a comparison of different plant expression systems is presented using aprotinin, a commercial phar- ceutical protein, as a test system. Although multiple books and monographs have been recently published on mol- ular pharming, there is a noticeable dearth of bench step-by-step protocols that can be used quickly and easily by beginners entering this new field.

The diversity and specialization in orchid floral morphology have fascinated botanists and collectors for centuries. In the past 10 years, the orchid industry has been growing substantially worldwide. This interesting book focuses on the recent advances in orchid biotechnology research since the last 10 years in Taiwan. To advance the orchid industry, enhancement of basic research as well as advanced biotechnology will provide a good platform to improve the flower quality and breeding of new varieties. Important topics covered include the new knowledge of basic genome, through floral morphogenesis, floral ontology, embryogenesis, micropropagation, to functional genomics such as EST, virus-induced gene silencing, and genetic transformation.

This book explains biosensor development fundamentals. It also initiates awareness in engineers and scientists who would like to develop and implement novel biosensors for agriculture, biomedicine, homeland security, environmental needs, and disease identification. In addition, the book introduces and lays the basic foundation for design, fabrication, testing, and implementation of next generation biosensors through hands-on learning.

This consolidated reference book addresses the various aspects of nano biomaterials used in ophthalmic drug delivery, including their characterization, interactions with ophthalmic system and applications in treatments of the ophthalmic diseases and disorders. In the last decade, a significant growth in polymer sciences, nanotechnology and biotechnology has resulted in the development of new nano- and bioengineered nano-bio-materials. These are extensively explored as drug delivery carriers as well as for implantable devices and scaffolds. At the interface between nanomaterials and biological systems, the organic and synthetic worlds merge into a new science concerned with the safe use of nanotechnology and nano material design for biological applications. For this field to evolve, there is a need to understand the dynamic forces and molecular components that shape these interactions. While it is impossible to describe with certainty all the bio physicochemical interactions at play at the interface, we are at a point where the pockets of assembled knowledge are providing a conceptual framework to guide this exploration, and review the impact on future product development. The book is intended as a valuable resource for academics and pharmaceutical scientists working in the field of polymers, polymers materials for drug delivery, drug delivery systems and ophthalmic drug delivery systems, in addition to medical and health care professionals in these areas.

The development of sustainable and renewable biofuels is attracting growing interest. It is vital to develop robust microbial strains for biocatalysts that are able to function under multiple stress conditions. This Microbiology Monograph provides an overview of methods for studying microbial stress tolerance for biofuels applications using a systems biology approach. Topics covered range from mechanisms to methodology for yeast and bacteria, including the genomics of yeast tolerance and detoxification; genetics and regulation of glycogen and trehalose metabolism; programmed cell death; high gravity fermentations; ethanol tolerance; improving biomass sugar utilization by engineered Saccharomyces; the genomics on tolerance of Zymomonas mobilis; microbial solvent tolerance; control of stress tolerance in bacterial host organisms; metabolomics for ethanologenic yeast; automated proteomics work cell systems for strain improvement; and unification of gene expression data for comparable analyses under stress conditions.