Variations in the Processing of DNA Double-Strand Breaks Along 60-MeV Therapeutic Proton Beams

PURPOSE: To investigate the variations in induction and repair of DNA damage along the proton path, after a previous report on the increasing biological effectiveness along clinically modulated 60-MeV proton beams.

METHODS AND MATERIALS: Human skin fibroblast (AG01522) cells were irradiated along a monoenergetic and a modulated spread-out Bragg peak (SOBP) proton beam used for treating ocular melanoma at the Douglas Cyclotron, Clatterbridge Centre for Oncology, Wirral, Liverpool, United Kingdom. The DNA damage response was studied using the 53BP1 foci formation assay. The linear energy transfer (LET) dependence was studied by irradiating the cells at depths corresponding to entrance, proximal, middle, and distal positions of SOBP and the entrance and peak position for the pristine beam.

RESULTS: A significant amount of persistent foci was observed at the distal end of the SOBP, suggesting complex residual DNA double-strand break damage induction corresponding to the highest LET values achievable by modulated proton beams. Unlike the directly irradiated, medium-sharing bystander cells did not show any significant increase in residual foci.

CONCLUSIONS: The DNA damage response along the proton beam path was similar to the response of X rays, confirming the low-LET quality of the proton exposure. However, at the distal end of SOBP our data indicate an increased complexity of DNA lesions and slower repair kinetics. A lack of significant induction of 53BP1 foci in the bystander cells suggests a minor role of cell signaling for DNA damage under these conditions.

Proton acceleration enhanced by a plasma jet in expanding foils undergoing relativistic transparency

Ion acceleration driven by the interaction of an ultraintense (2 × 10²⁰ W cm^-2) laser pulse with an ultrathin ( nm) foil target is experimentally and numerically investigated. Protons accelerated by sheath fields and via laser radiation pressure are angularly separated and identified based on their directionality and signature features (e.g. transverse instabilities) in the measured spatial-intensity distribution. A low divergence, high energy proton component is also detected when the heated target electrons expand and the target becomes relativistically transparent during the interaction. 2D and 3D particle-in-cell simulations indicate that under these conditions a plasma jet is formed at the target rear, supported by a self-generated azimuthal magnetic field, which extends into the expanded layer of sheath-accelerated protons. Electrons trapped within this jet are directly accelerated to super-thermal energies by the portion of the laser pulse transmitted through the target. The resulting streaming of the electrons into the ion layers enhances the energy of protons in the vicinity of the jet. Through the addition of a controlled prepulse, the maximum energy of these protons is demonstrated experimentally and numerically to be sensitive to the picosecond rising edge profile of the laser pulse.

Picosecond metrology of laser-driven proton bursts

Tracking primary radiation-induced processes in matter requires ultrafast sources and high precision timing. While compact laser-driven ion accelerators are seeding the development of novel high instantaneous flux applications, combining the ultrashort ion and laser pulse durations with their inherent synchronicity to trace the real-time evolution of initial damage events has yet to be realized. Here we report on the absolute measurement of proton bursts as short as 3.5±0.7 ps from laser solid target interactions for this purpose. Our results verify that laser-driven ion acceleration can deliver interaction times over a factor of hundred shorter than those of state-of-the-art accelerators optimized for high instantaneous flux. Furthermore, these observations draw ion interaction physics into the field of ultrafast science, opening the opportunity for quantitative comparison with both numerical modelling and the adjacent fields of ultrafast electron and photon interactions in matter.

Buffered high charge spectrally-peaked proton beams in the relativistic-transparency regime

Spectrally-peaked proton beams of high charge (E_p ≈ 8 MeV, ΔE ≈ 4 MeV, N ≈ 50 nC ) have been observed from the interaction of an intense laser (>10¹⁹Wcm^-2) with ultrathinCHfoils, as measured by spectrally-resolved full beam profiles. These beams are reproducibly generated for foil thicknesses 5-100 nm, and exhibit narrowing divergence with decreasing target thickness down to ≈8° for 5 nm. Simulations demonstrate that the narrow energy spread feature is a result of buffered acceleration of protons. The radiation pressure at the front of the target results in asymmetric sheath fields which permeate throughout the target, causing preferential forward acceleration. Due to their higher chargeto-mass ratio, the protons outrun a carbon plasma driven in the relativistic transparency regime.

