Proteins are still gaining importance in the pharmaceutical world, where they are used to improve our arsenal of therapeutic drugs and vaccines and as diagnostic tools. Proteins are different from "traditional" low-molecular-weight drugs. As a group, they exhibit a number of biopharmaceutical and formulation problems. These problems have drawn considerable interest from both industrial and aca demic environments, forcing pharmaceutical scientists to explore a domain previ ously examined only by peptide and protein chemists. Biopharmaceutical aspects of proteins, e.g., low oral bioavailability, have been extensively investigated. Although all possible conventional routes of ad ministration have been examined for proteins, no real, generally applicable alter native to parenteral administration in order to achieve systemic effects has yet been discovered. Several of these biopharmaceutical options have been discussed in Volume 4 of this series, Biological Barriers to Protein Delivery. Proteins are composed of many amino acids, several of which are notorious for their chemical instability. Rational design of formulations that optimize the native structure and/or bioactivity of a protein is therefore of great importance when long shelf life is required, as it is for pharmaceutical products. This issue has also been examined in two prior volumes of this series: Volume 2: Stability of Protein Pharmaceuticals (Part A) and Volume 5: Stability and Characterization of Protein and Peptide Drugs.

Signaling through antigen receptor initiates a complex series of events resulting in the activation of genes that regulate the development, proliferation and differentiation of lymphocytes. During the past few years, rapid progress has been made in understanding the molecular basis of signaling pathways mediated by antigen and cytokine receptors. These pathways involve protein tyrosine kinases which are coupled to downstream regulatory molecules, including small guanine nucleotide binding proteins (e. g. p21'OS), serine threonine kinases (e. g. , members of the ERK family), and a large group of transcription factors. More recently, there have been breakthroughs in elucidating the genetic defects underlying three X-linked primary immunodeficiency diseases in humans. This volume surveys aspects of these rapidly developing areas of research. The book is divided into 5 different sections. Section I deals with signaling pathways in B lymphocytes. It includes a contemporary assessment of B cell antigen receptor structures, and discussion of the role of Ig-a/lg-B polypeptides in linking the antigen receptor to intracellular signal transduction pathways. The role of accessory molecules in the regulation of signaling by the B cell antigen receptor is also considered. Section II adopts a similar approach to the analysis of the antigen receptor on T lymphocytes. The importance of specialized signaling motifs in the CD3 polypeptides, mechanisms whereby these motifs may interact with the lymphocyte-specific protein tyrosine kinases, and the downstream consequences of these interactions are reviewed. In addition, the role of antigen-induced apoptosis in the generation of immunological tolerance is discussed.

As we approach the twenty-first century the problems of industrialization are evident: we find there is a greenhouse effect, the ozone layer is being depleted, the rain is acidified, and there is a terrible problem of increasing C0 concentrations in the atmo 2 sphere. The carbonic anhydrases are a unique family of enzymes that solve these problems in the human body: they are responsible for converting C0 (a gas) to 2 HC0-, which is the biggest intracellular buffer, with a concomitant decrease in a 3 hydroxyl ion. Globally, the functions of the carbonic anhydrases in photosynthesis in rain forests and in the algae and plankton that cover our oceans indicate that they are also of utmost importance in the maintenance of the acid-base balance on our planet. Although the whole field of C0 metabolism is enormous and still rapidly 2 expanding, because of the research interests of the editors this book is mainly concerned with mammalian carbonic anhydrases. However, if the interested reader intends to purify carbonic anhydrases from nonmammalian sources, Dr. Cheg widden has provided the necessary information in Chapter 7. The carbonic anhydrases were first discovered in 1933; until1976 there were thought to be only two isozymes. Since then CA ill, IY, V, VI, and Vll have been discovered and well characterized. There is, of course, no reason to believe that we have found them all."

