Download or read book Superantigen Protocols written by Teresa Krakauer and published by Springer Science & Business Media. This book was released on 2008-02-05 with total page 259 pages. Available in PDF, EPUB and Kindle. Book excerpt: Leading researchers in the biological, chemical, and physical investigation of superantigens describe in step-by-step detail their best experimental techniques to assess the physical characteristics and biological effects of superantigens. Their protocols range from those for investigating the interactions of superantigens with cellular receptors to those for the analysis of their immunological and biological effects, including methods for using BIOcore to determine binding kinetics and establishing various lymphocyte cell culture systems. There are also accounts of such methods as the RNase protection assay, cytokine ELISA, FACS analysis, and cytokine production at the single cell level..
Download or read book MHC Protocols written by Stephen H. Powis and published by Springer Science & Business Media. This book was released on 2008-02-05 with total page 343 pages. Available in PDF, EPUB and Kindle. Book excerpt: The aim of MHC Protocols is to document protocols that can be used for the analysis of genetic variation within the human major histocompatibility complex (MHC; HLA region). The human MHC encompasses approximately 4 million base pairs on the short arm of chromosome 6 at cytogenetic location 6p21. 3. The region is divided into three subregions. The telomeric class I region contains the genes that encode the HLA class I molecules HLA-A, -B, and -C. The centromeric class II region contains the genes encoding the HLA class II molecules HLA-DR, -DQ, and -DP. In between is the class III region, originally identified because it contains genes encoding components of the complement pathway. The entire human MHC has recently been sequenced (1) and each subregion is now known to contain many other genes, a number of which have immunological functions. The study of polymorphism within the MHC is well established, because the region contains the highly polymorphic HLA genes. HLA polymorphism has been used extensively in solid organ and bone marrow transplantation to match donors and recipients. As a result, large numbers of HLA alleles have been identified, a process that has been further driven by recent interest in HLA gene diversity in ethnic populations. The extreme genetic variation in HLA genes is believed to have been driven by the evolutionary response to infectious agents, but relatively few studies have analyzed associations between HLA genetic variation and infectious disease, which has been difficult to demonstrate.
Download or read book Inflammation Protocols written by Paul G. Winyard and published by Springer Science & Business Media. This book was released on 2008-02-03 with total page 373 pages. Available in PDF, EPUB and Kindle. Book excerpt: Inflammation has been described as the basis of many pathologies of human disease. When one considers the updated signs of inflammation, they would be vasodilation, cell migration, and, in the case of chronic inflam- tion, cell proliferation, often with an underlying autoimmune basis. Gen- ally, inflammation may be divided into acute, chronic, and autoimmune, - though the editors believe that most, if not all, chronic states are often the result of an autoimmune response to an endogenous antigen. Thus, a proper understanding of the inflammatory basis may provide clues to new therap- tic targets not only in classical inflammatory diseases, but atherosclerosis, cancer, and ischemic heart disease as well. The lack of advances in classical inflammatory diseases, such as rh- matoid arthritis, may in part arise from a failure to classify the disease into different forms. That different forms exist is exemplified in patients with d- fering responses to existing antiinflammatory drugs, ranging from nonresponders to very positive responders for a particular nonsteroidal an- inflammatory drug (NSAID). Though researchers have progressively unr- eled the mechanisms, the story is far from complete. It should also be noted that the inflammatory response is part of the innate immune response, or to use John Hunter’s words in 1795, “inflammation is a salutary response.” That may be applied in particular to the defensive response to invading micro- ganisms.
