The third edition of this volume expands upon the previous two editions with new and up-to-date methods and protocols. Chapters include step-by-step procedures involved in quantifying cell growth, baculovirus infection and cell metabolism, methods to isolate new cell lines and develop your own serum-free medium, and routine maintenance and storage of insect cell lines and baculoviruses, small- and large-scale recombinant protein production with the BEVS in both insect and mammalian cell culture and in insect larvae, production and characterization of baculoviruses, green fluorescent protein, tubular reactors and RNAi, and baculovirus/insect cell system to study apoptosis and generating envelop-modified baculovirus for gene delivery into mammalian cells. 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 key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Baculovirus and Insect Cell Expression Protocols, Third Edition aims to not only aid the user in successfully completing the tasks described, but also stimulate the development of improved techniques and new applications of baculoviruses and insect cell culture.

Advances in Virus Research serial highlights new advances in the field with this new volume presenting interesting chapters. Each chapter is written by an international board of authors.

For a virus to invade a host cell it needs to penetrate the physical barrier imposed by the plasma membrane. Viruses have evolved specialized surface proteins to meet this challenge. These proteins facilitate delivery of the viral genetic information into the host cell by either fusing the viral envelope with a host cell membrane or by forming membrane pores. Membrane fusion and pore formation critically depend on the engagement of host cell receptors and receptor choice is a key determinant of viral tropism. The multi-faceted interplay between viral and cellular factors during virus entry is a fascinating field of study, which can provide important insight into viral pathogenesis and define new targets for intervention. This book provides a comprehensive overview of this exciting field of research.

The story of doctors who developed a safe and effective early treatment for COVID-19 and their battle with the Bio-Pharmaceutical Complex that suppressed it. At the beginning of 2020, Dr. Peter McCullough was a highly regarded practicing physician, program director, teacher, and clinical investigator at a major academic medical center in Dallas, Texas. When COVID-19 arrived in March, he felt a duty to find a treatment for the disease. He wasn't alone. Other doctors all over the world were also searching for a cure. They followed the longstanding principle that it's best to tackle a sickness early, before it becomes life threatening. This is the story of how Dr. McCullough and his colleagues developed an early treatment protocol of generic, repurposed drugs and supplements that has saved millions of COVID-19 patients from hospitalization and death. In spite of their success, their early treatment protocol was not welcomed by public health officials. On the contrary, the news of their promising results was dismissed as soon as it was reported. At first this seemed like conventional skepticism, but then fraudulent papers maligning the protocol's repurposed drugs were published in academic medical journals. This and other acts of fraud revealed that a coordinated smear campaign against early treatment was being waged. Dr. McCullough and his colleagues soon found themselves censured, censored, attacked in the media, and fired from their jobs. The greatest victims of the smear campaign were COVID-19 patients who were consequently deprived of early treatment. Hundreds of thousands needlessly died of the disease. At the same time early treatment was suppressed, the US government and mainstream media proclaimed that the cure to COVID-19 lay in a new generation of vaccines that were being developed at warp speed. These were heralded as a forthcoming panacea that would save mankind and restore normalcy. As soon as they were mass deployed, public health officials would lift the restrictions on social and economic life. While many observers were thunderstruck by this turn of events, there were historical precedents. In his 1961 Farewell Address, President Eisenhower warned: We must guard against the acquisition of unwarranted influence, whether sought of unsought, by the military-industrial complex. The potential for the disastrous rise of misplaced power exists and will persist. We must never let the weight of this combination endanger our liberties or democratic processes. As Dr. McCullough and his colleagues learned, Eisenhower's warning has become equally applicable to the Bio-Pharmaceutical Complex of multinational drug companies, the NIH and other federal agencies, research and virology labs, and the Gates Foundation. Since COVID-19 arrived, this Complex has obtained misplaced power over every aspect of our lives and taken our liberties. The Courage to Face Covid-19 recounts how Dr. McCullough and his colleagues began their work by fighting a novel infectious disease, and then became leaders in fighting the tyrannical regime that endangers our American way of life.