Laser driven MeV proton beam focussing by auto-charged electrostatic lens configuration

Significant reduction of inherent large divergence of the laser driven MeV proton beams is achieved by strong (of the order of 10^9 V/m ) electrostatic focussing field generated in the confined region of a suitably shaped structure attached to the proton generating foil. The scheme exploits the positively charging of the target following an intense laser interaction. Reduction in the proton beam divergence, and commensurate increase in proton flux is observed while preserving the beam laminarity. The underlying mechanism has been established by the help of particle tracing simulations. Dynamic focussing power of the lens, mainly due to the target discharging, can also be exploited in order to bring up the desired chromaticity of the lens for the proton beams of broad energy range.

Investigating the Implications of a Variable RBE on Proton Dose Fractionation Across a Clinical Pencil Beam Scanned Spread-Out Bragg Peak

Purpose: To investigate the clinical implications of a variable relative biological effectiveness (RBE) on proton dose fractionation. Using acute exposures, the current clinical adoption of a generic, constant cell killing RBE has been shown to underestimate the effect of the sharp increase in linear energy transfer (LET) in the distal regions of the spread-out Bragg peak (SOBP). However, experimental data for the impact of dose fractionation in such scenarios are still limited.

Methods and Materials: Human fibroblasts (AG01522) at 4 key depth positions on a clinical SOBP of maximum energy 219.65 MeV were subjected to various fractionation regimens with an interfraction period of 24 hours at Proton Therapy Center in Prague, Czech Republic. Cell killing RBE variations were measured using standard clonogenic assays and were further validated using Monte Carlo simulations and parameterized using a linear quadratic formalism.

Results: Significant variations in the cell killing RBE for fractionated exposures along the proton dose profile were observed. RBE increased sharply toward the distal position, corresponding to a reduction in cell sparing effectiveness of fractionated proton exposures at higher LET. The effect was more pronounced at smaller doses per fraction. Experimental survival fractions were adequately predicted using a linear quadratic formalism assuming full repair between fractions. Data were also used to validate a parameterized variable RBE model based on linear α parameter response with LET that showed considerable deviations from clinically predicted isoeffective fractionation regimens.

Conclusions: The RBE-weighted absorbed dose calculated using the clinically adopted generic RBE of 1.1 significantly underestimates the biological effective dose from variable RBE, particularly in fractionation regimens with low doses per fraction. Coupled with an increase in effective range in fractionated exposures, our study provides an RBE dataset that can be used by the modeling community for the optimization of fractionated proton therapy.

Interaction between glucocorticoid and adenosine A2A receptors in the hippocampus : implications for learning and memory

Tese de doutoramento, Ciências Biomédicas (Neurociências), Universidade de Lisboa, Faculdade de Medicina, 2014

Brain-derived neurotrophic factor and adenosine signalling on amyloid-β peptide induced toxicity : impact on hippocampal function

Tese de doutoramento, Ciências Biomédicas (Neurociências), Universidade de Lisboa, Faculdade de Medicina, 2014

Proton translocation by cytochrome c oxidase reconstituted into proteoliposomes

Cytochrome c oxidase .inserted into proteoliposomes translocates protons with a stoichiometry of approx-, imately 0.4-0.6 H+/e- in the presence of valinomycin plus pottasium. The existance .ofsuchproton translocation is .supportedby experiments with lauryl maltoside which abolished the pulses but~~d not inhibit cyt. c binding .or oxidase turnover. Pulses with K3FeCN6 did not induce acidification further supporting vectorial proton transport by cyt ..aa3 . Upon lowering the ionic strength and pulsing with ferrocytochrome c, H+/eratios increased. This increase is attributed to scaler proton release consequent upon cyt.c-phospholipid binding. Oxygen pulses at low ionic strength however did not exhibit this large scaler increase in H+/e- ratios.A-small increase was observed upon .02 pul'sing at·low ionic strengt.h. This increase was KeN and, ,pcep sensitive and thus possibly due to a redox linked scaler deprotonation. Increases in the H+/e- ratio also occurred ifp~lses ,were performed in the presence of nonactin rather.than valinomycin. The fluorescent pH indicator pyranine was internally trapped inaa3 conta~ning "proteoliposomes. Internal alkalinization, as mon,itored by pyranine fluorescence leads to a of approx.imately 0.35 units, which is proportional to electron flux. This internal alkalinization was also DCCD sensitive, being inhibited by approximately 50%. This 50% inhibition of internal alkalinization supports the existance of vectorial proton transport.

Handwritten sheet of paper detailing the cost of transporting boxes containing the Gravatt’s level

Handwritten sheet of paper detailing the cost of transporting boxes containing the Gravatt’s level from London to St. Catharines, April 1847.

