Molecular and Cell Biology Updates – serie

The basic knowledge of cell biology and molecular genetics in oncology is increasingly attracting the interest of clinical oncologists and is close to reaching a helpful application at the bedside. At present, it seems clear that the solution of the cancer problem lies within the comprehension of the intimate mechanisms leading to cell transformation and tumor progression as weIl as of the cancer-host relationship. According to this rationale every achievement in this context could drastically improve both diagnosis and therapy of neoplastic diseases. This={)ook represents the proceedings of the International Conference o~ Cancer: Biological Mechanisms and Clinical Applications, held in Rome on November 16-18, 1992. The meeting was organized by the Centro di Ricerche Oncologiche "Giovanni XXIII" of the Catholic University of Rome and the Consorzio Mario Negri Sud of S. M. Imbaro in collaboration with other colleagues from the Universities of Napoli and Bologna. As organizers and pmticipants to the Conference we think the meeting was a success, as was confirmed by the great interest raised in scientific and academic circles. The book collects contributions from leading scientists in all oncological m·eas ranging from molecular biology to immunology, diagnosis and therapy. The papers are organized into four sections: I - Molecular aspects of cell transformation and growth; II - Membrane receptors and signal transduction; III - Models for new therapeutical strategies; IV - Clinical Applications, following the strategy that chm·acterized the entire meeting: from molecule to bedside.

Adenine Nucleotides play a major role in cellular metabolism and functions, serving as high-potential phosphate transfer compounds in energy metabolism and as substrates and co-factors for proteins involved in signal transduction. During the last few years definite advancement has been made in elucidating the molecular and genetic aspects of the enzyme involved in oxidative phosphorylation, the so-called FoFl H~-ATP synthase. Non-invasive NMR technologies have been developed to monitor in vivo the energy level of tissues based on determination of the concentrations of adenine nucleotides, phosphate and phosphate esters. Thus it became clear that the capacity of oxidative phosphorylation adapts itself to the ATP demand which changes continuously with the physiological state in various tissues. This is achieved by regulation of the enzyme activity as well as by regulation of its biogenesis. The reversible phosphorylation of proteins is re- cognised as a major regulatory mechanism in eukaryotic as well as in prokaryotic cells for cellular signal transduction and control of gene expression, cell growth, differentation and oncogenesis. The same applies to the role played by cAMP.A further topic of growing interest concerns the discovery of the ATP binding cassette (ABC) superfamily of transport proteins which includes systems of primary importance in medicine such as the multi-drug resistance P glycoprotein, the cystic fibrosis transmembrane conductance regulator (CFTR) and the 70 kd peroxisomal membrane protein. Finally, much attention is being devoted in many laboratories to the molecular structure and role of ATP- modulated channels.

Oxidative stress and aging Over the past several years there has been an extraordinarily rapid growth in our knowledge of free radical chemistry and its possible involvement in both normal essential biology and age­ related disease and dysfunction. Much of this growth in the traditionally separate sciences of chemistry and molecular gerontology occurred independently, with little interaction or communi­ cation between the scientists working in these two fields. In view of the growing maturity of the two fields and the potential importance of advancing our knowledge in the area of oxidative stress and aging, we perceived a critical need to organize an international conference the "First Inter­ national Conference on Oxidative Stress and Aging" in Hawaii in 1994 to bring together the world's leading scientists in the fields of reactive oxygen species and molecular gerontology. The objective of this conference was to provide a unique opportunity for scholars working in these two related and rapidly growing fields to participate in the exchange, integration, and synthesis of new concepts and ideas, to engage in constructive criticism and to initiate new collaborative research projects. The conference focused on the molecular and cellular aspects of aging as related to oxidative stress. It was one of the largest and most comprehensive international conferences held in molecular gerontology. At this conference a call was made for submission of papers to be used in the publication of a book covering the major contributions of the meeting.

