Limb functional recovery is impaired in fibroblast growth factor-2 (FGF2) deficient mice despite chronic ischaemia-induced vascular growth.

FGF2 is a potent stimulator of vascular growth; however, even with a deficiency of FGF2 (Fgf2-/-), developmental vessel growth or ischaemia-induced revascularization still transpires. It remains to be elucidated as to what function, if any, FGF2 has during ischaemic injury. Wildtype (WT) or Fgf2-/- mice were subjected to hindlimb ischaemia for up to 42 days. Limb function, vascular growth, inflammatory- and angiogenesis-related proteins, and inflammatory cell infiltration were assessed in sham and ischaemic limbs at various timepoints. Recovery of ischaemic limb function was delayed in Fgf2-/- mice. Yet, vascular growth response to ischaemia was similar between WT and Fgf2-/- hindlimbs. Several angiogenesis- and inflammatory-related proteins (MCP-1, CXCL16, MMPs and PAI-1) were increased in Fgf2-/- ischaemic muscle. Neutrophil or monocyte recruitment/infiltration was elevated in Fgf2-/- ischaemic muscle. In summary, our study indicates that loss of FGF2 induces a pro-inflammatory microenvironment in skeletal muscle which exacerbates ischaemic injury and delays functional limb use.

Purified PEGylated human glucagon-like peptide-2 reduces the severity of irradiation-induced acute radiation enteritis in rats.

Radiation-induced acute intestinal injury after abdominal and pelvic irradiation is a common and serious problem in the clinical setting. Glucagon-like peptide-2 (GLP-2), a 33-amino acid peptide, exerts diverse effects related to the regulation of gastrointestinal growth and function. However, GLP-2 is relatively unstable in vivo. The aim of the present study was to improve GLP-2 stability in vivo and to evaluate its therapeutic effect on acute radiation enteritis. We generated long-lasting intestinal protection peptides by conjugating human GLP-2 (hGLP-2) peptides to polyethyleneglycol (PEG) to produce mPEGylation hGLP-2 (Mono-PEG-hGLP-2) through an enzymatic site-specific transglutamination reaction. Mono-PEG-hGLP-2 synthesized under optimal reaction conditions and separated by one-step ion-exchange chromatography was found to be resistant to degradation in vitro. Pretreatment with Mono-PEG-hGLP-2 reduced the severity of radiation-induced intestinal injury, oxidative stress, and the expression of NF-κB in rats with irradiation-induced acute radiation enteritis. The enhanced biological potency of Mono-PEG-hGLP-2 highlights its potential as a therapeutic agent for intestinal diseases.

Transarterial radioembolization with yttrium-90 of unresectable primary hepatic malignancy in children.

BACKGROUND: Primary malignant liver tumors are rare, accounting for 1% to 2% of all childhood cancers. When chemotherapy fails, transarterial radioembolization with yttrium-90 (TARE-Y90) may offer an alternative therapy as a bridge to surgical resection or liver transplant or for palliation. METHODS: We conducted a retrospective review of 10 pediatric patients with histologically confirmed primary liver malignancy who received treatment with TARE-Y90. RESULTS: The median age at treatment was 5.5 years (range, 2-18 years). Median patient survival from initial diagnosis was 12.5 months (range, 10-28 months), and median patient survival after TARE-Y90 was 4 months (range, 2-20 months). Retreatment was well tolerated in three of 10 patients, with these patients demonstrating the longest survival times (range, 17-20 months). One patient was transplanted 6 weeks after TARE-Y90. By RECIST 1.1 criteria of all target lesions, eight of nine patients had stable disease, and one of nine had progressive disease. By mRECIST criteria (requiring postcontrast arterial phase imaging), two of seven patients had a partial response, four of seven had stable disease, and one of seven had progressive disease. CONCLUSION: TARE-Y90 of unresectable primary liver malignancy is both technically feasible and demonstrates an anticancer effect, and retreatment is well tolerated. TARE-Y90 could be considered as adjunctive therapy in pediatric patients with unresectable hepatic malignancies and could be used as a bridge to surgical resection or liver transplant. More research is required to determine the efficacy of this treatment in children and to define the clinical scenarios where benefit is likely to be optimized.

