Assessment of space environmental factors by cytotoxicity bioassays

Cellular bioassays for detection of cyto- and genotoxicity are useful in the risk assessment of space environmental factors. Such bioassay systems have the potential complement the physical detector systems used in space, insofar as they yield intrinsically biologically weighted measures of cellular responses. The experiment Cellular Responses to Radiation in Space (CERASP) has been selected by NASA/ESA to be performed on the International Space Station. It will supply basic information on the cellular response to radiation applied in microgravity. One of the biological endpoints under investigation will be survival reflected by radiation-dependent reduction of constitutive expression of the enhanced variant of green fluorescent protein (EGFP), originally isolated from the bioluminescent jellyfish Aequorea victoria. In this ground based study, the usefulness of this approach in comparison to standard techniques (colony forming ability test, MTT test) is shown.

[1]  R. Muschel,et al.  The molecular basis for cell cycle delays following ionizing radiation: a review. , 1994, Radiotherapy and oncology : journal of the European Society for Therapeutic Radiology and Oncology.

[2]  L. Poulsen,et al.  New Unstable Variants of Green Fluorescent Protein for Studies of Transient Gene Expression in Bacteria , 1998, Applied and Environmental Microbiology.

[3]  B. Kramer,et al.  Evaluation of in vitro chemosensitivity using human lung cancer cell lines , 1996, Journal of cellular biochemistry. Supplement.

[4]  Giuseppe Schettino,et al.  Low-Dose Binary Behavior of Bystander Cell Killing after Microbeam Irradiation of a Single Cell with Focused CK X Rays , 2005, Radiation research.

[5]  W W Ward,et al.  Green fluorescent protein as a reporter of gene expression and protein localization. , 1995, BioTechniques.

[6]  O. Hug,et al.  Stochastik der Strahlenwirkung , 1966 .

[7]  D. Scudiero,et al.  New colorimetric cytotoxicity assay for anticancer-drug screening. , 1990, Journal of the National Cancer Institute.

[8]  J. G. Brewen,et al.  Cytogenetic effects of environmental mutagens in mammalian cells and the extrapolation to man. , 1974, Mutation research.

[9]  Elke Genschow,et al.  Embryotoxicity Screening Using Embryonic Stem Cells in vitro: Correlation to in vivo Teratogenicity , 1999, Cells Tissues Organs.

[10]  O. Shpilberg,et al.  Appraisal of the MTT-based Assay as a Useful Tool for Predicting Drug Chemosensitivity in Leukemia , 2003, Leukemia & lymphoma.

[11]  M. Chalfie,et al.  Green fluorescent protein as a marker for gene expression. , 1994, Science.

[12]  G. Horneck,et al.  On the Suitability of Red and Enhanced Green Fluorescent Protein (DsRed/EGFP) as Reporter Combination , 2002 .

[13]  R. Cox,et al.  Molecular mechanisms of radiation oncogenesis. , 1994, International journal of radiation biology.

[14]  G. Horneck,et al.  Fluorometric Analysis of DNA Unwinding (FADU) as a Method for Detecting Repair-induced DNA Strand Breaks in UV-irradiated Mammalian Cells¶ , 2000, Photochemistry and photobiology.

[15]  A. Kronenberg,et al.  Radiation-induced genomic instability. , 1994, International journal of radiation biology.

[16]  S. Kain,et al.  Optimized codon usage and chromophore mutations provide enhanced sensitivity with the green fluorescent protein. , 1996, Nucleic acids research.

[17]  S. Kain,et al.  Generation of Destabilized Green Fluorescent Protein as a Transcription Reporter* , 1998, The Journal of Biological Chemistry.

[18]  G. Barendsen.  The relationships between RBE and LET for different types of lethal damage in mammalian cells: biophysical and molecular mechanisms. , 1994, Radiation research.

[19]  D. Benford,et al.  Development of cytotoxicity tests for assessment of the toxicity of water samples taken from the environment. , 1987, Molecular toxicology.

[20]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[21]  G. Zlokarnik,et al.  Rapid fluorescence-based reporter-gene assays to evaluate the cytotoxicity and antitumor drug potential of platinum complexes. , 1999, Chemistry & biology.

[22]  F. Denizot,et al.  Rapid colorimetric assay for cell growth and survival. Modifications to the tetrazolium dye procedure giving improved sensitivity and reliability. , 1986, Journal of immunological methods.

[23]  U. Kristen,et al.  In vitro cytotoxicity testing for prediction of acute human toxicity , 1994, Cell Biology and Toxicology.

[24]  Suitability of enhanced green fluorescent protein as a reporter component for bioassays , 2001 .

[25]  J. Ward.  Radiation mutagenesis: the initial DNA lesions responsible. , 1995, Radiation research.

[26]  Petra Rettberg,et al.  Bacterial Lux-Fluoro test for biological assessment of pollutants in water samples from urban and rural origin , 2003 .

[27]  O. Flint,et al.  Prediction of phototoxic potential using human A431 cells and mouse 3T3 cells. , 1987, Molecular toxicology.

[28]  G. Horneck,et al.  Green Fluorescent Protein (GFP) as a Marker for Cell Viability After UV Irradiation , 2004, Journal of Fluorescence.

[29]  Christine E. Hellweg,et al.  Generation of Stably Transfected Mammalian Cell Lines as Fluorescent Screening Assay for NF- B Activation-Dependent Gene Expression , 2003, Journal of biomolecular screening.

[30]  T. Mosmann.  Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. , 1983, Journal of immunological methods.

[31]  G. Horneck,et al.  Enhanced green fluorescent protein as reporter protein for biomonitoring of cytotoxic effects in mammalian cells , 2001 .

[32]  G. Horneck,et al.  Mammalian cells as biomonitors of UV-exposure , 1999 .