The predecessor to this book was A Guide to the Laboratory Use of the Squid Loligo pealei published by the Marine Biological Laboratory, Woods Hole, Massachusetts in 1974. The revision of this long out of date guide, with the approval of the Marine Biological Laboratory, is an attempt to introduce students and researchers to the cephalopods and particularly the squid as an object of biological research. Therefore, we have decided to expand on its original theme, which was to present important practical aspects for using the squid as experimental animals. There are twenty two chapters instead of the original eight. The material in the original eight chapters has been completely revised. Since more than one method can be used for accomplishing a given task, some duplication of methods was considered desirable in the various chapters. Thus, the methodology can be chosen which is best suited for each reader's requirements. Each subject also contains a mini-review which can serve as an introduction to the various topics. Thus, the volume is not just a laboratory manual, but can also be used as an introduction to squid biology. The book is intended for laboratory technicians, advanced undergraduate students, graduate students, researchers, and all others who want to learn the purpose, methods, and techniques of using squid as experimental animals. This is the reason why the name has been changed to its present title. Preceding the chapters is a list of many of the abbreviations, prefixes, and suffixes used in this volume.

This volume describes the current state of our knowledge on the neurobiology of muscle fatigue, with consideration also given to selected integrative cardiorespiratory mechanisms. Our charge to the authors of the various chapters was twofold: to provide a systematic review of the topic that could serve as a balanced reference text for practicing health-care professionals, teaching faculty, and pre-and postdoctoral trainees in the biomedi cal sciences; and to stimulate further experimental and theoretical work on neurobiology. Key issues are addressed in nine interrelated areas: fatigue of single muscle fibers, fatigue at the neuromuscular junction, fatigue of single motor units, metabolic fatigue studied with nuclear magnetic resonance, fatigue of the segmental motor system, fatigue involving suprasegmental mechanisms, the task dependency of fatigue mechanisms, integrative (largely cardiorespiratory) systems issues, and fatigue of adapted systems (due to aging, under-and overuse, and pathophysiology). The product is a volume that provides compre of processes that operate from the forebrain to the contractile proteins.

In the last decade, research on platelet-activating factor (PAF) has expanded exponentially. Previous conferences on PAF in Paris, 1983, and the subsequent conferences in Gatlinburg, Ten nessee, Tokyo, Snowbird, Utah, and Berlin, at three-yearly intervals, have chronicled the devel opments in the field ofPAF. This volume records the proceedings of the Fifth International Con gress on PAF and Related Lipid Mediators, held at the Free University Medical Hospital Ben jamin Franklin in Berlin, from September 12-16, 1995. We are very much indebted to Free Uni versity Berlin for providing tremendous facilities and financial support. It was a great pleasure to have positive and generous input from the German Science Council (DFG), Bonn, Germany, and British.Biotech, Oxford, United Kingdom. Their support was crucial in making the congress a scientific success. Twenty other organizations provided additional financial support, for which we extend our deepest appreciation. The editors would like to thank all of those who participated in this congress and the authors for their contributions. The organization and planning of the Berlin Congress were carried out by an organizing committee. We gratefully acknowledge the support and assistance of the organizing commit tee members, especially Renate Nigam and Renate Roux for their untiring efforts to make the congress successful. Many colleagues also supported the congress with dedication, hard work, and expert input. We are grateful to them. We also wish to acknowledge the support of G. Sravan Kumar and Louis Kock for their efforts in producing this volume."

This volume contains the scientific papers and abstracts of posters presented at the International Symposium on Molecular Insect Science held in Tucson, Arizona, October 22-27, 1989. This meeting was organized by the Center for Insect Science at the University of Arizona in response to the growing need for a forum dedicated to the impact of modern biology on insect science. While scientific studies of a few insects, notably Drosophila melanogaster, have always had a central role in the development of biology, it is only recently that tools have become available to extend these studies to other insects, including those having economic and medical importance. The Tucson meeting was evidence of how far we have come in extending modern biological tools to the study of insects. It is also evident from the contents of this book that the study of insects is making an increasingly important contribution to the advancement of biology generally. Given the large impact of insects on human life, such a development has considerable importance for human welfare, and of the welfare of the ecosystem as a whole. It should be noted that several of the participants who presented posters were invited to prepare full length papers to ensure that the book covered the major areas of insect science. The financial support of the National Science Foundation and the Monsanto Corporation is gratefully acknowledged. Thanks are also due to Sharon Richards for her dedicated work on the manuscripts. Henry H.

