Histochemistry deals with the activities of chemical components in cells, and immunohistochemistry addresses the function of cell types in tissue or organs, such as those leading to acceptance or rejection of grafts or organs. This book is a methods volume focusing on antigen retrieval, particularly methods used in disease-related antigens. Because the book is a methods volume and a lab manual, it will have an audience of pathologists, biochemists, and lab technicians.

J. Oort and J. P. A. Baak This manual is written by pathologists and is meant for pathologists. It deals with a subject that became especially popular in pathology at the end of the last decade: morphometry. Morphometry is defined as the quantitative description of a structure (Weibel, 1969). This structure may be both of macroscopic and micro- scopic size. Strictly, the size of an organ is a morphometric feature. This type of morphometry has long been in use in pathology. Examples ofthis type in diagnostic pathology have been given by Roesle and Roulet (1932). In this manual, however, the term morphometry is used in its re- stricted microscopic meaning, i.e., the quantitative description of micro- scopic images and features. The quantitative analysis of microscopic pic- tures has for a long time been subordinated to the qualitative description. This may partly have been caused by the preference of the human mirld for pattern recognition rather than quantitative appraisal, which leads to judgements based on impressions subconsciously tested against the ob- server's (mental) image of the normal pattern. Another cause may have been the time-consuming difficulties of collecting and computing quanti- tative information. During the past decades the usefulness of quantitative methods in histo- and cytopathology has been stressed for many different subjects. In addition, minicomputers have become widely available and are becoming increasingly inexpensive; the more so since the introduction of micro com- puters.

We would like to take this opportunity of expressing our sincerest thanks to the many persons who have made adrenal tissue and related materials available to us for our work. Our especial gratitude is extended to Drs. J. J. Brown, A. Lever and J. I. S. Robertson of the M.R.C. Blood Pressure Unit, Glasgow, Dr. J. K. Grant, Royal Infirmary, Glasgow, Professor R. B. Welbourn and Dr. W. Kelly, Royal Postgraduate Medical School, Drs. D. B. Grant of Great Ormond Street, J. Ginsberg, Royal Free Hospital, D. C. Anderson, Hope Hospital, Salford, C. R. Edwards, St. Bartholomew's Hospital and Professor I. Doniach (for merly of the London Hospital) and Messrs. J.-c. Gazet, A. McKinna and P. Greening, Royal Marsden Hospital, London. The preparation and presentation of the material and the results would not have been possible without the help of Dr. P. Monaghan and his Electron Microscopy Unit, Ludwig Institute for Cancer Research (London Branch), Sutton, Mrs. Mitchell and her Histology Team, Royal Marsden Hospital, Sutton, Mr. K. Moreman of the Photographic Department of the Royal Marsden Hospital and Institute of Cancer Research, London and Mr. M. Hughes for graphics. Particular thanks are due for the untiring efforts and assistance ofMr. J. Ellis and Mrs. D. Corney of the Ludwig Institute for Cancer Research (London Branch), Sutton, for most of the photographic and secretarial work respectively. Professors G. Dhom and E. Mausle kindly provided material for Figs."

In commissioning this volume of Current Topics in Pathology, the editor of the series, Prof. Sir Colin Berry, asked me to produce something "which is aimed at the general pathologist but from the point of view of the neuropathologist who knows what is not being done well." This was easier said than done. Many aspects of neuropathology are encountered only infrequently in general pathology and are best dealt with by re ferral of material to a specialist unit. Others need more extensive investigation than is practicable in a general pathology setting. Yet neurological disorders in general are common and are encountered by general histopathologists, paediatric and forensic pathologists on a daily basis, particularly in autopsy practice. In inviting contributions to this volume of Current Topics in Pathology, I therefore biased the con tents towards disorders in which the conduct of the autopsy and the initial approach to neuropathological investigation are of critical importance in making the diagnosis. The authors all have a wealth of experience and expertise and I believe that this is reflected in their approach, which is both personal and practical. Indeed, the emphasis in this book is very much on the provision of practical advice. All of the chapters contain guidelines on the conduct of autopsies as well as infor mation on the macroscopic and basic histological assessment of the nervous system in the relevant disorders."

