This compendium presents some of the major applications of neutron scattering techniques to problems in biology. It is a record of the papers presented at the Neutrons in Biology Conference, the third in an occasional series held to highlight progress in the field and to provide a focus for future direction. The strength ofthe neutron scattering technique remains principally in the manipula tion of scattering density through hydrogen and deuterium atoms. The development ofad vanced detectors, innovative instrument and beamline components, and sophisticated data acquisition systems through the 1970s and early 1980s provided a sound foundation for the technique. With continued development, some of the exotic and expensive equipment has become affordable by the medium-sized facilities, thereby broadening the user base considerably. Despite problems with the major neutron sources in the late 1980s and early 1990s, some spectacular results have been achieved. Whilst the high and medium flux beam reac tors will continue to make a major impact in the field, the results from the first experi ments, and the planned developments on spallation neutron sources, clearly indicate that the technique has enormous potential.

This book is based upon a series of papers originally presented at a NATO Advanced Studies Workshop of the same title held at the study Center HI Cappuccini," a converted monastery in San Miniato, a small village located between Pisa and Florence, Italy, in October 1991. Authors were asked to submit their completed chapters by the following February; these were then scanned onto computer disks, edited and returned to the authors for final revision, and updating, with a final deadline of February 1993. The authors were encouraged to make whatever modifications they wanted, especially regarding updating their chapters, with an eye to making the final product both comprehensive and current. In this we succeeded beyond our expectations, with most chapters extensively altered and many completely re-written and significantly expanded. Thus, although the original meeting was held in 1991, the chapters in this volume may be regarded as current from at least February, 1993, with some of the final updated revisions received as late as July, 1993. This book, as agreed in our original contract, has been delivered to the publisher "camera-ready. " This means that all of the scanning, editing, proofing and typesetting were done here, by the office of the Department of Dermatology at the New Jersey Medical School. We essentially produced the book, which the publisher, for the most part, then photocopied. This has been an enormous burden, borne mainly by my colleague in this division, Robert A.

In mammalian cells many physiological processes rely on the dynamics of the organization of lipids and proteins in biological membranes. The topics in this volume deal with physicochemical methods in the study of biomembranes. Some of them have a long and respectable history in the study of soluble proteins and have only recently been applied to the study of membranes. Some have tradi tionally been applied to studies of model systems of lipids of well-defined com position, as well as to intact membranes. Other methods, by their very nature, apply to organized bilayers comprised of both protein and lipid. Van Meer and van Genderen provide us with an introduction to the field (Chapter I). From their personal perspective regarding the distribution, trans port, and sorting of membrane lipids, they formulate a number of biologically relevant questions and show that the physicochemical methods described in this book may contribute in great measure to solving these issues. The methods of analytical ultracentrifugation have served faithfully for 60 years in the study of water-soluble proteins. The use of detergent extraction of membrane proteins, and the manipulation of density with H20/D20 mixtures, has extended this technique to the study of proteins, and in particular their interactions, from biological membranes. As described by Morris and Ralston in Chapter 2, this technique can be used to determine a number of important properties of proteins.

In the ten years since the first publication on PCR (Saiki et al. , 1985), this in vitro method of nucleic acid replication and modification has grown to rival in popularity traditional microbiological, genetical und technical procedures for cloning, sequencing, gene detecting and related procedures. To date the PCR literature has emphasized six main areas of application: genetic mapping, detection of mutations, genetic polymorphism, transcriptional splicing and regulation, molecular virology and quantitative procedures. The overwhelming focus of quantification of DNA or RNA by PCR has been on human microbiology and oncological problems. The exquisite sensitivity of PCR gives this method the ability to detect extremely rare DNAs, mRNAs, mRNAs in small numbers of cells or in small amounts of tissue, and mRNAs expressed in mixed-cell populations. However, the exact and accurate quantification of specific nucleic acids in biological samples is in spite of numerous publications in that field still a general problem: during the peR process, an unknown initial number of target sequences are used as a template from which a large quantity of specific product can be obtained. Although the amount of product formed is easy to determine, it is difficult to deduce the initial copy number of the target molecule because the efficiency of the peR is largely unknown.

