Monitoring of surgical staff x-ray exposure in the operating room with DosiBadge.

Surgical procedures involving the use of x-rays in the operating room (OR) have increased in recent years, thereby increasing the exposure of OR staff to ionizing radiation. An individual dosimeter makes it possible to record the radiation exposure to which these personnel are exposed, but there is a lack of compliance in the wearing of these dosimeters for several practical reasons. This makes the dose results obtained unreliable. To try to improve the rate of dosimeter wearing in the OR, the Dosibadge project studied the association of the individual dosimeter with the hospital access badge, forming the Dosibadge. Through a study performed at the Tours University Hospital in eight different ORs for two consecutive periods of 3 months. The results show a significant increase in the systematic use of the dosimeter thanks to the Dosibadge, which improves the reliability of the doses obtained on the dosimeters and the monitoring of personnel. The increase is especially marked with clinicians. Following these results and the very positive feedback to this first single-centre study, we are then planning a second multicentre study to validate our proof of concept on different sites, with the three brands of individual dosimeters used in France i.e. dosimeters supplied by Dosilab; Landauer and IRSN.

Sample Preparation for X-Ray Fluorescence Microscopy of Iron Distribution in Biological Specimen.

Characterizing the two- and three-dimensional distribution of trace metals in biological specimens is key to better understand their role in biological processes. Iron (Fe) is of particular interest in these trace metals due to its widespread role in maintaining cellular health and preventing disease. X-ray fluorescence microscopy (XFM) is emerging as the method of choice for investigators to interrogate the cellular and subcellular distribution of Fe. XFM utilizes the intrinsic X-ray fluorescence properties of each element to produce quantitative 2D and 3D distributions of trace metals within a sample. Herein, methods for sample preparation of cells and tissue for the determination of Fe distribution by XFM are described.

Development of an Ex Vivo Functional Assay for Prediction of Irradiation Related Toxicity in Healthy Oral Mucosa Tissue.

Radiotherapy in the head-and-neck area is one of the main curative treatment options. However, this comes at the cost of varying levels of normal tissue toxicity, affecting up to 80% of patients. Mucositis can cause pain, weight loss and treatment delays, leading to worse outcomes and a decreased quality of life. Therefore, there is an urgent need for an approach to predicting normal mucosal responses in patients prior to treatment. We here describe an assay to detect irradiation responses in healthy oral mucosa tissue. Mucosa specimens from the oral cavity were obtained after surgical resection, cut into thin slices, irradiated and cultured for three days. Seven samples were irradiated with X-ray, and three additional samples were irradiated with both X-ray and protons. Healthy oral mucosa tissue slices maintained normal morphology and viability for three days. We measured a dose-dependent response to X-ray irradiation and compared X-ray and proton irradiation in the same mucosa sample using standardized automated image analysis. Furthermore, increased levels of inflammation-inducing factors-major drivers of mucositis development-could be detected after irradiation. This model can be utilized for investigating mechanistic aspects of mucositis development and can be developed into an assay to predict radiation-induced toxicity in normal mucosa.

Structure Function Studies of Photosystem II Using X-Ray Free Electron Lasers.

The structure and mechanism of the water-oxidation chemistry that occurs in photosystem II have been subjects of great interest. The advent of X-ray free electron lasers allowed the determination of structures of the stable intermediate states and of steps in the transitions between these intermediate states, bringing a new perspective to this field. The room-temperature structures collected as the photosynthetic water oxidation reaction proceeds in real time have provided important novel insights into the structural changes and the mechanism of the water oxidation reaction. The time-resolved measurements have also given us a view of how this reaction-which involves multielectron, multiproton processes-is facilitated by the interaction of the ligands and the protein residues in the oxygen-evolving complex. These structures have also provided a picture of the dynamics occurring in the channels within photosystem II that are involved in the transport of the substrate water to the catalytic center and protons to the bulk.

Methodology for shielding design and evaluation for container scanners.

