D L Stevens

University of Oxford, Oxford, England, United Kingdom

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Publications (43)89.49 Total impact

  • [Show abstract] [Hide abstract]
    ABSTRACT: While protracting exposures of low-LET radiations usually leads to a reduction in their effectiveness for a given dose, for high-LET radiation there is now substantial evidence for what has been called an inverse dose-rate effect, where under certain circumstances there is an increase in carcinogenesis or other biological effects, with decreasing dose rate. This study investigates the influence of dose rate on the induction of chromosome aberrations and gene mutations after irradiation of plateau phase V79-4 cells with high-LET alpha particles. The induction of chromosomal aberrations exhibited a linear relationship with dose and showed evidence of a small but significant conventional dose-rate dependence, with low-dose-rate exposures (0.28 Gy h(-1)) being less effective by about 20% (ratio 0.82 ± 0.04) compared to acute exposures. However no significant dose-rate effect was observed for cell survival or the induction of mutations in the HPRT gene for low-dose-rate exposure (8.0 × 10(-5) and 1.5 × 10(-2) Gy h(-1) for exposure of 0.36 and 0.69 Gy, respectively) when compared to acute exposures.
    Radiation research. 08/2014;
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    ABSTRACT: An important stage in tumorigenesis is the ability of precancerous cells to escape natural anticancer signals. Apoptosis can be selectively induced in transformed cells by neighboring normal cells through cytokine and ROS/RNS signaling. The intercellular induction of apoptosis in transformed cells has previously been found to be enhanced after exposure of the normal cells to very low doses of both low- and high-LET ionizing radiation. Low-LET ultrasoft X rays with a range of irradiation masks were used to vary both the dose to the cells and the percentage of normal cells irradiated. The results obtained were compared with those after α-particle irradiation. The intercellular induction of apoptosis in nonirradiated src-transformed 208Fsrc3 cells observed after exposure of normal 208F cells to ultrasoft X rays was similar to that observed for γ rays. Intercellular induction of apoptosis was stimulated by irradiation of greater than 1% of the nontransformed 208F cells and increased with the fraction of cells irradiated. A maximal response was observed when ∼10-12% of the cells were irradiated, which gave a similar response to 100% irradiated cells. Between 1% and 10%, high-LET α particles were more effective than low-LET ultrasoft X rays in stimulating intercellular induction of apoptosis for a given fraction of cells irradiated. Scavenger experiments show that the increase in intercellular induction of apoptosis results from NO(•) and peroxidase signaling mediated by TGF-β. In the absence of radiation, intercellular induction of apoptosis was also stimulated by TGF-β treatment of the nontransformed 208F cells prior to coculture; however, no additional increase in intercellular induction of apoptosis was observed if these cells were also irradiated. These data suggest that the TGF-β-mediated ROS/RNS production reaches a maximum at low doses or fluences of particles, leading to a plateau in radiation-stimulated intercellular induction of apoptosis at higher doses.
    Radiation Research 06/2011; 176(3):346-55. · 2.70 Impact Factor
  • M A Hill, C S Griffin, E L Pyke, D L Stevens
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    ABSTRACT: The importance of the spatial distribution of energy deposition through the nucleus in determining the resultant chromosome rearrangements was investigated using fluorescent in situ hybridisation technique following either uniform or partial irradiation of HF19 human fibroblast cells with low-LET 1.5 keV ultrasoft X-rays. Irradiations were performed with and without a copper irradiation mask with a Poisson distribution of micron-sized holes immediately below the irradiation dish and the results are compared with previous results obtained following exposure to a Poisson distribution of alpha particles. For the same radiation quality, the spatial distribution of energy deposition within the nucleus was found to be important in determining the ultimate biological response, with an increased ratio of complex-to-simple aberrations observed for partial compared to uniform irradiation. Comparisons between low-LET ultrasoft X-rays and high-LET alpha particles indicate that the sub-micron clustering of damage along the alpha particle track is more important than just the total number of double-strand breaks produced.