Plant Virology Protocols: New Approaches to Detect Viruses and Host Responses addresses recent developments in genome analyses and cytological technologies being used today to learn more about plant virology. Opening with chapters covering techniques relevant to the detection of unknown viruses and disease diagnosis, this detailed volume continues with chapters on the utilization of meta-genome sequencing and global gene expression analyses for the search and identification of viruses, as well as the elucidation of host responses to viral infection, construction methods of infectious cDNAs, and methods relevant to plant virus control. 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 practical, Plant Virology Protocols: New Approaches to Detect Viruses and Host Responses will be an invaluable guide to researchers working in the field of plant sciences.

One hundred years ago, when Martinus W. Beijerinck in Delft and Friedrich Loeffler on Riems Island discovered a new class of infectious agents in plants and animals, a new discipline was born. This book, a compilation of papers written by well-recognized scientists, gives an impression of the early days, the pioneer period and the current state of virology. Recent developments and future perspectives of this discipline are sketched against a historic background. With contributions by A. Alcami, D. Baulcombe, F. Brown, L. W. Enquist, H. Feldmann, A. Garcia-Sastre, D. Griffiths, M. C. Horzinek, A. van Kammen, H.-D. Klenk, F. A. Murphy, T. Muster, R. O'Neill, P. Palese, C. Patience, R. Rott, H.- P. Schmiedebach, S. Schneider-Schaulies, G. L. Smith, J. A. Symons, Y. Takeuchi, V. ter Meulen, P. J. W. Venables, V. E. Volchkov, V. A. Volchkova, R. A. Weiss, W. Wittmann, H. Zheng

The need for the development of techniques based on the characteristics of the viral proteins and genomic nucleic acids was realized in order to detect, identify, differentiate and quantify viruses in the infected plants/planting materials with or without symptoms of infection. Immunoassays have been successfully applied for the detection of viruses in crop and weed host plant species as well as in the vectors. Nucleic acid-based techniques have been demonstrated to be the most reliable and sensitive tests for detection, identification and differentiation of viruses and viroids present in plants and planting materials.. Inclusion of numerous protocols in appropriate chapters as appendix is a unique feature of this volume.

Although there have been many books on HIV and AIDS, surprisingly little has been published that focuses on the immunology of retroviral infections in general, and HIV in particular. Retroviral Immunology: Immune Response and Restoration is the first book of its kind to address the most important aspects of the immunology of retroviruses, including not only the virus-specific immune responses, but also genetic and virologic factors modulating these responses. The book also deals directly with the emerging concept of immune restora tion in retroviral infections, a particularly important subject to the thousands of clinicians who deal with this problem on a daily basis. With the advent of highly effective antiviral drug regimens to slow down the replication of HIV and the progression of AIDS, new challenges and opportunities are arising. Restoration of general immune function has brought with it not only complica tions of immune restoration-mediated disease, but also the realistic hope for meaningful restoration of the ability to control HIV replication with the immune system. Leading scientists in the field have summarized the most current informa tion regarding experimental and clinical aspects of retroviral infections. Retroviral Immunology: Immune Response and Restoration should prove an impor tant point of reference for basic scientists and clinicians in this area of research. We are indebted to all of our authors for their excellent contributions."

Epigenetic Regulation of Cancer in Response to Chemotherapy, Volume 158 of the Advances in Cancer Research series, highlights new advances in the field, with this new volume presenting interesting chapters. Each chapter is written by an international board of authors.