Handwritten sheet of paper detailing the cost of transporting boxes containing the Gravatt’s level

Handwritten sheet of paper detailing the cost of transporting boxes containing the Gravatt’s level from London to St. Catharines. This sheet is signed by S.D. Woodruff, Jan. 11, 1847.

Étude des collisions proton-proton dans l’expérience ATLAS avec les détecteurs ATLAS-MPX

Les seize détecteurs MPX constituant le réseau ATLAS-MPX ont été placés à différentes positions dans le détecteur ATLAS et sa averne au CERN dans le but de mesurer en emps réel les champs de radiation produits ar des particules primaires (protons des faisceaux) et des particules secondaires (kaons, pions, g, protons) issues des collisions proton-proton. Des films de polyéthylène (PE) et de fluorure de lithium (6LiF) recouvrent les détecteurs afin d’augmenter leur sensibilité aux neutrons produits par les particules primaires et secondaires interagissant avec les matériaux présents dans l’environnement d’ATLAS. La reconnaissance des traces laissées par les particules dans un détecteur ATLAS-MPX se fait à partir des algorithmes du logiciel MAFalda (“Medipix Analysis Framework”) basé sur les librairies et le logiciel d’analyse de données ROOT. Une étude sur le taux d’identifications erronées et le chevauchement d’amas a été faite en reconstruisant les activités des sources 106Ru et 137Cs. L’efficacité de détection des neutrons rapides a été mesurée à l’aide des sources 252Cf et 241AmBe (neutrons d’énergie moyenne de 2.13 et 4.08 MeV respectivement). La moyenne des efficacités de détection mesurées pour les neutrons produits par les sources 252C f et 241AmBe a été calculée pour les convertisseurs 6LiF et PE et donnent (0.8580 ± 0.1490)% et (0.0254 ± 0.0031)% pour LiF et (0.0510 ± 0.0061)% et (0.0591 ± 0.0063)% pour PE à bas et à haut seuil d’énergie respectivement. Une simulation du calcul de l’efficacité de détection des neutrons dans le détecteur MPX a été réalisée avec le logiciel GEANT4. Des données MPX correspondant aux collisions proton-proton à 2.4 TeV et à 7 TeV dans le centre de masse ont été analysées. Les flux détectés d’électrons et de photons sont particulièrement élevés dans les détecteurs MPX01 et MPX14 car ils sont plus près du point de collision. Des flux de neutrons ont été estimés en utilisant les efficacités de détection mesurées. Une corrélation avec la luminosité du LHC a été établie et on prédit que pour les collisions à 14 TeV dans le centre de masse et avec une luminosité de 10^34 cm-1*s-1 il y aura environ 5.1x10^8 ± 1.5x10^7 et 1.6x10^9 ± 6.3x10^7 particules détectées par les détecteurs MPX01 et MPX14 respectivement.

Elementary steps in aqueous proton transfer reactions : a first principles molecular dynamics study

La nature des acides dans un environnement aqueux est primordiale dans de nombreux aspects de la chimie et de la biologie. La caractéristique principale d'un acide est sa capacité à transférer un proton vers une molécule d'eau ou vers n'importe quelle base, mais ce procédé n'est pas aussi simple qu'il y paraît. Il peut au contraire être extrêmement complexe et dépendre de manière cruciale de la solvatation des différents intermédiaires de réaction impliqués. Cette thèse décrit les études computationnelles basées sur des simulations de dynamique moléculaire ab initio qui ont pour but d'obtenir une description à l'échelle moléculaire des divers procédés de transferts de proton entre acide et bases dans un milieu aqueux. Pour cela, nous avons étudié une serie de système, dont l'acide hydrofluorique aqueux, l'acide trifluoroacétique aqueux, et un système modèle constitué d'un phénol et d'une entité carboxylate reliés entre eux par une molécule d'eau en solution aqueuse. Deux états intermédiaires ont été identifiés pour le transfert d'un proton depuis un acide. Ces intermédiaires apparaissent stabilisés par un motif local de solvatation via des ponts H. Leurs signatures spectroscopiques ont été caractérisées au moyen de la spectroscopie infrarouge, en utilisant le formalisme de la dynamique moléculaire ab initio, qui inclut l'effet quantique nucléaire de manière explicite. Cette étude a aussi identifié trois chemins de réaction élémentaire, qui sont responsable pour le transfert d'un proton d'un acide à une base, ainsi que leurs échelles de temps caractéristiques. Les conclusions tirées de ces études sont discutées dans les détails, au niveau moléculaire, avec une emphase sur les comparaisons entre les résultats théoriques et les mesures expérimentales obtenues dans a littérature ou via des collaborateurs.