The hagfishes comprise a uniform group of some 60 species inhabiting the cool or deep parts of the oceans of both hemispheres. They are considered the most primitive representatives of the group of craniate chordates, which - apart from the hagfishes that show no traces of verte brae -includes all vertebrate animals. Consequently the hagfishes have played and still playa central role in discussions concerning the evolution of the vertebrates. Although most of the focus on hagfishes may be the result of their being primitive, it should not be forgotten that, at the same time, they are specialized animals with a unique way of life that is interesting in its own right. It is now more than 30 years since a comprehensive treatise on hagfishes was published. The Biology of Myxine, edited by Alf Brodal and Ragnar Fange (Universitetsforlaget, Oslo, 1963), provided a wealth of information on the biology of hagfishes, and over the years remained a major source of information and inspiration to students of hagfishes."

A NATO Advanced Study Institute on "New Developments in Lipid-Protein Interactions and Receptor Function" was held on the Island of Spetsai, Greece, from August 16-27, 1992. This Institute was organized to bring together researchers in the field of membrane organization and dynamics with those actively involved in studies on receptor function, signal transduction mechanisms and gene regulation. 2 Presentations and discussions focussed on the regulation of intracellular Ca +-levels, on the second messengers derived from inositol lipids and on the specific phospholipase C isozymes involved in these processes. A major focus was on G-proteins and the effect of lipid anchors on their function. These principles of regulation were further discussed in the context of receptors for acetylcholine, lysophosphatidic acid and low-density lipoproteins. In addition, various aspects of the genomic regulation of cell growth and differentiation by transcription factors were presented. These topics were put into perspective by discussing the most recent developments in lipid-protein interactions, protein insertion into membranes, membrane lipid organization and lipid dynamics as mediated by phospholipid transfer proteins. This book presents the content of the major lectures and a selection of the most relevant of the most important topics posters. These proceedings offer a comprehensive account presented during the course of the Institute. The book is intended to make these proceedings accessible to a large audience.

Genetic Engineering, Volume 24 contains discussions of contemporary and relevant topics in genetics, including: -Gene silencing: principles and applications, -Integrins and the myocardium, -Plant virus gene vectors: biotechnology and applications in agriculture and medicine, -Novel approaches to controlling transcription, -Use of DNA polymorphisms in genetic mapping, -Application of FLP/FRT site-specific DNA recombination system in plants. This principles and methods approach to genetics and genetic engineering is essential reading for all academics, bench scientists, and industry professionals wishing to take advantage of the latest and greatest in this continuously emerging field.

~he major theme of this book is the development of the vertebrate central nervous system. ~is volume contains summaries of most of the invited participants at the NA~ advanced study institute entitled "Development of central nervous system in vertebrates" held in Maratea, Italy, from June 23-July 5, 1991. In order to address this topic, we have drawn upon a selection of current studies dealing with molecular, cellular and system analysis which specifically pertain to the general principles of the development. ~he major aim of this institute was to bring together a select group of investigators who would present their views on the current issues in their respective fields and to foster extensive discussions amongst participants in smaller groups. Such interactions brought together the exchanges of ideas amongst participants and helped clarify the intricate details and formulate new vistas and collaborations. Since the study of nervous system development has focused mostly on the origin of neuron and glia cells, the area of current research was represented by talks on early cellular events including effects of growth factors, BOX and other gene expressions and cell lineage of specific cell type(s). Formation of specific cell types and the specific neuronal connections have been a major theme in the study of the nervous system development. Recent technical advances has resulted in new information at both cellular and molecular levels which have provided new details. Current research was represented by "selective" topics discussed at the meeting.

