This first volume in a projected series contains the proceedings of the first of the Keio University International Symposia for Life Sciences and Medicine under the sponsorship of the Keio University Medical Science Fund. As stated in the address by the President of Keio University at the opening of the 1996 symposium, the fund of Dr. Mitsunada Sakaguchi. The Keio was established by the generous donation University International Symposia for Life Sciences and Medicine constitute one of the core activities of the fund. The objective is to contribute to the international community by developing human resources, promoting scientific knowledge, and encouraging mutual exchange. Every year, the Executive Committee of the Interna tional Symposia for Life Sciences and Medicine selects the most interesting topics for the symposium from applications received in response to a call for papers to the Keio medical community. The publication of these proceedings is intended to publicize and distribute information arising from the lively discussions of the most exciting and current issues during the symposium. Weare grateful to Dr. Mitsunada Sakaguchi, who made the symposium possible, the members of the program committee, and the office staff whose support guaran teed the success of the symposium. Finally, we thank Springer-Verlag, Tokyo, for their assistance in publishing this work. Akimichi Kaneko, M. D. , Ph. D.

Research centering on blood flow in the heart continues to hold an important position, especially since a better understanding of the subject may help reduce the incidence of coronary arterial disease and heart attacks. This book summarizes recent advances in the field; it is the product of fruitful cooperation among international scientists who met in Japan in May, 1990 to discuss the regulation of coronary blood flow.

The objectives in this special issue are (1) to critically review current information on the mechanisms coupling extracellular regulatory signals to regulation of cross-bridge cycling and proliferation in smooth muscle, and (2) identify significant gaps or unresolved issues that are important topics for future research. The experimental and analytical difficulties discussed above are increasingly recognized and surmounted. Elucidation of the molecular and cellular events underlying the biologal properties of smooth muscle is in the midst of a period of rapid progress. While the reviews reveal many gaps to be filled and illustrate areas of contention, they also capture the excitement of new discoveries.

Resistance arteries have been recognized for some time as key factors in the regulation of vascular flow resistance, where they determine the regional and local distribution of blood and arterial pressure. Chapters provide an overview of the physiological, biochemical, and electrophysiological characteristics of these vessels, as well as a critical evaluation of the methodologies for studying small arteries and an examination of the membrane and neural mechanisms involved in the control of vascular tone.

The book combines general concepts and methods to investigate calcium signalling in cells ranging from molecular biology approaches to manipulation of calcium in living cells. The focus within these methods in on the broad range of fluorescence imaging technology, in particular on optical sectioning techniques and fast image acquisition. In addition to these general guidelines there are application examples in a context beyond calcium signalling in two major fields: investigations of isolated cardiac myocytes and red blood cell related research. While the cellular cardiology section provides snapshots of certain calcium signalling aspects, the red blood cell part presents an overview from the functional identification of calcium-channels to a concept of physiological and pathophysiological relevance.

Bone is a sex hormone-dependent organ, which has important implications for current therapeutic strategies in the prevention and possible treatment of bone loss in the elderly. This book contains the proceedings of the Schering Workshop on "Sex Steroids and Bone." The various chapters by leading experts give and overall view of current knowledge on the control of bone hemostasis by sex steroids and new ideas on how this controlcould be exerted. It was also of particular concern to integrate a general survey of in vivo experimentation and histomorphometric evaluation of bone tissue. The organizers of the workshop and editors of this volume hope that it will contribute to a better understanding of the role of sex steroids in bone and thus pave the way for a better experimental approach instudying drug effects on bone with the ultimate goal of improving therapy in bone diseases.

The papers ofthis Special Issue of Molecular and Cellular Nutraceuticals are also known as 'health foods' but they also Biochemistry are invited, peer-reviewed submissions from include any product derived from animal or plant sources speakers who attended the 2nd World Conference of the which ultimately can provide a medical benefit. F our symposia International Society for MolecuIar Nutrition & Therapy. This on various aspects concerning nutraceutical products and the Conference was held fromAugust 2--4th, 1997 in Winnipeg, marketing ofthese compounds was addressed. It is hoped that Canada. The goal of the Conference was to advance our these interactions represent the start of a new, highly interesting knowledge concerning the molecular events which link scientific venture uniting the nutraceutical industry with the nutrition to various disease processes in the body. Attending main stream academic medical research community. The the meeting were scientists from 18 countries ineluding nutraceutical industry is expected to be a multibillion dollar Canada, the United States, Japan, the United Kingdom, a year business world-wide by the turn of the century. The Franee, the Czeeh Republie, Germany, the Netherlands, identification of the active ingredients in any number of Bahrain, Hong Kong, India, Israel, China, Kuwait,Argentina, health foods reputed to be medieally beneficial would be just Sweden, Australia and Mexieo. There were 56 Distinguished one of the goals of such a scientific interaction.

