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REFLEX Final Report

REFLEX Final Report

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Published by Disiciencia
Risk Evaluation of Potential Environmental Hazards From Low Frequency Electromagnetic Field Exposure Using Sensitive in vitro Methods
Risk Evaluation of Potential Environmental Hazards From Low Frequency Electromagnetic Field Exposure Using Sensitive in vitro Methods

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Categories:Types, Research, Science
Published by: Disiciencia on Oct 03, 2011
Copyright:Attribution Non-commercial


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Risk Evaluation of Potential EnvironmentalHazards From Low Frequency ElectromagneticField Exposure Using Sensitive
in vitro
A project funded by the European Unionunder the programme
Quality of Life and Management of Living Resources
Key Action 4 "Environment and Health"Contract: QLK4-CT-1999-01574Start date: 01 February 2000End date: 31 May 2004Acronym: REFLEX
Table of contents
1.0 INTRODUCTION 72.0 MATERIAL AND METHODS 92.1 Exposure setups (Participant 10) 9
2.1.1 ELF-EMF exposure setup 92.1.2 RF-EMF exposure setup 10
2.2 Experiments with human fibroblasts, lymphocytes, monocytes, melanocytes and musclecells and with granulosa cells of rats (Participant 3) 12
2.2.1 ELF and RF-EMF exposure setups 122.2.2 Cell culture and exposure conditions 122.2.3 Comet assay 132.2.4 Micronucleus assay 132.2.5 Chromosomal aberrations 132.2.6 Fluorescence in situ hybridisation (FISH) 142.2.7 Changes in mitochondrial membrane potential (JC-1 staining) 142.2.8 Statistical analysis 15
2.3 Experiments with human HL-60 cells (Participant 2) 15
2.3.1 RF-EMF exposure setup 152.3.2 Cell culture and exposure conditions 152.3.3 in vitro genotoxicity tests 162.3.4 in vitro cytotoxicity testing 172.3.5 Preparation of nuclei suspensions from cells for flow cytometry analysis 172.3.6 Flow cytometric exclusion of apoptosis via Annexin V assay and TUNEL assay 182.3.7 Reactive oxygen species (ROS) and antioxidant enzyme activity 182.3.8 Analysis of cellular growth behaviour 202.3.9 Statistics 202.3.10 Proteomics 202.3.11 Gene expression profiling 21
2.4. Experiments with the human neuroblastoma cell line NB69 (Participant 5) 21
2.4.1 ELF-EMF exposure setup 212.4.2 RF-EMF exposure setup 222.4.3 Cell culture and EMF-Exposure 222.4.4 Immunocytochemical characterisation of NB69 Cells 232.4.5 Immunocytochemical characterisation of neural stem cells (NSC) 242.4.6 Immunocytochemical staining for the Cell Nuclear Antigen (PCNA). 242.4.7 5-bromo-2'-deoxyuridine (BrdU) labelling for identification of cells synthesising DNA. 24
QLK4-CT-1999-01574 / REFLEX / Final Report / I

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