Assessment of Cell Cytotoxicity and Comet Assay on HER2/neu Positive Cell Line Due to 111In Auger Electrons as DNA-Targeting Radioimmunoconjugate.

BACKGROUND: Breast cancer Auger electron therapy is a growing field of study in radioimmunotherapy and oncology research. Trastuzumab, a high affinity-binding monoclonal antibody against HER2/neu is which is over-expressed in breast tumors, is used in radiopharmaceutical development. OBJECTIVES: In this work, the lethal effects of 111In3+, 111In-DTPA-trastuzumab and 111In-trastuzumab coupled-nuclear localizing sequence peptide (111In-DTPA-NLS-trastuzumab) on malignant cells were studied in vitro. METHODS: DTPA-NLS-trastuzumab was prepared using sulfosuccinimidyl-4-(N-maleimidomethyl) cyclohexane-1-carboxylate (sulfo-SMCC) conjugation with NLS peptide in the first step, followed by conjugation with diethylenetriaminepentaacetic acid (DTPA). Both DTPA-trastuzumab and DTPA- NLS-trastuzumab were labeled with 111In followed by purification and quality control techniques. Sk-Br-3 (a HER2/neu+ cell line), was used in the cell viability assessment assay for 111In, 111In-DTPA-trastuzumab and 111In-DTPA-NLS-trastuzumab (3.7 MBq) at 37 ºC. The cytotoxicity of the three species was studied using MTT and comet assay was utilized DNA damage detection. RESULTS: A significant radiochemical purity for 111In-DTPA-NLS-trastuzumab (99.36% ± 0.30%, ITLC) at the DTPA:antibody ratio of 6.90 ± 0.34:1, was obtained. Significant cell viability difference was found for 111In-DTPA-NLS-trastuzumab compared to the other treatments at two-time points. In addition, comet assay demonstrated significant DNA damage at 144 h using 111In-DTPA- NLS-trastuzumab. CONCLUSION: The results of cell viability and cell death using MTT assay and comet assay, respectively, demonstrate the NLS-peptide effectively facilitates 111In-trastuzumab transport into the HER2/neu positive cancer cell nuclei to impose the radiotherapeutic effects of Auger electrons on DNA leading to cell death.

Simulated response of a multi-element thick gas electron multiplier-based microdosimeter to high energy neutrons.

The response of a microdosimeter for neutrons above 14 MeV is investigated. The mean quality factors and dose-equivalents are determined using lineal energy distributions calculated by Monte Carlo simulations (Geant4 toolkit). From 14 MeV to 5 GeV, the mean quality factors were found to vary between 6.00 and 9.30 and the dose-equivalents were in agreement with the true ambient dose-equivalent at the depth of 10 mm inside the ICRU sphere, H*(10). An energy-independent dose-equivalent response around a median value of 0.86 within 22% uncertainty was obtained. Therefore, the microdosimeter is appropriate for dose-equivalent measurement of high-energy neutrons.

The effect of 111In radionuclide distance and auger electron energy on direct induction of DNA double-strand breaks: a Monte Carlo study using Geant4 toolkit.

PURPOSE: In this study, the effect of 111In position and Auger electron energy on direct induction of DSBs was investigated. MATERIALS AND METHODS: The Geant4-DNA simulation toolkit was applied using a simple B-DNA form extracted from PDBlib library. First, the simulation was performed for electrons with energies of 111In and equal emission probabilities to find the most effective electron energies. Then, 111In Auger electrons' actual spectrum was considered and their contribution in DSB induction analysed. RESULTS: The results showed that the most effective electron energy is 183 eV, but due to the higher emission probability of 350 eV electrons, most of the DSBs were induced by the latter electrons. Also, it was observed that most of the DSBs are induced by electrons emitted within 4 nm of the central axis of the DNA and were mainly due to breaks with <4 base pairs distance in opposing strands. Whilst, when 111In atoms are very close to the DNA, 1.3 DSBs have been obtained per decay of 111In atoms. CONCLUSIONS: The results show that the most effective Auger electrons are the 350 eV electrons from 111In atoms with <4 nm distance from the central axis of the DNA which induce ∼1.3 DSBs per decay when bound to the DNA. This value seems reasonable when compared with the reported experimental data.

Spinning slithole collimation for high-sensitivity small animal SPECT: Design and assessment using GATE simulation.