There has been an increase in the use of high energy photon beam for container scanners in many countries for multi purposes such as detecting high atomic number materials which might be nuclear materials, drugs, high explosive materials and other contrabands etc. High energy photon beams generally 6 and 9 MV can be used for scanning such materials. However, it is important to ensure that radiation level beyond the container scanner installation is within the permissible dose limit specified by the national competent authority for the protection of public and radiation workers. In this paper, challenges in the biological shielding during the installation of high energy X-ray system for scanning vehicles containing suspected materials are discussed. The purpose of the present study is to develop a methodology for shielding design and evaluation for container scanner installations. The basic concept pertaining to shielding evaluation of radiotherapy installations provided in National Council on Radiation Protection and Measurements (NCRP)/International Atomic Energy Agency (IAEA) reports are referred, and appropriately used to calculate optimized shielding thicknesses requirements for container scanner installation. Workload is estimated based on number of containers scanned, machine ON time and dose rate at 1 m. The shielding evaluation includes use of beam stopper in the primary beam, scattering by heterogeneous metallic scrap materials or any other suspected materials contained in the vehicle and their impact on the thickness of shielding walls. A model lay out plan to be used for installation of container scanner is developed. A methodology for shielding evaluation for various protective walls and ceiling of this model is also discussed. The study provides basic requirement for designing a structural room for installing 9MV container scanner from radiological safety view point.

Theragnostic Gadolinium-Based Nanoparticles Safely Augment X-ray Radiation Effects in Patients with Cervical Cancer.

Activated guided irradiation by X-ray (AGuIX) nanoparticles are gadolinium-based agents that have the dual benefit of mimicking the effects of a magnetic resonance imaging (MRI) contrast agent used in a clinical routine and enhancing the radiotherapeutic activity of conventional X-rays (for cancer treatment). This "theragnostic" action is explained on the one hand by the paramagnetic properties of gadolinium and on the other hand by the generation of high densities of secondary radiation following the interaction of ionizing radiation and high-Z atoms, which leads to enhanced radiation dose deposits within the tumors where the nanoparticles accumulate. Here, we report the results of a phase I trial that aimed to assess the safety and determine the optimal dose of AGuIX nanoparticles in combination with chemoradiation and brachytherapy in patients with locally advanced cervical cancer. AGuIX nanoparticles were administered intravenously and appropriately accumulated within tumors on a dose-dependent manner, as assessed by T1-weighted MRI, with a rapid urinary clearance of uncaught nanoparticles. We show that the observed tumor accumulation of the compounds can support precise delineation of functional target volumes at the time of brachytherapy based on gadolinium enhancement. AGuIX nanoparticles combined with chemoradiation appeared well tolerated among the 12 patients treated, with no dose-limiting toxicity observed. Treatment yielded excellent local control, with all patients achieving complete remission of the primary tumor. One patient had a distant tumor recurrence. These results demonstrate the clinical feasibility of using theranostic nanoparticles to augment the accuracy of MRI-based treatments while focally enhancing the radiation activity in tumors.

X-Ray Film Gel Contact Print Method for Analyzing Differential Expression of Plant Protease Inhibitors Under Abiotic Stress.

The regulation of proteolytic enzymes by protease inhibitors is crucial for maintaining the balance between protein synthesis and degradation, preventing uncontrolled proteolysis and fine-tuning cellular processes essential for optimal function and survival of the plants. It is known that the plant protease inhibitors activities are induced in defense of biotic as well as abiotic stresses. Thus, beyond their fundamental physiological functions, their involvement in stress responses, such as drought, cold, and salinity, is of equally significant. The X-ray film contact print method is an effective method for assessing various protease inhibitors exposed to stress conditions. In this approach, initially plant protease inhibitors will be separated using electrophoresis, and then the gel is treated with trypsin, which inhibits protease inhibitors. This gel when placed on X-ray film, the trypsin will digest the gelatin layer present on the film and the gelatinolytic activity stalled at the premises of protease inhibitors. This will provide the impression of the differentially expressed protease inhibitors in stress-treated plants.

Cervical Spondylosis Diagnosis Based on Convolutional Neural Network with X-ray Images.