Download or read book RT PCR Protocols written by Nicola King and published by Springer Science & Business Media. This book was released on 2008-02-04 with total page 370 pages. Available in PDF, EPUB and Kindle. Book excerpt: Until the mid 1980s, the detection and quantification of a specific mRNA was a difficult task, usually only undertaken by a skilled molecular biologist. With the advent of PCR, it became possible to amplify specific mRNA, after first converting the mRNA to cDNA via reverse transcriptase. The arrival of this technique—termed reverse transcription-PCR (RT-PCR)—meant that mRNA suddenly became amenable to rapid and sensitive analysis, without the need for advanced training in molecular biology. This new accessibility of mRNA, which has been facilitated by the rapid accumulation of sequence data for human mRNAs, means that every biomedical researcher can now include measurement of specific mRNA expression as a routine component of his/her research plans. In view of the ubiquity of the use of standard RT-PCR, the main objective of RT-PCR Protocols is essentially to provide novel, useful applications of RT-PCR. These include some useful adaptations and applications that could be relevant to the wider research community who are already familiar with the basic RT-PCR protocol. For example, a variety of different adaptations are described that have been employed to obtain quantitative data from RT-PCR. Quantitative RT-PCR provides the ability to accurately measure changes/imb- ances in specific mRNA expression between normal and diseased tissues.
Download or read book Peptide Research Protocols written by Janet J. Maguire and published by Springer Science & Business Media. This book was released on 2008-02-04 with total page 241 pages. Available in PDF, EPUB and Kindle. Book excerpt: A panel of multidisciplinary experts describes in detail readily reproducible methods to investigate all aspects of the endothelin system from its synthesis and metabolism, to its function in health and disease. Theses methods use state-of-the-art molecular techniques to quantify the expression of mRNA for both endothelin receptors and the endothelin converting enzymes. They show how peptides, precursors, receptors, and synthetic enzymes can be localized and quantified in plasma, culture supernatants, tissue homogenate, and tissue sections using antibodies. Several in vivo protocols illustrate the role of the endothelin peptides in healthy human individuals and describe animal models that can be used to predict the therapeutic potential of cardiovascular drugs that manipulate endothelin synthesis or function.
Download or read book PCR Mutation Detection Protocols written by Bimal D. M. Theophilus and published by Springer Science & Business Media. This book was released on 2008-02-02 with total page 214 pages. Available in PDF, EPUB and Kindle. Book excerpt: 1Bimal D. Theophilus and Ralph Rapley provide biological and clinical investigators with a comprehensive collection of new, recent, and updated PCR-based screening methods suitable for detecting the presence of both known and novel mutations. The methods cover point mutations (e.g., ASO-PCR, SSCP, DGGE, chemical cleavage), deletions (multiplex PCR, FISH, blotting), non-sense mutations (PTT), and more. The new and exciting techniques of DNA array analysis, along with such recently developed experimental methods as conformation-sensitive gel electrophoresis, are also included. Each chapter explains the basic theory behind the technique and provides valuable notes essential for its successful execution.
Download or read book Protein Sequencing Protocols written by Bryan John Smith and published by Springer Science & Business Media. This book was released on 2008-02-02 with total page 489 pages. Available in PDF, EPUB and Kindle. Book excerpt: Determination of the protein sequence is as important today as it was a half century ago, even though the techniques and purposes have changed over time. Mass spectrometry has continued its recent rapid development to find notable application in the characterization of small amounts of protein, for example, in the field of proteomics. The “traditional” chemical N-terminal sequencing is still of great value in quality assurance of the increasing number of biopharmaceuticals that are to be found in the clinic, checking processing events of recombinant proteins, and so on. It is joined in the armory of me- ods of protein analysis by such techniques as C-terminal sequencing and amino acid analysis. These methods are continually developing. The first edition of Protein Sequencing Protocols was a “snapshot” of methods in use in protein biochemistry laboratories at the time, and this, the second edition, is likewise. Methods have evolved in the intervening period, and the content of this book has similarly changed, the content of some chapters having been superceded and replaced by other approaches. Thus, in this edition, there is inclusion of approaches to validation of methods for quality assurance work, reflecting the current importance of biopharmaceuticals, and also a guide to further analysis of protein sequence information, acknowledging the importance of bioinformatics.
