Reactive oxygen species-inducing titanium peroxide nanoparticles as promising radiosensitizers for eliminating pancreatic cancer stem cells.

BACKGROUND: Despite recent advances in radiotherapy, radioresistance in patients with pancreatic cancer remains a crucial dilemma for clinical treatment. Cancer stem cells (CSCs) represent a major factor in radioresistance. Developing a potent radiosensitizer may be a novel candidate for the eradication of pancreatic CSCs. METHODS: CSCs were isolated from MIA PaCa-2 and PANC1 human pancreatic cancer cell lines. Titanium peroxide nanoparticles (TiOxNPs) were synthesized from titanium dioxide nanoparticles (TiO2NPs) and utilized as radiosensitizers when added one hour prior to radiation exposure. The antitumor activity of this novel therapeutic strategy was evaluated against well-established pancreatic CSCs model both in vitro and in vivo. RESULTS: It is shown that TiOxNPs combined with ionizing radiation exhibit anti-cancer effects on radioresistant CSCs both in vitro and in vivo. TiOxNPs exhibited a synergistic effect with radiation on pancreatic CSC-enriched spheres by downregulating self-renewal regulatory factors and CSC surface markers. Moreover, combined treatment suppressed epithelial-mesenchymal transition, migration, and invasion properties in primary and aggressive pancreatic cancer cells by reducing the expression of proteins relevant to these processes. Notably, radiosensitizing TiOxNPs suppressed the growth of pancreatic xenografts following primary or dissociating sphere MIA PaCa-2 cell implantation. It is inferred that synergy is formed by generating intolerable levels of reactive oxygen species (ROS) and inactivating the AKT signaling pathway. CONCLUSIONS: Our data suggested the use of TiOxNPs in combination with radiation may be considered an attractive therapeutic strategy to eliminate pancreatic CSCs.

Elucidation of gastrointestinal dysfunction in response to irradiation using metabolomics.

Gastrointestinal toxicity is frequently observed secondary to accidental or therapeutic radiation exposure. However, the variation in the intestinal metabolites after abdominal radiation exposure remains ambiguous. In the present study, C57BL/6 mice were exposed to 0, 2, and 20 Gy irradiation dose. The Head and chest of each mouse were covered with a lead shield before x-ray irradiation. 24 h post-irradiation treatment, intestinal tissue of each mouse was excised and prepared for metabolites measurement using gas chromatography-mass spectrometry (GC-MS). Our comprehensive analysis of metabolites in the intestinal tissues detected 44 metabolites after irradiation, including amino acids, carbohydrates, organic acids, and sugars. Amino acid levels in the intestinal tissue gradually rose, dependent on the radiation dose, perhaps as an indication of oxidative stress. Our findings raise the possibility that amino acid metabolism may be a potential target for the development of treatments to alleviate or mitigate the harmful effects of oxidative stress-related gastrointestinal toxicity due to radiation exposure.

Real-time in vivo dosimetry system based on an optical fiber-coupled microsized photostimulable phosphor for stereotactic body radiation therapy.

PURPOSE: To develop an in vivo dosimeter system for stereotactic body radiation therapy (SBRT) that can perform accurate and precise real-time measurements, using a microsized amount of a photostimulable phosphor (PSP), BaFBr:Eu2+ . METHODS: The sensitive volume of the PSP was 1.26 × 10-5  cm3 . The dosimeter system was designed to apply photostimulation to the PSP after the decay of noise signals, in synchronization with the photon beam pulse of a linear accelerator (LINAC), to eliminate the noise signals completely using a time separation technique. The noise signals included stem signals, and radioluminescence signals generated by the PSP. In addition, the dosimeter system was built on a storage-type dosimeter that could read out a signal after an arbitrary preset number of photon beam pulses were incident. First, the noise and photostimulated luminescence (PSL) signal decay times were measured. Subsequently, we confirmed that the PSL signals could be exclusively read out within the photon beam pulse interval. Finally, using a water phantom, the basic characteristics of the dosimeter system were demonstrated under SBRT conditions, and the feasibility for clinical application was investigated. The reproducibility, dose linearity, dose-rate dependence, temperature dependence, and angular dependence were evaluated. The feasibility was confirmed by measurements at various dose gradients and using a representative treatment plan for a metastatic liver tumor. A clinical plan was created with a two-arc beam volumetric modulated arc therapy using a 10 MV flattening filter-free photon beam. For the water phantom measurements, the clinical plan was compiled into a plan with a fixed gantry angle of 0°. To evaluate the energy dependence during SBRT, the percent depth dose (PDD) was measured and compared with those calculated via Monte Carlo (MC) simulations. RESULTS: All the PSL signals could be read out while eliminating the noise signals within the minimum pulse interval of the LINAC. Stable real-time measurements could be performed with a time resolution of 56 ms (i.e., number of pulses = 20). The dose linearity was good in the dose range of 0.01-100 Gy. The measurements agreed within 1% at dose rates of 40-2400 cGy/min. The temperature and angular dependence were also acceptable since these dependencies had only a negligible effect on the measurements in SBRT. At a dose gradient of 2.21 Gy/mm, the measured dose agreed with that calculated using a treatment planning system (TPS) within the measurement uncertainties due to the probe position. For measurements using a representative treatment plan, the measured dose agreed with that calculated using the TPS within 0.5% at the center of the beam axis. The PDD measurements agreed with the MC calculations to within 1% for field sizes <5 × 5 cm2 . CONCLUSION: The in vivo dosimeter system developed using BaFBr:Eu2+ is capable of real-time, accurate, and precise measurement under SBRT conditions. The probe is smaller than a conventional dosimeter, has excellent spatial resolution, and can be valuable in SBRT with a steep dose distribution over a small field. The developed PSP dosimeter system appears to be suitable for in vivo SBRT dosimetry.