This book covers all aspects of oxygen delivery to tissue, including blood flow and its regulation as well as oxygen metabolism. Special attention will be paid to methods of oxygen measurement in living tissue and application of these technologies to understanding physiological and biochemical basis for pathology related to tissue oxygenation. This book is multidisciplinary and designed to bring together experts and students from a range of research fields including biochemical engineering, physiology, microcirculation, and hematology.

The human pancreas consists of two organs in one: the exocrine gland made up of pancreatic acinar cells and duct cells that produce digestive enzymes and sodium bicarbonate, respectively; the endocrine gland made up of four islet cells, namely alpha-, beta-, delta- and PP-cells that produce glucagon, insulin, somatostatin and pancreatic polypeptide, respectively. While the physiological role of exocrine p- creas is to secrete digestive enzyme responsible for our normal digestion, absorption and assimilation of nutrients, the endocrine pancreas is to secrete islet peptide h- mones maintaining our glucose homeostasis. The pancreatic functions are nely regulated by neurocrine, endocrine, paracrine and/or intracrine mechanisms. Thus, dysregulation of these pathways should have signi cant impacts on our health and disease. Nevertheless, the underlying mechanisms by which pancreatic functions are regulated remain poorly understood. Recent basic science and clinical studies con rm myriad physiological and pathophysiological roles of the tissue renin-angiotensin systems (RAS). Of parti- lar interest is the recent identi cation of a local and functional RAS in the pancreas, which in uences both its exocrine and endocrine function. Its role in the pat- genesis of pancreatic diseases including diabetes and pancreatitis is increasingly recognized, as is the therapeutic potential of RAS antagonism: RAS blockade l- its disease progression of type 2 diabetes mellitus and impaired glucose tolerance, and may also protect against pancreatic in ammation.

This volume explores all aspects of vascular biochemistry and includes chapters that provide an understanding of vascular function with descriptions of tissue components present in the vascular wall as well as an exploration of the hemodynamic and metabolic activities associated with this function. In addition, some chapters explore the vasculature under conditions which mimic various disease states. The information provided in this volume will provide new insights into the mechanisms that control vascular function as well as therapies designed to treat vascular disease.

Since 1959, the International Society of Arterial Chemoreception (ISAC) has organized in a variety of countries fifteen scientific meetings devoted to the mechanisms of peripheral arterial chemoreception and chemoreceptor reflexes. After the meeting held in Philadelphia with Sukhamay Lahiri as president, ISAC membership elected Lyon (CNRS, University Claude Bernard, France) as the site of the xv" ISAC Symposium. The Symposium was effectively held in Lyon from the 18th to the 22nd of November 2002 and Jean-Marc Pequignot was its president. The organizers were Jean-Marc Pequignot and Yvette Dalmaz Lyon (CNRS, University Claude Bernard, France) and the Scientific Committee was formed by John Carroll (University of Arkansas for Medical Sciences, USA), Constancio Gonzalez (University of Valladolid, Spain), Prem Kumar (University of Birmingham, U. K. ), Sukhamay Lahiri (University of Pennsylvania, Philadelphia, USA), Colin Nurse (McMaster University, Hamilton, Ontario, Canada), and Nanduri Prabhakar (Case Western University, Cleveland, Ohio, USA). The Symposium in Lyon intended to follow the path opened in Philadelphia gathering people working at the interface of cellular and molecular biology with researchers in the more classical topics of chemoreception pathways and reflexes. The aim was to join experts with different perspectives. Along these lines, some participants are engaged in the exploration of the intimate mechanisms of oxygen sensing and cellular responses, with their work centered in a great number of preparations covering a broad spectrum from bacteria, to chemoreceptor cells or to central nervous systems neurons.