In the past few years, the scientific community has witnessed significant progress in the study of ion channels. Technological advancement in biophysics, molecular biology, and immunology has been greatly ac celerated, making it possible to conduct experiments which were deemed very difficult if not impossible in the past. For example, patch-clamp techniques can now be used to measure ionic currents generated by almost every type of cell, thereby allowing us to analyze whole-cell and single channel events. It is now possible to incorporate purified ion channel components into lipid bilayers to reconstitute an "excitable membrane." Gene cloning and monoclonal antibody techniques provide us with new approaches to the study of the molecular structure of ion channels. A variety of chemicals have now been found to interact with ion channels. One of the classical examples is represented by tetrodotoxin, a puffer fish poison, which was shown in the early 1960s to block the voltage-activated sodium channel in a highly specific and potent manner.

This book is dedicated to the memory of two colleagues and friends, Amico Big- nami and Hendrick Vander Los. They were both pioneers in their fields: Bignami on on- togenesis and function of neuroglia, and Van der Los on brain plasticity and neuronal circuitry. Their ideas are further pursued by the authors in this book. Some of the chapters are products of a conference dedicated to these two scientists entitled "Recent Advances in Neurobiology: Plasticity and Regeneration." The conference was organized by the Insti- tute of Developmental Neuroscience and Aging and sponsored by the Region della Valle d' Aosta. Also, several chapters are written by colleagues who knew well either Amico or Hendrick and were invited to contribute to this dedication. The book is divided in four sections. The first part covers neurons, neuroglia includ- ing microglia, their plasticity and phenotypic expression, and specific functions and inter- actions. It is now established that neuroglia are an intimate component of the neuronal environment and thought to regulate several neuronal functions. More recently microglia have become prominent as the immune cells in the CNS. This part contributes new infor- mation for these cellular interactions. The second part deals with neuronal and glial cell plasticity as it relates to regeneration and neurodegeneration, more or less an extension of Part I. In recent years the role of transplantation in regeneration has become promising.

Here is the first effort in a single volume to cover all of the integrative functions of calcium signalling - how changes in intracellular calcium coordinate a variety of coherent cellular responses. Written by a team of internationally established researchers, Integrative Aspects of Calcium Signalling provides the latest experimental data and concepts, bringing together a detailed analysis of the events, processes, and functions regulated by calcium signalling. A unique resource for professionals and students of physiology, biophysics, neurobiology, biochemistry, and all related fields.

How does the brain code and process incoming information, how does it recog nize a certain object, how does a certain Gestalt come into our awareness? One of the key issues to conscious realization of an object, of a Gestalt is the attention de voted to the corresponding sensory input which evokes the neural pattern underly ing the Gestalt. This requires that the attention be devoted to one set of objects at a time. However, the attention may be switched quickly between different objects or ongoing input processes. It is to be expected that such mechanisms are reflected in the neural dynamics: Neurons or neuronal assemblies which pertain to one object may fire, possibly in rapid bursts at a time. Such firing bursts may enhance the synaptic strength in the corresponding cell assembly and thereby form the substrate of short-term memory. However, we may well become aware of two different objects at a time. How can we avoid that the firing patterns which may relate to say a certain type of move ment (columns in V5) or to a color (V 4) of one object do not become mixed with those of another object? Such a blend may only happen if the presentation times be come very short (below 20-30 ms). One possibility is that neurons pertaining to one cell assembly fire syn chronously. Then different cell assemblies firing at different rates may code different information."

