疾病及处理

Animal models are powerful tools to study the pathogenesis of diverse types of candidiasis. Murine models are particularly attractive because of cost, ease of handling, technical feasibility, and experience with their use. In this chapter, we describe methods for two of the most popular mur ...

The measurement of virulence using ex vivo and in vitro models is discussed in the context of the human pathogenic yeast, Candida albicans. The models described are of two types. First, reconstituted tissues of various sorts are used that are derived from human carcinomas. The tissues are grown in v ...

Development of Candida spp. biofilms on medical devices such as catheters and voice prosthesis has been recognized as an increasing clinical problem. Simple device removal is often impossible, while in addition, resulting candidal infections are difficult to resolve due to their incr ...

Biofilm formation is a common complication of the use of prosthetic devices. In clinical settings, biofilms can be comprised of one or more microbial species. In order to investigate the interaction between different species within a biofilm, a reproducible, reliable model system has to be u ...

A filter disk assay is described, which measures the penetration of antifungal agents through Candida biofilms. The technique involves forming a colony biofilm on a polycarbonate membrane filter, and capping it with a second, smaller membrane filter followed by a wetted paper disk of the type ...

Immunoprotection during most forms of candidiasis (oropharyngeal, invasive) is lacking since most candidiasis patients are immunosuppressed either as a result of their allogeneic transplant, cancer chemotherapy, or HIV infection. Consequently, immunization might be con ...

Candida albicans is a major fungal systemic pathogen in humans. Genetic manipulation of C. albicans is unwieldy. We report here a strategy that is useful and successful for large-scale genetic manipulation of C. albicans genes of interest: use of the UAU1 cassette on a Tn7 transposon. Streamlin ...

Genetic transformation is the primary method of genetic manipulation of Candida albicans. The lack of a complete sexual cycle prevents application of classical genetic analyses. However, transformation permits introduction into the genome of a wide variety of defined mutations i ...

Reporter systems are used in Candida albicans in three major experimental areas. These include gene expression, promoter analysis, and protein expression/localization. Heterologous expression in C. albicans is either not effective or inefficient due to the alternative codon us ...

Proteomics is the term for the large-scale analysis of proteins. Such studies have contributed greatly to our understanding of gene function in the postgenomic era. Besides identification, the characterization of protein function is also a key objective of proteomic research. One of the ...

Tandem-affinity purification (TAP) tagging systems, developed by the research group of Bertrand Seraphin and others, are a means of isolating physiologically relevant protein and protein–nucleic acid complexes. Where complete (or nearly complete) genome sequence data are ava ...

Upon uptake into a host cell, the intracellular bacterium Legionella pneumophila is not degraded on the lysosomal pathway but efficiently establishes a highly specialized replicative vacuole in which it readily multiplies. As many Icm/Dot type 4 secretion translocated bacterial ...

Those investigators who study the morphology of Legionella and Legionella-infected cells have greatly benefited from the superior resolution afforded by electron microscopy (EM). It can also be said with confidence that EM will continue to reveal as yet to be discovered features of this f ...

Legionella pneumophila replicates intracellularly in environmental and immune phagocytes within a unique membrane-bound compartment, the Legionella-containing vacuole (LCV). Formation of LCVs is strictly dependent on the Icm/Dot type IV secretion system and the translo ...

Legionella pneumophila the causative agent of Legionnaires’ disease, actively manipulates host cell �processes to establish a membrane-bound replication vacuole permissive for its replication. Establishment of such replication niche requires the Dot/Icm type IV secret ...

The translocation of effector proteins by the Dot/Icm type IV secretion system is central to the ability of Legionella pneumophila to persist and replicate within eukaryotic cells. The subcellular localization of translocated Dot/Icm proteins in host cells provides insight into th ...

Bacterial virulence proteins often mimic host eukaryotic proteins to modify or disturb host cellular �pathways. Increasing lines of evidence show that many bacterial effector proteins have E3 ubiquitin ligase activity. The effector protein LubX is one such bacterial E3 ubiquitin l ...

The intracellularly replicating lung pathogen Legionella pneumophila expresses a multitude of different phospholipases which are important virulence tools during host cell infection. To study the lipolytic properties including substrate specificities of potential L. ...

Although the study of protozoology has been active for centuries, very few current academic curricula incorporate requirements or even options for coursework on the study of protists; yet, protozoa are becoming widely recognized by investigators as organisms that play a significant ...

The professional phagocyte Dictyostelium discoideum is a simple eukaryotic microorganism, whose natural habitat is deciduous forest soil and decaying leaves, where the amoebae feed on bacteria and grow as separate, independent, single cells. In the last decade, the organism has been s ...