Radiation dose reduction through combining positron emission tomography/computed tomography (PET/CT) and diagnostic CT in children and young adults with lymphoma.

BACKGROUND: Both [F-18]2-fluoro-2-deoxyglucose positron emission tomography/computed tomography (18F-FDG PET/CT) and diagnostic CT are at times required for lymphoma staging. This means some body segments are exposed twice to X-rays for generation of CT data (diagnostic CT + localization CT). OBJECTIVE: To describe a combined PET/diagnostic CT approach that modulates CT tube current along the z-axis, providing diagnostic CT of some body segments and localization CT of the remaining body segments, thereby reducing patient radiation dose. MATERIALS AND METHODS: We retrospectively compared total patient radiation dose between combined PET/diagnostic CT and separately acquired PET/CT and diagnostic CT exams. When available, we calculated effective doses for both approaches in the same patient; otherwise, we used data from patients of similar size. To confirm image quality, we compared image noise (Hounsfield unit [HU] standard deviation) as measured in the liver on both combined and separately acquired diagnostic CT images. We used t-tests for dose comparisons and two one-sided tests for image-quality equivalence testing. RESULTS: Mean total effective dose for the CT component of the combined and separately acquired diagnostic CT exams were 6.20±2.69 and 8.17±2.61 mSv, respectively (P<0.0001). Average dose savings with the combined approach was 24.8±17.8% (2.60±2.51 mSv [range: 0.32-4.72 mSv]) of total CT effective dose. Image noise was not statistically significantly different between approaches (12.2±1.8 HU vs. 11.7±1.5 HU for the combined and separately acquired diagnostic CT images, respectively). CONCLUSION: A combined PET/diagnostic CT approach as described offers dose savings at similar image quality for children and young adults with lymphoma who have indications for both PET and diagnostic CT examinations.

Radiation Dose to the Breast by 64-slice CT: Effects of Scanner Model and Study Protocol.

RATIONALE AND OBJECTIVES: This work aimed to study the effects of scanner model and study protocol on radiation dose received by breast tissues from 64-slice computed tomography (CT) studies. MATERIALS AND METHODS: Four scanner models and three study protocols were used in scanning an anthropomorphic phantom with breast modules. Each protocol follows recommendations or guidelines from the American Association of Physicists in Medicine and the American College of Radiology. Twenty thermoluminescent dosimeters were placed inside the breast modules to measure breast tissue doses. Both the absolute and the normalized breast tissue doses were analyzed. RESULTS: The mean glandular doses of a lung cancer screening CT, a chest/abdomen/pelvis CT, and a virtual colonoscopy CT are equivalent to less than 1, 5-7, and 1-3 two-view digital mammograms, respectively, for a standard-sized patient. The normalized breast dose differs significantly (P < 0.01) between lung cancer screening CT and chest/abdomen/pelvis CT; however, it shows less than ±10% variation among scanner models for the same protocol. In virtual colonoscopy CT, breast tissue dose decreases with the distance between local tissues to the edge of the x-ray field, although the decreasing trend varies for different scanner models and protocol settings. CONCLUSIONS: When breasts are entirely included in the primary x-ray field, breast dose by 64-slice CT is mainly protocol dependent, with the normalized breast dose about 15% lower for protocols with modulated mA than for those with constant mA; when breasts are only partially included in the primary beam field, breast dose by 64-slice CT is dependent on both the scanner model and the protocol settings.

Recombinant covalently closed circular hepatitis B virus DNA induces prolonged viral persistence in immunocompetent mice.