In recent years experimental and numerical studies have shown that chaos is a widespread phenomenon throughout the biological hierarchy ranging from simple enzyme reactions to ecosystems. Although a coherent picture of the fundamental mechanisms responsible for chaotic dynamics has started to appear it is not yet clear what the implications of such dynamics are for biological systems in general. In some systems it appears that chaotic dynamics are associated with a pathological condi tion. In other systems the pathological condition has regular periodic dynamics whilst the normal non-pathological condition has chaotic dyna mics. Since chaotic behaviour is so ubiquitous in nature and since the phenomenon raises some fundamental questions about its implications for biology it seemed timely to organize an interdisciplinary meeting at which leading scientists could meet to exchange ideas, to evaluate the current state of the field and to stipulate the guidelines along which future research should be directed. The present volume contains the contributions to the NATO Advanced Research Workshop on "Chaos in Biological Systems" held at Dyffryn House, St. Nicholas, Cardiff, U. K., December 8-12, 1986. At this meeting 38 researchers with highly different backgrounds met to present their latest results through lectures and posters and to discuss the applica tions of non-linear techniques to problems of common interest. . In spite of their involvement in the study of chaotic dynamics for several years many of the participants met here for the first time."

I have been asked to write a brief foreword to this volume honoring Hisako Ikeda, providing a review of the accomplishments in our field over the past four decades, when Hisako was an active participant. This I am delighted to do. It has been a most exciting time in vision research and Hisako has been right in the middle of much of the excitement, publishing on a wide variety of topics and providing much new data and many new insights. Hisako's research career can be divided by decades into four quite distinct areas of inquiry. In the 1950s, as a student in Japan, her research interests were psychophysical in nature, and she was concerned with visual illusions, figural aftereffects, and motion detec tion. In the 1960s, after her move to London, she began electrophysiological studies. Much of her work in the 1960s was concerned with the electroretinogram (ERG), its components, and the use of this electrical response for evaluating spectral sensitivities of the eye and retinal degenerations. This work represented the beginning of her electrodiagnostic clinical work, which continued until her retirement."

William P. Cooney III, R. A. Berger, and K. N. An Orthopedic Biomechanics Laboratory Department of Orthopedic Surgery Mayo Clinic and Mayo Foundation Rochester, MN 55905, U. S. A. As surgeons struggle to find new insights into the complex diseases and deformities that involve the wrist and hand, new insights are being provided by applied anatomy, physiology and biomechanics to these important areas. Indeed, a fresh new interaction of disciplines has immersed in which anatomists, bioengineers and surgeons examine together basic functions and principles that can provide a strong foundation for future growth. Clinical interest in the hand and wrist are now at a peak on an international level. Economic implications of disability affecting the hand and wrist are recognized that have international scope crossing oceans, cultures, languages and political philosophies. As with any struggle, a common ground for understanding is essential. NATO conferences such as this symposium on Biomechanics of the Hand and Wrist provides such a basis upon which to build discernment of fundamental postulates. As a start, basic research directed at studies of anatomy, pathology and pathophysiology and mechanical modeling is essential. To take these important steps further forward, funding from government and industry are needed to consider fundamental principles within the material sciences, biomechanical disciplines, applied anatomy and physiology and concepts of engineering modeling that have been applied to other areas of the musculoskeletal system.

Fluorescence microscopy images can be easily integrated into current video and computer image processing systems. People like visual observation; they like to watch a television or computer screen, and fluorescence techniques are thus becoming more and more popular. Since true in vivo experiments are simple to perform, samples can be directly seen and there is always the possibility of manipulating the samples during the experiments; it is an ideal technique for biology and medicine. Images are obtained by a classical (now called wide-field) fluorescence microscope, a confocal scanning microscope, upright or inverted, with epifluorescence or transmission. Computerized image processing may improve definition, and remove glare and scattered light signal. It also makes it possible to compute ratio images (ratio imaging both in excitation and in emission) or lifetime imaging. Image analysis programs may supply a great deal of additional data of various types, starting with calculations of the number of fluorescent objects, their shapes, brightness, etc. Fluorescence microscopy data may be complemented by classical measurement in the cuvette yr by flow cytometry.

