A large number of chemical agents are known which affect blood and blood-forming organs. The purpose of this volume is to review the sig- nificant advances made over the past several years regarding such chemical agents. The purification, biological action, and therapeutic implications of several widely used hematopoietic growth factors such as interleukin 3 (IL-3 or multi-CSF), granulocyte/macrophage colony stimulating factor (GM-CSF), granulocyte colony stimulating factor (G-CSF), colony stimu- lating factor (CSF-I or M-CSF), thrombopoietin, and erythropoietin are included in this volume. These factors are important in regulating several hematopoietic cell lines such as neutrophils, monocytes, eosinophils, macrophages, megakaryocytes, platelets, and erythrocytes. People are exposed daily to numerous toxic chemical substances present in our environment which produce a suppression of erythropoiesis, myelo- poiesis, lymphocytopoiesis, and megakaryocytopoiesis. Attempts have been made in this volume to assess the therapeutic role of some of the hematopoietic factors such as erythropoietin in the anemia of end stage renal disease, as well as colony stimulating factors in other hematopoietic abnormalities. In addition, some of the chemical factors in our environment which suppress major hematopoietic lineages stimulated by erythropoietin, macrophage colony stimulating factor, granulocyte colony stimulating factor, interleukin I-alpha, interleukin I-beta, and interleukins 2, 3, 4, 5, 6, 7, and 9 are also included. An updating of the mechanism of action of each of these factors on the major hematopoietic lineages is covered.

The great strides made in the field of morphological methods during the past decades have perhaps found their most spectacular expression in the functional exploration of the nervous system. In comparison with other tissues, nerve tissue displays three kinds of specificity : structural, because of the unique organization of the neuronal networks ; chemical as shown, for example, by the informative molecules exchanged between the nerve cells, and of course functional, thanks to the particular metabolic and electrophysiological characteristics of the neurons. Although for a long time the structural properties of the nervous system were generally considered to constitute the only field to which morphological techniques could be applied, we are to-day justified in believing that they can also explore the nerve tissue through its specific chemical and functional aspects, thanks to the development of immunocytochemistry and in situ hybridization, to the elaboration of the deoxyglucose method, to the use of voltage or ion sensitive dyes, and to the progress made in the application of in vivo techniques like PET. These methods have evolved so fast, the technical and fundamental problems they raise are so numerous and stimulating, and the importance of the complementary data they provide is so obvious, that we thought it was a good time to organize a new meeting between distinguished specialists in the neurocytochemical field.

Harderian glands, first described by Jacob Harder in two species of deer in 1694, are large, intraorbital glands which, with but few exceptions, are found in all land vertebrates. Certainly, their relatively large size, their phylogenetic age, and persistent conservation in all groups as they emerged from an aqueous to an air/land environment lend strong support to the logical deduction that they probably play an important role in the physiological adaptation to terrestrial life. Yet, few biologists know even what or where these glands are. For a variety of reasons, the Harderian glands have not received the attention they deserve and, as a result, the published works available have been scarce and scattered throughout the world literature. The current situation is remarkably similar to that which existed in regard to the pineal gland prior to the mid-1960s, i. e. , scattered literature, unknown function, few investigators, and little interest. Yet, following a few key papers, interest in the pineal gland expanded and soon an explosive increase in the knowledge and understanding of the pineal gland took place and continues today. Will history repeat itself? Originally, a few of us discussed the desirability of an informal Symposium on the Harderian glands.

The recent interest in the pharmacology of the skin and the treatment of its diseases has come about for two reasons. The first is a realisation that many aspects of pharmacology can be studied as easily in human skin, where they may be more relevant to human physiology and diseases, as in animal models. Examples of this are the action of various vasoactive agents and the isolation of mediators of inflammation after UV irradiation and antigen-induced dermatitis. The second reason is the fortuitous realisation that a pharmacological approach to the treatment of skin disease need not always await the full elucidation of etiology and mechanism. For example, whilst the argument continued unresolved as to whether the pilo-sebaceous infection which constitutes acne was due to a blocked duct or to a simple increase in sebum production, 13-cis-retinoic acid was found quite by chance totally to ablate the disease; again, whilst cyclosporin, fresh from its triumphs in organ transplantation, has been found able to suppress the rash of psoriasis, it has resuscitated the debate on etiology. We are therefore entering a new era in which the pharmacology and clinical pharmacology of skin are being studied as a fascinating new way of exploring questions of human physiology and pharmacology as well as an important step in the development and study of new drugs, use of which will improve disease control and at the same time help to define pathological mechanisms.

