This volume includes a series of protocols focused on mitotic spindle assembly and function. The methods covered in this book feature a broad range of techniques from basic microscopy to the study of spindle physiologies relevant to cancer. These methods can be applied to diverse model systems that range from the cell-free Xenopus egg extract system to the moss Physcomitrella patens, in an effort to demonstrate the key contributions made by researchers using multiple model organisms. Chapters in The Mitotic Spindle: Methods and Protocols integrate cutting-edge technologies that have only become available due to the cross-disciplinary efforts, such as ATP analogue sensitive inhibition of mitotic kinases. 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. Thorough and informative, The Mitotic Spindle: Methods and Protocols, is a valuable resource for researchers who are new to mitosis or are already experts in the field.

This volume provides a wide range of protocols used in studying the nuclear envelope, with special attention to the experimental adjustments that may be required to successfully investigate this complex organelle in cells from various organisms. The Nuclear Envelope: Methods and Protocols is divided into five sections: Part I - Nuclear Envelope Isolation; Part II - Nuclear Envelope Protein Interactions, Localization, and Dynamics; Part III - Nuclear Envelope Interactions with the Cytoskeleton; Part IV - Nuclear Envelope-Chromatin Interactions; and Part V - Nucleo-Cytoplasmic Transport. Many of the modifications discussed in this book have only been circulated within laboratories that have conducted research in this field for many years. 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, The Nuclear Envelope: Methods and Protocols is a timely resource for researchers who have joined this dynamic and rapidly growing field.

This second edition expands on the first edition with new chapters describing methods for studying cell movement, molecular components involved in chemotaxis, spatiotemporal dynamics of signaling components, and quantitative modeling, as well as several updated chapters from the first edition. Various methods to investigate directional cell growth and movements are presented in Chapters 1-20. These chapters contains experimental procedures to visualize and measure migration behaviors of different kinds of organisms, including chemotropism in the budding yeast; cell growth and migration of D. discoideum; border cell migration in Drosophila; chemotaxis of mouse and human neutrophils; and HIV-induced T cell chemotactic response. Chemotaxis: Methods and Protocols, Second Edition also contains microscopy procedures for studying breast cancer cell migration, tumor cell invasion in vivo, and axon guidance. The book concludes with Chapters 21-27 describing methods that measure spatiotemporal dynamics of signaling components involved in chemotaxis; introduce imaging techniques, such as TRIF, BRET, FRET, and single-molecule microscopy; and mathematical models of experimentally generated chemoattractant gradients. 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, Chemotaxis: Methods and Protocols, Second Edition is a valuable resource for anyone who is interested in the diverse methodologies that are propelling chemotaxis research forward.

This second edition volume expands on the first edition by providing up-to-date protocols to characterize nanomaterials used as drug delivery agents. The chapters in this book are divided into 5 parts and cover topics such as: advances and obstacles in nanomedicine research; methods to test sterility and endotoxin, physicochemical features, immunological effects, drug release, and in vivo efficacy. 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 comprehensive, Characterization of Nanoparticles Intended for Drug Delivery, Second Edition is a valuable tool for researchers and pharmaceutical and biotechnology developers who are evaluating the clinical potential of nanomedicines in preclinical studies.>

Biological rhythms time the ebb and flow of virtually every physiological process, and their mutual coordination guarantees the integrity of the organism over space and time. Aging leads to the disintegration of this coordination, as well as to changes in the amplitude and/or frequency of the underlying rhythms. The results of this are accelerated loss of health during aging, and in experimental model systems curtailed lifespan occurs. This book will examine the machinery that constitutes circadian systems and how they impact physiologic processes. It will also discuss how disturbances of circadian rhythms can lead to complex diseases associated with aging. Much of this treatment will focus on metabolism and genome stability. Importantly, the chapters in this book will encompass work in several different models, in addition to human. The book will conclude with a discussion of modeling approaches to biologic cycles and chronotherapy, for future research and translation.