Upstream perturbation and floodplain formation effects on chute-cutoff-dominated meandering river pattern and dynamics.

A sustained dynamic inflow perturbation and bar-floodplain conversion are considered crucial to dynamic meandering. Past experiments, one-dimensional modelling and linear theory have demonstrated that the initiation and persistence of dynamic meandering require a periodic transverse motion of the inflow. However, it remains unknown whether the period of the inflow perturbation affects self-formed meander dynamics. Here, we numerically study the effect of the inflow perturbation period on the development and meander dynamics of a chute-cutoff-dominated river, which requires two-dimensional modelling with vegetation forming floodplain on bars. We extended the morphodynamic model Nays2D with growth and mortality rules of vegetation to allow for meandering. We tested the effect of a transversely migrating inflow boundary by varying the perturbation period between runs over an order of magnitude around typical modelled meander periods. Following the cutoff cascade after initial meander formation from a straight channel, all runs with sufficient vegetation show series of growing meanders terminated by chute cutoffs. This generates an intricate channel belt topography with point bar complexes truncated by chutes, oxbow lakes, and scroll-bar-related vegetation age patterns. The sinuosity, braiding index and meander period, which emerge from the inherent biomorphological feedback loops, are unrelated to the inflow perturbation period, although the spin-up to dynamic equilibrium takes a longer time and distance for weak and absent inflow perturbations. This explains why, in previous experimental studies, dynamic meandering was only accomplished with a sustained upstream perturbation in flumes that were short relative to the meander wavelength. Our modelling of self-formed meander patterns is evidence that scroll-bar-dominated and chute-cutoff-dominated meanders develop from downstream convecting instabilities. This insight extends to many more fluvial, estuarine and coastal systems in morphological models and experiments, which require sustained dynamic perturbations to form complex patterns and develop natural dynamics. © 2019 The Authors. Earth Surface Processes and Landforms Published by John Wiley & Sons Ltd.

Amelioration of Radiation Enteropathy by Dietary Supplementation With Reduced Coenzyme Q10.

PURPOSE: Effective methods to ameliorate radiation enteropathy have not been developed. To address this issue, we investigated the reduced form of coenzyme Q10 (rCoQ10) as a potential radioprotector in a mouse model. METHODS AND MATERIALS: rCoQ10 was added to a standard laboratory mouse diet at a final concentration of 1.0% 9 days before irradiation and 30 days thereafter or dissolved in corn oil and administered transorally. Accumulated amounts of coenzyme Q10 (CoQ10) or coenzyme Q9 in the intestine were measured by high-performance liquid chromatography. Reactive oxygen species (ROS), apoptosis, and morphologic changes in the intestine were assessed by immunohistochemistry after administration of 13 Gy of x-ray to the mouse abdomen. Body weight and survival were monitored for 30 days after irradiation. Cytotoxicity using 3 human cancer cell lines and the tumor growth-inhibiting effect in a xenograft were investigated to determine whether rCoQ10 interferes with radiation-specific cytotoxic effects on tumor growth. RESULTS: CoQ10 was greatly accumulated in all sections of the intestine after both massive transoral dosing and dietary administration, whereas coenzyme Q9 was not. Administration of rCoQ10 suppressed ROS production and inhibited apoptosis in the crypts, resulting in preservation of villi structures after irradiation. Notably, 92% of mice fed the rCoQ10-supplemented diet were healthy and alive 30 days after irradiation, whereas 50% of control mice died (P < .05). Moreover, rCoQ10 did not interfere with radiation-specific cytotoxic effects on tumors either in vitro or in vivo. CONCLUSIONS: Administration of rCoQ10 led to its accumulation in the intestine and induced radioprotective effects by inhibiting ROS-mediated apoptosis, thereby preserving intestinal structures. Our results indicated that rCoQ10 supplementation effectively ameliorated radiation enteropathy.

Evaluation of a Small Animal Irradiation System for Animal Experiments Using EBT3 Model GAFCHROMIC™ Film.