Muscarinic acetylcholine receptors have played a key role in the advancement of knowledge of pharmacology and neurotransmission since the inception of studies in these fields, and the effects of naturally occurring drugs acting on muscarinic receptors were known and exploited for both therapeutic and non-therapeutic purposes for hundreds of years before the existence of the receptors themselves was recognized. This volume presents a broad yet detailed review of current knowledge of muscarinic receptors that will be valuable both to long-time muscarinic investigators and to those new to the field. It describes the detailed insights that have been obtained on the structure, function, and cell biology of muscarinic receptors. This volume also describes physiological analyses of muscarinic receptors and their roles in regulating the function of the brain and of a variety of peripheral tissues. This volume shows how the study of muscarinic receptors continues to provide new and surprising insights not just to the cholinergic system but to the broad areas of neurobiology, cell biology, pharmacology, and therapeutics.

"What we call the beginning is often the end, and to make an end is to make a beginning. The end is where we start from." T. S. ELIOT, "Geroniton" The discovery of endothelin-l was both an end and a beginning. It is an end because this discovery has started a plethora of investigations as to endothe1in's sites of production, mode of action, it's normal and abnormal physiological role and its involvement in a variety of pathological conditions ranging from renal failure to myocardial infarction, cardiac hypertrophy, hyperten sion, cyclosporine induced toxicity, mental depression, renal failure, stress related illness, and even stroke and sudden death. It is a beginning because no one can foresee where the investigations will ultimately lead, or how they will end. The discovery of the endothelins was a very recent event and soon specialist books will start to appear dealing in detail with different aspects of their chemistry, physiological role and pathological importance. The present book is intended to act as a guide and primer for those who are becoming intrigued by the many endothelin related articles which are now appearing in the journals but who as yet have not had the time or opportunity to be caught up in the web of enthusiasm and excitement which is motivating those more directly involved in the field of research.

Stress, high blood pressure, smoking, pollution, fast foods, overweight, excessive travelling, surgery, less movement are common features in our modern life. These features are risky for blood clotting disorders. According to WHO, over 29% of the total mortalities worldwide are due to thrombosis. By the year, 2020 cardiovascular diseases (CVDs) may cause an estimated 25 million deaths per year, thus antithrombotic therapy is of great interest.

The available thrombolytic agents such as urokinase are highly expensive, antigenic, quite unspecific, pyretogenic and hemorrhagenic. Therefore, the production of fibrinolysing enzymes, which rapidly dissolute thrombi within the vascular tree, without the detriments by microorganisms, as described in this book, is the desirable aim of today s research. "

sCongenital heart disease is the leading cause of infant death and affects approximately one in every 100 babies born in the United States. The study of cardiovascular development has acquired new momentum in last twenty years due to the advancement of modern molecular biology and new available equipments and techniques. In Cardiovascular Development: Methods and Protocols expert researchers in the field in the field detail many of the methods which are now commonly used in the field of cardiovascular development research. These include methods and technique for using different organisms for cardiovascular developmental research, using cell and molecular biology methods to study cardiovascular development, as well as other available techniques for cardiovascular development research. 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 tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Cardiovascular Development: Methods and Protocols seeks to aid scientists in understanding new state-of-the-art techniques in the field of cardiovascular development research including in vivo imaging and Bioinformatics.

The methods of molecular biology, biochemistry, immunocytochem istry, and in-situ hybridization introduce new opportunities for the classification and functional characterization of cell receptors under normal conditions and for a better understanding of pathogenetic mechanisms in human diseases. The cellular localization and trans location of receptor proteins can be identified using morphological methods, and it is apparent that receptors and receptor defects play an important role in pathology, notably in genetic diseases, endocrine disorders, atherosclerosis, infections, and cancer. In this volume in ternational experts give a current review of the morphology and pathological aspects of cell receptors. The complex communication of multicellular organisms is coordi nated by two regulatory systems: neural and humoral. Both systems function via signaling substances (ligands) and signal-recognizing and -transmitting molecules, called receptors. The historical develop ment of the receptor concept is based upon Paul Ehrlich's theory of "receptors in the immune system," Langley's "receptive substances in postssynaptic membranes," and Earl Sutherland's discovery of "sec ond messengers" (cAMP and Ca2])."