Since the appearance of photosynthesis on our planet, all living organisms have been facing a new abundant, extremely reactive element, oxygen. This element is used for the synthesis of highly energetic compounds, but can also generate molecules capable of damaging constituents of living structures, including proteins, nucleic acids and lipids. It is therefore no surprise that all biological organisms have evolved multiple and sophisticated ways to reduce the detrimental effects of oxygen. For cells and tissues of highly organized animals, particularly cells of the immune system, superoxidation products play an important role, via oxidative stress, in activation, inflammation and viral and bacterial infections. We must be grateful to Drs. C. Pasquier, C. Auclair, L. Packer and R. Olivier for having brought together many specialists in an international meeting held in Paris in March 1993, at the Ministry of Research. This book comprises an impressive amount of recent knowledge, a real mine for the reader in this fast developing field of research. Furthermore, we should not forget that this basic research may lead in the future to new therapeutic approaches to the most important pathologies of the latter part of this century, including AIDS.

Antioxidants inhibit the formation and spread of free radicals which can be damaging in biological systems. Free radicals form in biological systems through metabolism, but it is also realized that exogenous environmental sources, such as radiation, food, and drugs, contribute significantly to the generation of free radicals in biological systems. Being reactive species, free radicals are short-lived and do not travel far from cellular targets. Their concentration in biological systems is very low and is difficult to detect directly by electron spin resonance spectroscopy (ESR). Indirect methods of reactions of radicals with specific biomolecules are also sufficiently sensitive to detect quantitatively their presence. Thus the response of antioxidant defenses which react with radical species, can serve as an indirect measure that free radicals have been formed. Redox-based antioxidants change their oxidation state and antioxidants become free radicals themselves. Often, however, the antioxidants give rise to more persistent free radicals, sometimes owing to delocalization of the lone electron around ring structures (in vitamin E, ubiquinones, and certain carotenes). Persistent free radicals react only rarely and the precursors often can be regenerated in biological systems. In recent years, it is becoming clearer from biochemical studies on how the major lipophilic antioxidants work. Particular attention has been given to vitamin E and quinones found in animal and plant membranes and in carotenoids, for the protection of membranes in lipoprotein systems. Flavonoids form another rich and varied source of natural antioxidants.

Rapid progress has been made in our understanding of the molecular mechanisms of cell growth and oncogenesis during the past decade. Special attention has been given to the presentation of the frequently neglected close correlation between changes in signal transduction and metabolic pathways during oncogenesis. This book advances the knowledge of mechanisms regulating metabolism and functioning of vitamin A and offers the most recent results of research on the clinical efficiency of retinoids in skin disorders and cancer. The book presents recent findings on the regulation of cell growth in normal and neoplastic tissues by growth factors including hormones, and by the activation and inactivation of oncogenes and tumor suppressor genes, respectively. It also offers a survey of the molecular and cell biochemistry of retinoids. Basic researchers in biochemistry, pharmacology and cell biology as well as clinicians will find this book very informative and up-to-date. This book advances the knowledge of mechanisms regulating metabolism and functioning of vitamin A and offers the most recent results of research on clinical efficiency of retinoids in skin disorders and cancer. Basic researchers in biochemistry, pharmacology, cell biology, and clinicians will find this book very informative and up-to-date. The chapters, organized in six sections, are contributed by leading scientists who have been working in the retinoid field for decades. Their experience and competence is aknowledged worldwide.

This book provides an account of the recent advances in our understanding of the role of proteases under physiological and pathological conditions. It reviews the contributions that have been made in the fields of biochemistry, molecular biology, medicine, agriculture and ecology. In addition there is an account of the growing number of practical applications in biotechnology, reflecting the fact that peptidases are major targets for medical and agricultural purposes. The book shows how the numerous protease structures are essential for drug design. The contributions place special emphasis on cysteine, aspartic and metalloproteases and their role in physiological and certain pathological states. Another focus is the classification and nomenclature of peptidases and a review of those proteases currently under the most intensive investigation. The book provides an informative introduction for teachers and newcomers to the field, such as graduate students, while providing a valuable source of material and ideas for the academic and industrial researcher in areas including biochemistry, medicine, agriculture and biotechnology.

