Suchergebnisse

Cover -- Copyright -- Acknowledgements -- Abstract and Benefits -- Table of Contents -- List of Tables -- List of Figures -- List of Acronyms -- Executive Summary -- Chapter 1.0: Introduction, Rationale and Pilot Studies for Microarray Development -- 1.1 Introduction -- 1.2 Rationale -- 1.2.1 Overview, Aquatic Toxicity Testing -- 1.2.2 Pharmaceutical Fate Distribution and Detection in the Aquatic Environment -- 1.2.3 Fate of Chemicals in Wastewater Treatment Processes -- 1.2.4 Gene Arrays -- 1.3 Hypothesis and Objectives -- 1.4 Materials and Methods -- 1.4.1 Animal Stocks -- 1.4.2 Exposures -- 1.4.3 Protein Analysis -- 1.5 Preliminary Studies and Project Preparation -- 1.5.1 IACUC Protocol and Developing Sufficient Fish Stock -- 1.5.2 Pilot Study: Range Finding -- 1.5.3 Pilot Study: PMP 70 Study -- 1.6 Study Plan -- Chapter 2.0: Suppressive Subtractive Hybridization: Acquisition of Medaka Specific cDNAs -- 2.1 Introduction -- 2.2 Materials and Methods -- 2.2.1 Animal Exposures -- 2.2.2 RNA Extraction -- 2.2.3 Subtractions (SSH) -- 2.2.4 DNA Sequencing -- 2.3 Results and Discussion -- 2.3.1 RNA Extraction -- 2.3.2 Subtraction Whole Fish -- 2.3.3 Subtractions Individual Tissues -- 2.3.4 Differential Screening: Selection of Positive and Negatively Regulated Clones -- 2.3.5 Clone Sequencing and Analysis -- 2.3.6 Tissue Specificity -- 2.3.7 Sequence Annotation -- 2.4 Concluding Remarks -- Chapter 3.0: Microarray Construction and Analysis -- 3.1 Introduction -- 3.2 Methods -- 3.2.1 Chemicals -- 3.2.2 Test Animals -- 3.2.3 Aqueous Exposures -- 3.2.4 RNA Isolation -- 3.2.5 cDNA Microarray Fabrication and Hybridizations -- 3.3 Results and Discussion -- 3.3.1 Maro/Microarray Array Design, Construction, Testing and Validation -- 3.3.2 Microarray Analysis of Fibrate Pharmaceuticals -- 3.4 Concluding Remarks.