The increase in Cervical Spondylosis cases and the expansion of the affected demographic to younger patients have escalated the demand for X-ray screening. Challenges include variability in imaging technology, differences in equipment specifications, and the diverse experience levels of clinicians, which collectively hinder diagnostic accuracy. In response, a deep learning approach utilizing a ResNet-34 convolutional neural network has been developed. This model, trained on a comprehensive dataset of 1235 cervical spine X-ray images representing a wide range of projection angles, aims to mitigate these issues by providing a robust tool for diagnosis. Validation of the model was performed on an independent set of 136 X-ray images, also varied in projection angles, to ensure its efficacy across diverse clinical scenarios. The model achieved a classification accuracy of 89.7%, significantly outperforming the traditional manual diagnostic approach, which has an accuracy of 68.3%. This advancement demonstrates the viability of deep learning models to not only complement but enhance the diagnostic capabilities of clinicians in identifying Cervical Spondylosis, offering a promising avenue for improving diagnostic accuracy and efficiency in clinical settings.

Mechanical properties of blood exosomes and lipoproteins after the rat whole blood irradiation with X-rays in vitro explored by atomic force microscopy.

Blood is a two-component system with two levels of hierarchy: the macrosystem of blood formed elements and the dispersed system of blood nanoparticles. Biological nanoparticles are the key participants in communication between the irradiated and non-irradiated cells and inducers of the non-targeted effects of ionizing radiation. The work aimed at studying by atomic force microscopy the structural, mechanical, and electrical properties of exosomes and lipoproteins (LDL/VLDL) isolated from rat blood after its exposure to X-rays in vitro. MATERIALS AND METHODS: The whole blood of Wistar rats fed with a high-fat diet was irradiated with X-rays (1 and 100 Gy) in vitro. The structural and mechanical properties (the elastic modulus and nonspecific adhesion force) of exosome and lipoprotein isolates from the blood by ultracentrifugation method were studied using Bruker Bioscope Resolve atomic force microscope in PF QNM mode, their electric properties (the zeta-potential) was measured by electrophoretic mobility. RESULTS: Lipoproteins isolated from non-irradiated blood were softer (Me(LQ; UQ): 7.8(4.9;12.1) MPa) compared to blood nanoparticles of its exosome fraction (34.8(22.6;44.9) MPa) containing both exosomes and non-membrane nanoparticles. X-ray blood irradiation with a dose of 1 Gy significantly weakened the elastic properties of lipoproteins. Exposure of the blood to 100 Gy X-rays made lipoproteins stiffer and their nonspecific adhesive properties stronger. The radiation effects on the mechanical parameters of exosomes and non-membrane nanoparticles in exosome fractions differed. The significant radiation-induced change in electric properties of the studied nanoparticles was detected only for lipoproteins in the blood irradiated with 1 Gy X-rays. The low-dose radiation-induced changes in zeta-potential and increase in lipoprotein size with the appearance of a soft thick surface layer indicate the formation of the modified lipoproteins covered with a corona from macromolecules of irradiated blood. CONCLUSION: Our data obtained using the nanomechanical mapping mode of AFM are the first evidence of the significant radiation-induced changes in the structural and mechanical properties of the dispersed system of blood nanoparticles after the X-ray irradiation of the blood.

Quantitative assessment of full-width at half-maximum and detector energy threshold in X-ray imaging systems.

BACKGROUND: The response function of imaging systems is regularly considered to improve the qualified maps in various fields. More the accuracy of this function, the higher the quality of the images. METHODS: In this study, a distinct analytical relationship between full-width at half-maximum (FWHM) value and detector energy thresholds at distinct tube peak voltage of 100 kV has been addressed in X-ray imaging. The outcomes indicate that the behavior of the function is exponential. The relevant cut-off frequency and summation of point spread function S(PSF) were assessed at large and detailed energy ranges. RESULTS: A compromise must be made between cut-off frequency and FWHM to determine the optimal model. By detailed energy range, the minimum and maximum of S(PSF) values were revealed at 20 keV and 48 keV, respectively, by 2979 and 3073. Although the maximum value of FWHM occurred at the energy of 48 keV by 224 mm, its minimum value was revealed at 62 keV by 217 mm. Generally, FWHM value converged to 220 mm and S(PSF) to 3026 with small fluctuations. Consequently, there is no need to increase the voltage of the X-ray tube after the energy threshold of 20 keV. CONCLUSION: The proposed FWHM function may be used in designing the setup of the imaging parameters in order to reduce the absorbed dose and obtain the final accurate maps using the related mathematical suggestions.