In 1898 Camillo Golgi reported his newly observed intracellular structure, the apparato reticolare interno, now universally known as the Golgi Apparatus. The method he used was an ingenious histological technique (La reazione nera) which brought him fame for the discovery of neuronal networks and culminated in the award of the Nobel Prize for Physiology and Medicine in 1906. This technique, however, was not easily reproducible and led to a long-lasting controversy about the reality of the Golgi apparatus. Its identification as a ubiquitous organelle by electron microscopy turned out to be the breakthrough and incited an enormous wave of interest in this organelle at the end of the sixties. In recent years immunochemical techniques and molecular cloning approaches opened up new avenues and led to an ongoing resurgence of interest. The role of the Golgi apparatus in modifying, broadening and refining the structural information conferred by transcription/translation is now generally accepted but still incompletely understood. During the coming years, this topic certainly will remain center stage in the field of cell biology. The centennial of the discovery of this fascinating organelle prompted us to edit a new comprehensive book on the Golgi apparatus whose complexity necessitated the contributions of leading specialists in this field. This book is aimed at a broad readership of glycobiologists as well as cell and molecular biologists and may also be interesting for advanced students of biology and life sciences.

On the basis of the agreement signed between UNESCO and the Government of the Republic of Poland the International Institute for Cell and Molecular Biology of UNESCO was officially inaugurated in October 1995 in Warsaw, Poland, as part of the activity of the Global Network for Molecular and Cell Biology (MCBN) of UNESCO. The occasion was marked by the bringing together in Warsaw of a broad spectrum of cell and molecular biologists from around the world under the auspices of the Global MCBN UNESCO. At the conclusion of that week-long celebration it became clear that Polish cell and molecular biology had come of age in terms of its depth, vigor and impact on the global scene. At the suggestion of Professor Angelo Azzi, chairman of Global MCBN UNESCO, we considered the challenge of compiling a volume in the Molecular and Cell Biology Updates tMCBU) Series that would address the molecular basis of cancer and its therapy, but one that would additionally serve to highlight Polish contributions to this field of research. We accepted the challenge presented to us by Professor Azzi and are grateful to all contributors of the present volume for making this a pleasant and stimulating project. We requested each contributor to present his personal perspective of respective topics. As a consequence, we hope that each contribution has a distinctive individual flavor which reflects the role played by individual research groups in advancing science.

Rapid progress has been made in our understanding of the molecular mechanisms of cell growth and oncogenesis during the past decade. This book comprises recent results on the regulation of cell growth in normal and neoplastic tissues by growth factors including hormones, and by the activation and inactivation of oncogenes and tumor suppressor genes, respectively. Special attention has been given to the presentation of the frequently neglected close correlation between changes in signal transduction and metabolism pathways during oncogenesis.

Skeletal muscle consumes significant amounts of oxygen, and its oxygen flux increases significantly under conditions of exercise and muscle contraction. This makes the muscle vulnerable to oxidative stress since concomitantly with the increase of oxygen flow there is an increase of free oxygen radicals which are a byproduct of muscle respiration. A number of studies in the last decade have documented the involvement of free oxygen radicals in exercising muscles. The consequences of muscle oxidative stress have resulted mainly in increased muscle protein oxidation, elevation of lipid peroxidation, and depletion of muscle antioxidants. The mechanisms of this oxidative stress are under extensive investigation in laboratories around the world and are topics of the chapters in this volume. This book is intended for professionals who are interested in muscle function, physiology, pathophysiology and well-being, such as therapists, trainers and medical professionals as well as for researchers in the field of muscle physiology.