It is a great honor and pleasure for me to introduce this book; an honor, because of the scientific renown and authority of the investigators who have edited the volume and contributed the chapters; a pleasure, because my own long-lasting interest in the baroreflexes has always gone in the same directions as those along which the authors of this book have conceived and organized their work. It is particularly meaningful, in my opinion, that the very title of this volume underlines the integrative functions and the clinical aspects of baroreceptor reflexes. Under the aspect of integration, it is more and more apparent that baroreceptor reflexes, though preponderantly influencing cardiovascular functions, are not limited to cardiovascular control. Their influence on respiration has been well known since the earliest studies on baroreflexes, and wider influences have more recently been shown, e. g. , on hormone release, on sleep and vigilance, and on emotional behavior. Even within the scope of cardiovascular regulation, the integrated action of baroreflexes is not only directly exerted on the heart and blood vessels, but is also exerted through more devious but no less important routes, such as renin release from juxtaglomerular cells and sodium and water reabsorption by the renal tubules.

The present book is the outcome of a symposium on magnesium and excitable membranes, held in September 1989 on the occasion of the 12th annual meeting of the European Neuroscience Associ- ation in Torino. Leading specialists from around the world met in order to discuss the physiological and clinical effects of magne- sium on excitable cells. Magnesium ions are known to playa key role in the regulation of a variety of cell functions and, as such, have been implicated in the control of neuronal excitability and cardiovascular diseases. In the last decade, the advent of new electro physiological techniques has provided important details on the modulatory action of mag- nesium on ion channels and the receptor proteins of neurotrans- mitters. Thus, in neuronal and muscle cells, extracellular and in- is shown to block ion passage through open tracellular magnesium channels in a voltage- and time-dependent manner. The mecha- nism appears to be common for voltage- and receptor-operated channels, and it is thought to be an important cofactor in the long-term potentiation of synaptic transmission, brain aging, and cardiac activity. Recent findings show that the action of internal magnesium has additional features. High and low affinity sites for internal magnesium are involved in the regulation of channel gat- ings and ion permeation. Biochemical studies suggest that internal magnesium interferes with G-protein subunits, GTP-ase activity, and second messengers involved in channel phosphorylation.

"Provides analytical chemists and biomedical scientists with an excellent summary of progress...This is a book that can be recommended to all analytical scientists interested in fluorimetry." (Analytical Chimica Acta) "This is a useful overview and gives the nonspecialist a feeling for the advantages and limitations of the methods. Overall this book is a worthwhile read and a good source of references." (TRAC)
The book is divided into chapters on new methods, new appli- cations, fluorescence immunoassays, fluorometric analysis and fluorescence spectroscopy in biomedical sciences. Specific topics are fluorescence spectroscopy using synchrotron radiation, picosecond fluorescence spectroscopy, fluorescence microscopy, fluorescence scattering by synthetic polymers, fluorescence immunoassays, fluorescence for environmental monitoring, fluorescence in flow injection analysis, hydro-geological studies, fluorescence of proteins, lipids and membranes, cell fluorescence, calcium transients.

During the last years much attention has been given to the investigation of the roles of specific growth modulators acting on liver cell populations. HGF functions as a pleotropic factor in the liver as well as in other extrahepatic tissues. The hepatic stem cell compartment is thought to play a major role in liver tissue restoration. Mechanisms of cell growth and cell death of the liver are described in detail, with particular emphasis on apoptosis. The functions of HBV-pX protein and the roles of hepatitis B virus gene expression relation to hepatocellular carcinoma are discussed. Techniques for obtaining human and nonhuman hepatocytes and other liver cell types and for efficient transfer and expression of genes in hepatocytes and liver-derived cell lines are presented.

Biological cell membranes regulate the transfer of matter and informa tion between the intracellular and extracellular compartments as basic survival and maintenance functions for an organism: This volume contains a series of reviews that are concerned with how biological plasma membranes regulate the transport of chloride between the intracellular and extracellular compartments of a cell. This book is also an attempt to analyze the molecular basis for the movement of chloride across a cell membrane. This volume is devoted to a diversity of electrogenic chloride transport mechanisms in representative cell membranes of all living things. The first section of the book (Chaps. 1-4) focuses on mech anisms of primary active chloride transport (i.e. photon or A TP-driven), while the second section (Chaps. 5-6) centers on secondary active chloride transporters (symport and antiport) in both plant and animal plasma membranes. The last section, which comprises seven chapters (Chaps. 7-13), deals with chloride channels in cell membranes of prokaryocytes and eukaryocytes of most every phylum. It is hoped that with this particular ordering the reader can glean a telescopic view of the evolutionary history of the various electrogenic chloride transporters."

