Biological Effects of Low-Dose Chest CT on Chromosomal DNA.

Background Although the National Lung Screening Trial reported a significant reduction in lung cancer mortality when low-dose (LD) CT chest examinations are used for a diagnosis, their biologic effects from radiation exposure remain unclear. Purpose To compare LD CT and standard-dose (SD) CT for DNA double-strand breaks and chromosome aberrations (CAs) in peripheral blood lymphocytes. Materials and Methods Between March 2016 and June 2018, 209 participants who were referred to a respiratory surgery department for chest CT studies were prospectively enrolled in this study. Individuals were excluded if they had undergone radiography examinations within the last 3 days or had undergone chemotherapy or radiation therapy. Peripheral blood samples were obtained before and 15 minutes after CT. The number of γ-H2AX foci and unstable CAs in lymphocytes was quantified by immunofluorescent staining of γ-H2AX and by fluorescence in situ hybridization by using peptide nucleic acid probes for centromeres and telomeres, respectively. The Wilcoxon signed rank test was used for statistical analysis. Bonferroni correction was applied for multiple comparisons. Results Of the 209 participants (105 women, 104 men; mean age, 67.0 years ± 11.3 [standard deviation]), 107 underwent chest LD CT and 102 underwent chest SD CT. Sex distribution, age, and body size metrics were similar between the two groups. The median effective dose of LD CT and SD CT was 1.5 and 5.0 mSv, respectively. The number of double-strand breaks and CAs increased after a SD CT examination (γ-H2AX, P < .001; CAs, P = .003); the number of double-strand breaks and CAs before and after LD CT was not different (γ-H2AX, P = .45; CAs, P = .69). Conclusion No effect of low-dose CT on human DNA was detected. In the same setting, DNA double-strand breaks and chromosome aberrations increased after standard-dose CT. © RSNA, 2020 Online supplemental material is available for this article. See also the editorial by Brenner in this issue.

Compensatory regulation of dopamine after ablation of the tyrosine hydroxylase gene in the nigrostriatal projection.

The tyrosine hydroxylase (TH; EC 1.14.16.2) is a rate-limiting enzyme in the dopamine synthesis and important for the central dopaminergic system, which controls voluntary movements and reward-dependent behaviors. Here, to further explore the regulatory mechanism of dopamine levels by TH in adult mouse brains, we employed a genetic method to inactivate the Th gene in the nigrostriatal projection using the Cre-loxP system. Stereotaxic injection of adeno-associated virus expressing Cre recombinase (AAV-Cre) into the substantia nigra pars compacta (SNc), where dopaminergic cell bodies locate, specifically inactivated the Th gene. Whereas the number of TH-expressing cells decreased to less than 40% in the SNc 2 weeks after the AAV-Cre injection, the striatal TH protein level decreased to 75%, 50%, and 39% at 2, 4, and 8 weeks, respectively, after the injection. Thus, unexpectedly, the reduction of TH protein in the striatum, where SNc dopaminergic axons innervate densely, was slower than in the SNc. Moreover, despite the essential requirement of TH for dopamine synthesis, the striatal dopamine contents were only moderately decreased, to 70% even 8 weeks after AAV-Cre injection. Concurrently, in vivo synthesis activity of l-dihydroxyphenylalanine, the dopamine precursor, per TH protein level was augmented, suggesting up-regulation of dopamine synthesis activity in the intact nigrostriatal axons. Collectively, our conditional Th gene targeting method demonstrates two regulatory mechanisms of TH in axon terminals for dopamine homeostasis in vivo: local regulation of TH protein amount independent of soma and trans-axonal regulation of apparent L-dihydroxyphenylalanine synthesis activity per TH protein.

Evaluating Individual Radiosensitivity for the Prediction of Acute Toxicities of Chemoradiotherapy in Esophageal Cancer Patients.