Medical imaging is an important topic and plays a key role in robust diagnosis and patient care. It has experienced an explosive growth over the last few years due to imaging modalities such as X-rays, computed tomography (CT), magnetic resonance (MR) imaging, and ultrasound. This book focuses primarily on model-based segmentation techniques, which are applied to cardiac, brain, breast and microscopic cancer cell imaging. It includes contributions from authors working in industry and academia, and presents new material.

The translation of Blood Smears Reinterpreted was begun when the French original was still undergoing revisions. I was accorded the oppor tunity to question any statement or turn of phrase that was unclear to me or appeared liable to misinterpretation. It is my hope that as a result, ambiguities-particularly those inherent in differences between American and Eu ropean usage-have been removed and that I have at least ap proached the ultimate goal of any translation: to reflect the author's intention accurately while remaining as readable as the original. Beyond the role of translator, I was encouraged to assume the role of critic. As a result, some pages or even single sentences were hotly debated, sometimes for hours, as Marcel Bessis insisted that any inter pretations on which we could not agree should be so indicated. In fact our discussion invariably ended in agreement, though they led to changes of a sentence here or a word there and, on occasion, to the addition of a footnote or a brief paragraph.

Sarcomas represent a group of rare tumors which arise from the largest tissue compartment of the body, but account only for about 1 % of all malignancies. Among this group of tumors, soft tissue sarcomas in adults are, in addition to osteosarcomas, the most important. In the diagnostic workup of soft tissue sarcomas modern radiological investigation methods such as CT and MRI techniques are being critically evaluated. While it is widely accepted that the conventional, stained microscopic specimen is still the cornerstone for properly planned treatment stra tegies, immunohistochemistry has come to play an indis pensable role in accurate classification. Reviewing the surgical techniques, the treatment of musculo-skeletal malignancies, especially of soft tissue sarcomas, has changed considerably over the past decades. While amputation used to be the surgical treatment of choice limb-saving procedures have now become an im portant therapeutic modality in treating such lesions, particularly in combination with percutaneous or interstitial radiotherapy. Despite adequate local treatment, many patients with soft tissue sarcomas will ultimately develop metastatic disease, usually in the lungs. For this reason, effective ad juvant systemic treatment should be given simultaneously with or soon after treatment of local disease. The results of several randomized trials evaluating adjuvant chemotherapy have been reported in the meanwhile, indicating im proved, disease-free survival with a trend towards improved overall survival. In disseminated disease, drug combi- VI Preface nations or single agents are used as palliative treatment and might help to improve survival in selected subgroups."

Pathology in general is closely linked with clinical medi- cardiographic changes may be desired and therefore a cine and in cardiovascular pathology this interdepend- short section has been included on cutting the heart ence is, perhaps, greater than in most other specialities. according to conventionally used echocardiographic In recent years great advances in investigatory proce- planes. dures have taken place, including the examination of The heart can only react to physiological changes or fresh endomyocardial tissue obtained by bioptome, per- damage in a limited way and therefore a separate chapter mitting not only clinico-pathological correlation but also on hypertrophy and dilatation and another chapter deal- examination at histochemical and ultrastructural levels. ing with changes in the endocardium, which reflect hae- Angiography has gained wider usage and two-dimen- modynamic alteration and may additionally show sional echocardiography is no longer the preserve of diagnostic features, are also included. Degeneration to- specialized units. Percutaneous coronary transluminal gether with changes in connective tissue and ischaemic angioplasty is a relatively new approach to relieving heart disease are separately presented. Atherogenesis, obstructive coronary artery disease. Cardiac pathology recognition of myocardial infarction and the earliest mor- has therefore moved apace, providing not only a back- phological changes discernible at histochemical and ground to clinical manifestations but also visual proof for ultrastructural levels of investigation, which are within other investigations such as immunology and virology. the scope of modern routine laboratories, are empha- Such investigations have played an essential role in sized.

