Legionellosis is a disease of significant medical and public interest. Legionella is commonly found in aquatic habitats where its ability to survive and to multiply within different protozoa equips the bacterium to be transmissible and pathogenic to humans. In addition, Legionella has become a favored model system to analyze the mechanisms of bacterial survival, acquisition of nutrients, and intracellular replication. Following the recent publication of the genome sequences of four L. pneumophila strains, it is now feasible to investigate the whole genome in silico, the transcriptome via micro arrays, and the proteome by two-dimensional gel electrophoresis. Research in the fields of clinical features, diagnosis, treatment, and epidemiology continues to generate new data. The topics covered by this volume range from the history of the identification of Legionella and clinical disease treatment, to the microbe's gene expression and secretion systems, as well as its strategies for intracellular multiplication and nutrient acquisition. The main focus of the book is the current state of many of the most critical features of Legionella. Internationally renowned authors have contributed chapters describing and discussing the latest research findings with an emphasis on molecular aspects. The editors and authors have produced an excellent book that will be an extremely useful reference source. This comprehensive publication is aimed at readers with teaching or research interests in microbiology, genetics, genomics, infectious diseases, or clinical research.

Direct cell-cell communication is a common property of multicellular organisms that is achieved through membrane channels which are organized in gap junctions. The protein subunits of these intercellular channels, the connexins, form a multigene family that has been investigated in great detail in recent years. It has now become clear that, in different tissues, connexins speak several languages that control specific cellular functions. This progress has been made possible by the availability of new molecular tools and the improvement of basic techniques for the study of membrane channels, as well as by the use of genetic approaches to study protein function in vivo. More important, connexins have gained visibility because mutations in some connexin genes have been found to be linked to human genetic disorders. Connexin Methods and Protocols presents in detail a collection of te- niques currently used to study the cellular and molecular biology of connexins and their physiological properties. The field of gap junctions and connexin research has always been characterized by a multidisciplinary approach c- bining morphology, biochemistry, biophysics, and cellular and molecular biology. This book provides a series of cutting-edge protocols and includes a large spectrum of practical methods that are available to investigate the fu- tion of connexin channels. Connexin Methods and Protocols is divided into three main parts.

Studies on stem cells have been attracting intense scientific and p- lic attention, not only because of controversies surrounding the use of embryonic stem cells but also because of very provocative data that have been emerging on adult stem cells. Much of the public attention and debate has been focused on the possibility that adult stem cells may be used as a substitute for human embryonic stem cells or as a justification for stopping work on them. This has somewhat dim- ished attention on very heated scientific debates that take us to the very heart of how the concept of stem cells is perceived. To this author, the latter debates have not been unlike certain philosophical debates of the last century. Since the seminal studies of Till and McCulloch in the 1960s, the popular paradigm on adult stem cells has been that lineage-restricted stem cells are derived from pluripotent stem cells very early during development. To many, and consistent with much data, the restriction to particular lineages was considered absolute. In other words, there was a sense of determinism in the stem quality of particular stem cells: once they were allocated, they were programmed to specific roles in a given tissue. Furthermore, some adult tissues were considered devoid of detectable stem cell presence or activity.

After a decade of dominance by recombinant DNA technology, the field of molecular and cell biology is witnessing a renewed interest in techniques and approaches that are not driven by DNA acrobatics. In hindsight, this is an inevitable outcome. Deoxyribonucleic acid is not the master; it is only a storage house. If one wishes to know how cells work, the secret is not to be found in DNA, but rather in everything outside DNA. Science based on DNA is useful but does not itself solve the problem. It is most fortunate that at the height of the DNA phenomenon, there remain scientists who continue to probe cells by non-DNA means. Suddenly, people with such expertise are in high demand.
In this volume, some truly original scientists take the time to tell us their stories of innovations-some almost iconoclastic. All of these researchers have pioneered approaches that were long neglected; moreover, each is now in the fruitful phase of great harvests. It is a wonderful lesson for graduate students and postdoctorates that, although not being in the pack might be risky, the reward of such work is sweeter. As in physics, biology needs more young people who think like Richard Feynman - the ultimate iconoclast.
On the surface, the eight chapters of this volume appear to be diverse, but they are not. If our purpose is to understand cells, we must stop the habit of constantly dissecting leaves. Once in a while we have to see which forest we are in. The contributions included herein cannot cover the whole cell, but they give a sufficient flavor to arouse a desire to think more globally about cells. At a time when our field is in danger of being buried by thousands of kinases and phosphatases, these chapters inform our intended audiences that there are other ways - other techniques, other approaches, other thinkings, and other stories. We need all of them to appreciate the holistic aspect of cells, which has been a taboo until now.