This invaluable resource discusses clinical applications with effects and side-effects of applications of stem cells in diabetes, kidney and wound treatment. All chapters are contributed by pre-eminent scientists in the field and covers such topics as stem cells and cell therapy in the treatment of diabetes mellitus, kidney failure, wound and other skin aging diseases, characteristics of some kinds of stem/progenitor cells for therapy, future directions of the discussed therapies and much more. Pancreas, Kidney and Skin Regeneration and the other books in the Stem Cells in Clinical Applications series will be invaluable to scientists, researchers, advanced students and clinicians working in stem cells, regenerative medicine or tissue engineering.

This detailed volume presents a wide range of techniques that go beyond the standard assays typically used to assess Myogenesis. The content included addresses assays to analyze skeletal muscle gene expression, proliferating muscle cells, the process of myoblast fusion, muscle development (in vivo), as well as muscle repair. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Skeletal Muscle Development serves as an ideal guide to the study of this highly complex yet carefully regulated process.>

This thesis describes the use of biophysical and biochemical methods to prove that calcium has a positive feedback effect on amplifying and sustaining CD3 phosphorylation and should enhance T-cell sensitivity to foreign antigens. The study presented shows that calcium can regulate the signal pathway in cells not only as a secondary messenger but also through direct interactions with the phospholipid bilayer. The approach used in the thesis also represents an important advance, as it couples the use of nuclear magnetic resonance (NMR) to the analysis of signaling phenomena in living cells. Moreover, the thesis optimizes the Nanodisc assembly protocol, which can broaden its range of applications in membrane protein studies. A preliminary study on the structure of dengue virus NS2B-NS3p in complex with aprotinin, which may help to develop new drugs against the dengue virus, is also included.

This book reviews the most recent developments in the field of osteochondral tissue engineering (OCTE) and presents challenges and strategies being developed that face not only bone and cartilage regeneration, but also establish osteochondral interface formation in order to translate it into a clinical setting. Topics include nanotechnology approaches and biomaterials advances in osteochondral engineering, advanced processing methodology, as well as scaffolding and surface engineering strategies in OCTE. Hydrogel systems for osteochondral applications are also detailed thoroughly. Osteochondral Tissue Engineering: Nanotechnology, Scaffolding-Related Developments and Translation is an ideal book for biomedical engineering students and a wide range of established researchers and professionals working in the orthopedic field.

Focusing on new methods and techniques developed to address longstanding questions about the Golgi complex, this volume explores a diverse set of chapters, ranging from live and fixed cell imaging techniques to in vitro biochemical reconstitution systems. Each chapter provides a detailed set of specific instructions, which should enable anyone to successfully complete the assays. Written for the highly successful Methods in Molecular Biology series program, chapters include brief introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips from the experts on troubleshooting and avoiding known pitfalls. Practical and detailed, The Golgi Complex: Methods and Protocols will aid both new and established researchers in the field by providing strong hands-on instructions that can be directly applied to their research programs.

This book on Candida albicans and similar pathogens provides a timely overview of the groundbreaking discoveries made in the areas of drug resistance, host-pathogen interactions, virulence, host immune system modulation, etc., in the last two decades. This comprehensive 2nd Edition includes chapters on fungal infections, hyphal morphogenesis, molecular mechanisms of antifungal resistance, antifungal agents, multidrug transporters, virulence mechanisms in Candida albicans, host-pathogen interactions, the cell wall, fungal biofilms, lipids and antifungal resistance, signaling mechanisms and last but not the least host-immune responses. As such, it offers an ideal reference guide for mycologists, researchers, pharmacists, clinicians, and undergraduate students engaged or interested in fungal research. It will also benefit clinicians, who are required to keep abreast of the current state of research on antifungal drug resistance and antifungal development.

Recent years have seen spectacular advances in the field of circadian biology. These have attracted the interest of researchers in many fields, including endocrinology, neurosciences, cancer, and behavior. By integrating a circadian view within the fields of endocrinology and metabolism, researchers will be able to reveal many, yet-unsuspected aspects of how organisms cope with changes in the environment and subsequent control of homeostasis. This field is opening new avenues in our understanding of metabolism and endocrinology. A panel of the most distinguished investigators in the field gathered together to discuss the present state and the future of the field. The editors trust that this volume will be of use to those colleagues who will be picking up the challenge to unravel how the circadian clock can be targeted for the future development of specific pharmacological strategies toward a number of pathologies.