New techniques make it possible for investigators to examine and sometimes quantify various aspects of nuclear morphology and function; now they can derive clinically and biologically useful information about the nucleus. This book draws together a series of techniques which have been successfully applied to the study of the nucleus of tumour cells. These are of fundamental importance to researchers in tumour histopathology and medical oncology. Detailed reviews are given of various aspects of the morphology, ploidy and karyotypic status and function of the nuclei in the cells of tumours and preneoplastic lesions. Topics covered include nuclear morphology in tumour diagnosis, the ultrastructure of the nucleus, karyotypic analyses of solid tumours, flow cytometric assessment of nuclear ploidy and other parameters, histomorphometry of the nucleus, and in situ hybridisation.

This book has been written from two points of view: firstly, from the viewpoint of those who are involved in the diagnosis and treatment of lymphoid malignancies, who must meet the challenge of integrating the new biological insights into their knowledge of these diseases; and secondly, from the viewpoint of those who are involved in basic biological approaches to malignancy and immunology, who wish to know more about the function of the lymphoid tissues and their malignant diseases. Neoplasia of lymphocytes is a focus for considering many of the most important biological advances impinging on cancer in the past two or three decades, because malignant lymphoproliferative diseases offer unequalled opportunities for studying many aspects of cancer. We probably know more about lymphocytes than other normal cells because of the ease with which they can be obtained. For the same reason we probably know more about malignant lymphocytes. One or other aspect of most of the momentous advances in biology of the past two or three decades has implications for lymphoid malignancies: hybridoma technology and the use of monoclonal antibodies, gene technology, the understanding of oncogenes and growth factors in the control of growth and differentiation, insights into causation of cancer by potent tumour promoters such as the phorbol esters and by viruses, and knowledge of the control of growth function of lymphocytes themselves. Conversely, many of the advances in understanding lym phocytic leukaemias and lymphomas have implications for other cancers."

There have been many advances in the field of gastrointestinal pa thology which are of considerable clinical significance during the 13 years since the last publication of a volume of Current Topics in Pathology devoted to this subject. Many have arisen from the app lication of new techniques of histochemistry, immunocytochemi stry, quantitative morphometry and molecular and cell biology to gastrointestinal diseases, but some, notably the recognition of the association of Campylobaeter pylori with the commonest type of chronic gastritis, have been achieved using such long established 'routine' histological procedures that one wonders how their signifi cance had escaped recognition for so long. The topics covered in this volume have been selected because they present advances of relevance to the diagnostic clinical pathologist. However, they re present the personal selection of the editor, and are in no way exhaustive. Many other examples of progress in our understanding of the pathophysiology of gastrointestinal diseases have been omit ted, either because of the confines of space or because they have been well reviewed recently in other publications. Most of the workload of the practising gastrointestinal patholo gist involves the diagnosis and assessment either of inflammation or of neoplasia in the alimentary tract, and this is reflected in the topics presented in this book."

The ability to diagnose cancer by simple measurement of a serum or tissue' 'marker" has been a goal of medical science for many years. There is ample evidence that tumor cells are different from normal cells and pro duce substances that can be detected by currently available immuno chemical or biochemical methods. These "cancer markers" may be se creted proteins, enzymes, hormones, fetal serum components, monoclonal immunoglobulins, cell surface components, or cytoplasmic constituents. The purpose of this book is to present the current status of our knowledge of such cancer markers. The first tumor marker identified by laboratory means was Bence Jones protein. In a series of lectures delivered to the Royal College of Phy sicians in London in 1846, Dr. H. Bence Jones described studies on a urine sample sent to him with the following note: "Dear Dr. Jones-The tube contains urine of very high specific gravity. When boiled it becomes slightly opaque . . . . etc. " Dr. Jones found that heating of the urine after addition of nitric acid resulted in formation of a heavy precipitate; acid ad dition may have been required to bring the urine to pH 4-6 at which Bence Jones proteins are more likely to precipitate when heated. This urinary pre cipitate was associated with a bone disease termed "mollities ossium. " H. Bence Jones, Papers on Chemical Pathology, Lecture III. Lancet 2, 269-274 (1847)]."

