RESUMEN
Understanding the mechanisms of X-ray radiation damage in biological systems is of prime interest in medicine (radioprotection, X-ray therapy ). Study of low-energy rays, such as soft-X rays and light ions, points to attribute their lethal effect to clusters of energy deposition by low-energy electrons. The first step, at the atomic or molecular level, is often the ionization of inner-shell electrons followed by Auger decay in an aqueous environment. We have developed an experimental set-up to perform electron coincidence spectroscopy on molecules in a water micro-jet. We present here the first results obtained on sodium benzoate solutions, irradiated at the oxygen and carbon K-edges.
RESUMEN
A combination of time-dependent density functional theory and Born-Oppenheimer molecular dynamics methods is used to investigate fragmentation of doubly charged gas-phase uracil in collisions with 100 keV protons. The results are in good agreement with ion-ion coincidence measurements. Orbitals of similar energy and/or localized in similar bonds lead to very different fragmentation patterns, thus showing the importance of intramolecular chemical environment. In general, the observed fragments do not correspond to the energetically most favorable dissociation path, which is due to dynamical effects occurring in the first few femtoseconds after electron removal.
Asunto(s)
Electrones , Gases/química , Simulación de Dinámica Molecular , Uracilo/química , Cinética , Conformación Molecular , Análisis EspectralRESUMEN
Theoretical simulations are particularly well suited to investigate, at a molecular level, direct and indirect effects of ionising radiations in DNA, as in the particular case of irradiation by swift heavy ions such as those used in hadron therapy. In the past recent years, we have developed the modeling at the microscopic level of the early stages of the Coulomb explosion of DNA molecules immersed in liquid water that follows the irradiation by swift heavy ions. To that end, Time-Dependent Density Functional Theory molecular dynamics simulations (TD-DFT MD) have been developed where localised Wannier orbitals are propagated. This latter enables to separate molecular orbitals of each water molecule from the molecular orbitals of the biomolecule. Our main objective is to demonstrate that the double ionisation of one molecule of the liquid sample, either one water molecule from the solvent or the biomolecule, may be in some cases responsible for the formation of an atomic oxygen as a direct consequence of the molecule Coulomb explosion. Our hypothesis is that the molecular double ionisation arising from irradiation by swift heavy ions (about 10% of ionisation events by ions whose velocity is about the third of speed of light), as a primary event, though maybe less probable than other events resulting from the electronic cascading (for instance, electronic excitations, electron attachments), may be systematically more damageable (and more lethal), as supported by experiments that have been carried out in our group in the 1990s (in studies of damages created by K holes in DNA). The chemical reactivity of the produced atomic oxygen with other radicals present in the medium will ultimately lead to chemical products that are harmful to DNA. In the present paper, we review our theoretical methodology in an attempt that the community be familiar with our new approach. Results on the production of atomic oxygen as a result of the double ionisation of water or as a result of the double ionisation of the Uracil RNA base will be presented.
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Daño del ADN , Modelos Teóricos , Radiación Ionizante , Agua/química , Biología Computacional , Iones Pesados , Inmersión , Simulación de Dinámica Molecular , Oxígeno/química , Uracilo/químicaRESUMEN
PURPOSE: To study the effect of hydration level and plasmid packing on strand break induction in DNA by ultrasoft X-ray. MATERIALS AND METHODS: Bluescript (pBS, tight packing) and pSP189 (pSP, loose packing) plasmids were irradiated by 250, 380, and 760 eV ultrasoft X-rays at the Laboratoire pour l'Utilisation du Rayonnement Electromagnétique synchrotron facility (Orsay, France). Single and double strand breaks (SSB and DSB) were quantified by gel electrophoresis. RESULTS: The number of DSB per Gray and per Dalton in pBS plasmids were (5.6 +/- 0.1), (6.3 +/- 0.1) and (8.5 +/- 0.4)x10(-12) at 250, 380 and 760 eV, respectively. They were respectively 1.4 +/- 0.1, 1.1 +/- 0.1 and 1.9 +/- 0.2 times larger for pSP plasmids. SSB/DSB ratios varied between 4.4 and 6.4. CONCLUSION: The observed dependency of strand break induction by ultrasoft X-rays on the hydration level of DNA in plasmids films may be associated with: (i) Damage transfer from the water shell to the DNA and/or (ii) change in packing. 760 eV photons which are more often absorbed in the hydration shell and yield longer range electrons than 250 and 380 eV photons, induce more DSB per Gray and per Dalton, especially for the looser plasmid (pSP).
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Daño del ADN/efectos de la radiación , ADN de Cadena Simple/efectos de la radiación , Plásmidos/efectos de la radiación , Agua/química , Rayos X , Relación Dosis-Respuesta en la Radiación , Electroforesis en Gel Bidimensional/métodos , FotonesRESUMEN
To test a possible specific effect of carbon K-shell ionizations in DNA, survival curves for Chinese hamster V79 cells were measured for X irradiations at energies below and above the carbon K-shell ionization threshold. Specific values of the X-ray energies (250 and 340 eV) were chosen to ensure isoattenuation of the two kinds of radiation within the cell. An enhancement of lethality by a factor of about 2 was found for X rays at 340 eV compared to below the threshold at 250 eV. This may be attributed to the production of highly efficient carbon K-shell ionizations located on DNA. A model of X-ray lethality (Goodhead et al., Radiat. Prot. Dosim. 52, 217-223, 1994) was extended to allow for a possible lethal effect from clusters of reactive species induced by K-shell photoionizations (K-shell clusters). Within this model, the increase in lethality above the carbon K-shell threshold may be explained by a value of 2% for the lethal efficiency of K-shell clusters overlapping the DNA. An extrapolation to the lethal effect of more complex ion-induced K-shell ionizations indicates that K-shell ionization may be a major process in the biological effectiveness of heavy ions.