The contributions of plant genetics to the production of higher yielding crops of superior quality are well documented. These successes have been realized through the application of plant breeding techniques to a diverse array of genetically controlled traits. Such highly effective breeding procedures will continue to be the primary method employed for the development of new crop cultivars; however, new techniques in cell and molecular biology will provide additional approaches for genetic modification. There has been considerable speculation recently concerning the potential impact of new techniques in cell and molecular biology on plant improvement. These genetic engineering techniques should offer unique opportunities to alter the genetic makeup of crops if applied to existing breeding procedures. Many questions must be answered in order to identify specific applications of these new technologies. This search for applications will require input from plant scientists working on various aspects of crop improvement. This volume is intended to assess the interrelationships between conventional plant breeding and genetic engineering.

Microbial toxins are secondary metabolites that accumulate in the organism and, to a large extent, are metabolically inactive towards the organism that produces them. The discovery of penicillin, a secondary metabolite of Penicillium notatum West (= P. chrysogenum Thom), in 1929 marked a milestone in the development of antibiotics (microbial toxins). In the intensive studies that followed this discovery, scientists chemically characterized several new molecules (toxins) from secondary metabolites of microbes, some having a definite function in causing pathogenesis in plants. Toxins are also known to playa significant role in inciting animal (human) and insect diseases and as plant growth regulators. Many common toxins have also been isolated from different microbes exhibiting a wide spectrum of biological activity. Toxins are broadly divisible into several characteristic groupings - polyketides, oxygen heterocyclic compounds, pyrons, terpenoidS, amino acids - diketopiperazines, polypeptides etc. Recent research has indicated that these toxins play an important role in plant pathogenesis, disease epidemics, plant breeding, biological control of plant pathogens and insect pests, induced resistance, plant-pathogen interactions etc. Toxins produced by weed pathogens are exploited as lead molecules in developing environmentally friendly herbicides.

There can be no doubt that some ofthe most spectacular advances made in science over the past few decades have been in the isolation, analysis, and manipulation of nucleicacids. Thishas ledtoamuchgreaterunderstandingofmechanismsandprocesses across many fields of bioscience, such as biochemistry, microbiology, physiology, pharmacology, and the medical sciences to name a few. It has also led to the growth of the biotechnology industry, which seeks to develop and commercialize many ofthese important processes and methods. Much ofthis has come about because ofthe devel opment of numerous molecular biology and genetic manipulation techniques. The discovery of restriction enzymes and the development of cloning vectors in the early 1970sopenedthedoortowaysofisolatingandmanipulatingnucleic acidsthathadnever been thought possible. Gene probe labeling and hybridization were developed and refined toprovidepowerfulmethodsofanalysis. These-togetherwiththedevelopment of DNA sequencing methods, protein engineering techniques, and PeR-have all continued to contribute substantially to the understandingofbiological processes at the molecular level. Theprotocols for these importantmethods are the focus ofThe Nucleic AcidProtocols Handbook, whose aim is to provide a comprehensive set oftechniques in onevolume thatwill enable the isolation, analysis, and manipulationofnucleic acids to be readily undertaken. The NucleicAcidProtocols Handbook is divided into 10 parts; within each there are approximately 10chapters. The first fourpartsfollow oneanotherlogically: nucleic acid extraction (Part I), basic separation and analysisofDNA (II), through probe design and labeling (III), and RNA analysis techniques (IV). The following three sections deal with gene libraryconstruction andscreening(V), DNA sequencing (VI), andthe polymerase chain reaction (VII)."

Increasing population and industrialization are the key pollutant contributors in water bodies. The wastes generated by industries are highly hazardous for humans and the ecosystem and require a comprehensive and effective treatment before being discharged into water bodies. Over the years, many up gradations have been introduced in traditional water treatment methods which were expensive and ineffective especially for removal of toxic pollutants. Phycoremediation has been gaining attention due to its mutual benefit in wastewater treatment and for valuable algae biomass production. Wastewater, especially sewage and industrial effluents, is rich in pathogenic organisms, organic and inorganic compounds and heavy metals that adversely affect human and aquatic life. Microalgae use these inorganic compounds and heavy metals for their growth. In addition, they also reduce pathogenic organisms and release oxygen to be used by bacteria for decomposition of organic compounds in a secondary treatment. In this book, the potential of microalgae in wastewater treatment, their benefits, strategies, and challenges are discussed. The increasing need of finding innovative, low-cost, low-energy, sustainable and eco-friendly solutions for wastewater treatment makes the publication of a book on phycoremediation timely and appropriate. Features: (1) Deals with the most emerging aspects of algal research with special reference to phycoremediation. (2) Studies in depth diversity, mutations, genomics and metagenomics study (3) An eco-physiology, culturing, microalgae for food and feed, biofuel production, harvesting of microalgae, separation and purification of biochemicals.