From humble beginnings over 25 years ago as a lipid kinase activity associated with certain oncoproteins, PI3K (phosphoinositide 3-kinase) has been catapulted to the forefront of drug development in cancer, immunity and thrombosis, with the first clinical trials of PI3K pathway inhibitors now in progress. Here we give a brief overview of some key discoveries in the PI3K area and their impact, and include thoughts on the current state of the field, and where it could go from here

Parameters such as membrane transport, metabolism and protein incorporation govern the fate of amino acids in living tissue. Is it possible to use positron tomography to measure some of them, and what is their meaning in normal and pathological situations? These questions have been addressed for a long time and no satisfactory answer has yet been given. This book, which derives from an EEC workshop organized in the frame of the Concerted Action on PET Investigation of Cellular Regeneration and Degeneration', held in Lyon in February 1992, gives the present state of knowledge in this field based on the most recent studies. Contributions from 24 leading European and American scientists are presented and discussed in the following four parts: biochemistry and animal studies; amino acids labelling with positron emittors, quality control and metabolites measurement; kinetic modelling of amino acids transport, metabolism, and protein incorporation; clinical use of amino acids. This book will aid and interest biochemists, radiochemists, pharmacologists, neurologists, oncologists and medical imaging scientists.

Diabetes is an autoimmune, inflammatory disease affecting many different organ systems and exhibiting both primary and secondary defects. Because diabetes affects a wide range of cellular systems, a multidisciplinary effort has been mounted over the past several decades using a wide range of investigative techniques and methodologies in order to identify molecular mechanisms responsible for cellular dysfunction. Because primary defects at various levels of sub-cellular signaling, intracellular calcium handling, protein expression and energy regulation are often a primary consequence of diabetes. This volume is a compilation of new multidisciplinary research that will broaden our current understanding of diabetes and cardiovascular disease as well as provide the basis for the development of novel therapeutic interventions.

This, the 148th volume of the Handbook of Experimental Pharmacology series, focuses on the very core of pharmacology, namely receptor theory. It is fitting that the originator of receptor pharmacology, A. J. CLARK, authored the fourth volume of this series 63 years ago. In that volume CLARK further developed his version of receptor theory first described four years earlier in his classic book The Mode of Action of Drugs. An examination of the topics covered in volume 4 reveals a striking similarity to the topics covered in this present volume; pharmacologists today are still as interested in unlocking the secrets of dose-response relationships to reveal the biological and che mical basis of drug action as they were over half a century ago. Sections in that 1937 volume such as "Curves relating exposure to drugs with biological effects" and "Implications of monomolecular theory" show Clark's keen insight into the essential questions that required answers to move pharma cology forward. With the advent of molecular biological cloning of human receptors has come a transformation of receptor pharmacology. Thus the expression of human receptors into surrogate host cells helped unlock secrets of receptor mechanisms and stimulus-transduction pathways. To a large extent, this elim inates the leap of faith required to apply receptor activity of drugs tested on animal receptor systems to the human therapeutic arena. However, a new leap of faith concerning the veracity of the effects found in recombinant systems with respect to natural ones is now required.

The study of plant development using molecular and genetic techniques is rapidly becoming one of the most active areas of research on flowering plants. Developmental Biology of Flowering Plants relates classical developmental work with the outstanding problems of the future in the study of plant development. An important feature of this book is the integration of results from molecular and genetic studies on various aspects of plant development in a cellular and physiological context.

The volume Appetite Control provides a comprehensive description of the mechanisms controlling food intake, and thereby energy balance, in the mammalian organism. During the last decade, research in this area has produced a remarkable wealth of information and has characterized the function of numerous peptides, transmitters, and receptors in appetite control. Dysfunction of these circuits leads to obesity, a growing health concern. However, the plethora of mechanistic information is in marked contrasts to an almost complete lack of anti-obesity drugs that meet the safety standards required for the chronic therapy of morbid obesity. Consequently, ongoing research aims to identify additional targets and agents for a pharmacological intervention. Thus, the mechanisms of appetite control as well as all agents interfering with its control are of considerable practical interest.

The authors of the volume are distinguished scientists who are leading experts in the field, and who have contributed important, original data to our understanding of the mechanisms of appetite control. They have quite different scientific backgrounds and, together, they represent all relevant disciplines. Thereby, the topics are presented from different points of view, not exclusively from that of pharmacology and neuroendocrinology. Thus, the volume addresses all scientists who are interested in the field of obesity research and the pathophysiology of appetite control."

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.