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Núcleo Celular/efectos de la radiación , ADN/efectos de la radiación , Iones Pesados , Animales , Carbono , Línea Celular , Supervivencia Celular/efectos de la radiación , Cricetinae , Cricetulus , Efectividad Biológica Relativa , Rayos XRESUMEN
The large RBE (approximately 7) measured for the killing of Chinese hamster V79 cells by 340 eV ultrasoft X rays, which preferentially ionize the K shell of carbon atoms (Hervé du Penhoat et al., Radiat. Res. 151, 649-658, 1999), was used to investigate the location of sensitive sites for cell inactivation and the physical modes of action of radiation. The enhancement of the RBE above the carbon K-shell edge either may indicate a high intrinsic efficiency of carbon K-shell ionizations (due, for example, to a specific physical or chemical effect) or may be related to the preferential localization of these ionizations on the DNA. The second interpretation would indicate a strong local (within 3 nm) action of K-shell ionizations and consequently the importance of a direct mechanism for radiation lethality (without excluding an action in conjunction with an indirect component). To distinguish between these two hypotheses, the efficiencies of core ionizations in DNA atoms (phosphorus L-shell, carbon K-shell, and oxygen K-shell ionizations) to induce damages were investigated by measuring their capacities to produce DNA double-strand breaks (DSBs). The effect of photoionizations in isolated DNA was studied using pBS plasmids in a partially hydrated state. No enhancement of the efficiency of DSB induction by carbon K-shell ionizations compared to oxygen K-shell ionizations was found, supporting the hypothesis that it is the localization of these carbon K-shell events on DNA which gives to the 340 eV photons their high killing efficiency. In agreement with this interpretation, cell inactivation and DSB induction, which do not appear to be correlated when expressed in terms of yields per unit dose in the sample, exhibit a rather good correlation when expressed in terms of efficiencies per core event in the DNA. These results suggest that core ionizations in DNA, through core-hole relaxation in conjunction with localized effects of spatially correlated secondary and Auger electrons, may be the major critical events for cell inactivation, and that the resulting DSBs (or a constant fraction of these DSBs) may be a major class of unrepairable lesions.
Asunto(s)
Daño del ADN/efectos de la radiación , ADN/efectos de la radiación , Fibroblastos/efectos de la radiación , Rayos X/efectos adversos , Animales , Carbono/efectos de la radiación , Línea Celular/efectos de la radiación , Núcleo Celular/efectos de la radiación , Supervivencia Celular/efectos de la radiación , Cricetinae , Cricetulus , ADN Bacteriano/efectos de la radiación , ADN Recombinante/efectos de la radiación , ADN de Cadena Simple/efectos de la radiación , ADN Superhelicoidal/efectos de la radiación , Relación Dosis-Respuesta en la Radiación , Electrones , Rayos gamma , Iones , Pulmón/citología , Modelos Biológicos , Oxígeno/efectos de la radiación , Fósforo/efectos de la radiación , Fotones , Plásmidos/efectos de la radiación , Efectividad Biológica RelativaRESUMEN
PURPOSE: To study the frequency of chromosome aberrations induced by soft X-rays. To see if the core ionization of DNA atoms is involved in this end-point as much as it appears to be in cell killing. MATERIALS AND METHODS: V79 hamster cells were irradiated by synchrotron radiation photons iso-attenuated in the cell (250, 350, 810eV). The morphological chromosome aberrations detected in the first post-irradiation cell division (dicentrics and centric rings) were studied by Giemsa staining. RESULTS: The chromosome aberrations at 350eV were, respectively, 2.6 +/- 0.8 and 2.1 +/- 0.8 times more numerous than at 250 and 810eV for the same average dose absorbed by the nucleus. These relative effectivenesses are comparable with the ones already measured for cell killing. Moreover, they roughly vary such as the relative numbers of core ionizations (including in the phosphorus L-shell) produced in DNA and its bound water (water being involved only at 810eV through the oxygen atoms). In particular, they reproduce the characteristic twofold enhancement at 350eV, above the carbon K threshold. CONCLUSIONS: Correlations suggest that the core ionization process is likely a common and essential mechanism initiating both chromosome aberration and cell killing end-points at these photon energies.
Asunto(s)
Aberraciones Cromosómicas , ADN/efectos de la radiación , Rayos X , Animales , Carbono/química , División Celular , Línea Celular , Núcleo Celular/metabolismo , Cricetinae , Relación Dosis-Respuesta en la Radiación , Rayos gamma , Iones , Oxígeno/metabolismo , Fotones , Radiación Ionizante , RadiometríaRESUMEN
In order to assess the lethal efficiency and other biological effects of inner shell ionisations of constituent atoms of DNA ('K' events), experiments were developed at the LURE synchrotron facility using ultrasoft X rays as a probe of K events. The lethal efficiency of ultrasoft X rays above the carbon K threshold was especially investigated using V79 cells and compared with their efficiency to induce double strand breaks in dry plasmid-DNA. A correlation between the K event efficiencies for these processes is shown. Beams at 340 eV were found to be twice as efficient at killing cells than were beams at 250 eV. In addition, a rough two-fold increase of the relative biological effectiveness for dicentric + ring induction has also been observed between 250 and 340 eV radiations.