X-ray activated near-infrared persistent luminescence nanoparticles for trimodality in vivo imaging.

As promising luminescence nanoparticles, near-infrared (NIR) persistent luminescence nanoparticles (PLNPs) have received extensive attention in the field of high-sensitivity bioimaging in recent years. However, NIR PLNPs face problems such as short excitation wavelengths and single imaging modes, which limit their applications in in vivo reactivated imaging and multimodal imaging. Here, we report for the first time novel Gd2GaTaO7:Cr3+,Yb3+ (GGTO) NIR PLNPs that integrate X-ray activated NIR persistent luminescence (PersL), high X-ray attenuation and excellent magnetic properties into a single nanoparticle (NP). In this case, Cr3+ is used as the luminescence center. The co-doped Yb3+ and coating effectively enhance the X-ray activated NIR PersL. At the same time, the presence of the high-Z element Ta also makes the GGTO NPs exhibit high X-ray attenuation performance, which can be used as a CT contrast agent to achieve in vivo CT imaging. In addition, since the matrix contains a large amount of Gd, the GGTO NPs show remarkable magnetic properties, which can realize in vivo MR imaging. GGTO NPs combine the trimodal benefits of X-ray reactivated PersL, CT and MR imaging and are suitable for single or combined applications that require high sensitivity and spatial resolution imaging.

Exploring the combined impact of cisplatin and copper-cysteamine nanoparticles through Chemoradiation: An in-vitro study.

Copper-Cysteamine nanoparticles (Cu-Cy NPs) have emerged as promising radiosensitizers in cancer treatment. This study aims to investigate the combined therapeutic effect of these nanoparticles and cisplatin using a clinical linear accelerator to enhance the efficacy of chemoradiation therapy for cervical cancer. Following successful synthesis and characterization of Cu-Cy NPs, the cytotoxicity effect of these nanoparticles and cisplatin in various concentrations was evaluated on HeLa cancer cells, individually and in combination. Additionally, the radiobiological effects of these agents were investigated under a 6MV linear accelerator. At a concentration of 25 mg/L, Cu-Cy NPs displayed no significant cytotoxicity toward HeLa cancer cells. However, when combined with 2Gy X-ray irradiation at this concentration, the nanoparticles demonstrated a potent radiosensitizing effect. Notably, cell viability and migration rate in the combination group (Cu-Cy NPs + cisplatin + radiation) were significantly reduced compared to the radiation-alone group. Additionally, the combination treatment induced a significantly higher rate of apoptosis compared to the radiation-alone group. Overall, Cu-Cy NPs exhibited a significant dose-dependent synergistic enhancement of radiation efficacy when combined with cisplatin under X-ray exposure, and may provide a promising approach to improve the therapeutic effect of conventional radiation therapy.

Autophagy is the main driver of radioresistance of HNSCC cells in mild hypoxia.

Hypoxia poses a significant challenge to the effectiveness of radiotherapy in head and neck squamous cell carcinoma (HNSCC) patients, and it is imperative to discover novel approaches to overcome this. In this study, we investigated the underlying mechanisms contributing to x-ray radioresistance in HPV-negative HNSCC cells under mild hypoxic conditions (1% oxygen) and explored the potential for autophagy modulation as a promising therapeutic strategy. Our findings show that HNSCC cells exposed to mild hypoxic conditions exhibit increased radioresistance, which is largely mediated by the hypoxia-inducible factor (HIF) pathway. We demonstrate that siRNA knockdown of HIF-1α and HIF-1ß leads to increased radiosensitivity in HNSCC cells under hypoxia. Hypoxia-induced radioresistance was not attributed to differences in DNA double strand break repair kinetics, as these remain largely unchanged under normoxic and hypoxic conditions. Rather, we identify autophagy as a critical protective mechanism in HNSCC cells following irradiation under mild hypoxia conditions. Targeting key autophagy genes, such as BECLIN1 and BNIP3/3L, using siRNA sensitizes these cells to irradiation. Whilst autophagy's role in hypoxic radioresistance remains controversial, this study highlights the importance of autophagy modulation as a potential therapeutic approach to enhance the effectiveness of radiotherapy in HNSCC.

