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T-Cell Cytotoxicity Assays for Studying the Functional Interaction Between the Superantigen Staphylococcal Enterotoxin A and T-Cell Receptors

The superantigens (SAgs) staphylococcal enterotoxins (SEs) are produced by certain strains of Staphylococcus aureus and comprise structurally related bacterial proteins, which are among the most potent mitogens known for murine and human T lymphocytes (1,2). T-cell activation ...

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In Vitro Physiological Studies on Clostridial Neurotoxins: Biological Models and Procedures for Extracellular and Intracellular Application of Toxins

Botulinum (BoNT, serotypes A-G, see also Chapter 2) and tetanus (TeNT) neurotoxins are known under the generic term of clostridial neurotoxins. These dichainal proteins comprise a light (Mr R~50) and a heavy (Mr R~100) chain that are disulfide linked. In mammals, these proteins are the causative a ...

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The Biology of Endotoxin

Approximately 100 yr ago Richard Pfeiffer, a co-worker of Robert Koch in Berlin, discovered that cholera bacteria produced, in addition to heat-labile exotoxin, another toxin (1). In contrast to the secreted exotoxins this new, heat-stable toxin was found to be a constituent of the bacterial ce ...

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Applications of Combined Capillary Electrophoresis-Electrospray Mass Spectrometry in the Characterization of Short-Chain Lipopolysaccharides: Haemophi

Lipopolysaccharide (LPS) is an essential component of the outer membrane of all Gram-negative bacteria (1). This complex class of lipoglycans can trigger a cascade of immunological responses in mammals including endotoxic effects and serum antibody production. LPSs have been found ...

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Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry of Lipopolysaccharides

The outer membrane of Gram-negative bacteria, which include many human pathogens, contains various proteins, polysaccharides, and glycolipids. Of these, lipopolysaccharides (LPS) are of particular microbiological, immuno-logical, and medical importance. As the major amp ...

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Deacylation of Lipopolysaccharides and Isolation of Oligosaccharide Phosphates

Two types of lipopolysaccharides (LPS) exist: smooth (S) and rough (R) forms (1–3). Both LPS forms are found in wild-type bacteria. They consist of a lipid moiety, lipid A, which comprises a (phosphorylated) disaccharide of glucosamine or 2,3-diamino-2,3-dideoxy-D-glucose that is acylated by ...

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Bacterial Protein Toxins: An Overview

This short overview attempts to highlight the current state of the art relevant to bacterial protein toxins. In particular we outline the major achievements in this field during the past decade and briefly describe some significant hallmarks of toxinological research since the advent of ...

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Molecular Diagnosis of Chlamydia trachomatis Infections by Probe Hybridization, PCR, LCR,TMA, and Q- Replicase

The use of cell cultures for the laboratory diagnosis of Chlamydia trachomatis infections was popularized during the 1970s and 80s (1–4). The techniques required live organisms and were restricted to specialized laboratories. During the 1980s the detection of chlamydia-specific a ...

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Hepatitis B Virus: Detection and Quantitation by Membrane and Liquid Hybridization, Branched DNA Signal Amplification, Hybrid Capture, and PCR Methods

Hepatitis B virus (HBV) is a member of the Hepadnaviridae family and has a (3200-bp) partially double-stranded circular DNA genome (1). This virus causes subclinical, acute self-limited, chronic, and fulminant hepatic infections. Chronic HBV infection is particularly important bec ...

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Isolation of Foamy Viruses From Peripheral Blood Lymphocytes

The isolation of a retrovirus from peripheral blood lymphocytes/monocytes can be a difficult task, requiring the fulfillment of three essential parameters. First, this viral agent must infect such cells in vivo. Second, these circulating cells should harbor wild-type proviruses. Fi ...

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Detection of Genital Mycoplasmas by PCR

Mycoplasmas are the smallest prokaryotes capable of self-replication. They belong to the class Mollicutes (meaning soft-skin) and have evolved regressively, by genome reduction, from Gram-positive bacterial ancestors, namely certain clostridia (1). The taxonomy of the class Mo ...

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Detection of Treponema pallidum, Haemophilus ducreyi, and Herpes Simplex Virus by Multiplex PCR

The three major causes of genital ulcer disease (GUD) are herpes simplex virus (HSV), Treponema pallidum, and Haemophilus ducreyi. Although techniques exist for the laboratory diagnosis of all three organisms, constraints of cost, availability of equipment and expertise, and the lack of ...

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Haemophilus ducreyi Detection by PCR

Genital ulcers are typically caused by one of three organisms, Haemophilus ducreyi, Treponema pallidum, or herpes simplex virus, which cause chancroid, syphilis, and genital herpes, respectively. Although traditionally these diseases have been differentiated by their clinic ...

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The Impact of Molecular Technology on STD Control: A Historical Perspective

Sexually transmitted diseases have afflicted humankind for millennia, based on references to apparent gonorrhea or nongonococcal urethritis in the Old Testament (Leviticus). For most of history there has been no means of specific diagnosis, and clinical diagnosis of syndromes was f ...

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Molecular Techniques for HIV and STDs: Implications for Research and Disease Control in the New Millennium

Molecular techniques have gradually shifted the paradigm in the laboratory diagnosis of sexually transmitted infections from biological to molecular amplification. It is now possible to combine the sensitivity and specificity of culture with all the convenience of nonculture ...

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Collection and Processing of Seminal Plasma for the Quantitation of HIV-1 RNA by NASBA and RT-PCR

Semen is the major vehicle for the sexual transmission of HIV-1. The ability to isolate infectious HIV from the semen and to quantitate viral burden in the form of cell-free or cell-associated HIV-1 RNA in semen are important for epidemiologic and public health aspects of the epidemic. Earlier stud ...

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Quantitation of HIV-1 RNA in Dried Plasma Spots (DPS): A Field Approach to Therapeutic Monitoring

The ability to measure accurately viral RNA in the plasma (1–3) and intracellular (4–7) compartments of HIV-1-infected persons has led to a dramatic improvement in our understanding of the natural history of HIV-1/AIDS. A number of recent studies have convincingly demonstrated that high le ...

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Human Papillomavirus Detection by PCR and Typing by Dot-Blot

The papillomaviruses form a nonenveloped virion with an icosahedral capsid structure and contain a double-stranded circular DNA genome of 7800–7900 bp. The HPV genome is organized into three major regions; two protein-coding regions (1) early and (2) late-expressing genes, and (3) a nonco ...

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Genotyping Chlamydia trachomatis by PCR

Strain identification of Chlamydia trachomatis has historically been accomplished using serotyping as a phenotypic marker to differentiate chlamydial isolates (1). The target for serotyping is the major outer membrane protein (MOMP) which is the most antigenically diverse and ...

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Neisseria gonorrhoeae: Detection and Typing by Probe Hybridization, LCR, and PCR

Neisseria gonorrhoeae, first described by Neisser in 1879, is a Gram-negative, nonmotile, nonspore-forming diplococcus, belonging to the family Neisseriaceae. It is the etiologic agent of gonorrhea. The other pathogenic species is Neisseria meningitidis, to which N. gonorrhoeae ...

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