前沿科技及其它

Histologically distinct noninvasive precursor lesions have been recognized in the pancreas for close to a century. The recent development of a consistent reproducible nomenclature and classification system for these lesions has been a major advance in the study of these noninvasi ...

Collagen-induced arthritis (CIA) is an animal model of rheumatoid arthritis (RA) that is widely used to address questions of disease pathogenesis and to validate therapeutic targets. Arthritis is normally induced in mice or rats by immunization with autologous or heterologous type II c ...

With the exponential increase in the number of genes identified by various genome projects, it has become imperative that efficient methods be developed for deciphering gene function. Genetically engineered strains of mice are now critical research tools for basic biomedical resea ...

Tumor necrosis factor α (TNFα) appears to be the main mediator causing the systemic manifestations of sepsis and septic shock associated with edema (1,2). In vitro it augments movement of permeability tracers across endothelial cell monolayers (3,4). Herein we describe a method which perm ...

In most cellular systems tumor necrosis factor (TNF) induces apoptotic cell death. However, in some particular cell lines, such as the L929sA fibrosarcoma, TNF induces necrotic cell death. This effect is not the result of an inability to die apoptotically, because triggering of Fas in L929sAh ...

The protocols for immunohistochemical localization of tumor necrosis factor α (TNFα), TNFRp55, and TNFRp75 in central nervous system (CNS) slices from laboratory animals are described. The procedure requires intracardiac perfusion of animals followed by fixation and cryostat ...

Polymorphisms in the promoter of the tumor necrosis factor alpha (TNFα) gene have been reported to affect the transcription rate and the release of this cytokine. Among the best studied, the −238 and −308 polymorphisms have been associated with an increased transcriptional activity and TN ...

Tumor necrosis factor (TNF) is a pleiotropic cytokine with a wide range of biological activities including cytotoxicity, immune-cell proliferation, and mediation of inflammatory responses. Mutational analysis of mature TNF has been facilitated by the high expression levels th ...

The finding that the two tumor necrosis factor receptors (TNFR) exist in soluble form in various body fluids not only has substantiated the paradigm of naturally existing soluble cytokine receptors but also has represented a milestone on the road to the biochemical and biological charact ...

Here we describe the methods for the expression of human and murine soluble tumor necrosis factor (hTNF and mTNF) in Escherichia coli cells and for their extraction, purification, and characterization. The expression and purification procedure takes about 2 wk. Human and murine TNF can be pu ...

Tumor necrosis factor (TNF) was originally described as a molecule with antitumor properties released by macrophages stimulated with bacterial products. Almost at the same time that TNF was cloned, it was found to be identical to cachectin, a mediator of cachexia. After the finding of this sec ...

A double-labeling assay for detecting apoptotic cells, using the terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end-labeling (TUNEL) assay, and antigens of interest, using immunofluorescence, is described. The assay has been used successfully on fixed, cult ...

Widespread inherent or acquired resistance to cytotoxic drugs is a major limitation to chemotherapy. There are many mechanisms that contribute to such resistance. In neuroblastomas there is evidence that acquired drug resistance may be associated with altered response to growth fa ...

The ability to examine gene expression differences during cancer progression is crucial to our understanding of the biological events that lead to uncontrolled growth of malignant cells. Differential expression screening of complementary DNA libraries is described in detail h ...

Oxidative stress has been shown to have a myriad of effects on cells. Treatment of cells with oxidants, such as hydrogen peroxide or agents that induce reactive oxygen intermediates, has been shown to induce many cellular signaling pathways and, in some cases, cell apoptosis. Many chemotherap ...

Gene targeting is one of the most powerful tools to define the role of signaling molecules in animal development and disease etiology. By using this technique, nearly 1000 knockout mice have been produced over the last two decades. Generating knockout mice, however, is a time-consuming proced ...

Molecular analysis of prostate cancer specimens is complicated by tumor heterogeneity and admixture of malignant cells with benign epithelium, stroma and inflammatory cells. Laser Capture Microdissection allows procurement of selected groups of cells from specific areas of t ...

Coimmunoprecipitation is a powerful tool to study protein-protein interactions and can be used to test for the physical association between a known protein kinase and its substrate. In this chapter, the author describes a protocol for the preparation of a cell lysate, the immunoprecipita ...

Acquiring functional knowledge from protein-protein interaction studies often necessitates the production of binding-impaired mutants and the study of their effects in biological systems. In many cases, multiple ligands compete for binding to the same protein domain and it beco ...

Development of sensitive methods to monitor and quantitatively assess the expression levels of endogenous genes and the association-interaction of proteins in living cells and whole organisms is a complex and challenging problem. In this chapter, we have described basic methods for ...