Genetic variability is an important parameter for plant breeders in any con ventional crop improvement programme. Very often the desired variation is un available in the right combination, or simply does not exist at all. However, plant breeders have successfully recombined the desired genes from cultivated crop gerrnplasm and related wild species by sexual hybridization, and have been able to develop new cultivars with desirable agronomie traits, such as high yield, disease, pest, and drought resistance. So far, conventional breeding methods have managed to feed the world's ever-growing population. Continued population growth, no further scope of expanding arable land, soil degradation, environ mental pollution and global warrning are causes of concern to plant biologists and planners. Plant breeders are under continuous pressure to improve and develop new cultivars for sustainable food production. However, it takes several years to develop a new cultivar. Therefore, they have to look for new technologies, which could be combined with conventional methods to create more genetic variability, and reduce the time in developing new cultivars, with early-maturity, and improved yield. The first report on induced mutation of a gene by HJ. Muller in 1927 was a major mi1estone in enhancing variation, and also indicated the potential applica tions of mutagenesis in plant improvement. Radiation sources, such as X-rays, gamma rays and fast neutrons, and chemical mutagens (e. g., ethyl methane sulphonate) have been widely used to induce mutations."

Chloroplasts are essential for the survival and flourishing of life on Earth. Over the years, chloroplast biology has been studied in a variety of different organisms, leading to the significant disadvantage that findings which were made by using different experimental systems or species were not always directly cross-comparable. The relatively recent adoption of Arabidopsis thaliana as the model organism of choice for plant science research, across the globe, has led to its emergence as a pre-eminent system for research on chloroplasts and other types of plastid. In Chloroplast Research in Arabidopsis: Methods and Protocols, expert researchers bring together some of the most important, modern techniques and approaches for chloroplast research, with the unifying theme of Arabidopsis as the model system. Volume II explores topics such as multiprotein complexes, protein-protein interactions, omics and large-scale analyses, proteomics and suborganellar fractionation, as well as photosynthesis and biochemical analysis. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and convenient, Chloroplast Research in Arabidopsis: Methods and Protocols serves as an ideal reference for all researchers with a general interest in chloroplasts, plastids, or related processes.

This second part in the two-volume work Microarrays details applications and data analysis. It includes insight into non-mammalian vertebrate systems, processes and protocols for high quality glass-based microarrays. Coverage includes applications in DNA, peptide, antibody and carbohydrate microarraying, oligonucleotide microarrays generated from hydrolysis PCR probe sequences, microarray platforms in clinical practice, and screening of cDNA libraries on glass slide microarrays. Authors in this volume also discuss protocols for predicting DNA duplex stability on oligonucleotide arrays and integrated analysis of microarray results.

Prior to 1974, the ~adrenergic receptors were known only in- directly as entities that responded to drugs in a selective manner to mediate a variety of physiologically important responses. During the intervening years, our view of ~adrenergic receptors has changed dramatically. The availability of high affinity 125I-labeled radioligands selective for these receptors presaged an explosion of experimenta- tion utilizing direct binding assays to establish the biochemical properties of the receptor protein. In the opening chapter, Stadel and Lefkowitz describe this development and its impact on our under- standing of the molecular basis of ~adrenergic receptor function. The availability of well-characterized receptor ligands, coupled with the development of efficient methods for detergent solubilization, formed the basis of receptor purification using affinity chromatography. The related technique of photoaffinity labeling provided a means to estimate the molecular mass of these receptors. The availability of substantial amounts of purified ~2-adrenergic receptor allowed determination of segments of its amino acid se- quence. This information led to the production of polynucleotide probes and eventually to cloning of the receptor gene and determi- nation of the complete primary sequence of the receptor protein. Caron and Lefkowitz review the investigations leading to this major development and discuss the methods involved. They analyze our current perception of the relation of receptor function to its structure and discuss the general features of the G protein-interacting receptor family, of which the ~-adrenergic receptors are prototypes.

This volume details methods that will aid in the selection of promoter sequences and vector components and methods for the assembly and testing of synthetic promoters with examples of their application. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Mammalian Synthetic Promoters: Methods and Protocols aims to aid researchers that are new to the field of synthetic promoters and inspire new developments.

