This volume of "The Enzymes" features high-caliber thematic articles on the topic of glycosylphosphatidylinositol (GPI) anchoring of proteins.
* Contributions from leading authorities
* Informs and updates on all the latest developments in the field

This volume of Methods in Enzymology covers the current methodology for the detection and assessment of constitutively active proteins. The chapters written by expert authors who are leaders in the field, provide hints and tricks not available in primary research publications.It is extensively referenced, with useful figures and tables throughout the volume.

A. Expert authors who are leaders in the field

B. Extensively referenced and useful figures and tables

C. Provides hints and tricks to facilitate reproduction of methods

In the past decade, researchers have made tremendous progress in the field of enzyme stabilization, opening up new opportunities for enzymes in molecular biology and for industrial applications. In Enzyme Stabilization and Immobilization: Methods and Protocols, expert researchers explore the latest developments through detailed laboratory protocols, which address many different theories and techniques in enzyme stabilization. Chapters outline protocols for enzyme stabilization in solutions, including: liposome formation, micelle introduction, crosslinking, and additives. Secondly, the book contain protocols for enzyme stabilization via enzyme immobilization, such as sol-gel encapsulation, polymer encapsulation, and single enzyme nanoparticle formation. 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, Enzyme Stabilization and Immobilization: Methods and Protocols is an essential handbook for all molecular biologists, biochemists, and biomedical and biochemical engineers.

Glycosaminoglycans (GAGs) are a family of linear polysaccharides that arefound in all animal tissues. Several are used as biomaterials, including heparin, heparin sulfate, keratan sulfate, dermatan sulfate, and chondroitin sulfate.

This volume discusses the role of GAGs in development, health and disease.

* This series provides a forum for discussion of new discoveries, approaches, and ideas * Contributions from leading scholars and industry experts * Reference guide for researchers involved in molecular biology and related fields"

In this 3 volume collection focusing on glycomics, readers will appreciate how such discoveries were made and how such methods can be applied for readers own research efforts

Each chapter has been designed so that enough scientific background will be given in each chapter for further development of methods by readers themselves. Useful for all levels of scientists starting from the last years of colleges, graduate students, postdoctoral fellows to professors and to all levels of scientists in research institutes including industry. "

This monograph describes metabolic and transport reactions of muscle cells using the laws of chemical thermodynamics. In particular, the thermodynamics of irreversible processes are used to formulate coupled reactions and their outcome on steady state cycling. The effects of ATP cycling on energy metabolism and heat production is described. The results of mathematical simulations of metabolism are used to underline theoretical approaches.

The editors invited selected authors who had participated in or observed developments in biochemistry and molecular biology, particularly in the second half of this century, to record their personal recollections of the times and circumstances in which they worked. Having been given free reign, both content and style of the contruibutions reflect the flavour of the personality of the author.

The book reflects the explosive development of biochemistry and molecular biology and related sciences that had led to the almost unique situation of these fields coming of age at a time when their founding fathers, or their scientific children, were alive and well.

The contributions in this volume encompass a wide variety of experiences in many different countries and in very different fields of biochemistry.

The field of oligonucleotide therapeutics research is ripe with the prospect of new discoveries. In "Therapeutic Oligonucleotides: Methods and Protocols," a selection of established and emerging methods for the application of oligonucleotides as therapeutics are presented, all providing the tools needed to inspire great changes in the field. Divided into twenty-one chapters, this detailed volume meticulously describes vital protocols for optimizing and improving cell uptake, such as photochemical internalization, modified cell penetrating peptides, antibody conjugates, and nanoparticles. Other chapters address quantitation of RNA therapeutics in cells, assaying gene knockdown, selecting the best target site and synthesis of various modified oligonucleotides. 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, "Therapeutic Oligonucleotides: Methods and Protocols "serves as a timely resource for both professionals and novices pursuing research in this exciting and pioneering field."

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.

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.

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.

