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.

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.

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.

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."

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.

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 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.

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.

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.

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.

Amino acid analysis is a technique that has become commonplace in biotechnology, biomedical, and food analysis laboratories. This book describes a variety of amino acid analysis techniques and how each technique can be used to answer specific biological questions. The first two chapters in Amino Acid Analysis Protocols introduce the concepts, basic theory, and practice of amino acid analysis. The following chapters give detailed instructions on various methods and their applications. As highlighted, there are many different approaches to amino acid ana- sis, but in all cases the results depend heavily on the quality of the sample. Therefore a new way to desalt samples prior to hydrolysis is covered as an introductory chapter (Chapter 3), and most authors have devoted a section to sample preparation, especially to the collection and storage of bodily fluids. Some of the amino acid analysis methods described in this book are based on HPLC separation and analysis after precolumn derivatization. The precolumn derivatization techniques described use (a) 6-aminoquinolyl-N-hydro- succinimidyl carbamate (AQC) (Chapters 4 and 8); (b) 1-fluoro-2- dinitrophenyl-5-L-alanine amide (Marfey's reagent), which allows sepa- tion and analysis of enantiomeric amino acids (Chapter 5); (c) O-phthalal- hyde (OPA) (Chapters 6 and 10); (d) butylisothiocyanate (BITC) and benzylisothiocyante (BZITC) (Chapter 11); (e) phenylisothiocyanate (PITC) (Chapters 12 and 13); (f) ammonium-7-fluorobenzo-2-oxa-1,3-diazole-4-s- fonate (SBD-F) (Chapter 17); and (g) 9-fluorenylmethyl-chloroformate (FMOC-Cl) (Chapter 10).

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.

Viruses and RNAi share an intricate relationship at many levels. RNAi is an important antiviral defense mechanism in plants and invertebrates, microRNAs of viral or cellular origin affect many aspects of virus biology, and replication of many, if not all, mammalian viruses can be suppressed by RNAi." Antiviral RNAi: Concepts, Methods, and Applications "provides a collection of protocols for the analysis of viral small RNAs and natural antiviral RNAi responses as well as for the development and optimization of RNAi-based antiviral drugs. As RNAi is a central regulatory mechanism in the cell, the methods in this volume can also be applied out of the context of a virus infection. Divided into five convenient parts, this detailed volume reviews important basic concepts in the field of antiviral RNAi, provides experimental and bio-informatic tools for the analysis of small silencing RNAs, covers methods to biochemically dissect RNAi-based antiviral defense and viral counter-defense mechanisms, describes methods for the design, expression, and delivery of therapeutic antiviral siRNAs, and finally presents genome-wide RNAi approaches for the identification of factors involved in virus replication. Written in the highly successful "Methods in Molecular Biology " series format, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls.

Authoritative and accessible, "Antiviral RNAi: Concepts, Methods, and Applications" serves as an ideal guide for both novice and experienced researchers alike striving to dissect the role of RNAi in the viral life cycle or to further boost the development of novel therapeutics and experimental tools based on RNAi technology.

The current book describes the chemical and physical behaviour of polymers and biopolymers that form highly associating structures in equilibrium solution. It summons the established results known of polymer complexes in solution, taking into account also the recent developments in biotechnology concerning this topic, in technological applications of polymer-protein interactions, in fluorescence and scattering techniques for the study of intra- and interpolymer association and in the study of ionomers in solution. The book covers the whole range from synthesis and fundamental aspects to applications and technology of associated polymers.

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.

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.