is"0l m 5-HT Ante-mortem (m = metabolite) . ". ~ Post-mortem m 0 Noradrenaline ,,-'-'-'-'0-J ,,-'-'-'-'-'-'- m Dopamine ~ . '-'-'-'-'- . " Somatostatin . '1 Plaques Tangles * . ". . ". . 'I Synapse loss * ChAT " ACh synthesis l"-,. '-'-'-'-'-'-'-'-'-'-'-'-'- ,,-'-'-'-'-'-'-'-'-'-'-'-'-'-'- * Neuronal loss ,,- ,,- . ". * o 0. 1 0. 2 0. 3 0. 4 0. 5 0. 6 0. 7 0. 8 0. 9 1. 0 Correlation coefficient with dementia rating Figure 2. 1 Various neurotransmitter and neuropathological measures on neocortex obtained either at diagnostic craniotomy (ante-mortem) or post-mortem and re- lationship to cognitive impairment (dementia rating). Asterisks identify significant correlations. Negative and positive correlations are not identified in this represen- tation of the data. Only the physiologically relevant measure of cholinergic neurones (acetylcholine synthesis) and indices of cortical pyramidal neurone degeneration (neuronal loss, synapse loss and tangles) correlate with severity of dementia. Note the lack of correlation with the structural marker of cholinergic neurones, ChAT activity. Loss of enzyme exceeded that of acetylcholine synthesis (Sims et al. , 1983a), suggesting a compensatory increase in synthesis in the few remaining neurones. Data derived from Palmer (1987) and Terry et al. (1991). emission tomography (PET) and magnetic resonance, may some day pro- vide routine insight into differences in brain functioning. Success in developing rapid and reliable diagnostic procedures, perhaps based on changes in body fluid that reflect specific alterations in brain protein metabolism, should ultimately play an important role in the clinical development of new therapeutic agents.

This book presents the contributions of the main speakers to the 12th Long Ashton InternationalSymposium, heldat theUniversityofBristolfrom 17thto20thSeptember 1991. Manyofthe 160delegates who attended presented posters, the abstractsofwhich have been published separately and reflected thevigour, excitementand originalityofthepresentations. The identity and molecular structure of solute of membrane proteins are of interest in of themselves but may be seen as only a step towards broader questions about the regulation their activity and their integration into the lifeofcells and organisms. This symposium was held at a time when exciting progress was being made on the molecular biology of some transporters, ion channels and hormone receptors. The contributions reflect this progress in different degrees. Some problems are less tractable than others and the mixture of success stories and hopeful aspirations was a deliberate choice by the organisers. At the symposium those who were still confronted with great difficulties were listened to sympathetically and with constructive interest. Itis hoped that their papers will be read in the same way. During the symposium we discussed the natureofmany transport proteinsand receptors, as well as the existence ofadditional regulatory proteins such as protein kinases, G-proteins and calmodulin. Wecould say that upto twentyproteins wereconsidered inany detail. While this may sound like a considerable achievement, the scaleofourignorance can be illustrated byconsideringthatonatypicalsilver-stained, two-dimensionalpolyacrylamidegelofpurified plant plasma membrane polypeptides, between 150 and 200 spots are routinely shown; radiolabelled membranes frequently giveeven more, indicating how much membraneprotein there is, the function of which we have no conceptual framework.

Scientific reviews are now of two complementary types: short, very up-to-date articles, as are found in the Trends series, and the more traditional longer re views, which are more comprehensive but take longer to publish. The Subcellu lar Biochemistry series belongs to the latter category where a number of reviews on a broad topic are collected together in the one volume. It has been the aim of this volume to summarize the present state of knowledge of membrane assembly. It is appreciated that some relevant topics have not been included, and an editor's selection is restricted by the many calls on potential authors who are unable to meet all requests made to them. The absence of a discussion of the roles of lipids is," however, a reflection of the fact that a recent volume in this series, edited by Dr. H. 1. Hilderson, has been devoted to this subject (Vol. 16, Intracellular Transfer of Lipid Molecules), and readers are recommended to this source.

American trypanosomiasis, or Chagas disease, is caused by the protozoan parasite, Trypanosoma cruzi. Sixteen to eighteen million people are currently infected with this organism, and 45,000 deaths are attributed to the disease each year. Infection with T. cruzi is life-long, and 10-30% of persons who harbor the parasite chronically develop cardiac and gastrointestinal problems associated with the parasitosis. Although major progress has been made in recent years in reducing vector-borne and transfusion-associated transmission of T. cruzi, the burden of disability and death in persons chronically infected with the organism continues to be enormous. Eight to ten million persons born in countries in which Chagas disease is endemic currently reside in the United States, and epidemiologic and census data suggest that 50,000-100,000 are chronically infected with T. cruzi. The presence of these infected persons poses a risk of transmission of the parasite in the USA through blood transfusion and organ transplantation and several such cases have now been documented. American Trypanosomiasis, volume seven of World Class Parasites is written for students of tropical medicine, parasitology and public health, for researchers and practitioners alike who wish to bring themselves abreast of the status quo with respect to this disease. It is intended to supplement formal textbooks, in order to broaden and illuminate current areas of scientific and public health concern. Uniquely for T. cruzi, this book addresses parasite, vector and host biology, the pathogenesis of Chagas disease and current and prospective therapeutics and control strategies in a single volume.