Structural genomics is the systematic determination of 3-D structures of proteins representative of the range of protein structure and function found in nature. The goal is to build a body of structural information that will predict the structure and potential function for almost any protein from knowledge of its coding sequence. This is essential information for understanding the functioning of the human proteome, the ensemble of tens of thousands of proteins specified by the human genome.
While most structural biologists pursue structures of individual proteins or protein groups, specialists in structural genomics pursue structures of proteins on a genome wide scale. This implies large-scale cloning, expression and purification. One main advantage of this approach is economy of scale.
Key Features
*Examines the three dimensional structure of all proteins of a given organism, by experimental methods such as X-ray crystallography and NMR spectroscopy
* Looks at structural genomics as a foundation of drug discovery as discovering new medicines is becoming more challenging and the pharmaceutical industry is looking to new technologies to help in this mission

This thesis reports on the development of a fully integrated and automated microsystem consisting of low-cost, disposable plastic chips for DNA extraction and PCR amplification, combined with a reusable glass capillary array electrophoresis chip, which can be employed in a modular-based format for genetic analysis. In the thesis, DNA extraction is performed by adopting a filter paper-based method, followed by an "in-situ" PCR carried out directly in the same reaction chamber of the chip without elution. PCR products are then co-injected with sizing standards into separation channels for detection using a novel injection electrode. The entire process is automatically carried out by a custom-made compact control and detection instrument. The author thoroughly tests the system's performance and reliability by conducting rapid genetic screening of mutations on congenital hearing loss and pharmacogenetic typing of multiple warfarin-related single-nucleotide polymorphisms. The successful development and operation of this microsystem establishes the feasibility of rapid "sample-in-answer-out" testing in routine clinical practice.

The book deals with various clinical aspects of cytochrome P450 2E1 (CYP2E1), which is a potent source for oxidative stress. Oxidative stress is critical for pathogenesis of diseases and CYP2E1 is a major contributor for oxidative stress. Several clinical disorders are associated with changes in regulation of CYP2E1 and the consequent abnormalities, which include alcoholic liver disease, alcoholic pancreatitis, carcinogenesis, non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, obesity, hepatitis C virus infection, reproductive organ toxicity, hepatocellular and cholestatic liver cirrhosis, inhibition of bone repair, cross- tolerance in smokers and people treated with nicotine, disorders of the central nervous system, changes in metabolism of protoxicants in the circulatory system and susceptibility to human papillomavirus infection. Hence, CYP2E1 emerges as a new and potent player in aggravating injury and furthering disease complications.

r ed Algae in Genome Age book most people reading this book have childhood memories about being enthralled at the beach with those rare and mysterious living forms we knew as seaweeds. We were fascinated at that time by their range of red hues and textures, and most of all, their exotic beauty. t o a scientist, red algae represent much more than apparent features. t heir complex forms have attracted morphologists for centuries; their intricate life cycles have brought more than one surprise to plant biologists familiar only with ferns and fowering plants; their unusual tastes have been appreciated for mill- nia, and their valuable chemical constituents have been exploited for nearly as long, most recently by biotech companies; their diversity in marine, freshwater, and t- restrial environments has offered centuries of engaging entertainment for botanists eager to arrange them in orderly classifcation systems; still, the red algae continue to teach us how many more challenges need to be overcome in order to understand their biodiversity, biological functions, and evolutionary histories.

In this 3 volume collection focusing on glycomics, readers will appreciate how such discoveries were made and how such methods can be applied for readers own research efforts

Each chapter has been designed so that enough scientific background will be given in each chapter for further development of methods by readers themselves. Useful for all levels of scientists starting from the last years of colleges, graduate students, postdoctoral fellows to professors and to all levels of scientists in research institutes including industry. "

In this 3 volume collection focusing on glycomics, readers will appreciate how such discoveries were made and how such methods can be applied for readers own research efforts

Each chapter has been designed so that enough scientific background will be given in each chapter for further development of methods by readers themselves. Useful for all levels of scientists starting from the last years of colleges, graduate students, postdoctoral fellows to professors and to all levels of scientists in research institutes including industry. "

Recent developments in stem cell biology have opened new directions in cell therapy. This book provides the state-of-the-art developments in using biomaterials as artificial niches for engineering stem cells, both for the purpose of better understanding their biology under 3D biomimetic conditions as well as for developing new strategies for efficient long term maintenance and directed differentiation of stem cells into various therapeutic lineages. Animal and human stem cells of both embryonic and adult origin are discussed with applications ranging from nerve regeneration, orthopedics, cardiovascular therapy, blood cell generation and cancer therapy. Both synthetic and natural biomaterials are reviewed with emphasis on how material-stem cell interactions direct specific signaling pathways and ultimately modulate the cell fate. This book is valuable for biomaterial scientists, tissue engineers, clinicians as well as stem cell biologists involved in basic research and applications of adult and embryonic stem cells.

