Unlike some other reproductions of classic texts (1) We have not used OCR(Optical Character Recognition), as this leads to bad quality books with introduced typos. (2) In books where there are images such as portraits, maps, sketches etc We have endeavoured to keep the quality of these images, so they represent accurately the original artefact. Although occasionally there may be certain imperfections with these old texts, we feel they deserve to be made available for future generations to enjoy.

Unlike some other reproductions of classic texts (1) We have not used OCR(Optical Character Recognition), as this leads to bad quality books with introduced typos. (2) In books where there are images such as portraits, maps, sketches etc We have endeavoured to keep the quality of these images, so they represent accurately the original artefact. Although occasionally there may be certain imperfections with these old texts, we feel they deserve to be made available for future generations to enjoy.

This volume is composed of chapters that review important fundamental aspects of HCV biology and disease pathogenesis including, for example, the discovery and identification of the HCV genome, early virus-cell interactions including identification of various cellular receptors, HCV gene expression studied using the HCV replicon system, identification and characterization of HCV structural- and non-structural HCV proteins, HCV replication in cultured cells, and host factors involved in viral replication. This volume also contains chapters dealing with immunity to HCV infection and pathogenesis. This is particularly important in understanding hepatitis C because HCV infection alone is not cell lytic. Mechanisms underlying the persistent nature of HCV infection are also discussed in these chapters. Many of the authors published articles that were listed among the "top 10 papers" published in the 24 years since HCV was discovered in 1989. Their citations are above 1,000 (Web of Science). The authors describe the background and significance of their contributions to the field in the context of findings from other research groups.

My Choice As I sit and think of each hurt and pain in my life over the years, I sure am glad that the choice I made to allow God to change me has only strengthened my relationship with him. I know that it was only God who came and rescued me because I was at the doorsteps of hell. He has guided me along with ordering my steps every day since I accepted him into my heart, my mind, and my soul. Jesus Christ is my personal Saviour. I listened to a sermon in church on Sunday, February 10, 2008, and I can't get it out of my head; I believe that it is because God is still healing me from that prophetic message. As I have learned to forgive others in order to be forgiven, I am learning to forgive myself daily. I thank God for each and every person who hurt me knowingly or unknowingly, along with the trials and tribulations, because if it had not happened just that way, I would not be praying all the time. I would not be so eager to serve on my ministry at church. I definitely wouldn't be reading the Bible as much as I do, nor would I attend church every Sunday. And yes, God knows I would have spent my money buying a pair of shoes or an outfit, but now I tithe faithfully just as the Lord asks. So he will work it all out if you allow his will to be done in your life. I didn't say that his road was easy, but I will inform anyone that I would not change my life for anything. I thank God for all the interruptions. And I most definitely thank him for all the people who pushed me closer to him. That is actually the people who I let hurt me. So thank you, God, for change. DeAngelia M. Walker April 1, 2008

John Walker and Ralph Rapley have collected a wide-ranging group of molecular and biochemical techniques that are the most frequently used in medical and clinical research, especially diagnostics. The authors-well-established investigators who run their own research programs and use the methods on a regular basis-outline the practical procedures for using them and describe a variety of pertinent applications. Among the technologies presented are southern and western blotting, electrophoresis, PCR, cDNA and protein microarrays, liquid chromatography, in situ hybridization, karyotyping, flow cytometry, bioinformatics, genomics, and ribotyping. The applications include assays for mutation detection, mRNA analysis, chromosome translocations, inborn errors of metabolism, protein therapeutics, and gene therapy.

Addressing the perennial question: why should we be moral? this book argues that we can only give a truly and morally satisfying answer to that question by radically reconfiguring our conception of the self and the way it relates to others.

Referendums—the direct popular vote on an issue—appear to be the most democratic of decision mechanisms because the voice of the people is directly heard rather than mediated through elected representatives in legislatures. But referendums can be manipulated by elites using tactics such as timing and wording of the question submitted to a popular vote. Leaders can orchestrate referendum campaigns to their benefit while still claiming the popular legitimacy granted. This takes place in long-established democracies like France, authoritarian regimes like Pinochet’s Chile, and new democracies like those among the Post-Soviet states where competition is raw, rules are new, and institutions weak. Mark Walker sheds light on the appeal and dangers of referendums and why democratic ideals are not always served.