Combination of proton- or X-irradiation with anti-PDL1 immunotherapy in two murine oral cancers.

Combining radiation therapy with immunotherapy is a strategy to improve both treatments. The purpose of this study was to compare responses for two syngeneic head and neck cancer (HNC) tumor models in mice following X-ray or proton irradiation with or without immune checkpoint inhibition (ICI). MOC1 (immunogenic) and MOC2 (less immunogenic) tumors were inoculated in the right hind leg of each mouse (C57BL/6J, n = 398). Mice were injected with anti-PDL1 (10 mg/kg, twice weekly for 2 weeks), and tumors were treated with single-dose irradiation (5-30 Gy) with X-rays or protons. MOC2 tumors grew faster and were more radioresistant than MOC1 tumors, and all mice with MOC2 tumors developed metastases. Irradiation reduced the tumor volume in a dose-dependent manner. ICI alone reduced the tumor volume for MOC1 with 20% compared to controls, while no reduction was seen for MOC2. For MOC1, there was a clear treatment synergy when combining irradiation with ICI for radiation doses above 5 Gy and there was a tendency for X-rays being slightly more biologically effective compared to protons. For MOC2, there was a tendency of protons being more effective than X-rays, but both radiation types showed a small synergy when combined with ICI. Although the responses and magnitudes of the therapeutic effect varied, the optimal radiation dose for maximal synergy appeared to be in the order of 10-15 Gy, regardless of tumor model.

Optimization of contrast and dose in x-ray phase-contrast tomography with a Talbot-Lau interferometer.

X-ray phase-contrast imaging has become a valuable tool for biomedical research due to its improved contrast abilities over regular attenuation-based imaging. The recently emerged Talbot-Lau interferometer can provide quantitative attenuation, phase-contrast and dark-field image data, even with low-brilliance x-ray tube sources. Thus, it has become a valid option for clinical environments. In this study, we analyze the effects of x-ray tube voltage and total number of images on the contrast-to-noise ratio (CNR) and dose-weighted CNR (CNRD) calculated from tomographic transmission and phase-contrast data of a phantom sample. Constant counting statistics regardless of the voltage was ensured by adjusting the image exposure time for each voltage setting. The results indicate that the x-ray tube voltage has a clear effect on both image contrast and noise. This effect is amplified in the case of phase-contrast images, which is explained by the polychromatic x-ray spectrum and the dependence of interferometer visibility on the spectrum. CNRD is additionally affected by the total imaging time. While submerging the sample into a water container effectively reduces image artefacts and improves the CNR, the additional attenuation of the water must be compensated with a longer exposure time. This reduces dose efficiency. Both the CNR and CNRD are higher in the phase-contrast images compared to transmission images. For transmission images, and phase-contrast images without the water container, CNRD can be increased by using higher tube voltages (in combination with a lower exposure time). For phase-contrast images with the water container, CNRD is increased with lower tube voltages. In general, the CNRD does not strongly depend on the number of tomographic angles or phase steps used.

Volatile flavor behavior characterization of Hericium erinaceus during postharvest storage using E-nose, HS-GC-IMS, and HS-SPME-GC-MS after treated with electron-beam generated X-ray irradiation.

