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Non-targeted effects, including bystander and systemic effects, play a crucial role during Auger targeted radionuclide therapy. Here, we investigated whether small extracellular vesicles (sEVs) produced by irradiated cells could contribute to the bystander cytotoxic effects in vitro and also to therapeutic efficacy in vivo, after their injection in tumor xenografts. B16F10 melanoma donor cells were exposed to radiolabeled antibodies (Auger radioimmunotherapy, RIT) for 48 h or to X-rays (donor cells). Then, donor cells were incubated with fresh medium for 2 h to prepare conditioned medium (CM) that was transferred onto recipient cells for bystander effect assessment, or used for sEVs enrichment. Resulting sEVs were incubated in vitro with recipient cells for determining bystander cytotoxicity, or injected in B16F10 melanoma tumors harbored by athymic and C57BL/6 mice. In vitro analysis of bystander cytotoxic effects showed that CM killed about 30–40% of melanoma cells. SEVs isolated from CM contributed to this effect. Moreover, the double-stranded DNA (dsDNA) content was increased in sEVs isolated from CM of exposed cells compared to control (not exposed), but the difference was significant only for the X-ray condition. These results were supported by immunodetection of cytosolic dsDNA in donor cells, a phenomenon that should precede dsDNA enrichment in sEVs. However, sEVs cytotoxicity could not be detected in vivo. Indeed, in athymic and in immunocompetent mice that received four intratumoral injections of sEVs (1/day), tumor growth was not delayed compared with untreated controls. Tumor growth was slightly (not significantly) delayed in immunocompetent mice treated with sEVs from X-ray-exposed cells, and significantly with sEVs purified from CM collected after 48 h of incubation. These results highlight the need to determine the optimal conditions, including radiation absorbed dose and sEVs collection time, to obtain the strongest cytotoxic effects. This study demonstrates that sEVs could play a role during Auger RIT through bystander effects in vitro. No systemic effects were observed in vivo, under our experimental conditions. However, X-rays experiments showed that sEVs collection time might be influencing the nature of sEVs, a parameter that should also be investigated during Auger RIT. [ABSTRACT FROM AUTHOR]

To examine the reliability to model cellular S-values for the Auger electron (AE) emitters, 111In, 197Hg and 197mHg with MCNP6 and their relative dose deposition in subcellular targets. A model cell was defined as four concentric spheres consisting of the nucleus (N), cytoplasm (Cy), cell and nuclear membranes (CM, NM) in which radionuclides distributed homogeneously. The transport of AE, conversion electrons and photons were simulated by MCNP6 to calculate cellular S values (SN←CM, SN←Cy, SN←NM, SN←N, SCM←CM, SNM←NM). SN←CM, SN←Cy and SN←N were also calculated with MIRDcell. MIRDcell and MCNP6-calculated SN←N were in excellent agreement, but a slight discrepancy on SN←Cy and SN←CM was observed. The ratios of SCM←CM or SNM←NM vs. SN←N were 9.7–51.0 or 10.5–37.4, 7.9–41.8 or 8.4–31.8 and 7.2–36.9 or 8.0–28.1 for 111In, 197Hg, 197mHg, respectively. The mean S(197Hg)/S(111In) and S(197mHg)/S(111In) were 2.5 ± 0.5 and 2.5 ± 0.6, respectively. Cellular S-values were reliably calculated with MCNP6. 197Hg and 197mHg deposit two-fold more doses than 111In at the subcellular scale. All AE emitters deposit a higher self-dose in the CM and NM than in the N, which warrants studies on the effects of targeting the CM and NM by AE emitters. [ABSTRACT FROM AUTHOR]

Auger electrons can be highly radiotoxic when they are used to irradiate specific molecular sites. This has spurred basic science investigations of their radiobiological effects and clinical investigations of their potential for therapy. Focused symposia on the biophysical aspects of Auger processes have been held quadrennially. This 9th International Symposium on Physical, Molecular, Cellular, and Medical Aspects of Auger Processes at Oxford University brought together scientists from many different fields to review past findings, discuss the latest studies, and plot the future work to be done. This review article examines the research in this field that was published during the years 2015–2019 which corresponds to the period since the last meeting in Japan. In addition, this article points to future work yet to be done. There have been a plethora of advancements in our understanding of Auger processes. These advancements range from basic atomic and molecular physics to new ways to implement Auger electron emitters in radiopharmaceutical therapy. The highly localized doses of radiation that are deposited within a 10 nm of the decay site make them precision tools for discovery across the physical, chemical, biological, and medical sciences. [ABSTRACT FROM AUTHOR]