Comparative endocrinology helps to find the roots of homeostatic regulation in organisms. In this context, many years ago a series of experiments were done, which demonstrated the hormonal regula tion also on the invertebrate level. The mechanisms are partly similar, partly different, from those found in vertebrates. The new receptor era of mammalian endocrinology stimulated research on invertebrate hormone receptors, and sophisticated methods are applied also to determine hormones. The experiments demonstrated the existence and even similar function of these structures and signaling molecules. However, data on hormones and receptors at the lowest level of metazoan life and the highest level of protozoan life were not at our disposal. About two decades ago, first observations on the presence of hormone receptors reacting to vertebrate hormones in protozoa were made. Since the early 1980s we know that hormone-like molecules similar to those of higher vertebrates are present also in unicellular organisms. The presence of some second messengers in Tetrahymena was recognized. Since then, the research has been extended and many structures - previously believed to be solely vertebrate characteristics, such as opiate receptors, similar to mammalian ones - were found in unicellular organisms. These observations justified the assumption of a complete endocrine system at protozoan level, where - considering the unicellularit- this seemed to be not required. However, it became clear that the roots of endocrine communication date back at least 2 billion years."

Contents Introduction .......................................... . 1 The Primate Eye ...................................... . 2 Embryology of Retina and Choroid ....................... . 4 Microscopic Anatomy .................................. . 4 Retina ............................................ . 4 Choroid ........................................... . 8 Material and Methods .................................. . 10 Fine Structure of the Retina 14 RetinaI Pigment Epithelium ............................. . 16 Photoreceptor Cells ................................... . 30 Outer Plexiform Layer and Horizontal Cells .................. . 64 Bipolar, Radial Clial, and Amacrine Cells .................... . 76 Canglion Cells and InternaI Limiting Membrane ............... . 98 Spatial Density of RetinaI Cells .......................... . 112 Fine Structure of the Choroid ........................... . 116 Choroidocapillaris and Its Fiber System ..................... . 118 Arteries, Veins, and Lymphatic Spaces ...................... . 134 Choroidal Nerves .................................... . 142 Cells of Choroidal Connective Tissue ....................... . 148 References ........................................... . 153 Index ................................................ . 157 vii This volume describes the morphology of the primate re tina as seen with the electron microscope. As it is an atlas, the electron micrographs are its most In trad lietian important part. The text accompanies the figures, highlighting selected topics either to explain structures or to point out structure-function relation ships. A scholarly review of the whole spectrum of research on the re tina and choroid is not feasible in a single volume. Thus, whenever available, review artides or monographs, rather than original work, are cited for reference."

Idiopathic cardiomyopathy has been categorized for over 10 years by the World Health Organization as dilated, hypertrophic, or restrictive. These categories have somewhat vague boundaries; however, idiopathic dilated cardiomyopathy (IDC) finds its general definition from unexplained four-chamber dilatation, prominent systolic dysfunction with depressed ejection fraction, a marked increase in total myocardial mass without wall thickening, myocyte hypertrophy and nuclear polyploidy, rarity of frank myocardial scars, but increased interstitial connective tissue and myocyte atrophy, all in the setting of anatomically normal heart valves and coronary arteries. Mural thrombi within cardiac chambers and atrioventricular valve regurgitation are common secondary con- comitants. Myocardial inflammation is rare, but immunological abnormalities have been suggested. Ultrastructural features in- clude increased mitochondrial numbers, lipid deposits, and myo- fibrillar loss. Progressive pump failure with or without rhythm disturbances typically leads to ever-intensified medical therapies, and ultimately to transplantation or early death. The need for better understanding of IDC is represented by incidence rates varying from 5 to 8/100000 population across several populations, 10 000-20000 IDC deaths annually in the United States, and the generally increasing prevalence of the disease. More than 50% of heart transplant recipients have IDC as their primary myocardial disease. The half-life of patient sur- vival without transplantation is generally less than 5 years. Treat- ment with antiarrhythmics, calcium channel blockers, ~-adrenergic blockers, and angiotensin-converting enzyme inhibitors is being evaluated in clinical trials, but the role of such treatments in the pathobiology of progressive disease is unclear.