PURPOSE: While traditional collimations are widely used in preclinical SPECT imaging, they usually suffer from possessing a low system sensitivity leading to noisy images. In this study, we are aiming at introducing a novel collimator, the slithole, offering a superior resolution-sensitivity tradeoff for small animal SPECT. METHODS: The collimator was designed for a molecular SPECT scanner, the HiReSPECT. The slithole is a knife-edge narrow long aperture extended across long-axis of the camera's head. To meet the data completeness requirement, the collimator-detector assembly spins at each regular SPECT angle. The collimator was modeled within GATE Monte Carlo simulator and the data acquisition was performed for NEMA Image Quality (IQ) phantom. In addition, a dedicated 3D iterative reconstruction algorithm based upon plane-integral projections was also developed. RESULTS: The mean sensitivity of the slithole is 285cps/MBq while the current parallel-hole collimator holds a sensitivity of 36cps/MBq at a 30mm distance. The slithole collimation gives rise to a tomographic resolution of 1.8mm compared to a spatial resolution of∼1.7mm for the parallel-hole one (even after resolution modeling). A 1.75 reduction factor in the noise level was observed when the current parallel-hole collimator is replaced by the slithole. Furthermore, quantitative analysis proves that 3 full-iterations of our dedicated image reconstruction lead to optimal image quality. For the largest rod in the NEMA IQ phantom, a recovery coefficient of∼0.83 was obtained. CONCLUSION: The slithole collimator outperforms the current parallel-hole collimation by exhibiting a better resolution-sensitivity compromise for preclinical SPECT studies.

Microdosimetry of DNA conformations: relation between direct effect of (60)Co gamma rays and topology of DNA geometrical models in the calculation of A-, B- and Z-DNA radiation-induced damage yields.

In order to obtain the energy deposition pattern of ionizing radiation in the nanometric scale of genetic material and to investigate the different sensitivities of the DNA conformations, direct effects of (60)Co gamma rays on the three A, B and Z conformations of DNA have been studied. For this purpose, single-strand breaks (SSB), double-strand breaks (DSB), base damage (BD), hit probabilities and three microdosimetry quantities (imparted energy, mean chord length and lineal energy) in the mentioned DNA conformations have been calculated and compared by using GEometry ANd Tracking 4 (Geant4) toolkit. The results show that A-, B- and Z-DNA conformations have the highest yields of DSB (1.2 Gy(-1) Gbp(-1)), SSB (25.2 Gy(-1) Gbp(-1)) and BD (4.81 Gy(-1) Gbp(-1)), respectively. Based on the investigation of direct effects of radiation, it can be concluded that the DSB yield is largely correlated to the topological characteristics of DNA models, although the SSB yield is not. Moreover, according to the comparative results of the present study, a reliable candidate parameter for describing the relationship between DNA damage yields and geometry of DNA models in the theoretical radiation biology research studies would be the mean chord length (4 V/S) of the models.

Calculation of DNA strand breaks due to direct and indirect effects of Auger electrons from incorporated 123I and 125I radionuclides using the Geant4 computer code.

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 (123)I and (125)I have been calculated by using the Geant4-DNA toolkit. We have performed and compared the calculations for several cases: (125)I versus (123)I, 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 (123)I has been considered to be in (123)IdUrd and three different locations for (125)I. 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 (123)I 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 (125)I and (123)I in (125)IdUrd and (123)IdUrd shows that (125)I is 1.5 times more effective than (123)I 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.

Lactose and sucrose aqueous solutions for high-dose dosimetry with 10-MeV electron beam irradiation.

In the present study, dosimetric characterisation of aqueous solutions of lactose and sucrose was analysed by UV spectrometry following irradiation using 10-MeV electron beam at doses between 0.5 and 10.5 kGy. As a dosimetric index, absorbance is selected at 256 and 264 nm for lactose and sucrose aqueous solutions, respectively. The intensity of absorbance for irradiated solutions depends on the pre-irradiation concentration of lactose and sucrose. The post-irradiation stability of both solutions was investigated at room temperature for a measurement period of 22 d.

Monte Carlo and experimental characterization of the first AMIRS 103Pd brachytherapy source.

TG-43U1 dosimetric parameters of a new brachytherapy (103)Pd source, including dose-rate constant, radial dose function, 2D anisotropy function, 1D anisotropy function and anisotropy constant, have been determined using MCNP4C code and have been verified by measurements in Perspex phantoms, using TLD-100 dosimeters calibrated in (60)Co radiation field. The comparison of calculated and measured dosimetric parameters showed the validity of Monte Carlo calculations and experimental results. The anisotropy constant was calculated as 0.87 in water and 0.88 in Perspex; and measured as 0.92 in Perspex. Comparing dosimetric parameters of the new source with other source models showed acceptable agreement.

Monte Carlo calculations and experimental measurements of dosimetric parameters of the IRA-103Pd brachytherapy source.