Found in all organisms, the alpha-keto acid dehydrogenase complexes have central roles in cellular metabolism and are major sites of regulation. The understanding of the organization, function and regulation of these quintessential multienzyme complexes has been greatly advanced by studies employing molecular biology and biophysical techniques. Although these enzyme systems have some features in common, their diversity in fulfilling unique organism - or tissue - specific roles is truly amazing. These systems have medical importance in areas ranging from defects in regulation (linked to diabetes, heart disease, obesity, nutrition defects), to inherited diseases (inborn errors, maple syrup urine disease) to acquired immune diseases (primary biliary cirrhosis). This book brings together wide-ranging recent findings on the structure(function relationships, gene regulation, and genetic defects of the alpha-keto acid dehydrogenase complexes, namely the pyruvate dehydrogenase, alpha-ketoglutarate dehydrogenase and the branched-chain alpha-keto acid dehydrogenase complexes. A wide variety of experimental approaches together with new results presented in this book should serve as a resource for beginning to established investigators in the field as well as scientists who are interested in mitochondria, dehydrogenases, kinases, phosphatases, lipoic acid, thiamine pyrophosphate, and enzyme complexes.

This book consists of a series of reviews on selected topics within the rapidly and vastly expanding field of membrane biology. Its aim is to highlight the most significant and important advances that have been made in recent years in understanding the structure, dynamics, and functions of cell membranes. The progress that is being made in research in this field is due to the application of integrated experimental ap- proaches, utilizing sophisticated and novel techniques in molecular biology, cell biology, biophysics and biochemistry. Due to the advances made, many problems have been or are being solved at the molecular level. With the help of an Editorial Advisory Board consisting of Jean-Pierre Changeux (Paris), Paolo Comoglio (Torino), Rainer Jaenicke (Regens- burg), Sten Orrenius (Stockholm), Lorenzo Pinna (Padova), Konrad Sandhoff (Bonn), and Gottfried Schatz (Basel), we have selected a number of topics in areas in which progress has been particularly rapid, and have invited internationally acknowledged experts in the field to review these topics.The areas covered in this monograph are: 1) Signal transduction; 2) Membrane traffic: Proteins and lipids; 3) Bioenergetics: Energy transfer and membrane transport; 4) Cellular ion homeostasis; 5) Growth factors and adhesion molecules; 6) Structural analysis of mem- brane proteins; and 7) Membranes and disease.

This book is based on two keywords: Bioradical and ESR. Bioradical is a newly coined word which encompasses paramagnetic species in biological systems, such as active oxygen radicals and transition metal ions. Research on the structure and function of bioradicals has been attrac­ ting growing attention in the field of biological science, and comprehensive investigations from many fields are helping to understand the real features of these species. ESR spectroscopy also has interdisciplinary features in that its techniques have been applied to many fields, ranging from physics to medicine. It was our hope, therefore, that this book would help to clarify many aspects of bioradicals and that significant progress would be achieved in combining basic research from many different fields. This book arises from the First International Conference on Bioradicals Detected by ESR Spectroscopy (ICBES), which was held in Yamagata, a city in the Yamagata Prefecture of Japan, in 1994. About 300 participants from 16 different countries attended this conference, and about 170 papers were presented. This book is a collection of contributions from the conference and also contains eleven chapters selected by the editorial board, based on sugges­ tions from the members of the international editorial board of ICBES. The Yamagata Technopolis Foundation is developing a biomedical technology for the 21st century based on life science fused with material and physical science. Based on such a techno­ logy, the Foundation plans to share its fruits all over the world.