It remains crucial to develop a laboratory model for studying hepatitis B virus (HBV) chronic infection. We hereby produced a recombinant covalently closed circular DNA (rcccDNA) in view of the key role of cccDNA in HBV persistence. A loxP-chimeric intron was engineered into a monomeric HBV genome in a precursor plasmid (prcccDNA), which was excised using Cre/loxP-mediated DNA recombination into a 3.3-kb rcccDNA in the nuclei of hepatocytes. The chimeric intron was spliced from RNA transcripts without interrupting the HBV life cycle. In cultured hepatoma cells, cotransfection of prcccDNA and pCMV-Cre (encoding Cre recombinase) resulted in accumulation of nuclear rcccDNA that was heat stable and epigenetically organized as a minichromosome. A mouse model of HBV infection was developed by hydrodynamic injection of prcccDNA. In the presence of Cre recombinase, rcccDNA was induced in the mouse liver with effective viral replication and expression, triggering a compromised T-cell response against HBV. Significant T-cell hyporesponsiveness occurred in mice receiving 4 µg prcccDNA, resulting in prolonged HBV antigenemia for up to 9 weeks. Persistent liver injury was observed as elevated alanine transaminase activity in serum and sustained inflammatory infiltration in the liver. Although a T-cell dysfunction was induced similarly, mice injected with a plasmid containing a linear HBV replicon showed rapid viral clearance within 2 weeks. Collectively, our study provides an innovative approach for producing a cccDNA surrogate that established HBV persistence in immunocompetent mice. It also represents a useful model system in vitro and in vivo for evaluating antiviral treatments against HBV cccDNA. Importance: (i) Unlike plasmids that contain a linear HBV replicon, rcccDNA established HBV persistence with sustained liver injury in immunocompetent mice. This method could be a prototype for developing a mouse model of chronic HBV infection. (ii) An exogenous intron was engineered into the HBV genome for functionally seamless DNA recombination. This original approach could be also extended to other viral studies. (iii) rcccDNA was substantially induced in the nuclei of hepatocytes and could be easily distinguished by its exogenous intron using PCR. This convenient model system affords the opportunity to test antivirals directly targeting HBV cccDNA.

Performance studies of four-dimensional cone beam computed tomography.

Four-dimensional cone beam computed tomography (4DCBCT) has been proposed to characterize the breathing motion of tumors before radiotherapy treatment. However, when the acquired cone beam projection data are retrospectively gated into several respiratory phases, the available data to reconstruct each phase is under-sampled and thus causes streaking artifacts in the reconstructed images. To solve the under-sampling problem and improve image quality in 4DCBCT, various methods have been developed. This paper presents performance studies of three different 4DCBCT methods based on different reconstruction algorithms. The aims of this paper are to study (1) the relationship between the accuracy of the extracted motion trajectories and the data acquisition time of a 4DCBCT scan and (2) the relationship between the accuracy of the extracted motion trajectories and the number of phase bins used to sort projection data. These aims will be applied to three different 4DCBCT methods: conventional filtered backprojection reconstruction (FBP), FBP with McKinnon-Bates correction (MB) and prior image constrained compressed sensing (PICCS) reconstruction. A hybrid phantom consisting of realistic chest anatomy and a moving elliptical object with known 3D motion trajectories was constructed by superimposing the analytical projection data of the moving object to the simulated projection data from a chest CT volume dataset. CBCT scans with gantry rotation times from 1 to 4 min were simulated, and the generated projection data were sorted into 5, 10 and 20 phase bins before different methods were used to reconstruct 4D images. The motion trajectories of the moving object were extracted using a fast free-form deformable registration algorithm. The root mean square errors (RMSE) of the extracted motion trajectories were evaluated for all simulated cases to quantitatively study the performance. The results demonstrate (1) longer acquisition times result in more accurate motion delineation for each method; (2) ten or more phase bins are necessary in 4DCBCT to ensure sufficient temporal resolution in tumor motion and (3) to achieve the same performance as FBP-4DCBCT with a 4 min data acquisition time, MB-4DCBCT and PICCS-4DCBCT need about 2- and 1 min data acquisition times, respectively.