This text attempts to introduce the molecular biology of cell membranes to students and professionals of diverse backgrounds. Although several membrane biology books are available, they do not integrate recent knowledge gained using modern molecular tools with more traditional membrane topics. Molecular techniques, such as cDNA cloning and x-ray diffraction, have provided fresh insights into cell membrane structure and function. The great excitement today, which I attempt to convey in this book, is that molecular details are beginning to merge with physiological responses. In other words, we are beginning to understand precisely how membranes work. This textbook is appropriate for upper-level undergraduate or beginning graduate students. Readers should have previous or concurrent coursework in biochemistry; prior studies in elementary physiology would be helpful. I have found that the presentation of topics in this book is appropriate for students of biology, biochemistry, biophysics and physiology, chemistry, and medicine. This book will be useful in courses focusing on membranes and as a supplementary text in biochemistry courses. Professionals will also find this to be a useful resource book for their personal libraries.

Although there is general agreement that exogenous electric and electromagnetic fields influence and modulate the properties of biological systems. there is no concensus regarding the mechanisms by which such fields operate. It is the purpose of this volume to bring together and examine critically the mechanistic models and concepts that have been proposed. We have chosen to arrange the papers in terms of the level of biological organization emphasized by the contributors. Some papers overlap categories. but the progression from ions and membrane surfaces. through macromolecules and the membrane matrix to integrated systems. establishes a mechanistic chain of causality that links the basic interactions in the relatively well understood simple systems to the complex living systems. where all effects occur simultaneously. The backgrounds of the invited contributors include biochemistry. biophysics. cell biology. electrical engineering. electrochemistry. electrophysiology. medicine and physical chemistry. As a result of this diversity. the mechanistic models reflect the differing approaches used by these disciplines to explain the same phenomena. Areas of agreement define the common ground. while the areas of divergence provide opportunities for refining our ideas through further experimentation. To facilitate the interaction between the different points of view, the authors have clearly indicated those published observations that they are trying to explain. i.e. the experiments that have been critical in their thinking. This should establish a concensus regarding important observations. In the discussion of theories.

Amino acid transport is a part of each of two larger subjects, amino acid metabolism and the biomembrane transport of various . small molecules and ions. Nevertheless in this volume we treat amino acid transport as more than a fragment of either of these two larger subjects. A more comprehensive approach is justified when we remember two historic and ongoing aspects of the title subject. First, amino acid transport had its beginning and acquired a distinct momentum (even if somewhat interrupted from 1913 until about 1945) as amino acid metabolism with the central and pioneer work of Van Slyke and Meyer in 1913. The reviews in this volume will show that it steadily becomes a larger aspect of amino acid metabolism, broadly perceived. These chapters will show for how many organelles, cells, tissues, organs and organ systems, the transmembrane compartmentations and flows of amino acids play very large parts in their fundamental biological relations. The authors here are tending collectively to evaluate an understanding of amino acid flows across biomernbranes, and the regulation of these flows, as necessary to an ultimate understanding of the full range of development and metabolism. Such an understanding goes far beyond the purely substrate-destabilizing contributions by enzymes, which have often been arbitrarily limited to that conceptual entity, "the cell," and which for so long a splendid time had most of biochemical research attention.

Since programmed cell death was first described in insects in 1964 and apoptosis was described in 1972, rapid progress has been made in understanding the basic mechanisms and genes regulating programmed cell death and apoptosis. In addition, defects in various genes regulating programmed cell death have been delineated in several experimental models of human diseases. This volume surveys various aspects of these rapidly developing areas of research in programmed cell death/apoptosis. This volume should be of interest to basic immunologists and molecular biologists. The volume begins with a historical perspective of cell death. The remainder of the volume is divided into four different parts. Part I deals with the signaling pathways in apoptosis, including cell cycle control of apoptosis, role of ceramide in apoptosis, role of antibody signaling, and biochemical regulation of apoptosis. The mechanisms for recognition of apoptotic lymphocytes by macrophages are also reviewed. Part II examines the role of various genes that regulate apoptosis, including the role ofFas, FasL, and other TNF family members in apoptosis and homeostatic regulation of immune response. Recently described splice variants and their influence on apoptosis are also reviewed, and the role of the members of the Bcl-2 family in apoptosis is discussed in detail. Part III reviews various aspects of apoptosis in B lymphocytes, including mechanisms that regulate apoptosis/survival of B lymphocytes and the regulation of Fas-mediated apoptosis in B lymphocytes.