It is nearly a decade since the first Male Mediated Developmental Toxicity conference was held in Pittsburgh. The continuing public/scientific interest, growing amounts of animal data, introduction of innovative technologies, and increasing quantity of human epidemiological studies all suggest that male-mediated developmental toxicity is of major concern. A number of researchers concluded that a Second International Confer- ence on Male Mediated Developmental Toxicity was necessary. The ensuing volume is particularly timely because it impacts on areas of special emphasis in many countries, with respect to children's and reproductive health, as well as to basic molecular mecha- nisms of environmental insult, and genetic susceptibility and predisposition. The Programme and Local Organizing Committee, composed of Barbara Hales (Chair, McGill University), BernardRobaire (McGill University), Daniel G. Cyr (INRS/ Armand Frappier), Jacquetta M. Trasler (McGill University), Andrew F. Olshan (Uni- versity of North Carolina), Sally Perreault Damey (US EPA), Donald R. Mattison (March of Dimes), and Jan M. Friedman (University of British Columbia), spent over two years identifying individuals who had made key contributions in this field over the past decade and planning various aspects of the meeting. The meeting was held in Montreal in June 2001. A total of 132 persons, coming from five continents and representing some 18countries, took an active role in the proceedings. The conference was considered by all attendees to be a rousing success. Important discussions were held in the four break-out sessions, with a preliminary set of recommendations for action being presented by each panel.

This volume contains the papers presented at the International Symposium on "Cirrhosis, Hyperammonemia and Hepatic Encephalopathy," held in Valencia, Spain, January 24th-27th, 1994. Liver cirrhosis and other hepatic dysfunctions such as fulminant hepatic failure and congenital defects of urea cycle enzymes can lead to hepatic encephalopathy, coma and death. Hepatic encephalopathy is one of the main causes of death in western countries. The ability to detoxify ammonia by its incorporation into urea is diminished by impaired liver function, resulting in increased ammonia levels in blood and brain. Hyperammonemia is considered one of the main factors in the mediation of hepatic encephalopathy and the classical clinical treatments are directed towards reducing blood ammonia levels. However, the molecular bases of the pathogenesis of hepatic encephalopathy and the role of hyperammonemia in this process remain unclear and several hypotheses have been proposed. To clarify the mechanisms involved in hepatic encephalopathy and hyperammonemia suitable animal models are necessary. The animal models available and the ideal features of an animal model are presented in the initial part of the book.

Sponges (phylum Porifera) are known to be very rich sources for bioactive compounds, mainly secondary metabolites. Main efforts are devoted to cell- and mariculture of sponges to assure a sustainable exploitation of bioactive compounds from biological starting material. These activities are flanked by improved technologies to cultivate bacteria and fungi which are associated with the sponges. It is the hope that by elucidating the strategies of interaction between microorganisms and their host (sponge), by modern cell and molecular biological methods, a more comprehensive cultivation of the symbiotic organisms will be possible. The next step in the transfer of knowledge to biotechnological applications is the isolation, characterization and structural determination of the bioactive compounds by sophisticated chemical approaches.

Corona- and related viruses are important human and animal pathogens that also serve as models for other viral-mediated diseases. Interest in these pathogens has grown tremendously since the First International Symposium was held at the Institute of Virology and Immunobiology of the University of Wiirzburg, Germany. The Sixth International Symposium was held in Quebec City from August 27 to September I, 1994, and provided further understanding of the molecular biology, immunology, and pathogenesis of corona-, toro-, and arterivirus infections. Lectures were given on the molecular biology, pathogenesis, immune responses, and development of vaccines. Studies on the pathogenesis of coronavirus infections have been focused mainly on murine coronavirus, and mouse hepatitis virus. Neurotropic strains ofMHV (e.g., JHM, A59) cause a demyelinating disease that has served as an animal model for human multiple sclerosis. Dr. Samuel Dales, of the University of Western Ontario, London, Canada, gave a state-of-the-art lecture on our current under standing of the pathogenesis of JHM-induced disease.

