This book covers the identification and role of endogenous lung stem cells in health and disease, particularly the most recent advances. In addition, it discusses the rapidly growing field of stem cells and cell therapy as it relates to lung biology and disease as well as ex vivo lung bioengineering. Such approaches may provide novel therapeutic approaches for lung diseases. Human pluripotent stem cell differentiation to model the pulmonary epithelium and vasculature is also discussed. World-recognized scientists who specialize in studying both the lung epithelium and pulmonary vasculature contribute the chapters. Topics covered include: stem cell niches in the lung, the role of progenitor cells in fibrosis and asthma, iPSC in modeling lung disease, vascular repair by endothelial progenitor cells and circulating fibrocytes in pulmonary vascular remodeling. This volume of the Stem Cell Biology and Regenerative Medicine series is essential reading for researchers and clinicians interested in stem cells, lung biology and regenerative medicine. It is also an invaluable resource for advanced students studying cell biology, regenerative medicine and lung physiology.

The purpose of these volumes is to provide a reference work for the methods of purifying many of the receptors we know about. This becomes increasingly important as full-length recep- tors are overexpressed in bacteria or in insect cell systems. A major problem for abundantly expressed proteins will be their purification. In addition to purification protocols, many other de- tails can be found concerning an individual receptor that may not be available in standard texts or monographs. No book of this type is available as a compendium of purification procedures. Receptor Purification provides protocols for the purification of a wide variety of receptors. These include receptors that bind: neurotransmitters, polypeptide hormones, steroid hormones, and ligands for related members of the steroid supergene family and others including receptors involved in bacterial motion. The text of this information is substantial so as to require its publica- tion in two volumes. Consequently, a division was made by grouping receptors depending upon the nature of their ligands. Thus, in volume 1 there are contributions on serotonin receptors, adrenergic receptors, the purification of GTP-binding proteins, opioid receptors, neurotensin receptor, luteinizing hormone re- ceptor, human chorionic gonadotropin receptor, follicle stimulat- ing hormone receptor, thyrotropin receptor, prolactin receptor, epidermal growth factor receptor, platelet derived growth factor receptor, colony stimulating factor receptor, insulinlike growth factor receptors, insulin receptor, fibronectin receptor, interferon receptor, and the cholecystokinin receptor.

Drawing especially on insights emerging from studies of the cellular networks formed by fungi, this book describes the fundamental indeterminacy that enables life forms to thrive in and create inconstant circumstances. It explains how indeterminacy arises from counteraction between associative and dissociative processes at the reactive interfaces between living systems and their surroundings. It stresses the relevance of these processes to understanding the dynamic contexts within which living systems of all kinds - including human societies-explore for, use up, conserve and recycle sources of energy.By focusing on dynamic boundaries, the book counterbalances the discretist view that living systems are assembled entirely from building-block-like units - individuals and genes - that can be freely sifted, as opposed to channeled, by natural selection. It also shows how the versatility that enables life forms to proliferate in rich environments, whilst minimizing losses in restrictive environments, depends on capacities for error and co-operation within a fluid, non-hierarchical power structure. Understanding this point yields a more compassionate, less competitive and less self-centred outlook on life's successes and failures.

For volume 2 alone:
A gold-standard collection of readily reproducible techniques for the molecular and genetic analysis of germ cells in a variety of different reproductive systems. Volume 2: Molecular Embryo Analysis, Live Imaging, Transgenesis, and Cloning focuses on molecular egg analysis, live egg imaging, nuclear cloning, oocyte cryopreservation, and nuclear transfer. Highlights include the identification and characterization of Oct-4, germline chromatin silencing by RNAi, cDNA subtraction and cloning in the field of trophoblast/placental development, and selective ablation. Each readily reproducible protocol is described in step-by-step detail and contains an introduction outlining the principle behind the technique, lists of equipment and reagents, and tips on troubleshooting and avoiding known pitfalls. Comprehensive and cutting-edge, Germ Cell Protocols offers both novice and established researchers a gold-standard collection of hard-to-find methods of high impact research, diverse procedures that are easy-to-follow, well-illustrated, and allow a cross-species transfer of knowledge from lower vertebrates to higher mammalian systems.
For both volumes:
A gold-standard collection of readily reproducible techniques for the molecular and genetic analysis of germ cells in a variety of different reproductive systems. The methods cover sperm and egg activation, motility, fertilization, nuclear development, nuclear cloning, the molecular characterization of specific events, and the imaging of cell structures. Volume 1: Sperm and Oocyte Analysis focuses on sperm cells, oocyte analysis, oocyte maturation, fertilization, and preparation techniques. Volume 2: MolecularEmbryo Analysis, Live Imaging, Transgenesis, and Cloning contains methods for molecular egg analysis, live egg imaging, nuclear cloning, oocyte preservation, and nuclear transfer.
Comprehensive and cutting-edge, Germ Cell Protocols offers both novice and established researchers a gold-standard collection of hard-to-find methods of high impact research, diverse procedures that are easy-to-follow, well-illustrated, and allow a cross-species transfer of knowledge from lower vertebrates to higher mammalian systems.