Since publication of the first edition of this Handbook, the usage of thenno plastic elastomers (TPEs) has doubled, with a compounded annual growth rate of approximately 9 percent. This second edition summarizes and documents the technological and commercial progress that has given rise to this phenomenal rate of growth. Over the past decade, numerous suppliers and users of ther moplastic elastomers have entered the field, and some have retired from it, a process that almost certainly will continue. This Handbook is intended to serve the broad spectrum of professionals ac tively engaged in the field of thennoplastic elastomers, which has seen a growth rate four to six times that of the rubber and plastics industries. As TPEs embrace both rubber and plastics technology, this book will be useful to rubber and plastics technologists with a broad variety of specific interests. This edition emphasizes commercial practice and practical application rather than research activity. Technology and innovation are stressed, with polymer science functioning as a basis for understanding and communication. We have focused on those TPEs that we consider to be of significant commercial impor tance-the ones now used in the fabrication of useful articles, or which probably will be so used in the foreseeable future."

The first volume in this Methods Molecular Biology series, Proteins (1984), concentrated on basic techniques for the analysis and purification of peptides and proteins. As the series developed, more specialized volumes on proteins were introduced, such as those on Immunochemical Protocols (vol. 10), Practical Protein Chro- tography (vol. 11), Analysis Glycoprotein Biomedicine (vol. 14), Protein-DNA Interactions (vol. 30), Biomembrane Protocols (vols. 19 and 27), Analyses and Methods (vol. 17), and Optical Spectroscopy, Microscopy, and Macroscopic Techniques (vol. 22). Further specialist volumes on peptides, monoclonal antibodies, immunoassays, ELISA, protein engineering, protein stability, mass spectrometry of proteins, automated sequence analysis, and protein NMR are currently in preparation. Since it is now a decade since the initial volume was published, it seems an especially appropriate moment to extensively reorganize, update, and revise the earlier volume. In an attempt to be more c- prehensive in our coverage, this current volume, Basic Protein and Peptide Protocols, is totally committed to basic analytical methods; a planned companion volume will later concentrate on preparative techniques. Those analytical techniques requiring expensive speci- ized instrumentation, such as NMR, mass spectrometry, X-ray cr- tallography, spectroscopy, and automated sequence analysis, are not described here, but in the appropriate specialized volumes listed above.

Hands-on researchers describe in step-by-step detail 73 proven laboratory methods and bioinformatics tools essential for analysis of the proteome. These cutting-edge techniques address such important tasks as sample preparation, 2D-PAGE, gel staining, mass spectrometry, and post-translational modification. There are also readily reproducible methods for protein expression profiling, identifying protein-protein interactions, and protein chip technology, as well as a range of newly developed methodologies for determining the structure and function of a protein. The bioinformatics tools include those for analyzing 2D-GEL patterns, protein modeling, and protein identification. All laboratory-based protocols follow the successful Methods in Molecular Biology (TM) series format, each offering step-by-step laboratory instructions, an introduction outlining the principle behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.

Plant Cell and Tissue Culture continues the high standards of Humana's Methods in Molecular Biology series. Its step-by-step approach (a hallmark of the series) is applied to a wide range of basic laboratory techniques and culture conditions appropriate to plant cells. Because of the diversity of cell types, species, and culture methods, much of this volume is devoted to the culture of particular cell types and to the regeneration of these cells into whole plants. Special attention is also given to the genetic modification of plants, as well as to the economic significance of plant products. Chapters cover a wide range of topics and techniques, including:* tissue culture media and selection * cryopreservation * callus culture techniques * organ culture * embryogenesis * batch culture * large-scale culture * hormonal control * fertilization techniques * gene transfer * cell immobilization * production systems * cell product purification * DNA expression * electrofusion of plant cells * mutant selection * mutagenesis techniques * automation * transfer of nuclei * protoplast culture * media analysis * micropropagation. A detailed appendix lists the formulas for the most commonly employed plant cell media. Comprehensive, easy to follow, and a pleasure to use, Pollard and Walker's Plant Cell and Tissue Culture is an essential tool for everyone--at all levels of proficiency and experience--involved in plant culture.