Apoptosis is a fascinating concept for the basic scientist. This is not only because of the multifaceted variety of proposed and discovered mechanisms, but because apoptosis represents a fundamental pathway for cell renewal. The study of apoptosis has resulted in an array of discoveries on signal transduc tion and downstream effects that have facilitated and advanced many fields in biology, including research on cancer and other diseases. Thus, the apoptotic process can be viewed as the largest effort of the scientific community to understand how cells work and tissues assemble or remodel. The most direct consequence of this accumulated knowledge is a greater understanding of disease and pathological mechanisms. The end result of these efforts will be significant contributions to health and the adoption of new, never anticipated, therapeutic approaches. This book represents the summation of considerable effort from a significant group of contributors from all over the world as well as from its editors. In this fashion, many viewpoints have been collected and SUbjected to thorough academic discussion. The concepts contained in this medically important volume will stimulate and renew the ideas of scientists and indeed, will generate additional work to advance biological knowledge even further. The emphasis of this volume cements what has been established, adds what has not been explored fully, and creates a fertile ground for further hypotheses that will lead to a more complete understanding of the apoptotic process."

A review of the current state of the clinical and basic science of the pharmacology of sleep. The information provided ranges from a historical perspective to current concepts of sleep mechanisms, including the interaction between pharmacology and sleep-wake regulation and between chronopahrmacology and sleep-wake rhythms. For the first time in one single volume, both the basic mechanisms of sleep, and the basic and clinical aspects of the pharmacology of sleep are dealt with in a thorough, comprehensive and authoritative manner. The chapters, written by the foremost scientific authorities in this field, integrate the latest information on the basic mechanisms of drugs as they relate to their effects on sleep.

Adopting an interdisciplinary approach to the chemistry and physics of materials, their biocompatibility, and the consequences of implantation of such devices into the human body, this text introduces readers to the principles of polymer science and the study of metals, ceramics and composites, and also to the basic biology required to understand the nature of the host-transplant interface. Topics covered include the macromolecular components of cells and tissues, self-assembly processes, biological cascade systems, microscopic structure of cells and tissues, immunology, transplantation biology, and the pathobiology of wound healing. The materials science section includes the structures and properties of polymers, metals, ceramics and composites, and the processes for forming materials as well as the pathobiology of devices. The final two chapters deal with tissue engineering and the relations between the biology of cells and tissue transplantation, and the engineering of tissue replacements using passaged cells.

A presentation of the most elementary form of pulsatile flow as an important prerequisite for the study of other flow applications in biological systems. The book provides in a single source a complete treatment of the fluid dynamics of flow with the required mathematics and emphasis on the basis mechanics. The style and level of this book make it accessible to students and researchers in biophysics, biology, medicine, bioengineering and applied mathematics working in theoretical and clinical work on the cardiovascular system, as well as in the design of new instrumentation, medical imaging systems, and artificial organs. With problems and exercises.