Grauzone and Completion of Meiosis During Drosophila Oogenesis describes the work behind a major, award winning discovery: the establishment of a new pathway that specifically regulates the female meiosis, a process essential for sexual reproduction. This book chronicles a new gene mapping method and the cloning and documentation of several types of genes that were proven to have significant influence on the cell cycle. It is of interest to anyone doing work with fruit flies, both graduate students and principal investigators.

The different aspects of muscle development are considered from cellular, molecular and genetic viewpoints, and the text is supported by black/white and color illustrations. The book will appeal to those studying muscle development and muscle biology in any organism.

This book discusses the role of oxidative stress in the reproductive system. The book reviews endogenous sources, methods of determining its levels in body fluid/tissues, the physiological roles of ROS, as well as its negative effects on the human reproductive processes. Also discussed are multiple extrinsic factors that could induce oxidative stress in the reproductive system. This volume covers various clinical pathologies related to the reproductive system that arise from or produce oxidative stress, both in the male and female. The use of antioxidants as a therapeutic measure to keep ROS levels in check are highlighted, describing the outcome of various clinical studies involving antioxidant supplementation in infertile patients. Infertility is a global disease that affects 15-25% of all couples, and oxidative stress arising from a multitude of sources has been implicated as one of the major contributing factors to the decline in human fertility. As such, this book provides an up-to-date review on the significance of ROS in human reproduction.

A much-needed work that provides an authoritative overview of the fundamental biological facts, theoretical models, and current experimental developments in this fascinating area. Cell motility is fundamentally important to a number of biological and pathological processes. The main challenge in the field of cell motility is to develop a complete physical description on how and why cells move. For this purpose new ways of modeling the properties of biological cells have to be found - and this volume is a major stepping-stone along the way.

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 volume covers methods for the analysis of extracellular vesicles (EV) that can be applied to isolated EVs from a wide variety of sources. This includes the use of electron microscopy, tunable resistance pulse sensing, and nanoparticle tracking analysis. The chapters in this book discuss EV cargoes containing proteins and genomic materials using a number of different approaches, and isolating EVs from platelets and neuronal cells and tissues. 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. Practical and comprehensive, Exosomes and Microvesicles: Methods and Protocols is a valuable resource containing methodologies for anyone interested in researching EVs.

Deficiencies in sperm function are usually the result of spermatogenic defects. Spermatogenesis is a biologically complex and essential process during which spermatogonia undergo meiotic recombination, reduction of the genome to a haploid state, and extensive cellular modifications that result in a motile cell capable of traversing the female reproductive tract, withstanding various potential assaults to viability, and finally successfully fertilizing a mature oocyte to give rise to an embryo. Defects in any step of spermatogenesis or spermatogenesis can lead to male infertility, a disease that affects approximately 5-7% of the population."Spermiogenesis and Spermatogenesis: Methods and Protocols"details protocols used in the study of spermatogenesis, clinical analytical protocols, and basic techniques used in clinical andrology laboratories, such as obtaining accurate results for a sperm count, and advanced procedures, such as genome-wide genetic study tools and evaluation of nuclear proteins.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, "Spermiogenesis and Spermatogenesis: Methods and Protocols"isunique in its breadth, and will be a useful reference for clinicians and researchers alike."