    Radiation Protection Dosimetry 12/2010; 143(2-4):172-6. · 0.91 Impact Factor
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    ABSTRACT: Cohesin, a hetero-tetrameric complex of SMC1, SMC3, Rad21 and Scc3, associates with chromatin after mitosis and holds sister chromatids together following DNA replication. Following DNA damage, cohesin accumulates at and promotes the repair of DNA double-strand breaks. In addition, phosphorylation of the SMC1/3 subunits contributes to DNA damage-induced cell cycle checkpoint regulation. The aim of this study was to determine the regulation and consequences of SMC1/3 phosphorylation as part of the cohesin complex. We show here that the ATM-dependent phosphorylation of SMC1 and SMC3 is mediated by H2AX, 53BP1 and MDC1. Depletion of RAD21 abolishes these phosphorylations, indicating that only the fully assembled complex is phosphorylated. Comparison of wild type SMC1 and SMC1S966A in fluorescence recovery after photo-bleaching experiments shows that phosphorylation of SMC1 is required for an increased mobility after DNA damage in G2-phase cells, suggesting that ATM-dependent phosphorylation facilitates mobilization of the cohesin complex after DNA damage.
    Experimental Cell Research 11/2010; 317(3):330-7. · 3.56 Impact Factor
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    ABSTRACT: Ultrasoft X-rays have been shown to be very efficient in inducing chromosomal aberrations in mammalian cells. The present study was aimed to evaluate the modifying effects of DMSO (a potent scavenger of free radicals) on the frequencies of chromosome aberrations induced by soft X-rays. Confluent held G1 Chinese hamster cells (V79) were irradiated with Carbon K ultrasoft X-rays in the presence and absence of 1M DMSO and frequencies of chromosome aberrations in the first division cells were determined. DMSO reduced the frequencies of exchange types of aberrations (dicentrics and centric rings) by a factor of 2.1-3.5. The results indicate that free radicals induced by ultrasoft X-rays contribute to a great extent to the induction of chromosome aberrations. The possible implications of these results in interpreting the mechanisms involved in the high efficiency of ultrasoft X-rays in the induction of chromosome aberrations are discussed.
    Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis 09/2010; 691(1-2):23-6. · 3.90 Impact Factor
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    ABSTRACT: To compare the induction of double-strand breaks (DSB) in cells irradiated by 250 and 350 eV ultrasoft X-rays and assess the residual yield of breaks 2 hours post irradiation in order to unravel the correlation between the sharp increase in cell-killing efficiency of ultrasoft X-rays above versus below the carbon-K threshold (284 eV) and the induction of core events in DNA atoms. V79-4 hamster cells were irradiated with synchrotron ultrasoft X-rays at isoattenuating energies of 250 eV and 350 eV. DSB were quantified using pulse field gel electrophoresis. A significant increase in DSB induction was observed for 350 eV ultrasoft X-rays above the carbon-K threshold, compared to 250 eV below the threshold, per unit dose to the cell. The DSB induced by the 350 eV ultrasoft X-rays were less repaired 2 h after irradiation. The increased DSB induction at 350 eV is attributed to the increase in the relative proportion of photon interactions in DNA resulting in significant dose inhomogeneity across the cell with a local increase in dose to DNA. It results from an increase in carbon-K shell interactions and the short range of the electrons produced. Core ionisations in DNA, through core-hole relaxation in conjunction with localised effects of spatially correlated low-energy photo- and Auger-electrons lead to an increase in number and the complexity of DSB.
    International Journal of Radiation Biology 03/2010; 86(3):205-19. · 1.84 Impact Factor
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    ABSTRACT: The cohesin protein complex holds sister chromatids together after synthesis until mitosis. It also contributes to post-replicative DNA repair in yeast and higher eukaryotes and accumulates at sites of laser-induced damage in human cells. Our goal was to determine whether the cohesin subunits SMC1 and Rad21 contribute to DNA double-strand break repair in X-irradiated human cells in the G2 phase of the cell cycle. RNA interference-mediated depletion of SMC1 sensitized HeLa cells to X-rays. Repair of radiation-induced DNA double-strand breaks, measured by gammaH2AX/53BP1 foci analysis, was slower in SMC1- or Rad21-depleted cells than in controls in G2 but not in G1. Inhibition of the DNA damage kinase DNA-PK, but not ATM, further inhibited foci loss in cohesin-depleted cells in G2. SMC1 depletion had no effect on DNA single-strand break repair in either G1 or late S/G2. Rad21 and SMC1 were recruited to sites of X-ray-induced DNA damage in G2-phase cells, but not in G1, and only when DNA damage was concentrated in subnuclear stripes, generated by partially shielded ultrasoft X-rays. Our results suggest that the cohesin complex contributes to cell survival by promoting the repair of radiation-induced DNA double-strand breaks in G2-phase cells in an ATM-dependent pathway.