TheobservationthatabloodclotspontaneouslydissolveswasfirstdescribedbyDenys in1889. Subsequently,thebloodclottingsystemwasshowntobeinvolvedintumor growth. Forexample,asearlyas1925,Fisherreportedthataviantissueexplantstrans- formedtomalignancybyvirusesgeneratedhighlevelsoffibrinolyticactivityundercon- ditionsinwhichculturesofnormalcellsdidnot. In1958,theconceptthatan equilibriumexistedbetweenthetendencyofbloodtoclotandtoremainfluidwaspro- posedbyAstrup. Atthattime,itwasbelievedthatthishemostaticbalancewasexplained bytheabilityofpolymerizingfibrintoorchestrateitsownclearancebystimulatingfib- rinolyticactivity. Sincethesepioneeringstudies,considerableinformationhasaccumu- latedthathasdefinedthecomponentsofthecoagulationandfibrinolyticsystemsand howtheyareinvolvedinphysiologicalandpathophysiologicalprocesses. Plasminogen: Structure, activation, and regulationfocusesonthebasicprinciplesandrecentdevelop- mentsintheplasminogen/plasminresearchfieldandhowtheseresultsprovideacon- ceptualframeworkforanunderstandingofthephysiologicalroleofplasminogenin healthanddisease. Theenzymaticcascadetriggeredbyactivationofplasminogenhasbeenimplicated inavarietyofnormalandpathologicaleventssuchasfibrinolysis,woundhealing,tis- sueremodeling,embryogenesis,angiogenesis,andtheinvasionandmetastasisoftumor cells. Thisimpressivelistofphysiologicalfunctionsforplasminogenreinforcesthewide diversityofrolesthatplasminogenplaysinvariousphysiologicalprocesses. Productive plasmingenerationrequirestheassemblyofbothplasminogenactivatorsandplasmino- genonasolidsupportsuchasthefibrinpolymerorthecellsurface. Theregulationof plasminproductioninvolvesacomplexinterplaybetweentheseplasminogenactivators, plasminogenactivatorinhibitors,andplasmininhibitors. Clearly,theexplosivegrowth inthisresearchfieldandthemanyexcitingdiscoveriessuggeststhattheresearchefforts inthenextdecadewillrevealthemechanismsbywhichthecomponentsoftheplas- minogensysteminteractandregulatebothplasminactivationandfunctionatacellular level. Plasminogen: Structure, activation, and regulationisdividedintotwosections. Thefirstsectiondealswiththestructureandregulationofplasminogen. Thechapters inthissectionrangefromdiscussionsofthestructureofplasminogenandtheregulation oftheplasminogengenetodiscussionsofthestructureandregulationofplasminogen activatorsandplasminogenactivatorinhibitors. Alsoexaminedistherelativelynewdata concerningthegenerationofanti-angiogenicmoleculesfromplasminogen. Thesecond sectiondealswiththephysiologicalandpathophysiologicalrolesofplasminogenaswell astheconsequencesofplasminogengeneknockout. Discussionsinthissectioninclude examinationoftheroleofplasminogeninhematopoieticmalignancies,tumorcell progression,angiogenesis,mammaryglandinvolution,woundhealing,andbone readsorption. xi xii Preface Inclosing,Iwouldliketothankmyadministrativeassistant,Ms. ViSommerfeld,for herinvaluableassistanceandtimelesseffortswiththeorganizationandeditingofthebook. Lastly,Iwouldliketoacknowledgetheeffortsoftheauthorsoftheindividualchapters, whoareauthorities inthisfield,foragreeingtotaketimefrombusyschedulestoprovide thesechaptersinatimelyfashion. DavidMortonWaisman Contents Part I. Plasminogen: Structure and Regulation 1. Human Plasminogen: Structure, Activation, and Function FrancisJ. Castellino and Victoria A. Ploplis 1. Introduction 3 2. StructureofHumanPlasminogen...3 2. 1. PrimaryProteinStructure...3 2. 2. GeneOrganization 5 3. ActivationofHumanPlasminogen...6 3. 1. ActivationbyPhysiologicalActivators 7 3. 1. 1. Urokinase-typePlasminogenActivator...7 3. 1. 2. Tissue-typePlasminogenActivator...8 3. 2. ActivationbyBacterial-derivedPlasminogenActivators...9 3. 2. 1. Streptokinase 9 3. 2. 2. Staphylokinase...9 4. TargetsforPlasminActivity...9 5. DysplasminogenemiasandPhenotypicManifestations 10 6. Conclusions 11 References...11 2. Plasminogen Activators: Structure and Function Vincent Ellis 1. Introduction ...19 2. SerineProteases...20 3. UrokinasePlasminogenActivator,uPA...21 3. 1. SerineProteaseDomain 22 3. 2. N-terminalDomains...24 3. 2. 1. KRModule 24 3. 2. 2. EGModule 24 4. MechanismsRegulatinguPAFunction...25 4. 1. ZymogenActivation...25 4. 2. ZymogenActivity...26 4. 3. ReciprocalZymogenActivation 27 4. 4. uPARStimulationofPlasminogenActivation...27 4. 4. 1. uPAandtheTemplateMechanism 28 4. 4. 2. PlasminogenandtheTemplateMechanism 29 4. 5. AvianuPA,aSpecialCase? 30 xiii xiv Contents 5. TissuePlasminogenActivator,tPA...30 5. 1. SerineProteaseDomain 31 5. 2. N-terminalDomains ,...33 5. 2. 1. KRModules ,. . ,. . ,...33 5. 2. 2. F1-EGSupermodule 33 6.

RNA technologies are the driving forces of modern medicine and biotechnology. They combine the fields of biochemistry, chemistry, molecular biology, cell biology, physics, nanotechnology and bioinformatics. The combination of these topics is set to revolutionize the medicine of tomorrow. After more than 15 years of extensive research in the field of RNA technologies, the first therapeutics are ready to reach the first patients. Thus we are witnessing the birth of a very exciting time in the development of molecular medicine, which will be based on the methods of RNA technologies. This volume is the first of a series. It covers various aspects of RNA interference and microRNAs, although antisense RNA applications, hammerhead ribozyme structure and function as well as non-coding RNAs are also discussed. The authors are internationally highly respected experts in the field of RNA technologies.