The past decade has witnessed a revolution in the attempts of scientists to under stand the molecular basis of dementia. Although dementia, as defined by global cogni tive decline involving gradual loss of memory, reasoning, judgment, and orientation, presents most commonly in the form of Alzheimer's disease (AD), an assortment of other less common disorders, such as prion and Pick's disease, can also lead to symp toms that are similar to those observed in patients with AD. The primary goal of Molecular Mechanisms of Dementia is to address the various mechanisms and multi faceted approaches currently being employed to more clearly delineate the etiological and pathogenic events responsible for the onset of dementia. Perhaps the greatest boon to obtaining a clearer understanding of the causes of AD has come from genetic and molecular biological studies carried out over the past decade. At the genetic level, it has become increasingly clear that AD is a heteroge neous disorder that can be broadly classified into two categories. "Late onset" (>60 yr) cases, which account for the vast majority of AD, genetically involve "susceptibility" genes representing risk factors for the disease (e. g. , inheritance of the 84 allele of the Apolipoprotein E gene). In many cases, the susceptibility gene can act as a "modifier" that modulates the pathogenic cascade occurring subsequent to a separate etiological event "initiating" or "causing" the disorder.

The 19 papers include discussions of constructing an integrated genetic and physical map of rice, commonalities and contrasts in the organization of the maize and sorghum nuclear genomes, prospects for comparative genome analyses among mammals, genome analysis in farm animals, sense suppression of p

Oxidation-reduction (i.e. redox) processes at the plasma membrane of any cell have been attracting more and more attention, both in basic and in applied research, since the first workshop dealing with the plasma membrane oxidoreductases was organized in Cordoba, Spain, in 1988. This evolution is evident considering the numerous cell functions performed by plasma membrane redox systems not only in healthy cells but also in cells that escaped from the normal metabolic control (e.g. cancer cells) and cells under attack by pathogens. Plasma membrane redox processes have now been demonstrated to play an essential role in growth control and defense mechanisms of these cells. The great importance of the plasma membrane redox systems originates in the fact that they are located in the membrane which is essentially the site of communication between the living cell and its environment. We may say that the plasma membrane can be considered as the "sensory part" of the cell. No chemical substance can enter the cell interior without interaction with the plasma membrane.

At the forefront of life sciences today is the emerging discipline of chembiomolecular science. This new term describes the integration of the frontier fields of chemical biology, chemistry, and pharmacology. Chembiomolecular science aims to elucidate new biological mechanisms as potential drug targets and enhance the creation of new drug therapies. This book comprises the proceedings of the Uehara Memorial Foundation Symposium 2011, which focused on the most recent advances in chembiomolecular science made by leading experts in the field. The book is divided into three main topics. The first is the chemical approach to understanding complex biological systems on a molecular level using chemical compounds as a probe. The second describes the biological approach used to develop new lead drug compounds. The third focuses on the biological system that serves as the potential drug target, the beginning step in the process of developing new drugs. Replete with the latest research, the book will draw the attention of all scientists interested in the synergies between chemistry and biology to elucidate life on a molecular level and to promote drug discovery. Ultimately, the book helps promote the understanding of biological functions at the molecular level and create new pharmaceuticals that can contribute to improving human health.

A comprehensive guide to the revolutionary area of systems biology and its application in cell culture engineering, this volume presents an overall picture of the current topics central to structural and functional genomics, proteomics, metabolomics and bioinformatics, including such hot topics as RNAi, metabolic engineering and unfolded protein response. It includes reviews of the cellular response of environmental modulation such as low temperature and osmolarity, critical assessments of the applications of metablomics and fluxomics approaches, examination of the utility of modulation of key genes and a presentation of a theory of chemical organisation which provides new view on the system's structure. The clearly written chapters by experts in the field describe methods applicable to investigating the unique facets of cell culture.