Flavor alteration is a crucial factor affecting the quality of mushrooms during preservation. The dynamic variations of volatile profiles of fresh Hericium erinaceus with electron-beam generated X-ray irradiation were investigated by combining E-nose, headspace-gas chromatography-ion mobility spectrometry (HS-GC-IMS), and headspace solid-phase microextraction gas chromatography-mass spectrometry (HS-SPME-GC-MS). E-nose analysis achieved rapid discrimination in all treatments over storage time. 65 and 73 volatile organic compounds (VOCs) were identified by HS-GC-IMS and HS-SPME-GC-MS, respectively. Thereinto, 1-octen-3-ol, 1-octen-3-one, and 2-octanone were screened out as the characteristic VOCs, which contents declined during storage. While the contents of (E)-2-octenal, (E)-2-nonenal, and 1-octanol increased. The flavor profile changes from distinct mushroom and floral odor to an intense alcohol and fatty odor. Notably, one-kGy irradiation remained more volatiles and denser mushroom odor after storage. Multivariate analysis further confirmed that 1.0 kGy irradiation contributed to the overall aroma retention during postharvest storage of H. erinaceus.

Stability testing of iomeprol and iopamidol formulations subjected to X-ray radiation.

INTRODUCTION: Exposure of iodinated contrast media (ICM) to X-rays is not uncommon, as contrast media are often stored in close proximity to radiological equipment. However, the interaction between X-rays and ICM is not widely investigated in literature. The present study aims to investigate the chemical stability of iomeprol and iopamidol, two commercial iodinated ICM commonly used in diagnostic imaging, under X-rays exposure. METHODS: Different formulations of iopamidol and iomeprol (iodine concentration 9 to 400 mgI/mL, volume 50-500 mL) were exposed to three different conditions of X-ray irradiation: i) 1 month storage in CT room (≈5-15 mGy); (ii) low-dose protocol (≈10 mGy); (ii) stressed protocol (≈100 mGy). Unexposed and exposed solutions were characterized by high-performance liquid chromatography in terms of concentration of active pharmaceutical ingredient (API), iodine species and by products. In addition, appearance and colour of the solutions were inspected and pH measured. RESULTS: API concentrations, appearance, colour and pH of the exposed formulations remained unaffected by X-rays. Measured concentrations of iodine species and by products were observed well within the acceptability criteria, i.e. values turned out to be lower than specifications limits established by the manufacturer, considering both release and shelf-life values. CONCLUSIONS: Up to 100 mGy X-ray exposure did not induce any alteration of iomeprol and iopamidol formulation, nor a detectable increase in the concentration of iodine species or by-products. IMPLICATIONS FOR PRACTICE: Our study strengthens the hypothesis that ICM are stable under X-rays exposure up to 100 mGy.

The impact of X-ray scatter correction software on abdomen radiography in terms of image quality and radiation dose.

INTRODUCTION: The conventional anti-scatter grid is widely used in X-ray radiography to reduce scattered X-rays, but it increases patient dose. Scatter-correction software offers a dose-reducing alternative by correcting for scattered X-rays without a physical grid. Grids and software correction are necessary to reduce scatter radiation and improve image quality especially for the large body parts. The scatter correction can be beneficial in situations where the use of grid is challenging. The implementation of grids and advanced software correction techniques is imperative to ensure that radiographic images maintain high levels of clarity, contrast, and resolution, and ultimately facilitating more accurate diagnoses. This study compares image quality and radiation dose for abdomen exams using scatter correction software and physical grids. METHODS: An anthropomorphic phantom (abdomen) underwent imaging with varying fat and lean tissue layers and body mass index (BMI) configurations. Imaging parameters included 70 kVp tube voltage, 110 cm SID, and Automatic Exposure Control (AEC) both lateral and central chambers. AP abdomen X-ray projections were acquired with and without an anti-scatter grid, and scatter correction software was applied. Image quality was assessed using contrast to noise ratio (CNR) and signal to noise ratio (SNR) metrics. The tube current mAs was considered an exposure factor that affected radiation dose and was used to compare the VG software and physical grid. Radiation dose was measured using Dose Area Products (DAP). The effective dose was estimated using Monte Carlo simulation-PCXMC software. Paired t-tests were used to investigate the image quality difference between the Gridless and VG software, Gridless and PG, and VG software and PG approaches. For the DAP and effective dose, paired t-test was used to investigate the difference between VG software and PG. RESULTS: Images acquired with a grid had the highest mean CNR (71.3 ± 32) compared to Gridless (50 ± 33.8) and scatter correction software (59.3 ± 37.9). The mean SNR of the grid images was (82.7.3 ± 38.9), which is 18% higher than the scatter correction software images (70.4 ± 36.7) and 29% higher than in the Gridless images (62.9.3 ± 34). The mean DAP value was reduced by 81% when the scatter correction software was used compared to the grid (mean: 65.4 µGy.m2 and 338.2 µGy.m2, respectively) with a significant difference (p = 0.001). Scatter correction software resulted in a lower effective dose compared to physical grid use, (mean difference± SD = -0.3 ± 0.18 mSv) with a significant difference (P = 0.02). CONCLUSION: Scatter correction software reduced the radiation dose required but images employing a grid yielded higher CNR and SNR. However, the radiation dose reduction might affect the image quality to a level that impacts the diagnostic information available. Thus, further research needs to be conducted to optimise the use of the scatter correction software. IMPLICATION FOR PRACTICE: Objectively, X-ray scatter correction software might be promising in conditions where a grid cannot be applied.