In this work, individual radiosensitivity was evaluated using DNA damage response and chromosomal aberrations (CAs) in peripheral blood lymphocytes (PBLs) for the prediction of acute toxicities of chemoradiotherapy (CRT) in esophageal cancer patients. Eighteen patients with esophageal cancer were enrolled in this prospective study. Prescribed doses were 60 Gy in 11 patients and 50 Gy in seven patients. Patients received 2 Gy radiotherapy five days a week. PBLs were obtained during treatment just before and 15 min after 2 Gy radiation therapy on the days when the cumulative dose reached 2, 20, 40 Gy and 50 or 60 Gy. PBLs were also obtained four weeks and six months after radiotherapy in all and 13 patients, respectively. Dicentric and ring chromosomes in PBLs were counted to evaluate the number of CAs. Gamma-H2AX foci per cell were scored to assess DNA double-strand breaks. We analyzed the association between these factors and adverse events. The number of γ-H2AX foci before radiotherapy showed no significant increase during CRT, while their increment was significantly reduced with the accumulation of radiation dose. The mean number of CAs increased during CRT up to 1.04 per metaphase, and gradually decreased to approximately 60% six months after CRT. Five patients showed grade 3 toxicities during or after CRT (overreactors: OR), while 13 had grade 2 or less toxicities (non-overreactors: NOR). The number of CAs was significantly higher in the OR group than in the NOR group at a cumulative dose of 20 Gy (mean value: 0.63 vs. 0.34, P = 0.02), 40 Gy (mean value: 0.90 vs. 0.52, P = 0.04), and the final day of radiotherapy (mean value: 1.49 vs. 0.84, P = 0.005). These findings suggest that number of CAs could be an index for predicting acute toxicities of CRT for esophageal cancer.

Diagnostic accuracy of in-stent restenosis using model-based iterative reconstruction at coronary CT angiography: initial experience.

OBJECTIVE: The purpose of our study was to compare the diagnostic performance of coronary CT angiography (CTA) subjected to model-based iterative reconstruction (IR) or hybrid IR to rule out coronary in-stent restenosis. METHODS: We enrolled 16 patients who harboured 22 coronary stents. They underwent coronary CTA on a 320-slice CT scanner. The images were reconstructed with hybrid IR (AIDR 3D) and model-based IR (FIRST) algorithms. We calculated the stent lumen attenuation increase ratio and measured the visible stent lumen diameter. Two blinded observers visually graded the likelihood of in-stent restenosis (lesions ≥ 50%) on hybrid IR and FIRST images. RESULTS: The stent lumen attenuation increase ratio on FIRST- was lower than on AIDR 3D images (0.20 vs 0.32). The ratio of the visible- compared to the true stent lumen diameter was higher on FIRST- than AIDR 3D images (52.5 vs 47.5%). Invasive coronary angiography identified five stents (22.7%) with significant in-stent restenosis. The use of FIRST improved the sensitivity (60 vs 100%), positive (75.0 vs 83.3%) and negative predictive value (88.9 vs 100%) and the accuracy (86.4 vs 95.5%) for the detection of in-stent restenosis. Specificity was 94.1% for both reconstruction methods. CONCLUSION: The model-based IR algorithm may improve diagnostic performance for the detection of in-stent restenosis. Advances in knowledge: Compared to hybrid IR, the new model-based IR algorithm reduced blooming artefacts and improved the image quality. It can be expected to improve diagnostic performance for the detection of in-stent restenosis on coronary CTA images.

Visualization of simulated small vessels on computed tomography using a model-based iterative reconstruction technique.

This article describes a quantitative evaluation of visualizing small vessels using several image reconstruction methods in computed tomography. Simulated vessels with diameters of 1-6 mm made by 3D printer was scanned using 320-row detector computed tomography (CT). Hybrid iterative reconstruction (hybrid IR) and model-based iterative reconstruction (MBIR) were performed for the image reconstruction.

Coronary Artery Stent Evaluation with Model-based Iterative Reconstruction at Coronary CT Angiography.