In recent years there has been a tremendous increase in our understanding of the functioning of the cell at the molecular level. This has been achieved in the main by the invention and development of new methodology, parti- larly in that area generally referred to as "genetic en- neering." While this revolution has been taking place in the field of nucleic acids research, the protein chemist has at the same time developed fresh methodology to keep pace with the requirements of present day molecular bi- ogy. Today's molecular biologist can no longer be content with being an expert in one particular area alone. He/she needs to be equally competent in the laboratory at h- dling DNA, RNA, and proteins, moving from one area to another as required by the problem he/she is trying to solve. Although many of the new techniques in molecular biology are relatively easy to master, it is often difficult for a researcher to obtain all the relevant information nec- sary for setting up and successfully applying a new te- nique. Information is of course available in the research l- erature, but this often lacks the depth of description that the new user requires. This requirement for in-depth pr- tical details has become apparent by the considerable - mand for places on our Molecular Biology Workshops held at Hatfield each summer.

In recent years there has been a tremendous increase in our understanding of the functioning of the cell at the molecular level. This has been achieved in the main by the invention and development of new methodology, parti- larly in that area generally referred to as "'genetic en- neering." While this revolution has been taking place in the field of nucleic acids research, the protein chemist has at the same time developed fresh methodology to keep pace with the requirements of present day molecular bi- ogy. Today's molecular biologist can no longer be content with being an expert in one particular area alone. He/she needs to be equally competent in the laboratory at h- dling DNA, RNA, and proteins, moving from one area to another as required by the problem he/she is trying to solve. Although many of the new techniques in molecular biology are relatively easy to master, it is often difficult for a researcher to obtain all the relevant information nec- sary for setting up and successfully applying a new te- nique. Information is of course available in the research l- erature, but this often lacks the depth of description that the new user requires. This requirement for in-depth pr- tical details has become apparent by the considerable - mand for places on our Molecular Biology Workshops held at Hatfield each summer.

Casinonomics provides a comprehensive analysis of the economic and social impacts of the casino industry. Examining the latest cutting-edge research, with a mix of theory and empirical evidence, Casinonomics informs the reader on the most important facets at the forefront of the public policy debate over this controversial industry. While the casino industry has continued to expand across the United States, and around the world, critics argue that casinos bring negative social impacts that offset any economic benefits. Casinonomics examines the evidence on the frequently claimed benefits and costs stemming from expansions in the casino industry, including the impact on economic growth, consumer welfare, and government tax revenues, as well as gambling disorders, crime rates, and the impact on other businesses. Readers will come away with a better-informed opinion on the merits of these arguments for and against public policies that would expand casino gambling.

For many years I performed tissue culture in large scientific insti- tions that had a great deal of infrastructure. When I set up a tissue l- oratory outside such an infrastructure, however, I found there was a shortage of easily accessible information about the basic needs, reagents, and techniques for establishing such a facility. Much had to be done by trial and error or gleaned from originalpapers. Consequently, I felt that a methods book covering a wide variety of techniques from basic culture to the most sophisticated cell analysis would be a very valuable addition to the scientific literature. In the interim, several useful books (listed in Chapter I of this volume) did appear, but none entirely fitted the bill and some are now somewhat dated. Then, in 1984, the first of the Methods in Molecular Biology volumes from Humana Press was published with its step-by-step recipe approach. This format appealed to me, and so I c- tacted John Walker, the series editor, about including cell culture in this series. The result was that we embarked upon a single volume covering both plant and animal cell culture. Such was the richness of the material that this project soon divided itself into separate volumes on animal cell (Volume 5) and plant cell (Volume 6) culture. In this volume (Volume 5), therefore, we have aimed to describe a variety of basic techniques and culture conditions for a range of cell types.

In this celebration of teachers and their art, Walker defies naysayers and affirms the importance of great educators and their work. Drawing on the experience of a career spanning decades, Walker uses poetry and prose to bear witness to the quiet accomplishments of men and women who make life-changing impacts on their students. He holds up these examples for the next generation, demanding that we acknowledge the dedication, talent, and artistry exemplified by these icons within the field.