Purpose: We set out to improve the accuracy of absorbed dose calculations for in vitro measurements of the relative biological effectiveness (RBE) of tritiated water (HTO) for the clonogenic cell survival assay, also considering the influence of the end-of-track linear energy transfer (LET) of low-energy electrons. Materials and methods: The COmputation Of Local Electron Release (COOLER) program was adopted to investigate the cell geometry and the tritium full beta-decay spectrum impact on the S-values and subsequently on the RBE of HTO for clonogenic cell survival at similar high dose rates (HDR). Results: S-values for cells growing in suspension are usually comparable to those for adherent cells. RBEs calculated at the 10% survival fraction through the use of the average energy are almost similar to those obtained with the beta-spectrum. For adherent cells, an RBE of 1.6 was found when HTO cell survival curves were compared to acute y-ray exposures. Irrespective of the geometrical configuration, the RBE was 2.0 when the comparison was made with similar dose rates. Conclusions: These results underline the importance of irradiating at equal dose rates and cell culture conditions when measuring in vitro RBE-values. [ABSTRACT FROM AUTHOR]

Purpose: A challenge for single-cell dosimetry of internalized Auger electron-emitting (AE) radiopharmaceuticals remains how best to elucidate their spatial distribution. To this end, a method, photoresist autoradiography (PAR), was previously developed to identify the lateral spatial distribution of AE-emitting radionuclides internalized in single cancer cells. In this paper, we present a simple mathematical model based on the radius and depth of radiation-induced patterns in photoresist material to identify the location in the z-plane of an 111In source capable of generating the pattern. Materials and methods: SQ20B cells, derived from a head and neck squamous cell carcinoma, were exposed to 111In-labeled epidermal growth factor (EGF) (8 MBq/μg). The integrated electron fluence after four half-lives from the internalized radionuclide-containing construct was detected by a photoresist layer that was placed in close proximity to the cells. The resultant latent patterns were chemically developed and analyzed by atomic force microscopy (AFM). The features in the patterns were matched to locations of electrons emitted from simulated point sources, thereby determining the likely locations of internalized radionuclides. Results: The modeling procedure was validated using simple patterns. The model relates the depth and radius (in the x-y plane) of a pattern to the location and fluence of the source giving rise to the pattern. This point source modeling method provided a good fit to experimental data and can be expanded to analyze more complex patterns. Conclusions: We have demonstrated the utility of the modelling technique to identify the location of internalized AE-emitting radionuclides. This methodology now needs to be extended to predict the source positions in more complex PAR patterns. [ABSTRACT FROM AUTHOR]

Purpose:To simulate the deceleration processes of secondary electrons produced by a high-energy Auger electron in water, and particularly to focus on the spatial and temporal distributions of the secondary electron and the collision events (e.g. ionization, electronic excitation, and dissociative electron attachment) that are involved in the multiplication of lesions at sites of DNA damage. Materials and methods:We developed a dynamic Monte Carlo code that considers the Coulombic force between an ejected electron and its parent cation produced by the Auger electron in water. Thus our code can simulate some return electrons to the parent cations. Using the code, we calculated to within the order of femtoseconds the temporal evolution of collision events, the mean energy, and the mean traveling distance (including its spatial probability distribution) of the electron at an ejected energy of 20 eV. Results:Some of the decelerating electrons in water in the Coulombic field were attracted to the ionized atoms (cations) by the Coulombic force within hundreds of femtoseconds, although the force did not significantly enhance the number of ionization, electronic excitation, and dissociative electron attachment collision events leading to water radiolysis. Conclusions:The secondary electrons are decelerated in water by the Coulombic force and recombined to the ionized atoms (cations). Furthermore, the some return electrons might be prehydrated in water layer near the parent cation in DNA if the electrons might be emitted from the DNA. The prehydrated electron originated from the return electron might play a significant role in inducing DNA damage. [ABSTRACT FROM PUBLISHER]

Purpose:To investigate an enhancement of DNA double-strand break (DSB) induction and cell killing effect by K-shell ionization of phosphorus atoms and Auger electrons on human cell lines. Materials and methods:Induction of DSB, DNA damage responses, cell cycle distributions, and cell killing effects were investigated after exposures of the cells with monochromatic synchrotron radiation soft X-rays of 2153 and 2147 eV, which were the resonance peak and off peak, respectively, of the K-shell photoabsorption of phosphorus. Results:Higher biological effects in the cells irradiated with soft X-rays at 2153 eV than at 2147 eV were observed in (i) the efficiency of 53BP1/γ-H2AX co-localized foci formation per dose and residual number of foci, (ii) prolonged phosphorylation levels of DSB repair and/or cell cycle checkpoint related proteins and G2 arrest, (iii) the cell killing effects at the 10% survival level of normal human fibroblasts, HeLa cells, and human glioblastoma M059K cells (1.2–1.5 times higher) and that of human ataxia telangiectasia mutated (ATM)-defective cells and glioblastoma DNA-dependent protein kinase catalytic subunit (DNA-PKcs)-defective cells (1.2 times). Conclusion:The yield of DSB and partly less-reparable complex DNA damage induction in human cells was enhanced by K-shell photoabsorption of phosphorus and low-energy Auger electrons. [ABSTRACT FROM PUBLISHER]