Recombinant viruses provide an efficient mechanism for the transfer and expression of DNA in eukaryotic cells. First, the transfer of DNA by viral infection-utilizing specific cell surface receptors and cellular intern- ization mechanisms-occurs much more readily than DNA transfer via uptake induced by such physical methods as calcium phosphate coprecipitation or electroporation. Second, the novel strategies employed by the virus to express its own genes can then be "hijacked" in the recombinant virus to express the researcher's gene of interest The purpose of Practical Molecular Virology isthus to compile a coll- tion of readily repeatable gene transfer and expression methods from wo- ers expert in the use of a variety of recombinant viral vectors . These include those designed for the production of recombinant antigens, such as pol- virus and yeast Ty-VLPs; those giving very high levels of recombinant protein expression, for example, baculovirus, vaccinia virus, and SV40; and finally viral vectors used for efficient, stable gene transfer to eu otic cells, such as retroviruses and herpesviruses . The first chapter describes the viral life cycle for each virus, and explains how this can be adapted to allow construction of recombinant vectors. Subsequent chapters deal with methods for producing and char- terizing recombinant viruses . I make no apology for the hyperproliferation of chapters dealing with recombinant retroviral methods and applications, since I believe this is clearly proportional to the recent expansion of interest in these techniques.

Structure-Function Relationships of Human Pathogenic Viruses provides information on the mechanisms by which viruses enter the cell, replicate, package their DNA into capsids and mature into new virions. The relation between structural features and the pathogenicity and oncogenicity of some of the most relevant human viral pathogens are demonstrated and the acquisition of defense mechanisms through virus-host interactions are presented. In contrast to textbooks, this volume combines timely research data to provide a holistic view of viral pathogenesis.

Furthermore Structure-Function Relationships of Human Pathogenic Viruses illustrates in a single volume the fundamental processes involved in viral life cycles using up-to-date information from research laboratories around the world. Knowledge of these processes is crucial to develop rationales for the design of future drugs. The timeliness of the data and the comprehensive yet concise approach this book takes in order to present the world of viral pathogens should make it a frontrunner in higher education and R&D.

Negative-strand RNA viruses, so named because of the polarity of their genomic RNA to mRNA, include important human and non-human pathogens. This volume covers major advances in reverse genetics techniques over the past decade, state-of-the-art basic science and the clinical implications of experimental findings. This should rekindle interest in negative-strand RNA viruses among readers, including those in other disciplines, leading to further progress in understanding these important viruses and in developing effective measures of control.

Advances in Applied Microbiology, Volume 120 continues the comprehensive reach of this widely read and authoritative review source in microbiology, providing invaluable references and information on a variety of areas relating to the topics of microbiology.

Advances in Applied Microbiology, Volume 123 continues the comprehensive reach of this widely read and authoritative review source in microbiology. Users will find invaluable references and information on a variety of areas relating to the topics of microbiology.

After three volumes on adenoviruses in 1995 the past years have seen rapid progress in the field of adenovirus research. Moreover, adenoviruses have attracted considerable interest as vectors in gene transfer regimens. After the first volume on virion and structure, viral replication and host-cell interaction this second volume deals with the immune system, oncogenesis and gene therapy.

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 history of research on hog cholera (HC)/classical swine fever (CSF) can be roughly divided into three phases which are characterized by the methods available at the time for demonstrati ng the causati ve agent. Phase covered the period before the viral etiology of HC was discovered by de Schweinitz and Dorset (1904)*. Thereafter (Phase II) the detection of HC virus (HCV) was accomplished by laborious, time-consuming and costly pig inoculation experiments. This explains the extensive search for methods not only for detection but also for accurate infectivity titration as well as for applicable serological techniques to solve urgent problems concerning the pathogenesis, diagnosis, epidemiology and prophylaxis of HC. It was not before the late fifties that HC research entered Phase III when fluorescent antibody techniques offered not only the means for detection and titration of HCV in porcine cell cultures but also for more intensive research on hog cholera and its virus. And yet, there are a number of questions to be answered, e. g. on the genetic and antigenic relation of HCV to bovine viral diarrhea (BVD) virus. There are indications that Phase IV of HC research will bear the stamp of biotechnology. In view of this development it appears appropriate to give an up-dating and summarizing account of HC/CSF including comparative aspects of infections caused by structurally related viruses. The editIon of the present volume would have been impossible without the cooperation of several known scientists who instantly agreed when asked for contribution.