This monograph contains the proceedings of a symposium entitled, "Tissue Hypoxia and Ischemia," which was held at the Annenberg Center of the University of Pennsylvania on August 13 and 14, 1976. The symposium was jointly sponsored by the following groups at the University of Pennsylvania: the Respiratory Physiology Group of the Department of Physio logy, the Cardiopulmonary Section of the Department of Medicine, the Johnson Research Foundation, the Cerebrovascular Research Center of the Department of Neurology, the Head Injury Center of the Department of Neurosurgery, the. Institute for Environ mental Medicine, and the International Society on Oxygen Transport to Tissues. Its purpose was to promote an interdisciplinary discussion of oxygen sensors in various tissues and their mechanism of action as well as to examine the deleterious effects of hypoxia and ischemia with special reference to the brain. There were four sessions, one on the biochemistry of physi ologic oxygen sensors, two on the mechanism of oxygen sensing in tissues and one on the circulatory and metabolic aspects of cerebral hypoxia and ischemia."

Cancer has become the most critical health problem in the United States. It is expected that 25% of the people will develop this dread disease, and many of these will die from the malady. The causes of cancer are varied, but the best estimate available is that 70--90% arise from environmental factors. These statistics have triggered widespread governmental action along two lines: (l) An effort to identify those chemicals and conditions that give rise to malignant processes has been mounted by the Carcino genesis Testing Program, the National Cancer Program, and subse quently, the National Toxicology Program. (2) Regulatory laws have been enacted that are administered by agencies such as TSCA, FIFRA, EPA, FDA, OSHA, and so on, whose mission is to minimize public ex posure to carcinogens. Since direct verification that specific chemicals induce cancer in hu of unanticipated expo mans is necessarily limited to known incidences sure and is therefore rare, most chemicals are identified as carcinogens only by laboratory experiments. At present, the only accepted procedure is long-term animal bioassay, and not only are these studies expensive and time-consuming, but current worldwide resources permit the evalua tion of only 300-400 chemicals per year, a miniscule amount compared to what is available in the commercial world: 30,000 existing chemicals, with approximately 700 new such materials being introduced every year."

A critical phase in coronary heart disease, unstable angina, is thoroughly discussed in contributions by well-known experts. Since myocardial infarction still belongs to the main causes of death, this topic is highly relevant for the present research and possible future treatment of coronary heart disease. The major recent advances made in the pathophysiology, diagnosis and therapy of unstable angina are reported in depth.

W. TH. DAEMS This volume contains the papers presented at a Boerhaave Course for Post graduate Education on The cell-biological aspects of disease: the plasma membrane and lysosomes. One of the purposes of this introduction is to explain the reasons for this choice of subject. The first question which might be asked - why a postgraduate course on the cell biological aspects of disease was considered neccessary - is not difficult to answer: the impact of the basic sciences on medicine is im measurably strong, and among these sciences cell biology has contributed immensely to the advances made in medicine during recent decades. It has provided clues leading to general insights into etiology and pathogenesis as well as to the development of diagnostic tools and a basis for therapeutic methods. These insights derived mainly from the still-increasing body of knowledge about the architecture of cells. Initially, this knowledge arose from the notion that cells are either simply bags full of enzymes or complex sponge like structures in which all organelles are permanently interconnected (Fig. 1). Later, this notion was replaced by a highly schematized picture of the cell as an essentially two-compartment structure (Fig. 2) in which cell organelles are discrete units separated from each other by membranes, enabling each organelle to maintain an internal microenvironment with optimal conditions for its specific metabolic pro cesses."