Purpose: In addition to gamma radiation, 99mTc emits low-energy Auger electrons with path-lengths of nanometers to micrometers that cannot be utilized for diagnostic procedures; however, they have frequently been discussed for therapeutic applications. We compared radiotoxicity of three 99mTc-labeled radiopharmaceuticals with differences in the subcellular distribution. Materials and methods: The intracellular radionuclide uptake and subcellular distribution of [99mTc]-pertechnetate (99mTc-pertechnetate), [99mTc]Tc-hexamethyl-propylene-aminoxime (99mTc-HMPAO) and [99mTc]Tc-hexakis-2-methoxyisobutylisonitrile (99mTc-MIBI) were quantified in rat thyroid FRTL-5 cells. Radiotoxicity was compared using late phosphorylated histone H2AX (γH2AX) foci as a marker for unrepaired DNA double-strand breaks (DNA-DSB) and clonogenic cell survival. Results:99mTc-HMPAO showed a substantially higher uptake into the nucleus and the membrane/organelles than 99mTc-pertechnetate or 99mTc-MIBI. The colony-forming assay showed that 99mTc-pertechnetate and 99mTc-HMPAO caused a similar reduction in cell survival. 99mTc-MIBI is less radiotoxic in terms of the estimated nucleus dose and induced the fewest number of γH2AX foci compared with the other 99mTc-tracers, and 99mTc-HMPAO induced a fewer number of γH2AX foci than 99mTc-pertechnetate. Conclusions: Our findings reveal that clonogenic cellular survival is not solely determined by the DNA-DSB response. This finding may suggest the involvement of extra-nuclear radiosensitive targets in cell inactivation. For example, the mitochondria or the cell membrane could be affected by 99mTc-HMPAO. [ABSTRACT FROM AUTHOR]

Purpose: We investigated the effect of incorporation of a nuclear localization signal (NLS) into a conjugate comprising the DNA binding ligand para-iodoHoechst (PIH) and octreotate on its DNA binding and affinity to the somatostatin receptor (SSTR). Confirmation of these properties would support development of similar conjugates labelled with Auger emitters for their potential in Auger endoradiotherapy. Materials and methods: We synthesized conjugates of PIH and octreotate (PO) or PIH and NLS (PN) and a conjugate comprising PIH, NLS and octreotate (PNO). DNA-binding characteristics of PIH and conjugates were assessed using synthetic DNA oligonucleotides employing spectrophotometric titration of ligand solutions with DNA. We used membranes from the type 2 SSTR (SSTR2) overexpressing human non-small cell lung cancer cell line A427-7 to investigate the binding affinity of PNO. Results: We demonstrated PN and PNO retain specific high affinity DNA-binding properties observed for PIH, and acquire an additional non-specific binding capacity. No DNA binding was observed for PO. PNO retains its binding affinity for SSTR. Conclusions: The DNA-binding properties of PNO and its affinity for SSTR suggests that it could potentially be used for tumour-specific delivery of PIH labelled with an Auger emitter in SSTR expressing tumours. [ABSTRACT FROM AUTHOR]

To benchmark a Monte Carlo model of the Auger cascade that has been developed at the Australian National University (ANU) against the literature data. The model is applicable to any Auger-electron emitting radionuclide with nuclear structure data in the format of the Evaluated Nuclear Structure Data File (ENSDF). Schönfeld's algorithms and the BrIcc code were incorporated to obtain initial vacancy distributions due to electron capture (EC) and internal conversion (IC), respectively. Atomic transition probabilities were adopted from the Evaluated Atomic Data Library (EADL) for elements with atomic number, Z = 1–100. Atomic transition energies were evaluated using a relativistic Dirac-Fock method. An energy-restriction protocol was implemented to eliminate energetically forbidden transitions from the simulations. Calculated initial vacancy distributions and average energy spectra of 123I, 124I, and 125I were compared with the literature data. In addition, simulated kinetic energy spectra and frequency distributions of the number of emitted electrons and photons of the three iodine radionuclides are presented. Some examples of radiation spectra of individual decays are also given. Good agreement with the published data was achieved except for the outer-shell Auger and Coster-Kronig transitions. Nevertheless, the model needs to be compared with experimental data in a future study. [ABSTRACT FROM AUTHOR]

To identify the emergence of the recognition of the potential of the Auger effect for clinical application, and after tracing the salient milestones towards that goal, to evaluate the status quo and future prospects. It was not until 40 years after the discovery of Auger electrons, that the availability of radioactive DNA precursors enabled the biological power, and the clinical potential, of the Auger effect to be appreciated. Important milestones on the path to clinical translation have been identified and reached, but hurdles remain. Nevertheless the potential is still evident, and there is reasonable optimism that the goal of clinical translation is achievable. [ABSTRACT FROM PUBLISHER]

Purpose: To understand the biological effect of external and internal exposure from 137Cs, DNA damage spectrum induced by directly emitted electrons (γ-rays, internal conversion electrons, Auger electrons) from 137Cs was compared with that induced by 137Cs γ-rays. Methods: Monte Carlo track simulation method was used to calculate the microscopic energy deposition pattern in liquid water. Simulation was performed for the two simple target systems in microscale. Radiation sources were placed inside for one system and outside for another system. To simulate the energy deposition by directly emitted electrons from 137Cs placed inside the system, the multiple ejections of electrons after internal conversion were considered. In the target systems, induction process of DNA damage was modeled and simulated for both direct energy deposition and the water radical reaction on the DNA. The yield and spatial distribution of simple and complex DNA damage including strand breaks and base lesions were calculated for irradiation by electrons and γ-rays from 137Cs. Results: The simulation showed that the significant difference in DNA damage spectrum was not caused by directly ejected electrons and γ-rays from 137Cs. Conclusions: The result supports the existing perception that the biological effects by internal and external exposure by 137Cs are equivalent. [ABSTRACT FROM AUTHOR]