    Nucleic Acids Research 11/2009; 38(2):477-87. · 8.81 Impact Factor
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    ABSTRACT: A direct comparison was carried out of the biological effectiveness of protons and α-particles of the same linear energy transfer (LET) under identical conditions with a variety of in vitro biologial systems. Monolayers of mammalian cells were irradiated with accelerated beams of protons (1·2 and 1·4 MeV) and α-particles (30 and 35 MeV) corresponding to LETs of 23 and 20 keV µm−1 for each particle type. For V79-4 cells it was observed that the linear term of the dose-response for cell inactivation by protons was significantly greater than that for α-particles of the same LET. For HeLa and HeLa S3 cells, also, the linear term appeared to be greater for protons, but this was not observed with more limited data for C3H 10T½ cells. The result for V79 cells is in agreement with the report of Belli et al. (1989) who observed that the biological effectiveness of protons rose sharply between 17 and 30 keV µm−1 in strong contrast to α-particles which reached a peak effectiveness at > 100 keV µm−1. These results place new constraints on the biologically relevant features of the microscopic structure of radiation tracks, and have implications for the mechanistic and practical comparison between radiations.
    International Journal of Radiation Biology 07/2009; 61(5):611-624. · 1.84 Impact Factor
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    ABSTRACT: In many radiotherapy situations patients are exposed to mixed field radiation. In particular in BNCT, as with all neutron beam exposures, a significant fraction of the dose is contributed by low LET gamma ray photons. The components of such a mixed field may show a synergistic interaction and produce a greater cell kill effect than would be anticipated from the independent action of the different radiation types. Such a synergy would have important implications for treatment planning and in the interpretation of clinical results. An irradiation setup has been created at the Medical Research Council in Harwell to allow simultaneous irradiation of cells by cobalt-60 gamma rays and plutonium-238 alpha-particles. The setup allows for variation of dose and dose rates for both sources along with variation of the alpha particle energy. A series of cell survival assays for this mixed field have been carried out using V79-4 cells and compared to exposures to the individual components of the field under identical conditions. In the experimental setup described no significant synergistic effect was observed.
    Applied radiation and isotopes: including data, instrumentation and methods for use in agriculture, industry and medicine 04/2009; 67(7-8 Suppl):S318-20. · 1.09 Impact Factor
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    ABSTRACT: The aim of this study was to assess the relative influence of the linear energy transfer (LET) of alpha particles on the complexity of chromosome aberrations in the absence of significant other differences in track structure. To do this, we irradiated human hemopoietic stem cells (CD34+) with alpha particles of various incident LETs (110-152 keV/microm, with mean LETs through the cell of 119-182 keV/microm) at an equi-fluence of approximately one particle/cell and assayed for chromosome aberrations by mFISH. Based on a single harvest time to collect early-division mitotic cells, complex aberrations were observed at comparable frequencies irrespective of incident LET; however, when expressed as a proportion of the total exchanges detected, their occurrence was seen to increase with increasing LET. Cycle analysis to predict theoretical DNA double-strand break rejoining cycles was also carried out on all complex chromosome aberrations detected. By doing this we found that the majority of complex aberrations are formed in single non-reducible cycles that involve just two or three different chromosomes and three or four different breaks. Each non-reducible cycle is suggested to represent "an area" of finite size within the nucleus where double-strand break repair occurs. We suggest that the local density of damage induced and the proximity of independent repair areas within the interphase nucleus determine the complexity of aberrations resolved in metaphase. Overall, the most likely outcome of a single nuclear traversal of a single alpha particle in CD34+ cells is a single chromosome aberration per damaged cell. As the incident LET of the alpha particle increases, the likelihood of this aberration being classed as complex is greater.