The articles in this volume provide a comprehensive overview of our current understanding of the roles played by cellular factors in the early steps of retroviral replication. A better understanding of these functions will provide critical new insights into retrovirus-host cell interactions and is likely to prove useful for the future development of effective antiretroviral therapies.

This book is an excellent, up-to-date reference on a relatively young area of research in which virology, cellular biology and molecular pathogenesis govern the principles of coinvestigation. Thus, the book will be of great interest to virologists, molecular immunologists and biologists, and biochemists but also to clinical pharmacologists in the long-term search for new antiviral agents. Ulrich Desselberger, Gif-sur-Yvette/Cambridge. Infection of a naive (non-immune) host with a virus elicits an immediate response which results in a cascade of changes in the host, including an interferon response (innate immunity). The outcome of this interaction is influenced by the genes of the virus as well as the genes of the host. Interestingly, different viruses do it in different ways. Not only is there a plethora of mechanisms used by the invading organisms, but the host has also evolved a great variety of redundant and robust countermeasures. This interplay of host and virus represents one of the most significant frontiers in biology today. A clearer understanding of the mechanisms involved will arm us with better strategies to deal with viruses, including emerging pathogens and potential bioterrorism agents. This book is sure to benefit students, scientists, and physicians working in the areas of virology, immunology, microbiology, and infectious diseases. Pharmaceutical industry professionals will also find interest in this illuminating look into virus/host interactions.

Baculovirus Expression Protocols, Second Edition, provides the detailed steps required to perform the techniques involved with the use of baculoviruses and insect cell culture and discusses problems that may be encountered. This newly-revised and updated edition provides a cutting edge step-by-step guide for biochemists, molecular biologists, biochemical engineers and others using the BEVS and/or insect cells for producing recombinant proteins. Furthermore, Baculovirus Expression Protocols, Second Edition, provides assistance to scientists and engineers interested in developing and producing baculovirus insecticides. Authors list all available insect cell lines and provide methods for isolating new cell lines. The procedures involved in producing products, both at laboratory scale and large scale, are discussed, as well as the production in insect larvae. It is hoped that this volume will not only aid the user in successfully completing the tasks described herein, but will also stimulate the development of improved techniques and new applications of baculoviruses and insect cell culture. Timely and thorough, this volume will become an essential reference for anyone using baculoviruses and insect cell culture.

Principles of Virology, the leading virology textbook in use, is an extremely valuable and highly informative presentation of virology at the interface of modern cell biology and immunology. This text utilizes a uniquely rational approach by highlighting common principles and processes across all viruses. Using a set of representative viruses to illustrate the breadth of viral complexity, students are able to under-stand viral reproduction and pathogenesis and are equipped with the necessary tools for future encounters with new or understudied viruses. This fifth edition was updated to keep pace with the ever-changing field of virology. In addition to the beloved full-color illustrations, video interviews with leading scientists, movies, and links to exciting blogposts on relevant topics, this edition includes study questions and active learning puzzles in each chapter, as well as short descriptions regarding the key messages of references of special interest. Volume I: Molecular Biology focuses on the molecular processes of viral reproduction, from entry through release. Volume II: Pathogenesis and Control addresses the interplay between viruses and their host organisms, on both the micro- and macroscale, including chapters on public health, the immune response, vaccines and other antiviral strategies, viral evolution, and a brand new chapter on the therapeutic uses of viruses. These two volumes can be used for separate courses or together in a single course. Each includes a unique appendix, glossary, and links to internet resources. Principles of Virology, Fifth Edition, is ideal for teaching the strategies by which all viruses reproduce, spread within a host, and are maintained within populations. This edition carefully reflects the results of extensive vetting and feedback received from course instructors and students, making this renowned textbook even more appropriate for undergraduate and graduate courses in virology, microbiology, and infectious diseases.