The main purpose of this book is to introduce black rice to a wider circle of people. Although there have been research on different aspects of black rice, the information is scattered and not easily accessible to laypersons. The book intends to cover all the aspects of black rice from research, history, to its development. As such, the book will be suitable for both rice researchers and non-professionals who want to know more about this unique rice crop. Black rice, also known as forbidden rice, is packed with high level of nutrients and antioxidants. The antioxidants found in black rice is higher than the blueberries (that contain highest amount of anthocyanins). Black rice is black due to anthocyanin content in the outer layer of its kernel. Legend tells that this rice was consumed only by royals in China and it was expected that this rice would increase life span of the king. Consumption of black rice without approval was hanged. Ordinary individuals were not allowed to consume black rice. Thus this rice is also known as forbidden rice and Emperor's rice. Now this black heirloom rice is widely available in different parts of the world. Researchers have found that black rice reduce Reactive Oxygen Species (ROS), the free radicals produced in the body which is the cause of many diseases. This rice also reduce diabetes, inflammation, heart attack, allergy and obesity; reduce the growth of cancer, improves digestive system and is panacea of many health problems. Thus this rice is also known as long life rice. Food nutritionists consider black rice as modern super foods. The cultivation method of black rice is similar to general rice cultivation practices. There are many varieties available in black rice which is of different Asian origin but Chinese black rice is the most famous among them. Black rice has a wide range of applications because its bran is used as a natural food colouring dye, and it is also used to prepare noodles, pasta, porridge, wine etc. This rice takes slightly longer time to cook than widely available white rice. In modern era, black rice serve as one of the best food materials available to us to maintain our health with regular physical exercise.

This book covers several aspects of perinatal tissue-derived stem cells, from theoretical concepts to clinical applications. Topics include functions and different sources, immunomodulatory properties, translational point of view, GMP facility design and manufacturing for clinical translation, therapeutic potentials, and finally ethical considerations. The text provides a brief review of each type of perinatal stem cells and then focuses on their multi- or pluripotent properties, regenerative capacity, and future therapeutic potential in regenerative medicine. Additionally, the book discusses GMP compliance in stem cell facilities and the manufacture of stem cells for clinical translation. The chapters are authored by world-renowned experts in the perinatal stem cell field. Perinatal Tissue-Derived Stem Cells: Alternative Sources of Fetal Stem Cells, part of Springer's Stem Cell Biology and Regenerative Medicine series, is essential reading for basic and clinical scientists, clinicians, and pharmaceutical experts working or conducting research in the fields of stem cell biology, molecular aspects of stem cell research, tissue engineering, regenerative medicine, and cellular therapy.

The book presents a new, powerful model of neuronal networks, consisting of a three-dimensional neuronal culture in which 3D neuronal networks are coupled to micro-electrode-arrays (MEAs). It discusses the main advantages of the three-dimensional system compared to its two-dimensional counterpart, and shows that the network dynamics, recorded during both spontaneous and stimulated activity, differs between the two models, with the 3D system being better able to emulate the in vivo behaviour of neural networks. The book offers an extensive analysis of the system, from the theoretical background, to its design and applications in neuro-pharmacological studies. Moreover, it includes a concise yet comprehensive introduction to both 2D and 3D neuronal networks coupled to MEAs, and discusses the advantages, limitations and challenges of their applications as cellular and tissue-like in vitro experimental model systems.