    Radiation Research 06/2007; 167(5):541-50. · 2.70 Impact Factor
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    ABSTRACT: Ionising radiation can induce responses within non-exposed neighbouring (bystander) cells which potentially have important implications on the estimates of risk from low dose or low dose rate exposures of ionising radiations. A range of strategies have been developed for investigating bystander effects in vitro for both high-LET alpha particles or low-LET ultrasoft X rays using either partial shielding (grids, half-shields and slits) or by using a co-culture system where two physically separated populations of cells can be cultured together, allowing one population of cells to be irradiated while the second population remains unirradiated. The techniques described provide a useful tool to study bystander effects and complement microbeam studies. Studies using these systems show significant increases in the unirradiated bystander cells for various end points including the induction of chromosomal instability in haemopoetic stem cells and transformation in CGL1 cells.
    Radiation Protection Dosimetry 02/2006; 122(1-4):260-5. · 0.91 Impact Factor
  • E L Pyke, D L Stevens, M A Hill
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    ABSTRACT: Ionising radiation can induce responses within non-exposed neighbouring (bystander) cells, which potentially have important implications on the estimates of risk at environmentally relevant doses. Using human skin fibroblasts (AG1522), a range of methods were used to investigate the nature of the signal(s) arising from the exposed cells. The signal(s) can be transmitted by direct cell-cell communication (investigated by using partial dish irradiations) or by medium-borne factors (a co-culture system where two monolayers share the same medium but only one monolayer is exposed to ionising radiation). CDKN1A was found to be up-regulated in both directly exposed and non-exposed cells. The data suggest that direct cell-cell communication dominates for these confluent cells, with medium-borne factors also contributing.
    Radiation Protection Dosimetry 02/2006; 122(1-4):266-70. · 0.91 Impact Factor
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    ABSTRACT: Complex chromosome aberrations (any exchange involving three or more breaks in two or more chromosomes) are effectively induced in peripheral blood lymphocytes (PBL) after exposure to low doses (mostly single particles) of densely ionising high-linear energy transfer (LET) alpha-particle radiation. The complexity, when observed by multiplex fluorescence in situ hybridisation (m-FISH), shows that commonly four but up to eight different chromosomes can be involved in each rearrangement. Given the territorial organisation of chromosomes in interphase and that only a very small fraction of the nucleus is irradiated by each alpha-particle traversal, the aim of this study is to address how aberrations of such complexity can be formed. To do this, we applied theoretical "cycle" analyses using m-FISH paint detail of PBL in their first cell division after exposure to high-LET alpha-particles. In brief, "cycle" analysis deconstructs the aberration "observed" by m-FISH to make predictions as to how it could have been formed in interphase. We propose from this that individual high-LET alpha-particle-induced complex aberrations may be formed by the misrepair of damaged chromatin in single physical "sites" within the nucleus, where each "site" is consistent with an "area" corresponding to the interface of two to three different chromosome territories. Limited migration of damaged chromatin is "allowed" within this "area". Complex aberrations of increased size, reflecting the path of alpha-particle nuclear intersection, are formed through the sequential linking of these individual sites by the involvement of common chromosomes.
    Cytogenetic and Genome Research 02/2006; 112(1-2):35-44. · 1.84 Impact Factor
  • Radiation Research. 01/2006; 166(4):686-687.
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    ABSTRACT: Bystander effects from ionizing radiation have been detailed for a number of cell systems and a number of end points. We wished to use a cell culture/ex vivo rat model of respiratory tissue to determine whether a bystander effect detected in culture could also be shown in a tissue. Examination by immunofluorescence techniques of tracheal cell cultures after exposure to very low doses of alpha particles revealed a large proportion of cells with proliferating cell nuclear antigen (PCNA) bound in their nuclei. PCNA was selected as an end point because it is involved in both DNA repair and the changes in cell cycle that are typical of many reported bystander effects. Maximum response can be detected in up to 28% of the cells in sub-confluent cultures with a dose of only 2 mGy. At this dose less than 2% of the cell nuclei have experienced a particle traversal and less than 6% of the cells have experienced an alpha-particle traversal through either their nucleus or some part of their cytoplasm. The hypothesis that this bystander response in nontargeted cells is mediated through secreted factor(s) is presented, and supporting evidence was found using partial irradiation and co-culture experiments. Examination of the effect with excised pieces of trachea demonstrated a response similar to that seen in culture.