Within Pyrosequencing Protocols, the protocols for utilizing Pyrosequencing technology are described in detail, including trouble-shooting tips and background information. Chapter 1 provides an introduction to the fascinating origins of the methodology. Chapter 2 provides a brief overview of some of the applications Pyrosequencing is used for. Chapters 3 and 4 describe primer selection and the basic technique. Chapters 5 through 7 provide methods for improving the throughput and decreasing the cost of Pyrosequencing. Detailed applications for the technique can be ound in Chapters 8-13, whilst the important aspect of data storage is discussed in Chapter 14.

A comprehensive state-of-the-art collection of the most frequently used techniques for plant cell and tissue culture. Readily reproducible and extensively annotated, the methods range from general methodologies, such as culture induction, growth and viability evaluation, and contamination control, to such highly specialized techniques as chloroplast transformation involving the laborious process of protoplast isolation and culture. Most of the protocols are currently used in the research programs of the authors or represent important parts of business projects aimed at the generation of improved plant materials. Two new appendices explain the principles for formulating culture media and the composition of the eight most commonly used media formulations, and list more than 100 very useful internet sites.

Cytomegaloviruses are members of the herpesvirus group and can infect humans and other primates. This text presents comprehensive reviews on every aspect of current research.

Plants are an important source of food and of valuable products for industry, agriculture and medicine. They are unique in many aspects of metabolic processes, development and reproduction. Most of these aspects can now be studied by the modern methods and technolo gies of molecular and cellular biology. Such studies are also encouraged as to improve plant yield and quality. During the past decade research in plant sciences has demonstrated the feasibility of plant cell and tissue culture techniques as major tools in biology and agriculture. These techniques are also essential in strategies for engineering of biological systems. The proceedings of the VII International Congress on Plant Tissue and Cell Culture in Amsterdam show that in recent years an impressive progress has been achieved. The papers of the congress, with more than 2000 participants, include the full text of plenary lectures, keynote lectures and presentations of speakers who have been selected out of more than 1400 abstracts. This combination, which provides readers with reviews as well as recent findings and future developments, captures an important part of the scientific exchange during the congress. The papers in these proceedings are a reflection of the role of plant cell and tissue culture in disciplines varying from plant breeding to molecular biology. Basic as well as applied studies in a variety of plant disciplines are presented in 4 sections: (1) Genetic manipulation and propagation, (2) Morphogenesis and metabolism, (3) Secondary metabolites and (4) Biotechnology and developing countries."

Techniques in the neurosciences are evolving rapidly. There are currently very few volumes dedicated to the methodology - ployed by neuroscrentists, and those that are available often seem either out of date or limited in scope. This series is about the methods most widely used by modern-day neuroscientrsts and is written by their colleagues who are practicing experts. Volume 1 will be useful to all neuroscientists since it concerns those procedures used routinely across the wrdest range of s- disciplines. Collecting these general techniques together in a single volume strikes us not only as a service, but will no doubt prove of exceptional utilitarian value as well. Volumes 2 and 3 describe all current procedures for the analyses of amines and their metabolites and of amino acids, respectively. These collections will clearly be of value to all neuroscientists working in or contemplating research in these fields. Similar reasons exist for Volume 4 on receptor binding techniques since experimental details are provided for all types of ligand-receptor binding, including chapters on general principles, drug discovery and development, and a most useful-appendix on computer programs for Scatchard, nonlinear, and competitive d- placement analyses. Volume 5 provides procedures for the asse- ment of enzymes involved in biogenic amme synthesis and catabolrsm. Volumes in the NEUROMETHODS series will be useful to neurochemists, -pharmacologists, -physiologists, -anatomists, psychopharmacologists, psychiatrists, neurologists, and chemists (organic, analyncal, pharmaceutical, medicinal); in fact, everyone involved in the neurosciences, both basic and clinical.

This thesis describes the inception, design, and implementation of stereoselective desymmetrization reactions in the total synthesis of the natural products pactamycin and paspaline. In the case of pactamycin, the author develops a novel asymmetric Mannich reaction and symmetry-breaking reduction strategy to enable facile construction of the complex core architecture in fifteen steps using commercially available materials - the shortest synthesis to date. He subsequently demonstrates the flexibility of this approach in SAR investigations by highlighting the preparation of twenty-five unique pactamycin structural congeners. For paspaline, the author develops a biocatalytic desymmetrization strategy that allows the highly controlled synthesis of core stereochemistry and provides a platform for the development of new conceptual disconnections in the synthesis of "steroid-like" natural products. This thesis offers a valuable resource for students embarking on a PhD in total synthesis.

