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Monitoring Microbial Activities Using Membrane Inlet Mass Spectrometry

Monitoring microbial activities in the environment is difficult owing to the lack of suitable methods. For a technique to be useful for monitoring in situ activities, it must possess the following properties: sensitivity, selectivity, stability, the ability to make continuous real-t ...

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Dielectrophoresis

The increasing trend toward quality assurance programs and hazard analysis, consumer demand for a wide variety of wholesome foods, and legislative pressures (e.g., the U.K. Food Safety Act) have increased the need for more rapid microbiological analyses. Although hazard analysis and cr ...

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Reporter Gene Expression for Monitoring Microorganisms in the Environment

Reporter gene is a generic term that is used to describe genes whose expression is indicative of the presence of a particular cell, a specific event occurring within that cell, or a reflection of the expression of other, usually targeted genes of interest. For example, the presence of such a gene may make an ...

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Pharmacogenetics of Resistance to Cisplatin and Other Anticancer Drugs and the Role of Sphingolipid Metabolism

Dictyostelium discoideum has proven to be a useful lead genetic system for identifying novel genes and pathways responsible for the regulation of sensitivity to the widely used anticancer drug cisplatin. Resistance to cisplatin is a major factor limiting the efficacy of the drug in treat ...

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N-Glycomic and N-Glycoproteomic Studies in the Social Amoebae

N-glycans modify the great majority of all secreted and plasma membrane proteins, which themselves constitute one-third to one-half of the proteome. The ultimate definition of the glycoproteome would be the identification of all the N-glycans attached to all the modified asparaginyl s ...

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Measuring Cheating, Fitness, and Segregation in Dictyostelium discoideum

Dictyostelium has become a model organism for the study of social evolution because of the stage in its life cycle where thousands of independent amoebae together form a fruiting body. Some individuals die to form a stalk that holds aloft the remaining cells for dispersal to new environments as sp ...

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The Application of the Cre-loxP System for Generating Multiple Knock-out and Knock-in Targeted Loci

Dictyostelium discoideum is an exceptionally powerful eukaryotic model to study many aspects of growth, development, and fundamental cellular processes. Its small-sized, haploid genome allows highly efficient targeted homologous recombination for gene disruption and k ...

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Investigation of DNA Repair Pathway Activity

DNA is constantly being damaged from endogenous and exogenous sources and efficient repair of different types of DNA lesions is essential for the survival of the organism. Dictyostelium is highly resistant to DNA damage and its genome sequence has revealed the presence of multiple repair p ...

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Isolation of Dictyostelium Nuclei for Light and Electron Microscopy

The significance of amoebae for studies of nuclear architecture has considerably increased in the recent years. The availability of a protocol for isolation of nuclei in a quality sufficient for high-resolution light and electron microscopy is a prerequisite for such studies. Here we pr ...

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Extrachromosomal Inducible Expression

Inducible expression systems are very convenient for proteins that induce strong side effects such as retardation of growth or development and are essential for the expression of toxic proteins. In this chapter we describe the doxycycline-inducible expression system, optimized f ...

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Mitochondrial Respiratory Complex Function and the Phenotypic Consequences of Dysfunction

Dictyostelium provides a well established model system for the study of mitochondrial biology and disease. Mitochondrial dysfunction in Dictyostelium has been generated by knockout of nonessential nuclear genes encoding mitochondrial proteins, by knockout of targeted mit ...

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Analysis of Mitochondrial Gene Expression

Dictyostelium provides a well-established model system for the study of mitochondrial biology and disease. A complete mitochondrial transcription and RNA-processing map has been generated, while the start site for transcription and the responsible RNA polymerase have also been ...

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Transcript Localization in Dictyostelium discoideum Cells by RNA FISH

Biogenesis of ribosomal RNA (rRNA) takes place preliminary in the nucleolus of eukaryotic cells, the site of rDNA transcription. Several processing steps of rRNA molecules have been implied to take place in the cytoplasm. To follow these processing events we have adapted protocols for fluo ...

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The Amoebozoa

The model organism Dictyostelium discoideum is a member of the Amoebozoa, one of the six major �divisions of eukaryotes. Amoebozoa comprise a wide variety of amoeboid and flagellate organisms with single cells measuring from 5 μm to several meters across. They have adopted many different li ...

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Quantitative Analysis of Phagocytosis and Phagosome Maturation

Phagocytosis and phagosome maturation lead to killing and digestion of bacteria by protozoans and innate immune phagocytes. Phagocytosis of particles expressing or coupled to various fluorescent reporters and sensors can be used to monitor quantitatively various parameters ...

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Micropipette Aspiration for Studying Cellular Mechanosensory Responses and Mechanics

Micropipette aspiration (MPA) is a widely applied method for studying cortical tension and deformability. Based on simple hydrostatic principles, this assay allows the application of a specific magnitude of mechanical stress on cells. This powerful method has revealed insights ab ...

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Setting Up and Monitoring an Infection of Dictyostelium discoideum with Mycobacteria

Mycobacterium marinum is the causative agent of fish and amphibian tuberculosis in the wild. It is a genetically close cousin of Mycobacterium tuberculosis, and thereby the infection process remarkably shares many of the hallmarks of M. tuberculosis infection in human, at both the cellul ...

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Isolation of Pathogen-Containing Vacuoles

Dictyostelium discoideum cells are “professional phagocytes,” as they ingest a large variety of bacteria, yeast, and inert particles. Several bacterial pathogens are able to survive intracellularly within specialized vacuoles of D. discoideum by interfering with host signali ...

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Monitoring Autophagy in Dictyostelium

Autophagy is an intracellular degradation mechanism essential for cell survival and maintenance of cellular homeostasis, differentiation, and development. Recent research has highlighted the impact of autophagy in neurodegenerative diseases and aging. We are still far from ...

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Secretory Lysosomes in Dictyostelium: Visualization, Characterization, and Dynamics

Dictyostelium discoideum is a unicellular eukaryotic model largely used for the study of the endocytic pathway, mainly due to its high resemblance to the mammalian pathway. Over the last years, Dictyostelium has been increasingly employed for investigating the biogenesis and secre ...

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