This article presents a brachytherapy source having 103Pd adsorbed onto a cylindrical silver rod that has been developed by the Agricultural, Medical, and Industrial Research School for permanent implant applications. Dosimetric characteristics (radial dose function, anisotropy function, and anisotropy factor) of this source were experimentally and theoretically determined in terms of the updated AAPM Task group 43 (TG-43U1) recommendations. Monte Carlo simulations were used to calculate the dose rate constant. Measurements were performed using TLD-GR200A circular chip dosimeters using standard methods employing thermoluminescent dosimeters in a Perspex phantom. Precision machined bores in the phantom located the dosimeters and the source in a reproducible fixed geometry, providing for transverse-axis and angular dose profiles over a range of distances from 0.5 to 5 cm. The Monte Carlo N-particle (MCNP) code, version 4C simulation techniques have been used to evaluate the dose-rate distributions around this model 103Pd source in water and Perspex phantoms. The Monte Carlo calculated dose rate constant of the IRA-103Pd source in water was found to be 0.678 cGy h(-1) U(-1) with an approximate uncertainty of +/-0.1%. The anisotropy function, F(r, theta), and the radial dose function, g(r), of the IRA- 103Pd source were also measured in a Perspex phantom and calculated in both Perspex and liquid water phantoms.

Measurement of photoneutron dose produced by wedge filters of a high energy linac using polycarbonate films.

Radiotherapy represents the most widely spread technique to control and treat cancer. To increase the treatment efficiency, high energy linacs are used. However, applying high energy photon beams leads to a non-negligible dose of neutrons contaminating therapeutic beams. In addition, using conventional linacs necessitates applying wedge filters in some clinical conditions. However, there is not enough information on the effect of these filters on the photoneutrons produced. The aim of this study was to investigate the change of photoneutron dose equivalent due to the use of linac wedge filters. A high energy (18 MV) linear accelerator (Elekta SL 75/25) was studied. Polycarbonate films were used to measure the dose equivalent of photoneutrons. After electrochemical etching of the films, the neutron dose equivalent was calculated using Hp(10) factor, and its variation on the patient plane at 0, 5, 10, 50 and 100 cm from the center of the X-ray beam was determined. By increasing the distance from the center of the X-ray beam towards the periphery, the photoneutron dose equivalent decreased rapidly for the open and wedged fields. Increasing of the field size increased the photoneutron dose equivalent. The use of wedge filter increased the proportion of the neutron dose equivalent. The increase can be accounted for by the selective absorption of the high energy photons by the wedge filter.

Experimental measurements and Monte Carlo calculations of dosimetric parameters of the IRA1-103Pd brachytherapy source.

This work presents a brachytherapy source having (103)Pd adsorbed onto a cylindrical silver rod that has been developed by Agricultural, Medical and Industrial Research School for permanent implant applications. Dosimetric characteristics (dose-rate constant, radial dose function, anisotropy function and anisotropy factor) of this source were experimentally and theoretically determined in terms of the updated AAPM Task Group 43 (TG-43U1) recommendations. Measurements were performed using TLD-GR200A circular chip dosimeters using standard methods employing thermoluminescent dosimeters in a Perspex phantom. Precision machined bores in the phantom located dosimeters and source in a reproducible fixed geometry providing for transverse-axis and angular dose profiles over a range of distances from 0.5 to 5 cm. The Monte Carlo N-Particle (MCNP) code, version 4C was used to evaluate the dose-rate distributions around this model (103)Pd source in water and Perspex phantoms. The Monte Carlo calculated dose-rate constant of the IRA1-(103)Pd source in water was found equal to Lambda=0.669 cGy/h/U with approximate uncertainties of +/-0.1%. The anisotropy function, F(r, theta), and the radial dose function, g(L)(r), of the IRA1-(103)Pd source were also measured in Perspex phantom and calculated in both Perspex and liquid water phantom.

The effect of field modifier blocks on the fast photoneutron dose equivalent from two high-energy medical linear accelerators.

High-energy linear accelerators (linacs) have several advantages, including low skin doses and high dose rates at deep-seated tumours. But, at energies more than 8 MeV, photonuclear reactions produce neutron contamination around the therapeutic beam, which may induce secondary malignancies. In spite of improvements achieved in medical linac designs, many countries still use conventional (non-intensity-modulated radiotherapy) linacs. Hence, in these conventional machines, fitting the beam over the treatment volume may require using blocks. Therefore, the effect of these devices on neutron production of linacs needs to be studied. The aim of this study was to investigate the effect of field shaping blocks on photoneutron dose in the treatment plane for two high-energy medical linacs. Two medical linacs, a Saturn 43 (25 MeV) and an Elekta SL 75/25 (18 MeV), were studied. Polycarbonate (PC) films were used to measure the fluence of photoneutrons produced by these linacs. After electrochemical etching of the PC films, the neutron dose equivalent was calculated at the isocentre and 50 cm away from the isocentre. It was noted that by increasing the distance from the centre of the X-ray beam towards the periphery, the photoneutron dose equivalent decreases rapidly for both the open and blocked fields. Increasing the energy of the photons causes an increase in the amount of photoneutron dose equivalent. At 25 MeV photon energy, the lead blocks cause a meaningful increase in the dose equivalent of photoneutrons. In this research, a 30% increase was seen in neutron dose contribution to central axis dose at the isocentre of a 25 MeV irregular field shaped by lead blocks. It is concluded that lead blocks must be considered as a source of photoneutron production when treating irregular fields with high-energy photons.