Understanding the molecular basis of complex biological processes has been a major goal of biological chemistry from early on. Inflammation is one such entity, and recent years have seen exciting progress in the under- standing of molecular interactions; there has been a long way from dolor, rubor, calor, and tumor as a fundamental description of the phenomenon to current knowledge, e.g., on the control of the respiratory burst of the granulocyte, the atomic details of protease regulation, or the interaction of cytokines. We were glad to have been given the opportunity by the Gesellschaft fUr Biologische Chemie to call upon experts in this bustling field of research for this Colloquium. The sessions were organized in the order of increasing complexity, starting with the key phenomena of the inflamma- tory response and its modulation by cytokines to intravascular events and shock and sepsis; thus, the current attempts to apply basic knowledge on mediators of inflammation to the clinical situation were also considered. In-depth chapters presenting the state of the art in these areas are collected in this book, and we thank the authors for their efforts. We also thank the Chairmen of the sessions, Profs. C. Sorg (Munster), D. Roos (Amsterdam), S. Bhakdi (Mainz), H. J. Muller-Eberhard (Hamburg), H. G. Schwick (Marburg), K. Resch (Hannover), W. Schaper (Bad Nauheim), D. Keppler (Heidelberg), and O. Trentz (Zurich) for their input, and it is a great pleasure to acknowledge the support by Drs.

Stability of the internal environment in which neuronal elements are situated is unquestionably an important prerequisite for the effective transmission of information in the nervous system. During the past decade our knowledge on the microenvironment of nerve cells has expanded. The conception that the microenvironment of neurones comprises a fluid with a relatively simple and stable composition is no longer accepted; the microenvironment is now envisaged as a dynamic structure whose composition, shape, and volume changes, thereby significantly influencing neuronal function and the trans mission of information in the nervous system. The modern conception of the neuronal microenvironment is based on the results of research over the last 20 years. The extracellular space (ECS) is comprehended not only as a relatively stable microenvironment containing neurones and glial cells (Bernard 1878), but also as a channel for communica tion between them. The close proximity of the neuronal elements in the CNS and the narrowness of the intercellular spaces provides a basis not only for interaction between the elements themselves, but also between the elements and their microenvironment. Substances which can cross the cell membranes can easily find their way through the microenvironment to adjacent cellular elements. In this way the microenvironment can assure non-synaptic com munication between the relevant neurones. Signalization can be coded by modulation of the chemical composition of the ECS in the vicinity of the cell membrane and does not require classic connection by axones, dendrites, and synapses.

The details of the receptor mechanism are not yet fully understood for any sensory system. However, sufficient data are available (for the vestibular system and for other systems) to permit meaningful tracking of the sensory messages through the nervous system and via conscious experience. The reception, process ing, storage and output of information in man and other animals, as done by means of receptors, neurons, secretory cells and muscle fibers, are collectively referred to as mind. Sensory physiologists tend to disbelieve in extrasensory perception. Sensory physiology in general is an area upon which different sciences and methods converge. Anatomists, physiologists, psychologists, physicists, chemists, and engineers have made important contributions to sensory physiology. What is special about vestibular physiology is the fact that many research workers are clinicians, living under the constant pressure of their patient's demands. This is a disadvantage when it comes to writing handbooks, but an advantage for the pa tient, since research is guided by clinical practice and can be quickly applied. Modern methods, such as recording from single nerve units and the correlation of electrophysiological and psychophysical data, have greatly contributed to our knowledge, yet the study of lesions is still important, especially in the vestibular field."

Traditionally, cardiac hypertrophy is regarded as an adaptation of the heart to permanent mechanical overload. Regardless of the fact that many different and often unknown primary causes can result in heart failure, mechanical overload and myocardial hypertrophy is found in almost all forms of manifest chronic heart failure (apart from failure due to extramyocardial hindrances to inflow or to relaxation). However, the reactive enlargement of myocardial mass in response to an enhanced hemodynamic burden appears to be a double-edged sword. Obviously, the hypertrophy helps to reduce the enhanced ventricular wall stress in heart failure by adding contractile units to the overdistended chamber wall. However, in recent years it became clear that this adaptive hypertrophic process is rather complex and may include problematic facets. The adaptive hypertrophy includes proliferation of the nonmyocyte cardiac cells as well as substantial alterations in the phenotype of the growing myocytes due to differential changes in gene expression.

The authors, all accomplished and world-renowned experts in their topic areas, have provided controversial and up-to-date accounts of the current status of research in calcium and bone metabolism by reviewing major areas of basic science which have an impact on the understanding of bone diseases and related disorders.