In cancer research, small animal models, for example, mice, rats, or rabbits, facilitate the in-depth study of biological processes and the effects of radiation treatment that can lead to breakthrough discoveries. However, the physical quality of small animal irradiation systems has not been previously evaluated. In this study, we evaluate the quality of a small animal irradiation system using GAFCHROMIC™ film and a Tough Water Phantom. The profiles and percentage depth dose curves for several irradiation conditions were measured to evaluate the quality of the irradiation system. The symmetry ratios when the table was rotated were 1.1 (no filter), 1.0 (0.5 mm Al filter), 1.0 (1.0 mm Al filter), 1.1 (2 mm Al filter), and 1.0 (filter consisting of 0.5 mm Al combined with 0.1 mm Cu). The results of measuring the percentage depth dose curve showed that the relative doses were 17.5% (10 mm depth), 12.4% (20 mm depth), 9.5% (30 mm depth), and 7.4% (40 mm filter) with no filters inserted, 78.0% (10 mm depth), 61.1% (20 mm depth), 46.9% (30 mm depth), and 35.3% (40 mm depth) when a 1.0 mm Al filter was inserted, and 94.4% (10 mm depth), 81.7% (20 mm depth), 68.1% (30 mm depth), and 54.7% (40 mm depth) when a filter consisting of 1.0 mm Al combined with 0.2 mm Cu was inserted. These physical assessments seem to be necessary especially in vivo experiments because those increase reliability of data obtained from small animal irradiation systems.

Sparing of tissue by using micro-slit-beam radiation therapy reduces neurotoxicity compared with broad-beam radiation therapy.

Micro-slit-beam radiation therapy (MRT) using synchrotron-generated X-ray beams allows for extremely high-dose irradiation. However, the toxicity of MRT in central nervous system (CNS) use is still unknown. To gather baseline toxicological data, we evaluated mortality in normal mice following CNS-targeted MRT. Male C57BL/6 J mice were head-fixed in a stereotaxic frame. Synchrotron X-ray-beam radiation was provided by the SPring-8 BL28B2 beam-line. For MRT, radiation was delivered to groups of mice in a 10 × 12 mm unidirectional array consisting of 25-µm-wide beams spaced 100, 200 or 300 µm apart; another group of mice received the equivalent broad-beam radiation therapy (BRT) for comparison. Peak and valley dose rates of the MRT were 120 and 0.7 Gy/s, respectively. Delivered doses were 96-960 Gy for MRT, and 24-120 Gy for BRT. Mortality was monitored for 90 days post-irradiation. Brain tissue was stained using hematoxylin and eosin to evaluate neural structure. Demyelination was evaluated by Klüver-Barrera staining. The LD50 and LD100 when using MRT were 600 Gy and 720 Gy, respectively, and when using BRT they were 80 Gy and 96 Gy, respectively. In MRT, mortality decreased as the center-to-center beam spacing increased from 100 µm to 300 µm. Cortical architecture was well preserved in MRT, whereas BRT induced various degrees of cerebral hemorrhage and demyelination. MRT was able to deliver extremely high doses of radiation, while still minimizing neuronal death. The valley doses, influenced by beam spacing and irradiated dose, could represent important survival factors for MRT.

The possible interplanetary transfer of microbes: assessing the viability of Deinococcus spp. under the ISS Environmental conditions for performing exposure experiments of microbes in the Tanpopo mission.

To investigate the possible interplanetary transfer of life, numerous exposure experiments have been carried out on various microbes in space since the 1960s. In the Tanpopo mission, we have proposed to carry out experiments on capture and space exposure of microbes at the Exposure Facility of the Japanese Experimental Module of the International Space Station (ISS). Microbial candidates for the exposure experiments in space include Deinococcus spp.: Deinococcus radiodurans, D. aerius and D. aetherius. In this paper, we have examined the survivability of Deinococcus spp. under the environmental conditions in ISS in orbit (i.e., long exposure to heavy-ion beams, temperature cycles, vacuum and UV irradiation). A One-year dose of heavy-ion beam irradiation did not affect the viability of Deinococcus spp. within the detection limit. Vacuum (10(-1) Pa) also had little effect on the cell viability. Experiments to test the effects of changes in temperature from 80 °C to -80 °C in 90 min (± 80 °C/90 min cycle) or from 60 °C to -60 °C in 90 min (± 60 °C/90 min cycle) on cell viability revealed that the survival rate decreased severely by the ± 80 °C/90 min temperature cycle. Exposure of various thicknesses of deinococcal cell aggregates to UV radiation (172 nm and 254 nm, respectively) revealed that a few hundred micrometer thick aggregate of deinococcal cells would be able to withstand the solar UV radiation on ISS for 1 year. We concluded that aggregated deinococcal cells will survive the yearlong exposure experiments. We propose that microbial cells can aggregate as an ark for the interplanetary transfer of microbes, and we named it 'massapanspermia'.