Download or read book Liposome Methods and Protocols written by Subhash C. Basu and published by Springer Science & Business Media. This book was released on 2008-02-04 with total page 250 pages. Available in PDF, EPUB and Kindle. Book excerpt: In vitro utilization of liposomes is now recognized as a powerful tool in many bioscience investigations and their associated clinical studies, e.g., liposomes in drug targeting; liposomes in gene transport across plasma and nuclear membranes; liposomes in enzyme therapy in patients with genetic disorders. However, before these areas can be effectively explored, many basic areas in liposome research require elucidation, including: (a) attachment of liposomes to cell surfaces; (b) permeation of liposomes through the plasma membranes; and (c) stability of liposomes in cell or nuclear matrices. None of these areas have been exhaustively explored and liposome researchers have ample opportunities to contribute to our knowledge. The aim of Liposome Methods and Protocols is to bring together a wide range of detailed laboratory protocols covering different aspects of liposome biology in order to assist researchers in those rapidly advancing medical fields mentioned earlier. With this goal in mind, in each protocol chapter we have detailed the materials to be used, followed by a step-by-step protocol. The Notes section of each protocol is also certain to prove particularly useful, since the authors include troubleshooting tips straight from their benchtops, valuable information that is seldom given in restricted methods sections of standard research journals. For this reason we feel that the book will prove especially useful for all researchers in the liposome field.
Download or read book Protein Kinase C Protocols written by Alexandra C. Newton and published by Springer Science & Business Media. This book was released on 2008-02-03 with total page 565 pages. Available in PDF, EPUB and Kindle. Book excerpt: Since the discovery that protein kinase C (PKC) transduces the ab- dance of signals that result in phospholipid hydrolysis, this enzyme has been at the forefront of research in signal transduction. Protein Kinase C Protocols covers fundamental methods for studying the structure, function, regulation, subcellular localization, and macromolecular interactions of PKC. Protein Kinase C Protocols is divided into 11 sections representing the major aspects of PKC regulation and function. Part I contains an introduction and a historical perspective on the discovery of PKC by Drs. Yasutomi Nishizuka and Ushio Kikkawa. Part II describes methods to purify PKC. Part III describes the standard methods for measuring PKC activity: its enzymatic activity and its stimulus-dependent translocation from the cytosol to the membrane. Part IV describes methods for measuring the membrane interaction of PKC in vivo and in vitro. Part V provides methodologies and techniques for measuring the ph- phorylation state of PKC, including a protocol for measuring the activity of PKC’s upstream kinase, PDK-1. Novel methods for identifying substrates are described in Part VI. Part VII presents protocols for expressing and analyzing the membrane targeting domains of PKC. Part VIII provides a comprehensive c- pilation of methods used to identify binding partners for PKC. Part IX describes pharmacological probes used to study PKC. The book ends with a presentation of genetic approaches to study PKC (Part X) and a discussion of approaches used to study PKC in disease (Part XI).
Download or read book E coli Gene Expression Protocols written by Peter E. Vaillancourt and published by Springer Science & Business Media. This book was released on 2008-02-02 with total page 347 pages. Available in PDF, EPUB and Kindle. Book excerpt: Peter E. Vaillancourt presents a collection of popular and emerging methodologies that take advantage of E. coli's ability to quickly and inexpensively express recombinant proteins. The authors focus on two areas of interest: the use of E. coli vectors and strains for production of pure, functional protein, and the use of E. coli as host for the functional screening of large collections of proteins and peptides. Among the cutting-edge techniques demonstrated are those for rapid high-level expression and purification of soluble and functional recombinant protein and those essential to functional genomics, proteomics, and protein engineering.
Download or read book Lentivirus Gene Engineering Protocols written by Maurizio Federico and published by Springer Science & Business Media. This book was released on 2008-02-03 with total page 310 pages. Available in PDF, EPUB and Kindle. Book excerpt: Cell gene engineering is emerging as a field with outstanding impact, not only in medicine/biology, but also, and perhaps most importantly, in agriculture and in all those food sciences involved in the fight against world hunger. Lentivirus vector-based technologies represent the last frontier in the development of powerful and reliable methods for both in vitro and in vivo gene transfer in eukaryotic animal cells. Although the design of lentivirus vectors is closely reminiscent of those already successfully applied to the construction of oncoretroviral vectors, some unique features, e.g., the ef- ciency in transducing both postmitotic and stem cells, render the use of lentivirus vectors invaluable. It has been a great pleasure to edit Lentivirus Gene Engineering Pro- cols, owing in part to the high level of enthusiasm that the authors dem- strated in contributing to this book. The fact that so many outstanding scientists engaged in lentivirus vector research have provided articles renders it so- thing more than a technical handbook. In addition to detailed descriptions of the most innovative methodologies, the reader may find very informative ov- views concerning both theoretical and practical aspects of the origin and the development of diverse lentivirus vector types. This, in my opinion, rep- sents a unique added value of this volume, which should help our work resist the passage of time, to which books such as this are particularly sensitive.