With a focus on food safety, this book highlights the importance of microbes in sustainable agriculture. Plants, sessile organisms that are considered as primary producers in the ecosystem and communicate with above- and below-ground communities that consist of microbes, insects, and other vertebrate and invertebrate animals, are subjected to various kinds of stress. Broadly speaking, these can be subdivided into abiotic and biotic stresses. Plants have evolved to develop elaborate mechanisms for coping with and adapting to the environmental stresses. Among other stresses, habitat-imposed biotic stress is one serious condition causing major problems for crop productivity. Most plants employ plant-growth-promoting microorganisms (PGPMs) to combat and protect themselves from stresses and also for better growth. PGPMs are bacteria associated with plant roots and they augment plant productivity and immunity. They are also defined as root-colonizing bacteria that have beneficial effects on plant growth and development. Remarkably, PGPMs including mycorrhizae, rhizobia, and rhizobacteria (Acinetobacter, Agrobacterium, Arthrobacter, Azospirillum, Bacillus, Bradyrhizobium, Frankia, Pseudomonas, Rhizobium, Serratia, Thiobacillus) form associations with plant roots and can promote plant growth by increasing plants' access to soil minerals and protecting them against pathogens. To combat the pathogens causing different diseases and other biotic stresses, PGPMs produce a higher level of resistance in addition to plants' indigenous immune systems in the form of induced systemic resistance (ISR). The ISR elicited by PGPMs has suppressed plant diseases caused by a range of pathogens in both the greenhouse and field. As such, the role of these microbes can no longer be ignored for sustainable agriculture. Today, PGPMs are also utilized in the form of bio-fertilizers to increase plant productivity. However, the use of PGPMs requires a precise understanding of the interactions between plants and microbes, between microbes and microbiota, and how biotic factors influence these relationships. Consequently, continued research is needed to develop new approaches to boost the efficiency of PGPMs and to understand the ecological, genetic and biochemical relationships in their habitat. The book focuses on recent research concerning interactions between PGPMs and plants under biotic stress. It addresses key concerns such as - 1. The response of benign microbes that benefit plants under biotic stress 2. The physiological changes incurred in plants under harsh conditions 3. The role of microbial determinants in promoting plant growth under biotic stress The book focuses on a range of aspects related to PGPMs such as their mode of action, priming of plant defence and plant growth in disease challenged crops, multifunctional bio-fertilizers, PGPM-mediated disease suppression, and the effect of PGPMs on secondary metabolites etc. The book will be a valuable asset to researchers and professionals working in the area of microbial-mediated support of plants under biotic stress.

This book introduces key fundamentals of microarray bioprinting, including the required chip platforms and associated instruments/devices, experimental protocols for cell printing and biochemical- and cell-based assays, and several example applications. Various bioprinting approaches that allow for the rapid testing of hundreds of different cell culture conditions in combinations on a single chip are discussed in detail. Also covered is high-content, 3D cell-based imaging assays of tissue functions on miniaturized tissue constructs for high-throughput, predictive screening of drug efficacy and toxicity. This is an ideal book for graduate and postgraduate students in the field of biomedical engineering as well as scientists in the pharmaceutical industry. This book also: Broadens readers' understanding of the principles of microarray bioprinting, chip platforms and associated instruments/devices, and surface chemistry for micropatterning of cells on the chip platform Covers the latest developments in printing cells in hydrogels and methods of gelation as well as printing other biological samples in aqueous solutions Illustrates the complete process for cell staining and high-content imaging of 3D cells on the chip and predicting human metabolism and toxicology on the chip

This volume provides a state-of-art-report on the new methodologies in tissue engineering and developments in the biomaterials field based on the extracellular matrix-relevant discovery. Extracellular Matrix for Tissue Engineering and Biomaterials opens with an overview of the latest extracellular matrix research and in Part I, focuses on its biology and its role on cell behavior and cell fate relevant for the design of biomimetic surfaces. Part II details issues regarding the strategies currently applied in the research of biologically inspired materials and material systems for the replacement, repair and regeneration of tissues and organs. Part III presents the latest development methods applying knowledge from biology towards nanotechnology, to promote the restoration of the functionality of a living tissue. The book ranges from fundamental biology associated with tissue regeneration for the development of biomimetic approaches to controlling tissue formation, cell function, differentiation and angiogenesis using factors involved in normal tissue development and function. With the breadth and depth of the coverage of this topic, this book will serve as a valuable reference for anyone working in tissue engineering or biomaterials - from scientists, chemists and biologists through physicists, bioengineers and clinicians.