Free radical-mediated reactions have been well known in chemistry and physical chemistry for many years. Applying this knowledge to living organisms, biochemists have shown that reactive free radicals are formed at many intracellular sites during normal metabolism, and they have started to suggest possible roles in various pathological processes and conditions, for example in radiation damage, in the metabolism of xenobiotics, in carcinogenesis and in metabolic disorders. At present, a large and relevant mass of experimental evidence supports the view that reactive free radicals are involved in the pathogenesis of several diseases and syndromes. This literature has captured the attention and interest of people involved in the biomedical field. Exciting developments in radical research are probable in the near future, establishing a greater interaction between basic science research and medicine. While the task of defining the involvement of free radicals in human pathology is difficult, it is nonetheless extremely important that such interaction be fulfilled as soon as possible. These were the considerations motivating us during the organization of the VI Biennial Meeting of the International Society for Free Radical Research held in Torino, Italy, in June 1992, and also during the preparation of this book. Experts in the various aspects of free radical research were invited to participate in the Torino Meeting and to contribute chapters for this volume.

Increasing interest has been emerging in the last decade in the field of signal recognition and transduction. This is particularly true for animal systems where an impressive amount of literature is appearing and where many important pathways have been clarified at a molecular level. In the elucidation of the functions of single components of a given pathway, gene cloning has played a major role and opened the field to the genetic engineering of these complex systems. At variance with this situation, plant systems are less well elucidated, even if in recent years exciting research of developments have been initiated especially with the view toward the most promising role plants in biotechnology. Recent studies have elucidated some of the events involved in the perception of the plant hormone signals and some steps concerning its transduction. Only for three of the five hormones in plants, namely auxin, ethylene and cytokinins, have specific receptors been isolated. The use of classical molecular approaches, together with the more recently isolated mutants, have produced crucial information on receptors and shed light on possible transduction pathways. As in the case of red light, more than one pathway can be triggered by one specific signal. Many systems involved in animal signaling are now shown to be present also in plants, and in view of the fast progress in this area, it will be possible in the near future to fully describe the content of the "black boxes" in the reaction chain specifically triggered by a signal.

Increasing interest has been emerging in the last decade in the field of signal recognition and transduction. This is particularly true for animal systems where an impressive amount of literature is appearing and where many important pathways have been clarified at a molecular level. In the elucidation of the functions of single components of a given pathway, gene cloning has played a major role and opened the field to the genetic engineering of these complex systems. At variance with this situation, plant systems are less well elucidated, even if in recent years exciting research of developments have been initiated especially with the view toward the most promising role plants in biotechnology. Recent studies have elucidated some of the events involved in the perception of the plant hormone signals and some steps concerning its transduction. Only for three of the five hormones in plants, namely auxin, ethylene and cytokinins, have specific receptors been isolated. The use of classical molecular approaches, together with the more recently isolated mutants, have produced crucial information on receptors and shed light on possible transduction pathways. As in the case of red light, more than one pathway can be triggered by one specific signal. Many systems involved in animal signaling are now shown to be present also in plants, and in view of the fast progress in this area, it will be possible in the near future to fully describe the content of the "black boxes" in the reaction chain specifically triggered by a signal.

Highlights recent progress in the molecular, cellular and immunological mechanisms that contribute to the pathophysiology of cancer and the design of therapeutic modalities based upon these molecular insights. The work combines a research-oriented approach together with a review style approach.

Rapid progress has been made in our understanding of the molecular mechanisms of cell growth and oncogenesis during the past decade. This book comprises recent results on the regulation of cell growth in normal and neoplastic tissues by growth factors including hormones, and by the activation and inactivation of oncogenes and tumor suppressor genes, respectively. Special attention has been given to the presentation of the frequently neglected close correlation between changes in signal transduction and metabolism pathways during oncogenesis.

This volume provides an account of advances in the understanding of the role of proteases under physiological and pathological conditions. It reviews the contributions made in the fields of biochemistry, molecular biology, medicine, agriculture and ecology.

This is a compilation of the current research findings in the field of oxidative stress in skeletal muscle. It is designed for professionals who are interested in muscle function, physiology, pathophysiology and well-being, as well as researchers in the field of muscle physiology.