Download or read book Transgenic Mouse Methods and Protocols written by Marten H. Hofker and published by Springer Science & Business Media. This book was released on 2008-02-04 with total page 382 pages. Available in PDF, EPUB and Kindle. Book excerpt: Marten Hofker and Jan van Deursen have assembled a multidisciplinary collection of readily reproducible methods for working with mice, and particularlyfor generating mouse models that will enable us to better understand gene function. Described in step-by-step detail by highly experienced investigators, these proven techniques include new methods for conditional, induced knockout, and transgenic mice, as well as for working with mice in such important research areas as immunology, cancer, and atherosclerosis. Such alternative strategies as random mutagenesis and viral gene transduction for studying gene function in the mouse are also presented.
Download or read book Oxidative Stress Biomarkers and Antioxidant Protocols written by Donald Armstrong and published by Springer Science & Business Media. This book was released on 2008-02-04 with total page 320 pages. Available in PDF, EPUB and Kindle. Book excerpt: The first protocols book, Free Radical and Antioxidant Protocols (1) was published in late 1998. Sections were divided into three parts, covering selected biochemical techniques for measuring oxidative stress, antioxidant (AOX) activity, and combined applications. In choosing the 40 methods to be included in that book, I realized there were considerably more of equal value than that which we could have presented in a single volume. To produce a comprehensive resource, this book and a third are being compiled to expand coverage of the field. A summary of papers (2) published on this important subject emphasizes the continuing rapid growth in oxidative stress investigations relating to our understanding of biochemical reactions, their relevance to pathophysiological mechanisms, how disease may arise, and how therapeutic intervention may be achieved(3). Although there is some overlap between the categories, the ana- sis shown below illustrates where current studies are concentrated and are almost evenly distributed between free radicals and AOX. Over the last 4 yr, there has been a 55% increase in the number of papers published in the area.
Download or read book Septic Shock Methods and Protocols written by Thomas J. Evans and published by Springer Science & Business Media. This book was released on 2008-02-01 with total page 206 pages. Available in PDF, EPUB and Kindle. Book excerpt: Septic shock remains a serious medical condition with high mortality. Despite many advances in intensive care medicine and antibiotic devel- ment, this has not changed appreciably in the last 20 years. Frustratingly, over the same period of time, enormous advances have been made in understa- ing the underlying pathogenic mechanisms of this condition. This has resulted in the development of several novel therapies for septic shock, which, despite excellent theoretical grounds for their efficacy, have failed in altering mort- ity attributable to sepsis. The reasons for these failures are multiple, but it is clear that further research is required aimed at increasing our understanding of the basic pat- physiological processes that occur following infection. Research into septic shock draws upon a number of different disciplines, ranging from molecular and cellular biology to physiological measurements on whole animals. Septic Shock Methods and Protocols is an attempt to draw together into one volume a number of protocols that are of use in the investigation of the mechanisms of septic shock. I have divided the book into five sections. The first deals with endotoxin, the lipopolysaccharide component of the Gram-negative cell m- brane that can mimic many of the features of septic shock. Gram-positive organisms are found increasingly as causes of septic shock, and several - pects of toxins produced from these bacteria are considered in the second s- tion.