Grauzone and Completion of Meiosis During Drosophila Oogenesis describes the work behind a major, award winning discovery: the establishment of a new pathway that specifically regulates the female meiosis, a process essential for sexual reproduction. This book chronicles a new gene mapping method and the cloning and documentation of several types of genes that were proven to have significant influence on the cell cycle. It is of interest to anyone doing work with fruit flies, both graduate students and principal investigators.

Progress in Cell Cycle Research is a new annual series designed to be the source for up-to-date research on this rapidly expanding field. Review articles by international experts examine various aspects of cell division regulation from fundamental perspectives to potential medical applications. Researchers as well as advanced undergraduate and graduate students in cell biology, biochemistry, and molecular biology will benefit from this series.

Myofibrillogenesis has been studied extensively over the last 100 years. Until recently, we have not had a comprehensive understanding of this fundamental process. The emergence of new technologies in molecular and cellular biology, combined with classical embryology, have started to unravel some of the complexities of myofibril assembly in striated muscles. In striated muscles, the contractile proteins are arranged in a highly ordered three dimensional lattice known as the sarcomere. The assembly of a myofibril involves the precise ordering of several proteins into a linear array of sarcomeres. Multiple isoforms in many of these proteins further complicate the process, making it difficult to define the precise role of each component. This volume has been compiled as a comprehensive reference on myofibrillogenesis. In addition, the book includes reviews on myofibrillar disarray under various pathological conditions, such as familial hypertrophic cardiomyopathy (FHC), and incorporates a section on the conduction system in the heart. Much of the information in this volume has not been described elsewhere. Presented in a manner to be of value to students and teachers alike, "Myofibrillogenesis" will be an invaluable reference source for all in the fields of muscle biology and heart development.

It is widely accepted that vaccination still renains the best answer to ITDst infectious diseases. Recently, vaccine developnent has been greatly facilitated by advances in ITDlecular and cell biology which have laid the foundations of a new generation of vaccines. '!hese are exemplified by submit vaccines produced through gene cloning and synthetic peptides mimicking snall regions of proteins on the outer coat of viruses and capable of eliciting virus neutralizing antibodies. However, submit and peptide vaccines are only weakly or non-inmmogenic in the absence of immunological adjuvants. The latter are a diverse array of agents that augment specific cell-mediated immune responses to the antigens and the formation of protective antibodies. '!his book contains the proceedings of the 3rd NATO Advanced Studies Institute (ASI) "New-Generation vaccines: '!he Role of Basic Irrmmology" held at Cape Sounion Beach, Greece, during 24 June-5 July, 1992. It deals with recent developnents in the understanding of inmmity at the ITDlecular and cellular levels and the application of such knowledge in the search for novel inmmological adjuvants and the fonnulation of new-generation vaccines for experimental and clinical use. We express our appreciation to Professor K. Dalsgaard and H. Snippe for their cooperation in planning the ASI and to Mrs. Concha Perring for her excellent production of the manu- scripts. '!he ASI was held tmder the sponsorship of NATO Scientific Affairs Division and generously co-sponsored by SrnithKline Beecham Pharmaceuticals (Fhiladelp,. ia). Financial assistance was also provided by Pasteur Merieux (Marcy L'Etoile), British Biotechnology Ltd.

The last decades have witnessed a radical change in our views on central nervous system damage and repair. This change is not only due to the emergence of new powerful tools for the analysis of the brain and its reactions to insults, but it also reflects a conceptual change in the way we approach these problems. As an illustration to this development, it is instructive to go back to the proceedings of a meeting at the NIH in 1955 edited by William F. Windle, which summarizes the disillusioned and pessimistic view on CNS regeneration prevailing at the time. While this generation of researchers were well aware of the issues at stake, they felt they had reached the end of the road; the approaches they had pursued had got stuck and the tools available could not take them any further. I can very well imagine that the participants, most of them leaders in the field, left that conference feeling they had heard their field being sentenced to death.

