Parasite infections are very widespread; however, effective therapeutics are not yet available. This manual with detailed protocols for analysing parasitological properties, pathways or interactions with hosts should help to develop new diagnostic tests, therapeutic approaches or even vaccination.

Klappentext

This laboratory manual with detailed background information and practical protocols will be a useful guide for researchers engaged in all areas of parasitology. Most techniques described here are suitable for helminths as well as protozoa, but also included are protocols for particular parasite species. Parasite infections are very widespread. However, effective therapeuticals are not yet available. The analysis of the life cycle and parasite host interactions at the molecular level may help in the search of the "Achilles heel" of a parasite and thus promote the development of new therapeutic approaches. Parasite molecules such as surface antigens, excretory proteins or metabolic enzymes may serve as targets for new diagnostics tests, chemo- or immunotherapeutics or even vaccination.

Inhalt
1 Biochemical Pathways in Parasites.- 1.1 Introduction.- 1.2 Preparation of Enzyme Fractions.- 1.3 Enzyme Assays.- 1.4 Metabolite Measurements.- 1.5 Use of Inhibitors.- References.- 2 Electrophoresis of Parasite Proteins.- 2.1 Introduction.- 2.2 Lysis of Parasites and Extraction of Proteins.- 2.3 SDS-PAGE.- 2.4 Modifications of Standard Method.- 2.5 Applications of SDS-PAGE to the Characterisation of Individual Polypeptides.- References.- 3 Molecular Techniques in Analytical Parasitology.- 3.1Introduction.- 3.2 Isolation of Parasite Nucleic Acid.- 3.3 Gel Electrophoresis and Blotting of Nucleic Acid.- 3.4 Labelling and Hybridisation of DNA Probes.- References.- 4 The Production and Analysis of Helminth Excretory-Secretory (ES) Products.- 4.1 Introduction.- 4.2 Collection of Excretory-Secretory (ES) Products.- 4.3 Acetylcholinesterase.- 4.4 Proteolytic Enzymes.- 4.5 Anticoagulants, Blood Coagulation Assays.- 4.6 Potential Immunomodulators.- 4.7 Concluding Remarks.- References.- 5 Parasite Proteinases.- 5.1 Introduction.- 5.2 Sample Preparation.- 5.3 Substrates and Proteinase Assays.- 5.4 Proteinase Inhibitors.- 5.5 Electrophoretic Methods.- 5.6 Purification Methods.- 5.7 Endogenous Inhibitors.- 5.8 Final Comments.- References.- 6 Neurobiology of Helminth Parasites.- 6.1 Introduction.- 6.2 Immunocytochemical Localisation of Neuroactive Substances in Helminth Parasites.- 6.3 Characterisation and Purification of Neuropeptides.- 6.4 Physiological Action of Transmitters and Drug Assessment In Vitro.- References.- 7 Electron Microscopy in Parasitology.- 7.1 Introduction.- 7.2 Preparation for Transmission Electron Microscopy (TEM).- 7.3 Cytochemistry and Staining Protocol in TEM.- 7.4 Preparation for Scanning Electron Microscopy (SEM).- 7.5 Immunogold-Labelling Techniques.- References.- 8 Studies of the Surface Properties, Lipophilic Proteins and Metabolism of Parasites by the Use of Fluorescent and Caged Compounds.- 8.1 Introduction.- 8.2 Incorporation of Fluorescent Compounds into Membranes.- 8.3 Using Fluorescent Techniques to Measure Membrane Fluidity.- 8.4 Distribution of Actin.- 8.5 Assessment of Parasite Membrane Damage.- 8.6 Localisation of Probes in Organelles.- 8.7 Measurement of pH at the Surface of Parasites and Intracellular pH.- 8.8 The Use of Caged Compounds.- 8.9 Use of Fluorescent Probes in the Analysis of Parasite Proteins.- References.- 9 Detection and Identification of Parasite Surface Carbohydrates by Lectins.- 9.1 Introduction.- 9.2 Lectin-Mediated Trypanosomatid Parasite Agglutination.- 9.3 Fluorescein-Isothiocyanate (FITC)-Conjugated Lectins.- 9.4 Peroxidase-Labelled Lectins.- 9.5 Biotinylated Lectins.- 9.6 Fixatives and Enzymatic Treatments.- 9.7 Histological Sections.- 9.8 Controls.- References.- 10 Immunological Analysis of Parasite Molecules.- 10.1 Introduction.- 10.2 Parasite Extracts.- 10.3 Production of Antibodies to Parasite Molecules.- 10.4 Enzyme Linked Immunosorbent Assays (ELISA).- 10.5 Immunoblotting.- 10.6 Affinity Chromatography.- References.