Extraction of tumor motion trajectories using PICCS-4DCBCT: a validation study.

PURPOSE: As a counterpart of 4DCT in the treatment planning stage of radiotherapy treatment, 4D cone beam computed tomography (4DCBCT) method has been proposed to verify tumor motion trajectories before radiation therapy treatment delivery. Besides 4DCBCT acquisition using slower gantry rotation speed or multiple rotations, a new method using the prior image constrained compressed sensing (PICCS) image reconstruction method and the standard 1-min data acquisition were proposed. In this paper, the PICCS-4DCBCT method was combined with deformable registration to validate its capability in motion trajectory extraction using physical phantom data, simulated human subject data from 4DCT and in vivo human subject data. METHODS: Two methods were used to validate PICCS-4DCBCT for the purpose of respiratory motion delineation. The standard 1-min gantry rotation Cone Beam CT acquisition was used for both methods. In the first method, 4DCBCT projection data of a physical motion phantom were acquired using an on-board CBCT acquisition system (Varian Medical Systems, Palo Alto, CA). Using a deformable registration method, the object motion trajectories were extracted from both FBP and PICCS reconstructed 4DCBCT images, and compared against the programmed motion trajectories. In the second method, using a clinical 4DCT dataset, Cone Beam CT projections were simulated by forward projection. Using a deformable registration method, the tumor motion trajectories were extracted from the reconstructed 4DCT and PICCS-4DCBCT images. The performance of PICCS-4DCBCT is assessed against the 4DCT ground truth. The breathing period was varied in the simulation to study its effect on motion extraction. For both validation methods, the root mean square error (RMSE) and the maximum of the errors (MaxE) were used to quantify the accuracy of the extracted motion trajectories. After the validation, a clinical dataset was used to demonstrate the motion delineation capability of PICCS-4DCBCT for human subjects. RESULTS: In both validation studies, the RMSEs of the extracted motion trajectories from PICCS-4DCBCT images are less than 0.7 mm, and their MaxEs are less than 1 mm, for all three directions. In comparison, FBP-4DCBCT shows considerably larger RMSEs in the physical phantom based validation. PICCS-4DCBCT also shows insensitivity to the breathing period in the 4DCT based validation. For the in vivo human subject study, high quality 3D motion trajectory of the tumor was obtained from PICCS-4DCBCT images and showed consistency with visual observation. CONCLUSIONS: These results demonstrate accurate delineation of tumor motion trajectory can be achieved using PICCS-4DCBCT and the standard 1-min data acquisition.

Radiation dose efficiency comparison between differential phase contrast CT and conventional absorption CT.

PURPOSE: The superior radiation dose efficiency of a newly implemented differential phase contrast CT imaging method compared to the conventional absorption CT method is demonstrated. METHODS: A differential phase contrast CT imaging method has recently been implemented using conventional x-ray sources with a grating interferometer consisting of three gratings. This approach offers the possibility of simultaneous reconstruction of both attenuation contrast and phase contrast images from a single acquisition. This enables a direct comparison of radiation dose efficiency of both types of reconstructed images under identical conditions. Radiation dose efficiency was studied by measuring the change in contrast-to-noise ratio (CNR) with different exposure levels. A physical phantom of 28.5 mm diameter was constructed and used for measurement of CNR in both the absorption and phase contrast CT images, which were reconstructed from the same data set. RESULTS: For three of the four materials studied, at any given exposure level, the CNR of the differential phase contrast CT images was superior to that of the corresponding absorption contrast CT images. The most dramatic improvement was noted in the contrast between PMMA and water, where the CNR was improved by a factor of approximately 5.5 in the differential phase contrast CT images. Additionally, the CNR of phase contrast CT is empirically shown to have the same square root dependence on exposure, as is the case for absorption contrast CT. CONCLUSIONS: The differential phase contrast CT method provided higher CNR than conventional absorption CT at equivalent dose levels for most of the materials studied, and so may enable achievement of the same object visibility as conventional absorption CT methods at a lower exposure level.