实验基础

Hemocytes of the horseshoe crab contain a family of arthropod peptide antibiotics, termed the tachyplesin family, and an antibacterial protein, called anti-LPS factor, of which the former is located in the small (S) granules and the latter in the large (L) granules of the hemocytes (1–5). In our ongo ...

The human mitochondrial genome was completely sequenced in 1981 by Anderson and co-workers (1) and consists of a closed circular supercoiled DNA molecule of 16,569 base pairs. Mammalian cells characteristically contain a few hundred to several thousand mitochondria, each with 2-10 cop ...

The endosymbiotic hypothesis proposes that mitochondria were once freeliving organisms that colonized another cell. Millions of years of evolutionary pressure created the present-day situation in which the mitochondrion is a semiautonomous organelle that is fully integra ...

The neutral/neutral two-dimensional (2D) agarose gel technique has proved to be a powerful tool for analyzing DNA structure. It was initially adapted and developed by Brewer and Fangman to localize and define bidirectional DNA replication origins on plasmids in yeast (1). Since then, neut ...

Although a variety of degenerative diseases are now known to be caused by two mutations in mitochondrial genes, the pathophysiology of these diseases remains poorly understood. As a consequence, relatively little progress has been made in developing new therapies for mitochondrial d ...

The human mitochondrial genome (see Fig. 1) is a 16,569-bp (base pair) circle of double-stranded DNA (1). It contains genes encoding 2 ribosomal RNAs, 22 transfer RNAs, and 13 structural genes, all of which are subunits of the respiratory chain complexes. Of the 13 structural genes, 7 encode subunits of c ...

Recently, clear evidence has been obtained in our laboratory of a large accumulation of specific point mutations at the same critical control sites for replication in mitochondrial DNA (mtDNA) of human fibroblasts (1) and skeletal muscles (2). This demonstration was based on the use of a very se ...

The ability to measure rare mutational events in mitochondrial genomes from human blood and tissues without resorting to phenotypic selection is invaluable. It is essential for the study of the cause(s) of mitochondrial mutation and for the identification of mutations associated with ...

Resistance of bacteria to antibiotics has become one of the main problems in human health. In addition, in agronomy, microbial diseases are largely responsible for the decrease in agricultural production. The discovery of new antibiotic families is a way to circumvent such problems and ant ...

The advent of high performance liquid chromatography (HPLC) has greatly accelerated the discovery, purification, and characterization of antimicrobial peptides. Virtually every modern study of an antimicrobial peptide includes or was preceded by a description of its purific ...

Peptide antibiotic research, which in the larger sense includes protein antibiotic research, actually began during the late 19th century with the work of Ehrlich, Metchnikov, Kanthack, and Petterson. Now it has been absorbed into the fields of microbiology, immunology, histochemist ...

It has been established that at least three of the neutrophil-derived cationic antibiotic proteins, in addition to their antibiotic activity, can bind lipopolysaccharide. It is generally accepted that the binding to lipopolysaccharide, an outer membrane component of Gram-negat ...

The traditional view of the polymorphonuclear leukocyte (PMN) or neutrophil has been a cell whose primary function was to ingest and kill bacteria. It is becoming increasingly clear that a number of these granule constituents that were originally studied because of their antibiotic acti ...

All living organisms are constantly confronted by invading microbes. In mammals, host defense against microbial invasion requires specialized cellular and extracellular elements in blood and a complex signalling machinery that triggers their mobilization to extravascul ...

Since the development of methods for in vitro cultivation of the asexual blood stages of P.faZciparum (1) and the production of mature gametocytes from such cultures capable of infecting mosquitoes (2), it has been possible to conduct a wide range of studies on the sexual stages of P. filciparum. The ...

This chapter deals with relatively simple ways to use control charts to monitor the performance of ELISAs. A rinderpest competition ELISA, for the estimation of antibodies in serum samples, is used to demonstrate the methods. This assay is available in a kit form. Constant evaluation of the use of t ...

The aim of this chapter is to illustrate the principles of ELISA by (1) showing worked examples of each assay, including diagrams of plates and representational data from assays, (2) analyzing such data in terms of important rules that are learned at each stage, and (3) providing full working instruc ...

This chapter examines in detail the necessary practical skills needed to facilitate the development and sustainability of ELISAs. Basic methodologies to set up all systems are discussed so that the reader can both investigate the possibilities of tests using his or her reagents and use with ...

This chapter defines the terms and examines the configurations used for most applications of ELISA. Such a chapter is important because the possibilities inherent in the systems of ELISA must be understood in order to maximize their versatility in assay design. All heterogeneous systems h ...

This chapter consists of a test concerning ELISA and the sciences needed to perform assays. The test can be used to gage knowledge on ELISA. Suggested points are given for the benefit of educators. Of course, the test can be adapted. All the answers can be obtained by reading this book. It may be useful to test ones ...