, 1m Kleinsten die wirkliche Wahrheit gibt graBen Gedanken erst Klarheit" KARL THOMAS Each year sees the publication of hundreds of reports of experimental work on the lymphatic tissue, yet morphological studies of the cells involved can be counted on the fingers of one hand. Furthermore, anyone who tries to identify these cells by morphological criteria is accused of sophistry and hair splitting, whereas it is accounted scientifically correct and unbiased to speak of "lymphoid cells," "blast cells" etc. Not so many years ago things were different: there were too many names and too many classifi cations and everyone backed his particular fancy. People thought of cells in terms of rigid classes, nothing then being known about the transformability of mononuclear blood cells. Today we must look for the middle way: cells should be named and defined according to morpho logical criteria but their transformation potential should be borne in mind. Once the cells are analysed and subdivided, it will be simple enough to set up proper classifications afterwards. This book arose out of the conviction that there should be more criteria and more information available on the morphology of human lymphoreticular cells; previously such information had been restricted by the difficulty of the special hematological and cytochemical staining methods."

Previous volumes in this series have discussed the current progression have identified a variety of targets and strategies state of our knowledge concerning the pathophysiology of to allow these goals to be realized. This volume critically cancer growth and progression. The complexity of the in reviews approaches towards cancer management in man at teraction of malignant neoplasms and the host, the the levels of: detection, diagnosis, surgery, radiology, heterogeneity of malignant cell subpopulations, and the chronobiology and endocrine treatment. existence of metastatic tumor cells resistant to drug thera Several chapters review selected methods of cancer diag pies remain as significant clinical challenges to clinical on nosis. In addition, a variety of on-going and novel ap cologists. Indeed, conventional treatment regimens of che proaches for cancer treatment are also presented in this volume. Progress in the early detection of malignant neo motherapy, surgery and radiology are often ineffective for the therapy of a large variety of established metastatic can plasms, coupled with novel approaches for the therapy of cer in patients. When one considers the insidiousness of such neoplasms, may ultimately yield safe and well-tolerated agents for the selective therapy of solid malignancies. New progressive neoplastic growth and the emergence of con tinuously more aggressive and malignant cellular subpop therapeutic approaches, directed towards the biochemical ulations one is overwhelmed with the challenges inherent in and molecular targets identified in the earlier volumes of this series, may ultimately lead to the generation of new mo attempting to control malignant neoplasms."

Human Lymphoma: The Clinical Implications of the REAL Classification is a unique volume. It is based on the recent developments in classification and overall understanding of human lymphoid neoplasms which are relatively common neoplasms and which epidemiological evidence suggests are increasing in frequency. This field has been the cause of confusion in the past as a result of conficting ideas on the classification of lymphoma and related diseases. However a new vision of the field has emerged and the is encapsulated in the pioneering REAL classification and in a forthcoming WHO scheme, both of which are covered in the book. The volume will appeal to hematologists, pathologists and oncologists and will, thanks to a diverse and expert authorship, serve to increase the working knowledge of all three groups.

Spinal Cord Injury or disease can happen to anyone at any time and the effects can be devastating. I found this out personally when I was thrown from the back of a pick up truck at age 15 was left paralyzed from the waist down. It was during my recuperation as a young teenager that I first gained insight into the importance of rehabilitation. My family, doctors, nurses, fellow patients and researchers who were dedicated to helping me over come my personal tragedy helped me pull through. Today, rehabilita tion medicine is taking great strides and empowering the person with the injury to take control of their future, overcome their setbacks and, through collaborative support, reach their personal goals and potential. Since 1987 the Legacy raised by my Man in Motion World Tour (24901 miles wheeled around the world March 1985-May 1987) has pro $13 million dollars to research and rehabilitation in the areas vided over of spinal cord injury. I hope that in some small way this funding has contributed to the development of the vital programmes that supported me and many others. The effects of spinal cord injury are traumatic and life-shattering and require a skilled interdisciplinary approach. I congratulate those who have contributed to this book and challenge each one of you to never give up on your dreams to find the answers to the optimum treatment of spinal cord injury and disease."