Somuchofwhatweknowaboutthepathogenesisofhumandiseasehascomefromthe systematic and careful study of histological material. Indeed, every internal medicine discipline has its landmark papers describing the clinico-pathological correlations. However, increasingly, it is molecular and cellular biology that provides the necessary mechanistic insights. For many years, it was thought that the two skill sets were mutually exclusive, but we hope that this book shows that this is not necessarily so. Implicitinthescienceofhistologyisthepreservationandarchivingoftissue.PartIof the book concentrates on the preparation of tissue, providing an overview of fixation, embedding, and processing (Chapter 1), and in Chapters 2 and 3, the required techniques for the retrieval of RNA from histological sections. Both routine and specialist histological staining techniques are provided in Part II. These include pro- cols for immuno (Chapters 4-7), lectin (Chapter 8), and hybridization (Chapter 9) histochemistry, histologicalstaining (Chapters10and11), aswellasspecificmethods for the in situ identification of hypoxia (Chapter 12) and apoptosis (Chapter 13). Finally, Part III details advances in imaging (Chapters 14-16) and image analysis (Chapter 17). It is hoped that this volume will provide molecular biologists with the basic his- chemical techniques and histologists with the molecular techniques to realise the potential of their resource. We are indebted to the authors for their generosity in sharing these protocol

tRNA and AminoacyltRNA Synthetases: Charging of RNA Microhelices and Decoding Genetic Information; P. Schimmel. rRNA and mRNA: Regulation, Processing, and Assembly: Genetic Approaches to the Study of Eukaryotic Ribosomes; J.R. Warner, et al. Translational Initiation and Termination: Mechanisms of Translational Initiation and Repression in Prokaryotes; D.E. Draper. The Elongation Process: Sites, Factors, Nascent Chain: The Allosteric ThreeSite Model and the Mechanism of Action of Both Elongation Factors EFTu and EFG; K.H. Nierhaus, et al. Accuracy in Translation: Mutants of tRNA, Ribosomes, and mRNA Affecting Frameshifting, Hopping, or Stop Codon Read-Through; J.F. Atkins, et al. Quaternary Structure, Functional Centers, and Domains in the Ribosome. Translation in Cell Organelles. The Translational Apparatus and Evolution. 64 additional articles. Index.

A major mechanism by which cells regulate protein function is to place phosphate groups on serine and threonine residues. Though the steady-state level of protein phosphorylation depends on the relative activities of both kinases and phosphatases, a much greater effort has previously gone into the study of the former that the latter . Today, however, there is an increasing appreciation for the role that protein phosphatases play in the dynamic p- cess of protein phosphorylation . To date, there are four major types of protein serine/threonine phosphatase catalytic subunits, designated protein phosphatase type 1, 2A, 2B, and 2C . Each has been identified by the techniques of protein chemistry and enzymology and can be distinguished from one another by their preference for specific substrates as well as their sensitivity to certain acti- tors and inhibitors . Protein Phosphatase Protocols has been assembled in response to the growing interest these enzymes are receiving . The goal of this compilation is to provide a "how-to" experimental guide to aid newcomers as well as s- soned veterans in their research endeavors, thus further contributing towards our ever increasing knowledge of serine/threonine phosphatases . What you have before you contains contributions by many of the current and emerging leaders in the field . To highlight just a few, these chapters c- tain step-by-step information on how to isolate novel phosphatases and re- latory subunits, assay for activity, and generate immunological reagents for both biochemical and biological characterization of these enzymes .

Updating and building upon previous editions, "Hematopoietic Stem Cell Protocols, Third Edition" provides up-to-date protocols from leading stem cell researchers. This in-depth volume presents a clear view of the landscape of assays available to the stem cell researcher working in the growing hematopoietic stem cell (HSC) field. A robust and active field, it is supported by an abundance of innovative mouse models and molecular tools for analysis of phenotypes and functions in mouse and human cells. Understanding more about hematopoietic stem cell biology is integral if these versatile cells are to be applied effectively to treat and cure a wide range of blood diseases.An introductory chapter puts the major contributions of the book into the proper perspective. 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.

Essential for the laboratory-based researcher, "Hematopoietic Stem Cell Protocols, Third Edition" is a much needed technical resource in the critically important field of hematopoietic stem cell investigation."

