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Apoptosis Methods and Protocols
The most fundamental question facing each and every cell within an org- ism is to survive or to die. Cell death is required for normal function; some estimates suggest that as many as one million cells undergo cell death every second in the adult human body. Almost all cells undergoing physiological, or programmed, cell death, independent of cell type, manifest a stereotypic p- tern of morphological changes termed apoptosis. Typically, apoptotic cells d- play shrinkage, membrane blebbing, chromatin condensation, and nuclear fragmentation. The integrity of the cell membrane is not lost during apoptosis and so avoids eliciting the inflammatory response that would have been caused by the spilla...
Germ Cell Protocols
The study of germ cells has undergone enormous advances in recent years and has entered into an explosive phase of new discoveries with the introd- tion of transgenic technologies and nuclear cloning. Basic knowledge and te- niques developed for lower vertebrate and invertebrate systems have facilitated the study of higher vertebrates, including humans. Many experiments that have first been performed on lower vertebrates provided the tools and strategies that could later be applied to other less readily available mammalian systems. The discovery of centrosomes in ascidians and sea urchin eggs now benefits st- ies of fertility and infertility in mammals including humans. External in vitro fer...
Bacterial Artificial Chromosomes
Several developmental and historical threads are woven and displayed in these two volumes of Bacterial Artificial Chromosomes, the first on Library Construction, Physical Mapping, and Sequencing, and the second on Fu- tional Studies. The use of large-insert clone libraries is the unifying feature, with many diverse contributions. The editors have had quite distinct roles. Shaying Zhao has managed several BAC end-sequencing projects. Marvin Stodolsky during 1970–1980 contributed to the elucidation of the natural b- teriophage/prophage P1 vector system. Later, he became a member of the Genome Task Group of the Department of Energy (DOE), through which s- port flowed for most clone library re...
Neuroprotection Methods and Protocols
This book examines current research into the role of neuronal death in cell signaling pathways, and its role in neurodegenerative diseases, such as Alzheimer's and Parkinson's. After introducing neurodegenerative, traumatic, and ishemic disorders, the authors cover in vitro and animal systems, and cellular and molecular mechanisms.
Parasite Genomics Protocols
Parasitic diseases remain a major health problem throughout the world, for both humans and animals. For many of us, our technologically advanced lifestyle has decreased the prevalence and transmission of parasitic diseases, but for the majority of the world’s population, they are ever present in homes, domestic animals, food, or the environment. The study of parasites and parasitic disease has a long and distinguished history. In some cases, it has been driven by the great importance of the presence of the parasite to the community, for example, those that affect our livestock. In other cases, it is clear that applied research has suffered for lack of funding because the parasite affects p...
Ribozymes and siRNA protocols
In this completely updated and expanded edition of a classic bench manual, hands-on experts take advantage of the latest advances in ribozyme, DNAzyme, hammerhead ribozymes and derivatives, and RNA interference technologies to describe in detail the exciting and successful methods now available for gene inactivation in vitro and in vivo. Their optimized techniques employ hairpin ribozymes, DNAzymes, hammerhead ribozymes and derivatives, group I intron ribozymes, RNase P ribozymes, and siRNAs, as well as general methods for RNA structure analysis, delivery of oligonucleotides, and gene therapy. Also provided are novel methods for identifying accessible cellular mRNA sites; group I intron and ...
Gene Expression Profiling
Leading scientists in gene expression methodology and bioinformatics data analysis describe readily reproducible methods for measuring RNA levels in cells and tissues. The techniques presented include new methods for applying the Affymetrix GeneChip®, SAR-SAGE, StaRT-PCR, SSH, the Invader Assay®, and ADGEM. The authors also provide critical bioinformatics insight and resources for data analysis and management. By distilling the basic underlying principles of many methods to a few straightforward concepts, investigators can easily choose the method most appropriate to their application.
Protein Arrays
Protein Arrays: Methods and Protocols is an introduction to protein array technology and its application to the multiplexed detection of proteins. Although protein array technology has some roots in gene array technology, it can only be described as a distant relative. Unlike DNA, with its established rules of base pairing, and therefore predictable biochemical behavior, proteins are rich with diversity. Proteins can be large or small, compact or extended, basic or acidic, hydrophobic or hydrophilic, and so on. Just as importantly, their behavior is determined by the environment in which they reside, and so the composition of the buffer in which experiments are performed has a dramatic impac...
Vaccinia Virus and Poxvirology
The Right Book at the Right Time The poxviruses comprise a family of complex DNA viruses that replicate in the cytoplasm of vertebrate or invertebrate cells. Of the eight recognized g- era of vertebrate poxviruses, those belonging to the orthopoxvirus genus have been most intensively studied. This group includes variola virus, the agent of smallpox, as well as cowpox virus and vaccinia virus. Jenner’s original sma- pox vaccine, described in 1798, consisted of live cowpox virus, but vaccinia virus later replaced it (1). There has been speculation as to the origin of v- cinia virus; the most likely idea is that it is a separate species, possibly ori- nally isolated from a horse, and is now e...
Signal Transduction Protocols
In 1995, Signal Transduction Protocols, edited by David A. Kendall and Stephen J. Hill, was published in the Methods in Molecular Biology series. This second edition represents an update to that previous work with an emp- sis on new methodologies that have developed in the last few years. The goal, then and now, is to provide procedures written by experts with first-hand ex- rience in a detail that goes far beyond what is generally encountered in the “methods” section of most journals and thus actually permits a particular p- cedure to be replicated. In addition, we have had as a secondary goal the id- tification of protocols for the assay of general classes of signal transduction compon...