The past decade has seen a steady increase in studies oflemur behavior and ecology. As a result, there is much novel information on newly studied populations, and even newly discovered species, that has not yet been published or summarized. In fact, lemurs have not been the focus of an international symposium since the Prosimian Biology Conference in London in 1972. Moreover, research on lemurs has reached a new quality by addressing general issues in behavioral ecology and evolutionary biology. Although lemurs provide important comparative information on these topics, this aspect of research on lemurs has not been reviewed and compared with similar studies in other primate radiations. Thus, as did many in the field, we felt that the time was ripe to review and synthesize our knowledge of lemur behavioral ecology. Following an initiative by Gerry Doyle, we organized a symposium at the XIVth Congress of the International Primatological Society in Strasbourg, France, where 15 contributions summarized much new information on lemur social systems and their ecological basis. This volume provides a collection of the papers presented at the Strasbourg symposium (plus two reports from recently completed field projects). Each chapter was peer-reviewed, typically by one "lemurologist" and one other biologist. The first three chapters present novel information from the first long-term field studies of three enigmatic species. Sterling describes the social organization of Daubentonia madagascariensis, showing that aye-aye ranging patterns deviate from those of all other nocturnal primates.

During the past few decades, much research has been reported on the formation of insoluble monomolecular films of lipids and biopolymers (synthetic polymers and proteins) on the surface of water or at the oil-water interface. This interest arises from the fact that monomolecular film studies have been found to provide much useful information on a molecular scale, information that is useful for understanding many industrial and biological phenomena in chemical, agricultural, pharmaceutical, medical, and food science applications. For instance, information obtained from lipid monolayer studies has been useful in determining the forces that are known to stabilize emulsions and biological cell membranes. The current texts on surface chemistry generally devote a single chapter to the characteristics of spread monolayers of lipids and biopolymers on liquids, and a researcher may have to review several hundred references to determine the procedures needed to investigate or analyze a particular phenomenon. Furthermore, there is an urgent need at this stage for a text that discusses the state of the art regarding the surface pheqomena exhibited by lipids and biopolymers, as they are relevant to a wide variety of surface and interfacial processes.

This book focuses on sensing and the evolution of animals. Using the five senses (visual, auditory, and olfactory perception, and taste and touch), animals can receive environmental stimuli and respond to them. Changes in these sensitivities might cause changes in aspects of animals lives such as habitat, activity timing, and diet and vice versa. Recent advances in genome and molecular analysis enable us to investigate certain changes in the receptors or mechanisms involved in sensing and provide clues for understanding the evolution of animals related to those changes. The first chapter deals with the molecular evolution of opsins. In addition to the well-known function of opsins as visual receptors, opsins can be related to non-visual photoreception such as photoentrainment of circadian rhythm, photoperiodism, and background adaptation. Molecular phylogenic studies reveal that all opsin genes have evolved from one ancient opsin gene. The evaluation of the functions of each extant opsin protein based on the molecular features enables us to predict the molecular evolution and diversification of opsins during the evolution of animals. These studies shed light on which amino-acid substitutions cause the functional diversification of opsins and how they have influenced the evolution of animals. The second chapter has to do with bitter taste perception, a key detection mechanism against the ingestion of bioactive substances. Genetic and behavioral evidence reveal the existence of "non-taster" Japanese macaques for specific bitter compounds, which originated in a restricted region of Japan. This finding might provide a clue for elucidating the ecological, evolutionary, and neurobiological aspects of bitter taste perception of primates. The third chapter presents an extreme example of the evolution of olfaction, namely, that fully aquatic amniotes have generally reduced their olfactory capacity considerably compared to their terrestrial relatives. Interestingly, the remaining olfactory abilities are quite different among three fully aquatic amniotes investigated: toothed whales have no nervous system structures that mediate olfaction, but baleen whales can smell in air, and it has been suggested that sea snakes smell underwater."

Methods in Bone Biology is unique in being devoted to describing the methodology used by bone researchers. This book describes in detail the techniques of cell and organ culture used in the study of bone and bone cell function and the techniques used to monitor the skeleton and skeletal remodelling both in clinical and experimental settings.