Purpose DNA ligands labelled with 125I induce cytotoxic DNA double-strand breaks (DSB), suggesting a potential for Auger endoradiotherapy. Since the 60-day half-life of 125I is suboptimal for therapy, we have investigated another Auger-emitter 124I, with shorter half-life (4.18 days), and the additional feature of positron-emission, enabling positron emission tomography (PET) imaging. The purpose of this study was to compare the two radionuclides on the basis of DNA DSB per decay. Materials and methods Using a 124I- (or 125I)-labelled minor groove binding DNA ligand, we investigated DNA breakage using the plasmid DNA assay. Biodistribution of the conjugate of the labelled ligand with transferrin was investigated in nude mice bearing a K562 human lymphoma xenograft. Results The probability of DSB per decay was 0.58 and 0.85 for 124I and 125I, respectively, confirming the therapeutic potential of the former. The crystal structure of the ligand DNA complex shows the iodine atom deep within the minor groove, consistent with the high efficiency of induced damage. Biodistribution studies, including PET imaging, showed distinctive results for the conjugate, compared to the free ligand and transferrin, consistent with receptor-mediated delivery of the ligand. Conclusions Conjugation of 124I-labelled DNA ligands to tumor targeting peptides provides a feasible strategy for Auger endoradiotherapy, with the advantage of monitoring tumor targeting by PET imaging. [ABSTRACT FROM AUTHOR]

Purpose: In this work the number of DNA single-strand breaks (SSB) and double-strand breaks (DSB) due to direct and indirect effects of Auger electrons from incorporated 123I and 125I have been calculated by using the Geant4-DNA toolkit. We have performed and compared the calculations for several cases: 125I versus 123I, source positions and direct versus indirect breaks to study the capability of the Geant4-DNA in calculations of DNA damage yields. Materials and methods: Two different simple geometries of a 41 base pair of B-DNA have been simulated. The location of 123I has been considered to be in 123IdUrd and three different locations for 125I. Results: The results showed that the simpler geometry is sufficient for direct break calculations while indirect damage yield is more sensitive to the helical shape of DNA. For 123I Auger electrons, the average number of DSB due to the direct hits is almost twice the DSB due to the indirect hits. Furthermore, a comparison between the average number of SSB or DSB caused by Auger electrons of 125I and 123I in 125IdUrd and 123IdUrd shows that 125I is 1.5 times more effective than 123I per decay. Conclusions: The results are in reasonable agreement with previous experimental and theoretical results which shows the applicability of the Geant-DNA toolkit in nanodosimetry calculations which benefits from the open-source accessibility with the advantage that the DNA models used in this work enable us to save the computational time. Also, the results showed that the simpler geometry is suitable for direct break calculations, while for the indirect damage yield, the more precise model is preferred. [ABSTRACT FROM AUTHOR]

Purpose: The paper gives an introduction into Interatomic and Intermolecular Coulombic Decay (ICD). ICD is an autoionization process, which contrary to Auger decay involves neighbouring sites of the initial vacancy as an integral part of the decay transition. As a result of ICD, slow electrons are produced which generally are known to be active in radiation damage. The author summarizes the properties of ICD and reviews a number of important experiments performed in recent years. Materials and methods: Intermolecular Coulombic Decay can generally take place in weakly bonded aggregates in the presence of ionizing particles or ionizing radiation. Examples collected here mostly use soft X-rays produced by synchrotron radiation to ionize, and use rare-gas clusters, water clusters or solutes in a liquid jet to observe ICD after irradiation. Results: Intermolecular Coulombic Decay is initiated by single ionization into an excited state. The subsequent relaxation proceeds via an ultra-fast energy transfer to a neighbouring site, where a second ionization occurs. Secondary electrons from ICD have clearly been identified in numerous systems. ICD can take place after primary ionization, as the second step of a decay cascade which also involves Auger decay, or after resonant excitation with an energy which exceeds the ionization potential of the system. Conclusions: ICD is expected to play a role whenever particles or radiation with photon energies above the ionization energies for inner valence electrons are present in weakly bonded matter, e.g., biological tissue. The process produces at the same time a slow electron and two charged atomic or molecular fragments, which will lead to structural changes around the ionized site. [ABSTRACT FROM AUTHOR]