Viral Genomic RNA as a Single Translation Unit 100 Initiation Codon Choice and Translation in Plants 100 Conventional Scanning and Initiation 100 Non-AUG Initiation Codons in Plant Virus RNAs 101 Role of5'- and 3'- Noncoding Regions in Initiation of Translation 101 Access to Internal Genes 103 Divided versus Monopartite Genomes 104 Transcriptional Control: Subgenomic RNAs 104 Non-Orthodox Mechanisms of Initiation 107 Leaky Scanning 107 Internal Ribosome Entry Sites (IRES Elements) 107 Non-Orthodox Mechanisms of Elongation and Termination 108 Readthrough of Leaky Stop Codons 108 Ribosomal Frameshifiting 109 Proteolytic Processing 110 Chymotrypsin-like Serine or Cysteine Proteinases 112 Papain-like Cysteine Proteinases 112 Conclusions 113 References 114 5. Molecular Basis of Genetic Variability in RNA Viruses 121-141 Jozef J. Bujarski Introduction 121 Genetic Mechanisms of Variability 121 Mutation 121 Point Mutations 122 Insertions and Deletions 123 Frameshift Mutations 123 Mutant Stability 124 Effects of Mutations on Host-Virus Interactions 124 RNA Recombination 125 Natural Sequence Rearrangement 126 Recombination between Viral and Host RNAs 126 Role of RNA Structure 127 Role of Replicase Proteins 130 Pseudorecombination 131 Defective Interfering RNAs 132 Conclusions 134 Acknowledgements 136 References 136 viii 6. Genetic Variability and Evolution 143-159 F. Garcia-Arenal, A. Fraile, and J. M. Malpica Introduction 143 Variability under Experimental Conditions 143 Variability under Natural Conditions 145 Factors Determining Genetic Structure of Virus Populations 149 Founder Effects 149 Selection 150 Complementation 152 Conclusions 153 Acknowledgements 155 References 155 7. Molecular Basis of Virus Transport in Plants 161-182 Scott M.

Years ago when we were asked to write a book on the present-day knowledge of the molecular biology of poliovirus, we did not expect that such an apparently simple task could involve so much time and effort. Our writing was hampered by the fact that both of us are full time "workers," so that this monograph is mainly a spare time expedience. The main attention of this book focuses on a detailed review of the molecular biology of poliovirus and especially on the advances of the last decade; medical and environmental aspects are only briefly mentioned. Observations from older studies are considered in view of more recent information. Some of the older ob servations provided fundamental insights and paved the way for present day research; too often such data has been neglected or independently rediscovered. Today, poliovirus research has again attracted considerable interest. High points gained within the last few years were the elucidation of the complete nucleotide sequences of the RNAs of the three poliovinls serotypes and the corresponding vaccine strains, the demonstration of genome evolution during transmission of poliovirus in an epidemic, further characterization of the antigenic sites on the virus particle and of the antigenic drift, characterization of alternate conforma tional states of the virion capsid, the development of monoclonal antibodies against some of the virus proteins, observations on the role of the plasma membrane, cytoskeleton, and cytoplasmic membranes as mediators in the virus induced redirection of the synthetic machinery of the host cell, and characteriza"

Cann's Principles of Molecular Virology, Seventh Edition provides an easily accessible introduction to modern virology, presenting principles in a clear and concise manner. The new edition provides the history of virology and the fundamentals of the molecular basis of how viruses work. It discusses the interactions which control the structure of virus particles, the ways viruses infect cells, how viruses replicate themselves, and the consequences and pathogenesis of virus infection for host organisms. This fully updated edition also reflects advances made in the field and includes new content on phage therapy, CRISPR as a phage defense / offense system, new ideas about evolution, and giant viruses. With the addition of ancillary resources, Principles of Molecular Virology, Seventh Edition is an essential foundational reference for academics, graduate students, and advance undergraduates in virology, molecular biology, and microbiology as well as researchers entering virology, infectious disease, and immunology research.