* Several in vivo assays are currently used in the study of angiogenesis and antiangiogenesis * The chick embryo chorioallantoic membrane is one of the most common and versatile assay to study angiogenesis and antiangiogenesis in vivo * Angiogenesis plays a critical role in tumor progression and metastasis * Antiangiogenesis is one of most promising approach to the treatment of cancer and metastasis The chick embryo chorioallantoic membrane (CAM) is an extraembryonic membrane which serves as a gas exchange surface and its function is supported by a dense capillary network. Because of its extensive vascularization and easy accessibility, the CAM has been broadly used to study the morpho-functional aspects of the angiogenesis process in vivo and to investigate the efficacy and mechanisms of action of pro-angiogenic and anti-angiogenic natural and synthetic molecules. The CAM is a suitable site for transplanting tissues, which can survive and develop in the CAM by peripheral anastomoses between graft and original CAM vasculature or by new angiogenic vessels grown from the CAM that invade the graft. While the formation of peripheral anastomoses between host and pre-existing donor vessels is the main, and the most common, mechanism involved in the revascularization of embryonic grafts, the growth of CAM-derived vessels into the graft is only stimulated in tumor grafts. The CAM has long been a favored system for the study of tumor angiogenesis and metastasis, because at this stage the chick immunocompetence system is not fully developed and the conditions for rejection have not been established. Tumors remain avascular for 72 h, after which they are penetrated by new blood vessels and begin a phase of rapid growth. Also, delivery of tumor cells onto the CAM allows the fine study of the effects of tumor derived angiogenic growth factors on blood vessel structure and functionality. The CAM may also used to verify the ability to inhibit th

This volume represents the proceedings of the 9th International Proton Transport Conference, "Mechanisms and Consequences of Proton Transport" held in Leura, Australia, August 19 -21, 200 I. This conference has been held since 1971 every 3 to 4 years with few exceptions in association with the Congress of the International Union of Physiological Sciences. The first meeting was held in Frankfurt, Germany, at the Max-Planck'Institute. Countries hosting the conference have been Germany (twice), Sweden (twice), India, Canada, USA, Great Britain, and now Australia. Over the past 30 years participants at these Proton Transport Conferences have been principal contributors to the major discoveries in the physiology, biochemistry and pharmacology of gastric acid secretion. These include development of the H2-receptor blockers, defining the signaling pathways for the regulation of acid secretion, identifying the gastric proton pump, discovery and development of proton pump inhibitory drugs, and elucidating the physiology and biochemistry of Helicobacter pylori.

Cutting-edge articles review our current understanding of lipid microdomain signaling mechanisms and their physiological and pathological importance. The book describes the role of lipid rafts in learning, memory, and cancer, presents the emerging evidence that lipid rafts play critical roles in signaling pathways and the regulation of synaptic function in the nervous system, and shows how alterations in lipid raft metabolism are implicated in the pathogenesis of neurodegenerative disorders. Techniques are also described for the isolation of lipid rafts, the analysis of the lipid and protein components of lipid rafts, the imaging of lipid rafts in living cells, and the analysis of signal transduction in lipid rafts.

Since the epochal discovery of the radical and highly toxic gas nitric oxide (NO) as a signaling molecule, two other no less toxic gases - carbon monoxide (CO) and hydrogen sulfide (H2S) - have been found to also be involved in a plethora of physiological and pathophysiological functions. The gases termed gasotransmitters play an increasingly important role in understanding how signalling into and between cells is modulated and fine-tuned. The advent of gasotransmitters has profoundly changed our way of thinking about biosynthesis, liberation, storage and action mechanisms in cellular signaling. In recent years an impressive amount of new data, distributed throughout the existing literature, has been generated. For this book the editors have recruited distinguished colleagues in the field to summarize and review important biological, pharmacological and medical functions and their implications, as well as methods for the detection of gasotransmitters.

Death receptors play a central role in directing apoptosis in mammalian cells. This process of active cell death is important for a number of biological processes, e.g. for the regulation of the immune system. Death receptors are cell surface receptors that transmit apoptotic signals initiated by corresponding death ligands. Many complex signaling pathways are activated and apoptosis is the final result of a complex biochemical cascade of events.

Besides their role in the induction of cell death, evidence now exists that death receptors are able to activate several non-apoptotic signaling pathways which, depending on cellular context, may lead to apoptosis resistance, secretion of pro-inflammatory proteins, proliferation and invasive growth of cancer cells.

This book looks at the molecular basis of death receptor signaling and the role of death receptors in cancer development.