There is considerable interest in thermophile microorganisms, in their environments, their ability to survive at temperatures which normally denature proteins, but more importantly, as a valuable resource for bio technology. The first reported isolation of Thermus by Tom Brock was in 1969. This initiated the present era of thermophilic research with the realization that where liquid water is available, there may be no limits to the temper ature at which microorganisms can grow. Considerable research into the ecology, physiology, metabolism, and thermostable enzymes of thermo philes has led to their evaluation for a range of industrial and commercial processes. The past fifteen years have been an explosive period of dis covery of many new genera and species, including the descriptions of a new fundamental kingdom-the Archaea. Much of the current research has been focused on the Archaea; but it is significant that during this period, the original type strain YT-l of Thermus aquaticus described by Brock has provided a major step forward in molecular biology. DNA polymerase from strain YT-I has proved to be the major success in the commercialization of enzymes from thermophilic microorganisms to date. The ease with which Thermus strains can be handled in laboratories without specialized equipment, together with the large investment in de scribing their structure, metabolism, and genetics, should ensure a con tinuing effort in Thermus research.

This volume summarizes the proceedings of the Eighth International Symposium on the Immunobiology of Proteins and Peptides which was held on November 16-20 in Rio Rico, Arizona. The articles represent papers by invited speakers as well as papers selected by the Scientific Council, from among those submitted by the participants, on the basis of quality and timeliness. This symposium series was established in 1976 for the purpose of bringing together, once every two or three years, active investigators in the forefront of contemporary immunology, to present their findings, discuss their significance in the light of current concepts and identify important new directions of investigation. The founding of the symposium was stimulated by the achievement of major breakthroughs in the understanding of the immune recognition of proteins and peptides. We believed that these breakthroughs would lead to the creation of a new generation of peptide reagents, which could have enormous potential in biological, therapeutic, and basic applications. This anticipated explosion has since occurred and many applications ofthese peptides are now being realized. The eighth symposium focused on the manipulation or modulation of the immune response. This volume broadly covers the areas of adjuvants, cytokines, vaccines, and the use of intravenous immunoglobulins for disease management. There is a clear need to identify methods for improving vaccine efficacy and guiding the host to respond with a particular type of immune response.

As we assume the editorship of Current Ornithology, it seems useful to review the objectives of the series. We cannot improve on the state ments of our predecessors when they began their service as editors. In his preface to Volume 1 (1983), Richard F. Johnston wrote: The appearance of the first volume of a projected series is the occasion for comment on scope, aims, and genesis of the work. The scope of Current Or nithology is all of the biology of birds. Ornithology, as a whole-organism sci ence, is concerned with birds at every level of biological organization, from the molecular to the community, at least from the Jurassic to the present time, and over every scholarly discipline in which bird biology is done; to say this is merely to expand a dictionary definition of "ornithology. " The aim of the work, to be realized over several volumes, is to present reviews or position statements concerning the active fields of ornithological research. Dennis M. Power, who edited Volumes 6-12 (1989-1995), began his preface to Volume 6 (1989) as follows: This edited series has three principal goals. The first is to provide information in a relatively concise way for researchers needing an overview of specific disciplines. The second is to provide an update on specific schools of thought, bringing together ideas from colleagues whose works often appear in a variety of journals. And the third is to stimul&te and suggest directions for new re search."

The genus Aspergillus has a worldwide distribution and is one of the most common of all groups of fungi. They are possibly the greatest contami nants of natural and man-made organic products, and a few species can cause infections in man and animals. The aspergilli are also one of the most important mycotoxin-producing groups of fungi when growing as contaminants of cereals, oil seeds, and other foods. Not all aspergilli are viewed as troublesome contaminants, however, as several species have had their metabolic capabilities harnessed for commercial use. The aspergilli have long been associated in the Far East with the koji stage of several food fermentations, particularly soy sauce and miso, and subsequently as a source of useful enzymes. The ability of these fungi to produce several organic acids, especially citric acid, has created major industrial complexes worldwide. Traditional methods of strain develop ment have been extensively studied with the industrial strains, while more recently, recombinant DNA technology has been applied to the aspergilli with emphasis on heterologous protein production. In compiling this book, I have been fortunate to have the full enthu siastic involvement of the authors, and to them I extend my very grateful thanks for mostly being on time and for producing such readable and authoritative chapters. Collectively, we hope that our efforts will strengthen the scientific understanding of this intriguing group of filamentous fungi and further their use in the field of biotechnology.