Purpose: A comparative study of the effects of different radiation modalities on cell death was performed. Materials and methods: Radiation modalities included γ-rays, fast neutrons, a mixed energy neutron beam called the modified enhanced thermal neutron beam and the mixed beam including Auger electron irradiation by gadolinium neutron capture. U87 (human brain tumor cells) cell survival curve data were modeled to predict how cells died. Transmission electron microscopy (TEM) images were assembled into a morphology of cell death (MCD) database and used to determine the fraction of necrotic or autophagic cells. Results: Linear energy transfer (LET) differences for the different radiation modalities were revealed by modeling. All radiation modalities induced autophagy but only fast neutrons induced significant levels of necrosis. No necrosis, above control levels, was found in cells irradiated with mixed beam irradiation including Auger electrons. The number of autophagosomes increased with increasing time after exposure to all radiation modalities indicating progression of autophagy but only cells irradiated with the mixed beam plus Auger electrons exhibited extreme autophagy. Conclusions: Mixed neutron beam irradiation plus Auger electron irradiation from gadolinium neutron capture is a moderately high LET modality that kills U87 cells without the induction of necrosis and with progression of autophagy to an extreme state. [ABSTRACT FROM AUTHOR]

Purpose: Iodine-125 decay induces localized DNA damage by three major mechanisms: (1) Direct damage by the emitted Auger electrons, (2) indirect damage by diffusible free radicals, and (3) charge neutralization of the residual, highly positively charged, tellurium daughter atom by stripping electrons from neighboring residues. The charge neutralization mechanism of 125I-induced DNA damage is poorly understood. Charge transport along a DNA molecules can occur by either a hopping mechanism initiated by charge injection into DNA and propagated by charge migration through DNA bases along the DNA length, or by a tunneling mechanism in which charge transfers directly from a donor to an acceptor residue. In the first case additional damage in DNA nucleotides can be inflicted by the traveling charge; therefore, it is important to learn if charge hopping plays a role in 125I-decay-induced DNA damage. In our previous work, we determined that at 193K the charge hopping mechanism was not an appreciable component of the mechanism of 125I-induced DNA damage. However, the question whether this is also the case at higher temperatures remained open. Methods: In the current study we used a well-known chemical barrier for charge hopping, 8-oxo-7, 8,-dihydroguanine (8-oxo-G), to assess the role of this mechanism in 125I-decay-induced DNA damage at the following temperatures: 198, 253, 277 and 298 K. Results: We found that varying the temperature had little effect on the distribution of 125I-induced DNA breaks, as well as on the breaks found at the 8-oxo-G probe both with and without piperidine treatment. Conclusions: We thus conclude that charge transport by the hopping mechanism is not a major factor in 125I-decay-induced DNA damage at biologically relevant temperatures. [ABSTRACT FROM AUTHOR]

Purpose: To review recent Auger emitter research with an emphasis on a review of findings on targeting and accumulation of Auger emitters in tumor cells. Conclusion: Significant progress can be reported for targeting methods and improvements in methods to accumulate Auger emitters in the target cells, as well as advances in dose calculations. These studies further our understanding of how Auger emitters induce cell death at a cellular and molecular level, supporting the targeted radiomedical applications of Auger emitters. [ABSTRACT FROM AUTHOR]

Purpose: Triplex-forming oligonucleotides (TFO) bind to the DNA double helix in a sequence-specific manner. Therefore, TFO seem to be a suitable carrier for Auger electron emitters to damage exclusively targeted DNA sequences, e.g., in tumor cells. We studied the influence of I-125 labeled TFO with regard to cell survival and induction of DNA double-strand breaks (DSB) using TFO with different genomic targets and target numbers. Furthermore, the ability of TFO to alter the gene expression of targeted genes was examined. Materials and methods: TFO were labeled with I-125 using the primer extension method. DNA triplex formation and sequence-specific DSB were demonstrated in vitro. Cell survival was analyzed by colony-forming assay and DNA damage was assessed by microscopic quantification of protein 53 binding protein 1 (53BP1) foci in the human squamous carcinoma cell line II (SCL-II). Quantitative real-time polymerase-chain-reaction (qRT-PCR) was performed to analyze gene expression alterations. Results: The sequence-specific induction of a single DSB in a 1695 bp long DNA double stranded fragment was demonstrated in vitro. I-125-labeled TFO binding to single and multiple targets were shown to induce a pronounced decrease in cell survival and an increase of DSB. TFO targeting multiple sites differing in the total target number showed a significant different cell killing per decay that is also in good accordance with the observed induction of DSB. Single gene targeting I-125-labeled TFO significantly decreased cell survival and altered gene expression in the targeted gene. Conclusions: I-125-labeled TFO enable specific targeting of DNA in vitro as well as in a cellular environment and thus induce sequence-specific complex DNA lesions. Therefore I-125-labeled TFO might be a very useful tool for basic DNA repair research. [ABSTRACT FROM AUTHOR]

Purpose: To further elucidate the mechanisms behind the strong biological effectiveness of DNA-incorporated Auger electron emitters 123I and 125I, which are mostly attributed to the shower of low-energy electrons released during the decay. A second, frequently mentioned cause can be seen in the charges accumulated during the Auger cascade on the decaying nuclide and its subsequent intra-molecular redistribution leading to a Coulomb explosion. Methods: To assess the size of the charge and the dimensions of DNA damage thus determined, the first Auger cascade was simulated by Monte Carlo methods. The consequences of intra-molecular charge transfer in terms of structural molecular alterations were estimated by density functional theory (DFT) calculations and folding with the results of the Monte Carlo studies. Results: Charge distributions of 123I and 125I were found to be very similar with values between + 1 and + 15 and a mean value of + 6.4. The molecules could tolerate charges up to + 5 (base), + 2 (nucleoside) and + 7 (nucleotide) without being destroyed. Conclusions: The strong molecular DNA damage after 123I and 125I decay depends very much on the size of the DNA molecule involved in the calculation. In general, not every decay can be expected to lead to a Coulomb explosion. [ABSTRACT FROM AUTHOR]