This edited volume concerns a group of devastating neurological disorders that share a common pathological mechanism, namely the aggregation and deposition of insoluble, proteinaceous lesions, termed 'amyloid'. Examples of cerebral amyloid disorders include common neurodegenerative diseases like Alzheimer's disease-related dementia and Parkinson's disease, as well as other less prevalent conditions like Huntington's disease, cerebral amyloid angiopathy and the transmissible prion disorders. A disease-modifying therapeutic agent is still lacking for all these diseases, and there are no approved therapies that target amyloid formation directly. Nevertheless, a large and complex group of natural aromatic compounds known as polyphenols are rapidly emerging as potentially potent anti-amyloidogenic agents. This book collectively presents a considerable body of experimental and epidemiological evidence from peer-reviewed scientific publications that support a role for natural compounds and herbal extracts in the chemoprevention and therapy of amyloidogenic disorders. Each contribution is written by scientific experts in the relevant field; chapters are devoted to Mediterranean diet and olive oil phenols, traditional Chinese medicine, herbal extracts, polyphenols (with a particular emphasis on epigallocatechin-3-gallate) and bi-flavonoids, amongst others. The topic of this book is relevant to a wide audience, from academia and university students in the biological and chemical sciences, to physicians and allied health professionals, as well as people working in the nutraceutical industry.

The major theme of this book is analytical approaches to trace metal and speciation analysis in biological specimens. The emphasis is on the reliable determination of a number of toxicologically and environmentally important metals. It is essentially a handbook based on the practical experience of each individual author. The scope ranges from sampling and sample preparation to the application of various modern and well-documented methods, including quality assessment and control and statistical treatment of data. Practical advice on avoiding sample contamination is included.

In the first part, the reader is offered an introduction into the basic principles and methods, starting with sampling, sample storage and sample treatment, with the emphasis on sample decomposition. This is followed by a description of the potential of atomic absorption spectrometry, atomic emission spectrometry, voltammetry, neutron activation analysis, isotope dilution analysis, and the possibilities for metal speciation in biological specimens. Quality control and all approaches to achieve reliable data are treated in chapters about interlaboratory and intralaboratory surveys and reference methods, reference materials and statistics and data evaluation.

The chapters of the second part provide detailed information on the analysis of thirteen trace metals in the most important biological specimens. The following metals are treated in great detail: Aluminium, arsenic, cadmium, chromium, copper, lead, selenium, manganese, nickel, mercury, thallium, vanadium and zinc.

The book will serve as a valuable aid for practical analysis in biomedical laboratories and for researchers involved with trace metal and species analysis in clinical, biochemical and environmental research.

MILS-14 provides a most up-to-date view of the exciting biogeochemistry of gases in our environment as driven mostly by microorganisms. These employ a machinery of sophisticated metalloenzymes, where especially transition metals (such as Fe, Ni, Cu, Mo, W) play a fundamental role, that is, in the activation, transformation and syntheses of gases like dihydrogen, methane, carbon monoxide, acetylene and those of the biological nitrogen and sulfur cycles. The Metal-Driven Biogeochemistry of Gaseous Compounds in the Environment is a vibrant research area based mainly on structural and microbial biology, inorganic biological chemistry and environmental biochemistry. All this is covered in an authoritative manner in 11 stimulating chapters, written by 26 internationally recognized experts and supported by nearly 1200 references, informative tables and about 100 illustrations (two thirds in color). MILS-14 also provides excellent information for teaching. Peter M. H. Kroneck is a bioinorganic chemist who is exploring the role of transition metals in biology, with a focus on functional and structural aspects of microbial iron, copper and molybdenum enzymes and their impact on the biogeochemical cycles of nitrogen and sulfur. Martha E. Sosa Torres is an inorganic chemist, with special interests in magnetic properties of newly synthesized transition metal complexes and their reactivity towards molecular oxygen, applying kinetic, electrochemical and spectroscopic techniques.

