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Validation of endpoints as biomarkers of low-dose radiation damage
Author(s)
Rossouw, Maria Susanna
Date Issued
2004
Type
Thesis
Publisher
Cape Technikon
Abstract
The need for radiobiological research was bom from the discovery that high doses of
radiation could cause cancer and other health effects. However, recent
developments in molecular biology uncovered the effects of low doses of radiation on
different biological systems and as a result new techniques have been developed to
measure these effects.
The aim of this study was thus to validate biomarkers of initial DNA strand breaks,
micronucleus formation, and the different pt ;ases of apoptosis as biological indicators
of low-dose radiation damage. Furthermore, the difference in response of blood cells
to different qualities and doses of radiation was investigated by irradiating cells with
low- and high-LET radiation simultaneously.
Blood from one donor was irradiated with doses between 0 and 4 Gy gamma- and
neutron radiation. The alkaline single-cell gel electrophoresis (comet) assay was
performed on different cell preparations directly after irradiation for the detection of
initial DNA strand breaks. Radiation-induced cytogenetic damage was investigated
using the cytokinesis-blocked micronucleus assay while different features of
apoptosis were investigated by measuring caspase activation, enzymatic DNA
fragmentation, and cellular morphology.
The comet assay was sensitive enough to detect DNA strand breaks above 0.25 Gy
and showed that the Iymphocyte isolation process induced some endogenous
damage in cells, detected by the formation of highly damaged cells and hedgehogs in
isolated cell preparations only.
radiation could cause cancer and other health effects. However, recent
developments in molecular biology uncovered the effects of low doses of radiation on
different biological systems and as a result new techniques have been developed to
measure these effects.
The aim of this study was thus to validate biomarkers of initial DNA strand breaks,
micronucleus formation, and the different pt ;ases of apoptosis as biological indicators
of low-dose radiation damage. Furthermore, the difference in response of blood cells
to different qualities and doses of radiation was investigated by irradiating cells with
low- and high-LET radiation simultaneously.
Blood from one donor was irradiated with doses between 0 and 4 Gy gamma- and
neutron radiation. The alkaline single-cell gel electrophoresis (comet) assay was
performed on different cell preparations directly after irradiation for the detection of
initial DNA strand breaks. Radiation-induced cytogenetic damage was investigated
using the cytokinesis-blocked micronucleus assay while different features of
apoptosis were investigated by measuring caspase activation, enzymatic DNA
fragmentation, and cellular morphology.
The comet assay was sensitive enough to detect DNA strand breaks above 0.25 Gy
and showed that the Iymphocyte isolation process induced some endogenous
damage in cells, detected by the formation of highly damaged cells and hedgehogs in
isolated cell preparations only.
Additional information
Thesis (MTech (Biomedical Technology))--Cape Technikon, Cape Town, 2004
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