RATIONALE AND OBJECTIVES: This study aims to compare the image quality of coronary artery stent scans on computed tomography images reconstructed with forward projected model-based iterative reconstruction solution (FIRST) and adaptive iterative dose reduction 3D (AIDR 3D). MATERIALS AND METHODS: Coronary computed tomography angiography scans of 23 patients with 32 coronary stents were used. The images were reconstructed with AIDR 3D and FIRST. We generated computed tomography attenuation profiles across the stents and measured the width of the edge rise distance and the edge rise slope (ERS). We also calculated the stent lumen attenuation increase ratio (SAIR) and measured visible stent lumen diameters. Two radiologists visually evaluated the image quality of the stents using a 4-point scale (1 = poor, 4 = excellent). RESULTS: There was no significant difference in the edge rise distance between the two reconstruction methods (P = 0.36). The ERS on FIRST images was greater than the ERS on AIDR 3D images (325.2 HU/mm vs 224.4 HU/mm; P <0.01). The rate of the visible stent lumen diameter compared to the true diameter on FIRST images was higher than that on AIDR 3D images (51.4% vs 47.3%, P <0.01). The SAIR on FIRST images was lower than the SAIR on AIDR 3D images (0.19 vs 0.30, P <0.01). The mean image quality scores for AIDR 3D and FIRST images were 3.18 and 3.63, respectively; the difference was also significant (P <0.01). CONCLUSION: The image quality of coronary artery stent scans is better on FIRST than on AIDR 3D images.

Coronary CT angiography in patients with implanted cardiac devices: initial experience with the metal artefact reduction technique.

OBJECTIVE: Single-energy metal artefact reduction (SEMAR), a new technique that can now be used in routine CT examinations, has recently become applicable to volume data acquired with electrocardiography gating. We evaluated the effect of this technique on the visualization of the coronary arteries in patients harboring cardiac devices. METHODS: We subjected 8 patients (7 males, 1 female; mean age 65.5 ± 11.3 years) with implanted cardiac devices to coronary CT angiography on a 320-slice CT scanner (Aquilion ONE Vision™; Toshiba Medical Systems Corp., Tokyo, Japan). Image data sets were reconstructed with and without SEMAR. Two radiologists visually evaluated the image quality based on metal artefacts from the electronic device leads using a four-point scale (1 = vessel not visible to 4 = minimal or no metal artefacts). Images with a score of 3 or 4 were considered diagnostic. RESULTS: In both SEMAR and non-SEMAR data sets, 94 coronary artery segments were available for evaluation. Without SEMAR, 11 segments (11.7%) were rated as non-diagnostic; SEMAR improved the image quality of 9 of the 11 segments (81.8%), and the images became diagnostic. CONCLUSION: SEMAR reduced metal artefacts from the electronic device leads and improved the image quality of the coronary arteries in patients with cardiac devices. Advances in knowledge: SEMAR has recently become applicable to volume data acquired with electrocardiography gating. SEMAR reduces metal artefacts elicited by electronic device leads and improves the image quality of the coronary arteries in patients with cardiac devices.

Visualization and genetic analysis of alternative splicing regulation in vivo using fluorescence reporters in transgenic Caenorhabditis elegans.

Transgenic multicolor fluorescence reporters enable the visualization of alternative splicing patterns at a single-cell resolution in living organisms and facilitate further genetic analyses to identify cis-elements and trans-acting factors involved in splicing regulation. In this paper, we describe a method of generating fluorescence alternative splicing reporters for the nematode Caenorhabditis elegans. We describe strategies for designing minigene reporters and methods for constructing them; DNA fragments ('modules', such as promoter/3' cassettes, a genomic fragment of interest and a fluorescent protein cassette) that exist in separate vectors are assembled using site-directed recombination. We also describe strategies and methods for mutant screening and single-nucleotide polymorphism mapping using fluorescence reporters. This is the first detailed description of the design and construction of fluorescence alternative splicing reporters for C. elegans and their use in subsequent genetic analyses. It takes 2-4 months to construct minigenes and generate extrachromosomal lines for visualizing spatiotemporal distribution of alternative splicing events in vivo. Identification of regulators by integration of transgenes, mutant screening and mapping of the responsible genes takes a further 6-12 months. The fluorescence-reporter construction described here can also be applied to the vertebrate cell culture system.