Download or read book Recombinant Antibodies for Cancer Therapy written by Martin Welschof and published by Springer Science & Business Media. This book was released on 2008-02-05 with total page 476 pages. Available in PDF, EPUB and Kindle. Book excerpt: Since the advent of hybridoma technology more than two decades ago, numerous antibodies have entered the clinical setting as potent therapeutic agents. Their repeated application in humans, however, is limited by the development of human antimouse antibodies (HAMA) in the recipient, leading to allergic re- tions against the foreign murine protein and rapid neutralization. To circumvent these limitations many new antibodies have recently been tailored through recombinant antibody technology. The initial clinical data show encouraging results, thus demonstrating the potential of these new therapeutic agents. The purpose of Recombinant Antibodies for Cancer Therapy is to present a collection of detailed protocols in recombinant antibody technology. It is pri- rily addressed to scientists working on recombinant antibodies as well as cli- cians involved with antibody-based therapies. As with other volumes of this series, we placed the main focus on providing detailed protocols describing procedures step-by-step. Moreover, each protocol supplies a troubleshooting guide containing detailed information on possible problems and hints for pot- tial solutions. Antibody technology is a subject of constant and rapid change. This volume, therefore, does not attempt to cover all possible current experimental approaches in the field. Rather, we present carefully selected protocols, written by competent authors who have successfully verified the particular method described. Given our own professional backgrounds and interest in oncology, we chose to conc- trate chiefly on therapeutic agents for cancer patients.
Download or read book Molecular Cytogenetics written by Yao-Shan Fan and published by Springer Science & Business Media. This book was released on 2008-02-05 with total page 409 pages. Available in PDF, EPUB and Kindle. Book excerpt: The new techniques of molecular cytogenetics, mainly fluorescence in situ hybridization (FISH) of DNA probes to metaphase chromosomes or interphase nuclei, have been developed in the past two decades. Many FISH techniques have been implemented for diagnostic services, whereas some others are mainly used for investigational purposes. Several hundreds of FISH probes and hybridization kits are now commercially available, and the list is growing rapidly. FISH has been widely used as a powerful diagnostic tool in many areas of medicine including pediatrics, medical genetics, maternal–fetal medicine, reproductive medicine, pathology, hematology, and oncology. Frequently, a physician may be puzzled by the variety of FISH techniques and wonder what test to order. It is not uncommon that a sample is referred to a laboratory for FISH without indicating a specific test. On the other hand, a cytogeneticist or a technologist in a laboratory needs, from case to case, to determine which procedure to perform and which probe to use for an informative result. To obtain the best results, one must use the right DNA probes and have reliable protocols and measures of quality assurance in place. Also, one must have sufficient knowledge in both traditional and molecular cytogenetics, as well as the particular areas of medicine for which the test is used in order to appropriately interpret the FISH results, and to correlate them with clinical diagnosis, treatment, and prognosis.
Download or read book Combinatorial Library written by Lisa B. English and published by Springer Science & Business Media. This book was released on 2008-02-04 with total page 380 pages. Available in PDF, EPUB and Kindle. Book excerpt: The continued successes of large- and small-scale genome sequencing projects are increasing the number of genomic targets available for drug d- covery at an exponential rate. In addition, a better understanding of molecular mechanisms—such as apoptosis, signal transduction, telomere control of ch- mosomes, cytoskeletal development, modulation of stress-related proteins, and cell surface display of antigens by the major histocompatibility complex m- ecules—has improved the probability of identifying the most promising genomic targets to counteract disease. As a result, developing and optimizing lead candidates for these targets and rapidly moving them into clinical trials is now a critical juncture in pharmaceutical research. Recent advances in com- natorial library synthesis, purification, and analysis techniques are not only increasing the numbers of compounds that can be tested against each specific genomic target, but are also speeding and improving the overall processes of lead discovery and optimization. There are two main approaches to combinatorial library production: p- allel chemical synthesis and split-and-mix chemical synthesis. These approaches can utilize solid- or solution-based synthetic methods, alone or in combination, although the majority of combinatorial library synthesis is still done on solid support. In a parallel synthesis, all the products are assembled separately in their own reaction vessels or microtiter plates. The array of rows and columns enables researchers to organize the building blocks to be c- bined, and provides an easy way to identify compounds in a particular well.