Biobanking is considered to be one of the ten ideas changing the world with an estimated value of $45 billion by 2025. Despite the challenges, as the climate for innovation in the biobanking industry continues to flourish around the world, it is certain that amazing discoveries will emerge from this large-scale method of preserving and accessing human samples; biobanking is no longer just a place for collecting and storing samples. This book will cover a wide variety of subjects from across the future biobanking spectrum including scientific strategies, personalized medicine, regenerative medicine and stem cell challenges, disease surveillance, population genetics and innovative methods of biobanking.

This book spans diverse aspects of modified nucleic acids, from chemical synthesis and spectroscopy to in vivo applications, and highlights studies on chemical modifications of the backbone and nucleobases. Topics discussed include fluorescent pyrimidine and purine analogs, enzymatic approaches to the preparation of modified nucleic acids, emission and electron paramagnetic resonance (EPR) spectroscopy for studying nucleic acid structure and dynamics, non-covalent binding of low- and high-MW ligands to nucleic acids and the design of unnatural base pairs. This unique book addresses new developments and is designed for graduate level and professional research purposes.

This invaluable resource discusses clinical applications with effects and side-effects of applications of stem cells in liver, lung and heart regeneration. All chapters are contributed by pre-eminent scientists in the field and covers such topics as cell therapy in the treatment of cirrhosis and other liver, heart and lung diseases, characteristics of hepatic progenitor cells, future directions of the discussed therapies and much more. Liver, Lung and Heart Regeneration and the other books in the Stem Cells in Clinical Applications series will be invaluable to scientists, researchers, advanced students and clinicians working in stem cells, regenerative medicine or tissue engineering.

This book presents regenerative strategies for the treatment of knee joint disabilities. The book is composed of four main sections totaling 19 chapters which review the current knowledge on the clinical management and preclinical regenerative strategies. It examines the role of different natural-based biomaterials as scaffolds and implants for addressing different tissue lesions in the knee joint. Section one provides an updated and comprehensive discussion on articular cartilage tissue regeneration. Section two focuses on the important contributions for bone and osteochondral tissue engineering. Section three overview the recent advances on meniscus repair/regeneration strategies. Finally, section four further discusses the current strategies for treatment of ligament lesions. Each chapter is prepared by world know expert on their fields, so we do firmly believe that the proposed book will be a reference in the area of biomaterials for regenerative medicine.

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

This detailed volume provides a single, valuable reference source for methods that definitively identify and accurately quantify apoptosis. The book begins with common methods utilized to detect and quantitate apoptosis, as well as apoptosis signaling pathways in toxicological and other related research. It continues with multi-parametric and phased apoptosis assays for detecting early and late apoptosis or distinguishing apoptosis from necrosis and autophagy. Subsequent chapters focus on recent advances in real time and high-throughput assays that detect and quantitate apoptosis and apoptosis signaling pathways. Final chapters focus on recent developments in preclinical anticancer therapeutics targeting apoptosis. Written for the Methods in Pharmacology and Toxicology series, chapters feature step-by-step descriptions of the methodologies, as well as expert tips and implementation advice. Vital and authoritative, Apoptosis Methods in Toxicology serves novice scientists as well as experts, utilizing a range of instruments from common laboratory equipment to high-end expensive and automated machinery capable of performing real time apoptotic measurements.

This book discusses properties of apoptosis and other cell death modalities in cancer pathogenesis and treatment. Its nine chapters discuss modulation of anti-tumor inflammatory and immune responses, effects on the tumor microenvironment, to strategies for improving pro-apoptotic therapies, mechanisms and implications for disease pathogenesis, axl and mer receptor tyrosine kinases, immunogenic apoptotic cell death and anti-cancer immunity and cancer cell death-inducing radiotherapy. This book places the onco-biology of apoptosis in clear and objective perspective through an expertly synthesized series of reviews. Apoptosis in Cancer Pathogenesis and Anti-cancer Therapy is a deft and thorough exploration of cutting-edge research in apoptosis and anti-cancer mechanisms from basic biology to oncology. It highlights a rapidly growing field within cancer research and is essential reading for oncologists, biochemists and advanced graduate students alike.