It is difficult for a teacher to accept the fact that not all of the information in his or her field is a necessary part of the armamentarium of the student. This holds for histologists, and for that matter, pathologists, biochemists, and ophthalmologists. It may especially hold for those who were trained during exciting periods in the history of their discipline. In the past two decades, information in every basic medical science has increased greatly, perhaps exponentially, and the basic science curriculum has been compressed to the point where the medical student can reasonably be expected to acquire only a superficial knowledge of anyone of these sciences. Hence the information provided must be carefully selected, effectively presented in retainable doses, and useful for understanding material to be presented later in the curriculum and for the solution of clinical problems. In the compact courses of today, the presentation of only enough data to support basic working concepts of structure and function is appropriate. Additional factual information consti tutes trivia, which in our opinion interferes with effective development, reten tion, and use of the concepts. This philosophy has been the basis for the preparation of this book. Consistent with this philosophy, the text is brief, and the content has been carefully examined for its appropriateness to the needs of the medical student."

This is a book about renal cysts and cystic kidneys. Its contributors have created a resource of current information in a field that once aroused only curiosity, but that now stands at the leading edge of molecular nephrology. Its authorship includes 'oldtimers', who bring the wisdom of experience, and 'newcomers', whose presence attests to the contributions made by the investigative and technological advances of the past decade. Its text is organized to carry the reader from renal cyst to cystic renal disease. Each of its chapters defines or explores a challenge or an advance. Cells that line renal cysts are diverse in structure, type, and perhaps function. The cysts themselves lie within an interstitium that is not normal and may influence cyst development and growth. Experimental analogs of human disease offer increasing opportunities to basic researchers to examine, in sequence and under controlled circumstances, those events that favor nephron dilation, cyst growth and ultimate renal failure.

Over the last decade, high performance Capillary electrophoresis (HPCE) has emerged as a powerful and versatile separation technique that promises to rival high performance liquid chromatography when applied to the separation of both charged and neutral species. The high speed and high separation efficiency which can be attained using any of the various modes of HPCE has resulted in the increased use of the technique in a range of analytical environments. The procedures are, however, still in the early stages of development and several barriers remain to their adoption as the technique of choice for a range of analytical problems. One such barrier is the selection and optimization of the conditions required to achieve reproducible separations of analytes and it is in this area that this new book seeks to give assistance. The book is written by an international team of authors, drawn from both academic and industrial users, and the manufacturers of instruments. At its heart are a number of tables, divided into specific application areas. These give details of published separations of a wide range of archetypal analytes, the successful separation conditions and the matrix in which they were presented. These tables are based on separations reported since 1992 and are fully referenced to the original literature. The tables are supported by discussions of the problems that a particular area presents and the strategies and solutions adopted to overcome them. The general areas covered are biochemistry, pharmaceutical science, bioscience, ion analysis, food analysis and environmental science.

The last volume dealing with lung cancer in this series in Cancer Treatment and Research was published in 1986 and entitled Lung Cancer: Basic and Clinical Aspects. The present book continues the outline of the previous volume by presenting up-to-date information on lung cancer in critical reviews of new important basic and clinical concepts of lung cancer. The present volume has broadened the scope by also including chapters dealing with issues such as epidemiology, prophylaxis, and histopathology of lung cancer. The content of the book thus reflects the increasing awareness of a global disease that is more and more in focus, not only scientifically but also politically. The latter fact results increasingly in changes in health legisla tion, with prevention measures influencing everyday life. The great interest in the disease is natural, considering that more than one patient dies from lung cancer every minute globally. The first chapter is from the Cancer Unit, WHO, Geneva, and describes in detail the epidemiologic features of lung cancer, which is the second most frequent cancer in the world with 660,500 new cases annually; it will soon surpass stomach cancer as the leader. Thirty-one percent of the cases occur in developing countries, where the increase is especially dramatic."