    Radiation Research 02/2005; 163(1):36-44. · 2.70 Impact Factor
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    ABSTRACT: The clonogenic survival of cells of the radiation-sensitive hamster cell lines irs1, irs2, irs3 and xrs5, representing different DNA repair pathways, was compared to that of their parent lines after alpha-particle irradiation. Measurements of nuclear area were made to calculate the probability of surviving a single alpha-particle traversal, the average number of lethal lesions per track and per unit dose, along with the "intrinsic radiosensitivity" of these cells, allowing for the potential of multiple lethal lesions per traversal. For all cell lines studied, alpha particles were found to be more biologically effective per unit absorbed dose than X rays at inducing cell inactivation. The repair-deficient cells showed an enhanced sensitivity to alpha particles compared to their parent line, but the degree of enhancement was less than for X rays. The reduction in additional sensitivity for alpha-particle irradiation was shown not to be due predominantly to differences in cell geometry limiting the probability of a cell nucleus being traversed. The results suggest that both the nonhomologous end-joining pathway and to a lesser extent the homologous recombination repair pathway play a role in successful repair of alpha-particle-induced damage, although a large proportion of damage is not repaired by either pathway.
    Radiation Research 01/2005; 162(6):667-76. · 2.70 Impact Factor
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    ABSTRACT: In radiobiology, low doses of high-LET radiation correspond to a few particle traversals through the cell population. Therefore, for studies on cell monolayers irradiated with a low dose of α-particles, it is extremely useful if the number and position of particle traversals can be determined. In this study we describe a new method, based on UV-curing, to obtain a thick CR-39 grafted onto a thick PolyEthylene Terephtalate (PET). This thin double polymeric layer, used as a dish base, has a regular and reproducible detector thickness which can be traversed by 3.5 MeV α-particles, with a sufficient residual energy to traverse mammalian cells attached to the base. The recording properties of a PET-CR-39 dish, together with a demonstration of its use for radiobiological experiments, are presented. This new tool allows the precise determination of single-track impact parameters at a sub-cellular level.
    Radiation Measurements. 01/2005; 40:279-282.
  • Radiation Research - RADIAT RES. 01/2005; 163(1):36-44.
  • Radiation Research - RADIAT RES. 01/2004; 162(6):667-676.
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    ABSTRACT: To investigate the biological effect of single, isolated, short electron tracks (<70 nm) relevant to practical human exposures to low-linear energy transfer radiation. An irradiation rig was constructed that allowed environmentally controlled, protracted irradiations with an individually prescribed dose to up to 20 samples over a period of days. Inactivation of V79-4 mammalian cells by Al(K) ultrasoft X-rays was studied at high and low dose-rates with a maximum exposure time of 42 h. A significant increase in clonogenic survival was observed at the higher doses when the exposure time was increased from <6 min to 21 h, with no further increase observed for 42-h exposures. Despite the short range of the low-energy electrons produced (<70 nm), significant cell inactivation was observed for these low dose-rate exposures. The results are consistent with the hypothesis that even individual tracks can be biologically effective.
    International Journal of Radiation Biology 12/2002; 78(11):967-79. · 1.84 Impact Factor

Publication Stats

755 Citations
89.49 Total Impact Points

Institutions

  • 2011–2014
    • University of Oxford
      • Gray Institute for Radiation Oncology and Biology
      Oxford, England, United Kingdom
  • 2010
    • Pierre and Marie Curie University - Paris 6
      • Institut de Minéralogie et de Physique des Milieux Condensés (IMPMC)
      Paris, Ile-de-France, France
  • 2009
    • University of Birmingham
      • School of Physics and Astronomy
      Birmingham, ENG, United Kingdom
  • 2006–2007
    • Brunel University
      अक्सब्रिज, England, United Kingdom
  • 2002–2006
    • Mrc Harwell
      Oxford, England, United Kingdom
  • 1990–2005
    • Medical Research Council (UK)
      Londinium, England, United Kingdom
  • 1999
    • Hiroshima University
      • Research Institute for Radiation Biology and Medicine (RIRBM)
      Hiroshima-shi, Hiroshima-ken, Japan
  • 1997
    • University of St Andrews
      Saint Andrews, Scotland, United Kingdom