This book addresses the possibilities and challenges in mimicking biological membranes and creating membrane-based sensor and separation devices. Recent advances in developing biomimetic membranes for technological applications will be presented with focus on the use of integral membrane protein mediated transport for sensing and separation. It describes the fundamentals of biosensing as well as separation and shows how the two processes are working in a cooperative manner in biological systems. Biomimetics is a truly cross-disciplinary approach and this is exemplified using the process of forward osmosis will be presented as an illustration of how advances in membrane technology may be directly stimulated by an increased understanding of biological membrane transport. In the development of a biomimetic sensor/separation technology, both channels (ion and water channels) and carriers (transporters) are important. An ideal sensor/separation device requires the supporting biomimetic matrix to be virtually impermeable to anything but the solute in question. In practice, however, a biomimetic support matrix will generally have finite permeabilities to water, electrolytes, and non-electrolytes. These non-protein mediated membrane transport contributions will be presented and the implications for biomimetic device construction will be discussed. New developments in our understanding of the reciprocal coupling between the material properties of the biomimetic matrix and the embedded proteins will be presented and strategies for inducing biomimetic matrix stability will be discussed. Once reconstituted in its final host biomimetic matrix the protein stability also needs to be maintained and controlled. Beta-barrel proteins exemplified by the E. Coli outer membrane channels or small peptides are inherently more stable than alpha-helical bundle proteins which may require additional stabilizing modifications. The challenges associated with insertion and stabilization of alpha-helical bundle proteins including many carriers and ligand and voltage gated ion (and water) channels will be discussed and exemplified using the aquaporin protein. Many biomimetic membrane applications require that the final device can be used in the macroscopic realm. Thus a biomimetic separation device must have the ability to process hundred of liters of permeate in hours - effectively demanding square-meter size membranes. Scalability is a general issue for all nano-inspired technology developments and will be addressed here in the context biomimetic membrane array fabrication. Finally a robust working biomimetic device based on membrane transport must be encapsulated and protected yet allowing massive transport though the encapsulation material. This challenge will be discussed using microfluidic design strategies as examples of how to use microfluidic systems to create and encapsulate biomimetic membranes. The book provides an overview of what is known in the field, where additional research is needed, and where the field is heading.

Recent work has begun to elucidate at the molecular level how albumin is handled by the kidney and how albuminuria develops in various proteinuric diseases including minimal change disease and focal segmental glomerulosclerosis. This volume provides a comprehensive overview of the renal handling of albumin - from basic mechanisms to the pathophysiology of proteinuric diseases. In describing the basic mechanisms of albuminuria, a particular highlight will be the focus on advanced imaging techniques such as intravital microscopy that have allowed a detailed "window" into albumin transit through the kidney. The volume will cover the epidemiological studies which show that albuminuria is a strong and independent marker of kidney disease progression and cardiovascular events, the molecular details of albumin handling in the kidney at the level of the glomerulus and the proximal tubule and the pathophysiology of proteinuric diseases including minimal change disease, membranous nephropathy, focal segmental glomerulosclerosis and diabetic nephropathy.

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.

Progress in functional proteomics has been limited for a long time, partially caused by limitations in assay sensitivity and sample capacity; however, protein microarrays have the ability to overcome these limitations so that a highly parallel analysis of hundreds of proteins in thousands of samples is attainable. In Protein Microarrays: Methods and Protocols, expert researchers in the field present an up-to-date collection of robust strategies in the field of protein microarrays and summarize recent advantages in the field of printing technologies, the development of suitable surface materials, as well as detection and quantification technologies. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key notes on troubleshooting and avoiding known pitfalls. Comprehensive and cutting-edge, Protein Microarrays: Methods and Protocols aims to stimulate the application and further advancement of this powerful technology in labs worldwide.