Evaluation of Cell Cycle-Dependent Migration Activity after X-ray Exposure: A Radiobiological Approach for Optimization of Radiotherapy with Cell Cycle-Targeting Agents.

Radiotherapy with cell cycle-specific anticancer agents has become an important option in the control of both primary tumors and metastases. Here, we used image analysis algorithms that enable quick segmentation and tracking to describe a radiobiological approach for the optimized selection of cell cycle-targeting anticancer drugs for radiotherapy. We confirmed cell cycle-synchronization using human cervical cancer HeLa cells expressing a fluorescent ubiquitination-based cell cycle indicator (FUCCI) as a cell cycle-monitoring probe. Cells synchronized in the G1 and G2 phases were irradiated with X rays at 0.5-2 Gy. Each cell was identified using Cellpose, a deep learning-based algorithm for cellular segmentation, and the velocity and direction of migration were analyzed using the TrackMate plugin in Fiji ImageJ. G1 phase synchronized cells showed a dose-dependent decrease in velocity after irradiation, while G2 cells tended to increase their velocity. The migration pattern of all cells appeared to be a random walk model, regardless of the exposure dose. In addition, we used cisplatin to arrest the cell cycle. HeLa-FUCCI cells arrested at the G2 phase via cisplatin treatment showed enhanced cell migration after X-ray exposure. These results indicated that anticancer agents that arrest the cell cycle of cancer cells in a specific phase may enhance cell migration after radiotherapy. Our approach, using cellular segmentation and tracking algorithms, could enhance the radiobiological assessment of cell cycle-specific migration after irradiation to aid in optimizing radiotherapy using cell cycle-targeting agents.

Mechanism of the Radioresistant Colorectal Cancer Cell Line SW480RR Established after Fractionated X Irradiation.

Radioresistant cancer cells are risk factors for recurrence and are occasionally detected in recurrent tumors after radiotherapy. Intratumor heterogeneity is believed to be a potential cause of treatment resistance. Heterogeneity in DNA content has also been reported in human colorectal cancer; however, little is known about how such heterogeneity changes with radiotherapy or how it affects cancer radioresistance. In the present study, we established radioresistant clone SW480RR cells after fractionated X-ray irradiation of human colorectal cancer-derived SW480.hu cells, which are composed of two cell populations with different chromosome numbers, and examined how cellular radioresistance changed with fractionated radiotherapy. Compared with the parental cell population, which mostly comprised cells with higher ploidy, the radioresistant clones showed lower ploidy and less initial DNA damage. The lower ploidy cells in the parental cell population were identified as having radioresistance prior to irradiation; thus, SW480RR cells were considered intrinsically radioresistant cells selected from the parental population through fractionated irradiation. This study presents a practical example of the emergence of radioresistant cells from a cell population with ploidy heterogeneity after irradiation. The most likely mechanism is the selection of an intrinsically radioresistant population after fractionated X-ray irradiation, with a background in which lower ploidy cells exhibit lower initial DNA damage.