Purpose: We studied the response of human embryonic stem cells (hESC) to the β-emitter 131I, which affects the entire cell and to the Auger electron emitter 125I-deoxyuridine (125I-dU), primarily affecting the deoxyribonuleic acid (DNA). The effects were also studied in keratinocytes as a prototype for somatic cells. Methods: HESC (H1) and human keratinocytes (HaCaT, human) were exposed to 125I-dU (5 × 10−5 - 5 MBq/ml) and 131I-iodide (5 × 10−5 - 12.5 MBq/ml) and apoptosis was measured by DNA-fragmentation. Cell morphology was studied by light microscopy and electron microscopy. Transcriptional profiling was done on the Agilent oligonucleotide microarray platform. Results: Auger-process induced no apoptosis but a strong transcriptional response in hESC. In contrast, HaCaT cells showed a pronounced induction of apoptosis but only a moderate transcriptional response. Transcriptional response of hESC was similar after 125I-dU and 131I treatments, whereas HaCaT cells expressed a much more pronounced response to 125I-dU than to 131I. A striking radiation-induced down-regulation of pluripotency genes was observed in hESC whereas in keratinocytes the enriched gene annotations were related primarily to apoptosis, cell division and proliferation. Conclusions: Human embryonic stem cells respond to ionizing radiation by 125I-dU and 131I in a different way compared to keratinocytes. Transcriptional response and gene expression appear to facilitate an escape from programmed cell death by striking a new path which probably leads to cell differentiation. [ABSTRACT FROM AUTHOR]

Purpose: To develop a method for simulating the dynamics of the photoelectrons and Auger electrons ejected from DNA molecules irradiated with pulsed monochromatic X-rays. Materials and methods: A 30-base-pair (bp) DNA molecule was used as the target model, and the X-rays were assumed to have a Gaussian-shaped time distribution. Photoionization and Auger decay were considered as the atomic processes. The atoms from which the photoelectrons or Auger electrons were emitted were specified in the DNA molecule (or DNA ion) using the Monte Carlo method, and the trajectory of each electron in the electric field formed around the positively charged DNA molecule was calculated with a Newtonian equation. The kinetics of the electrons produced by irradiation with X-rays at an intensity ranging from 1 × 1012 to 1 × 1016 photons/mm2 and energies of 380 eV (below the carbon K-edge), 435 eV (above the nitrogen K-edge), and 560 eV (above the oxygen K-edge) were evaluated. Results: It was found that at an X-ray intensity of 1 × 1014 photons/mm2 or less, all the produced electrons escaped from the target. However, above an X-ray intensity of 1 × 1015 photons/mm2 and an energy of 560 eV, some photoelectrons that were ejected from the oxygen atoms were trapped near the target DNA. Conclusions: A simulation method for studying the trajectories of electrons ejected from a 30-bp DNA molecule irradiated with pulsed monochromatic X-rays has been developed. The present results show that electron dynamics are strongly dependent on the charged density induced in DNA by pulsed X-ray irradiation. [ABSTRACT FROM AUTHOR]

Purpose: To investigate differences in energy depositions and microdosimetric parameters of low-energy electrons in liquid and gaseous water using Monte Carlo track structure simulations. Materials and methods: KURBUC-liq (Kyushu University and Radiobiology Unit Code for liquid water) was used for simulating electron tracks in liquid water. The inelastic scattering cross sections of liquid water were obtained from the dielectric response model of Emfietzoglou et al. ( Radiation Research 2005;164:202-211). Frequencies of energy deposited in nanometre-size cylindrical targets per unit absorbed dose and associated lineal energies were calculated for 100-5000 eV monoenergetic electrons and the electron spectrum of carbon K edge X-rays. The results for liquid water were compared with those for water vapour. Results: Regardless of electron energy, there is a limit how much energy electron tracks can deposit in a target. Phase effects on the frequencies of energy depositions are largely visible for the targets with diameters and heights smaller than 30 nm. For the target of 2.3 nm by 2.3 nm (similar to dimension of DNA segments), the calculated frequency- and dose-mean lineal energies for liquid water are up to 40% smaller than those for water vapour. The corresponding difference is less than 12% for the targets with diameters ≥ 30 nm. Conclusions: Condensed-phase effects are non-negligible for microdosimetry of low-energy electrons for targets with sizes smaller than a few tens of nanometres, similar to dimensions of DNA molecular structures and nucleosomes. [ABSTRACT FROM AUTHOR]