This volume contains the papers presented in the NATO Advanced Research Workshop "Activation of Hormone and Growth Factor Receptors: Molecular Mechanisms and Consequences" held in Nafplion, Greece on September 25-30, 1988. The objective of NATO ARW is to assess the state of-the-art in a given scientific area and to formulate recommendations for future research in emerging areas of science by promoting international scientific contacts. In the Nafplion meeting this objective was reached by an international group of speakers, senior Greek scientists and graduates involved in relevant research areas. The Workshop was made possible by the generous support of the Scientific Council of NATO. We thank Drs. G. Sinclair and L.V. daCunha, Directors of the NATO ARW's and ASI's (Advanced Study Institutes) respectively, for their wholehearted support and advice. The International Union of Biochemistry awarded additional travel grants leading to increased international participation. Furthermore, the Secretariat of Science and Technology, the Ministry of Culture and Sciences and the National Hellenic Research Foundation contributed financially and by supporting personnel. We sincerely thank all these organizations for their support. Our heartful thanks are also extended to the Mayor of the Municipality of Nafplion, Mr.

The First Asia --- Pacific Conference on Agricultural Biotechnology was held in Beijing, China on 20-24, August, 1992. Over half the population in the world is in the Asian and Pacific Region. With an increasing population and decreasing farming lands, it is important to develop agricultural biotechnology for improvement of the productivity, profitability and stability of the farming system. The Conference's main objectives were to bring together scientists working in different fields of agricultural biotechnology to stimulate discussion on this important process and to have an appraisal of the most recent studies concerning genetic manipulation of plants, plant cell and tissue culture, plant gene regulation, plant-microbe interaction, animal biotechnology etc. The Conference was attended by 391 scientists from different countries and regions. This volume presents the contributions of the lectures and a selected number of posters, which are an up-to-date account of the state of knowledge on agricultural biotechnology. The book provides a valuable reference source not only for specialists in agricultural biotechnology, but also for researchers working on related aspects of agronomy, biochemistry, genetics, molecular biology, microbiology and animal sciences. It is with great pleasure to acknowledge the contributions of the authors in assuring the prompt publication of this volume. We would also extend our sincere thank to Kluwer Academic Publishers for the publication of these proceedings.

Leading drosophilists describe in step-by-step detail all the essential techniques for studying Drosophila chromosomes and suggest new avenues for scientific exploration. The chapters emphasize specimen preparation (from dissection to mounting) and cover both polytene and mitotic/meiotic chromosomes in depth. Each fully tested and readily reproducible protocol offers a background introduction, equipment and reagent lists, and tips on troubleshooting and avoiding pitfalls. A cutting-edge FISH and immunolocalization technique will be important for discovering how DNA sequence influences higher-order chromosome architecture and ultimately gene expression.

Recent advances in large scale DNA sequencing technology have made it possible to sequence the entire genome of an organism. Attention is now turning to the analysis of the product of the genome, the proteome, which is the set of proteins being expressed by a cell. Mass spectrometry is the method of choice for the rapid large-scale identification of these proteomes and their modifications. This is the first book to extensively cover the applications of mass spectrometry to proteome research.

Techniques in the neurosciences are evolving rapidly. There are currently very few volumes dedicated to the methodology - ployed by neuroscrentists, and those that are available often seem either out of date or limited in scope. This series is about the methods most widely used by modern-day neuroscientists and 1s written by their colleagues who are practicing experts. Volume 1 will be useful to all neuroscientists since it concerns those procedures used routinely across the widest range of s- drsciplines. Collecting these general techniques together in a single volume stnkes us not only as a service, but will no doubt prove of exceptional utilitarian value as well. Volumes 2 and 3 describe all current procedures for the analyses of ammes and theirmetabolites and of amino acrds, respectively. These collections will clearly be of value to all neuroscientists working in or contemplating research in these fields. Similar reasons exist for Volume 4 on receptor binding techniques since experimental details are provided for many types of ligand-receptor binding, including chapters on general prin- ples, drug discovery and development, and a most useful app- dix on computer programs for Scatchard, nonlinear, and compe- tive displacement analyses. Volume 5 provides procedures for the assessment of enzymes involved in biogenic amine synthesis and catabolism. Volumes in the NEUROMETHODS series will be useful to neurochemists, -pharmacologists, -physrologists, -anatomists, psychopharmacologists, psychiatrists, neurologists, and chemists (organic, analytical, pharmaceutical, medicinal); in fact, everyone involved in the neurosciences, both basic and clinical.