Purpose: To explore poly(methyl methacrylate) (PMMA950) as an autoradiography substrate. Materials and methods: PMMA950 was spin coated onto a silicon substrate. Resists were exposed to either a 25 or 50 keV electron beam (e-beam) with fluences of 0.1-33.6 μC/cm2. The resulting patterns were analyzed by atomic force microscopy (AFM). The dependence of pattern sensitivity and resolution on resist thickness, development time and electron energy was evaluated and correlated with Monte Carlo (MC) modeling. Conventional micro-autoradiography (MAR) images were compared to AFM images of photoresist patterns obtained following exposure from 111In-diethylenetriaminepentaacetic acid (DTPA)-human epidermal growth factor (hEGF) (4-6 MBq/μg, 40 nM DTPA-hEGF)-treated human breast cancer cells MDA-MB-468. Results: MC simulation results confirmed the similarity of particle transport in PMMA950 exposed to either an 111In point source or a 25 keV e-beam. Sensitivity was inversely related to resist thickness. Development conditions of the resists greatly affected image quality. Sensitivity of PMMA950 was similar to the UVIII™ resist (consisting of a copolymer of 4-hydroxystyrene and t- butylacrylate) at low electron fluence for both 25 and 50 keV e-beam exposure. AFM evaluation of the exposure patterns from 111In-DTPA-hEGF treated cells and nuclei provides more detailed information in comparison with that from MAR. Conclusions: Photoresist autoradiography can provide information on both the distribution of radiation sources and their strengths within a biological sample; however, the choice of photoresist material and processing conditions greatly affects the outcome. [ABSTRACT FROM AUTHOR]

Purpose: To determine the possible effects of 125I-to-DNA distance on the magnitude and mechanism of Auger-electron induced-double-strand break (DSB) production. Materials and methods: We have synthesized a series of 125I-labeled Hoechst (H) derivatives (125IE-H, 125IB-H, 125I-C8-H and 125I-C12-H). While all four molecules share a common DNA minor groove binding bis-benzimidazole motif, they are designed to position 125I at varying distances from the DNA helix. Each Hoechst derivative was incubated at 4°C in phosphate buffered saline (PBS) together with supercoiled (SC) 3H-pUC19 plasmid DNA (ratio 3:1) ± the •OH scavenger dimethyl sulfoxide (DMSO) (0.2 M). Aliquots were analyzed on agarose gels over time and DSB yields per decay of 125I atom were determined. Docking of the iodinated compounds on a DNA molecule was carried out to determine the distance between the iodine atom and the central axis of DNA. Results: In the absence of DMSO, the results show that the DSB yields decrease monotonically as the 125I atom is distanced - by 10.5 Å to 13.9 Å - from the DNA helix (125IEH: 0.52 ± 0.01; 125IB-H: 0.24 ± 0.03; 125I-C8-H: 0.18 ± 0.02; 125I-C12-H: 0.10 ± 0.00). In the presence of DMSO, DSB yields for 125IEH (0.49 ± 0.02) and 125IB-H (0.26 ± 0.04) remain largely unchanged indicating that DSB are entirely produced by direct effects. Strikingly, 125I-C8-H or 125I-C12-H, did not produce detectable DSB in the presence of DMSO under similar conditions suggesting when 125I atom is positioned > 12 Å from the DNA, DSB are entirely produced by indirect effects. Conclusion: These results suggest that at a critical distance between the 125I atom and the DNA helix, DSB production switches from an 'all' direct to an 'all' indirect mechanism, the latter situation being comparable to the decay of 125I free in solution. These experimental findings were correlated with theoretical expectations based on microdosimetry. [ABSTRACT FROM AUTHOR]

Purpose: To calculate the absorbed fraction (AF) of low energy electrons in small tissue-equivalent spherical volumes by Monte Carlo (MC) track structure simulation and assess the influence of phase (liquid water versus density-scaled water vapor) and of the continuous-slowing-down approximation (CSDA) used in semi-analytic calculations. Methods: An event-by-event MC code simulating the transport of electrons in both the vapor and liquid phase of water using appropriate electron-water interaction cross sections was used to quantify the energy deposition of low-energy electrons in spherical volumes. Semi-analytic calculations within the CSDA using a convolution integral of the Howell range-energy expressions are also presented for comparison. Results: The AF for spherical volumes of radii from 10-1000 nm are presented for monoenergetic electrons over the energy range 100-10,000 eV and the two Auger-emitting radionuclides 125I and 123I. The MC calculated AF for the liquid phase are found to be smaller than those of the (density scaled) gas phase by up to 10-20% for the monoenergetic electrons and 10% for the two Auger-emitters. Differences between the liquid-phase MC results and the semi-analytic CSDA calculations are up to ∼ 55% for the monoenergetic electrons and up to ∼ 35% for the two Auger-emitters. Conclusions: Condensed-phase effects in the inelastic interaction of low-energy electrons with water have a noticeable but relatively small impact on the AF for the energy range and target sizes examined. Depending on the electron energies, the semi-analytic approach may lead to sizeable errors for target sizes with linear dimensions below 1 micron. [ABSTRACT FROM AUTHOR]

Purpose: The frequency of DNA strand breaks produced by the decay of Auger electron-emitting radionuclides is inversely proportional to the distance of DNA nucleotides from the decay site; and thus is very sensitive to changes in the local conformation of the DNA. Analysis of the frequency of DNA breaks, or radioprobing, gives valuable information about the local DNA structure. More than 10 years ago, we demonstrated the feasibility of radioprobing using a DNA-repressor complex with a known structure. Herein, we used radioprobing to study the conformation of DNA in complex with the tumor suppressor protein 53 (p53). Several structures of p53-DNA complexes have been solved by X-ray crystallography. These structures, obtained with the p53 DNA binding domain, a truncated form, laid the groundwork for understanding p53-DNA interactions and their relation to p53 functions. However, whether all observed stereochemical details are relevant to the native p53-DNA complex remains unclear. A common theme of the crystallographic structures is the lack of significant bending in the central part of the DNA response element. In contrast, gel electrophoresis and electron microscopy data showed strong DNA bending and overtwisting upon binding to the native p53 tetramer. Methods: To analyze DNA in complex with p53, we incorporated 125I-dCTP in two different positions of synthetic duplexes containing the consensus p53-binding site. Results: The most significant changes in the break frequency distributions were detected close to the center of the binding site, which is consistent with an increase in DNA twisting in this region and local DNA bending and sliding. Conclusions: Our data confirm the main results of the studies made in solution and lay a foundation for systematic examination of interactions between DNA and native p53 using 125I radioprobing. [ABSTRACT FROM AUTHOR]

Purpose: The nanodosimetric description of the radiation action of Auger electrons on nitrogen targets of nanometric size is presented. Materials and methods: Experimental microdosimetry at nanometer scale for Auger electrons has been accomplished with the set-up called Jet Counter. This consists of a pulse-operated valve which injects an expanding nitrogen jet into an interaction chamber where a gaseous sensitive volume of cylindrical shape is created. The ionization cluster size distributions (ICSD) created by Auger electrons emitted by 125I while crossing a nanometer-sized volume have been measured. Results: The ICSD for the sensitive volumes corresponding to 3 and 12 nm in diameter (in unit density 1 g/cm3) irradiated by electrons emitted by a 125I source were collected and compared with the corresponding Monte Carlo (MC) simulation. The preliminary results of the experiments with Auger electrons of 125I interacting with a nitrogen jet having nanometric size comparable to a deoxyribonucleic acid (DNA) and nucleosome, showing the discrete spectrum of ICSD with extended cluster size, are described. Conclusions: The presented paper describes for the first time the nanodosimetric experiments with Auger electrons emitted by 125I. A set of the new descriptors of the radiation quality describing the radiation effect at nanometer level is proposed. The ICSD were determined for the first time for an Auger emitter of 125I. [ABSTRACT FROM AUTHOR]

Purpose: To determine the mode of killing of B-lymphoma cell lines by 111In-conjugated antibodies (Ab). Materials and methods: Cells were treated with a range of radiolabeled Ab concentrations to produce a level of killing from 80 - 99.9%, as determined by a long-term proliferation assay. On days 1, 2, 3 and 4, apoptosis was evaluated in individual cells by two assays: (i) Mitochondrial function by JC-1 staining; and (ii) presence of cleaved PARP (poly-ADP-ribose-polymerase) by immunofluorescence. Five cell lines were tested, including two non-Burkitt lines, RL and FSCCL. Results: In preliminary experiments, it was realized that the standard JC-1 assay, which is widely used, does not distinguish between apoptotic and necrotic cells. The JC-1 assay becomes useful if it is combined with a stain for lysed cells, Sytox green, and then is able to distinguish between viable, apoptotic and necrotic cells. Of the 5 cell lines tested, only Ramos had a large population of apoptotic cells detected by the modified JC-1 assay, but all four of the other cell lines showed evidence of cleaved PARP, albeit to markedly different degrees. The caspase inhibitor, Z-VAD (N-benzyloxycarbonyl-Val-Ala-Asp(O-Me) fluoromethyl ketone), significantly inhibited the killing of Daudi cells. Conclusions: Apoptosis was activated in at least four of the five cell lines tested, and participated in the killing of the cells. Staining for cleaved PARP was much more sensitive than the modified JC-1 assay, for most cell lines, probably because it can detect cells that have already lysed, but which died via apoptosis. In addition, the methodological issues discussed may lead to a better understanding of the role of apoptosis in tumor cell cytotoxicity. [ABSTRACT FROM AUTHOR]

The measurements of the K x-ray intensity ratio I(Kß)/I(Ka) for the 16 elements, Ag, Cd, Sn, Sb, Cs, Ba, La, Ce, Pr, Sm, Eu, Gd, Tb, Dy, Ho, and Er, have been done following ionization by 59.5 keV ?-rays from a 241 Am point source. Ratios of emission probabilities Auger electrons and the vacancy transfer coefficients have been extracted in terms of the intensity ratios. The obtained results agree with previously published experimental results. [ABSTRACT FROM AUTHOR]

A correction to the article "Auger Processes in the 21st Century" that was published in the 2008 issue is presented.