Identification of genomic alterations with clinical impact in canine splenic hemangiosarcoma.

Canine hemangiosarcoma (HSA) is an aggressive cancer of endothelial cells with short survival times. Understanding the genomic landscape of HSA may aid in developing therapeutic strategies for dogs and may also inform therapies for the rare and aggressive human cancer angiosarcoma. The objectives of this study were to build a framework for leveraging real-world genomic and clinical data that could provide the foundation for precision medicine in veterinary oncology, and to determine the relationships between genomic and clinical features in canine splenic HSA. One hundred and nine dogs with primary splenic HSA treated by splenectomy that had tumour sequencing via the FidoCure® Precision Medicine Platform targeted sequencing panel were enrolled. Patient signalment, weight, metastasis at diagnosis and overall survival time were retrospectively evaluated. The incidence of genomic alterations in individual genes and their relationship to patient variables including outcome were assessed. Somatic mutations in TP53 (n = 44), NRAS (n = 20) and PIK3CA (n = 19) were most common. Survival was associated with presence of metastases at diagnosis and germline variants in SETD2 and NOTCH1. Age at diagnosis was associated with somatic NRAS mutations and breed. TP53 and PIK3CA somatic mutations were found in larger dogs, while germline SETD2 variants were found in smaller dogs. We identified both somatic mutations and germline variants associated with clinical variables including age, breed and overall survival. These genetic changes may be useful prognostic factors and provide insight into the genomic landscape of hemangiosarcoma.

Shared hotspot mutations in oncogenes position dogs as an unparalleled comparative model for precision therapeutics.

Naturally occurring canine cancers have remarkable similarities to their human counterparts. To better understand these similarities, we investigated 671 client-owned dogs from 96 breeds with 23 common tumor types, including those whose mutation profile are unknown (anal sac carcinoma and neuroendocrine carcinoma) or understudied (thyroid carcinoma, soft tissue sarcoma and hepatocellular carcinoma). We discovered mutations in 50 well-established oncogenes and tumor suppressors, and compared them to those reported in human cancers. As in human cancer, TP53 is the most commonly mutated gene, detected in 22.5% of canine tumors overall. Canine tumors share mutational hotspots with human tumors in oncogenes including PIK3CA, KRAS, NRAS, BRAF, KIT and EGFR. Hotspot mutations with significant association to tumor type include NRAS G61R and PIK3CA H1047R in hemangiosarcoma, ERBB2 V659E in pulmonary carcinoma, and BRAF V588E (equivalent of V600E in humans) in urothelial carcinoma. Our findings better position canines as a translational model of human cancer to investigate a wide spectrum of targeted therapies.

HOPE: A Pilot Study of Psilocybin Enhanced Group Psychotherapy in Patients With Cancer.

CONTEXT/OBJECTIVES: Psilocybin-assisted psychotherapy shows promise in treating depression and existential distress in people with serious medical illness. However, its individual-based methodology poses challenges for scaling and resource availability. The HOPE trial (A Pilot Study of Psilocybin Enhanced Group Psychotherapy in Patients with Cancer) is an Institutional Review Boards-approved open-label feasibility and safety pilot study examining psilocybin-assisted group therapy in cancer patients with a DSM-5 depressive disorder (including major depressive disorder as well as adjustment disorder with depressed mood). We report here the safety and clinical outcome measures including six-months follow up data. METHODS: Outcome measures were collected at baseline, two-weeks and 26-weeks postintervention. The study involved three group preparatory sessions, one high-dose (25 mg) group psilocybin session, and three group integration sessions with cohorts of four participants over a three-week intervention. RESULTS: Twelve participants completed the trial. no serious adverse events attributed to psilocybin occurred. The primary clinical outcome measures of change in symptoms of depression on the clinician administered 17-item-HAM-D showed clinically substantial decrease in HAM-D scores from baseline to the two-week timepoint (21.5-10.09, P < 0.001) and the 26-week timepoint (21.5-14.83, P = 0.006). Six out of 12 participants met criteria for remission at two weeks, as defined by HAM-D < 7, three out 12 demonstrated a clinically significant change (4-6 points), and eight out of twelve demonstrated a clinically substantial change (7-12 points). CONCLUSION: This pilot study demonstrated the safety, feasibility, and possible efficacy of psilocybin-assisted group therapy for cancer patients dealing with depressive symptoms. Based on demonstrated efficacy and significant reductions in therapist time, future investigations with the group therapy model are warranted.

Evaluating motion of pancreatic tumors and anatomical surrogates using cine MRI in 0.35T MRgRT under free breathing conditions.

Treatment tolerability is a significant limitation to pancreatic cancer treatment with radiotherapy due to proximity to highly radiosensitive organs and respiratory motion necessitating expanded target margins. Further, pancreatic tumors are difficult to visualize on conventional radiotherapy systems. Surrogates are often used to locate the tumor but are often inconsistent and do not provide strong positional relations throughout the respiratory cycle. This work utilizes a retrospective dataset of 45 pancreatic cancer patients treated on an MR-Linac system with cine MRI acquired for real-time target tracking. We investigated intra-fraction motion of tumors and two abdominal surrogates, leading to prediction models between the tumor and surrogate. Patient specific motion evaluation and prediction models were generated from 225 cine MRI series acquired during treatment. Tumor contours were used to evaluate the pancreatic tumor motion. Linear regression and principal component analysis (PCA) based models were used to predict tumor position from the anterior-posterior (AP) motion of the abdominal surface, the superior-inferior (SI) motion of the diaphragm, or a combination. Models were evaluated using mean squared error (MSE) and mean absolute error (MAE). Contour analysis showed the average pancreatic tumor motion range was 7.4 ± 2.7 mm and 14.9 ± 5.8 mm in the AP and SI directions, respectively. The PCA model had MSE of 1.4 mm2 and 0.6 mm2 , for the SI and AP directions, respectively, with both surrogates as inputs for the models. When only the abdomen surrogate was used, MSE was 1.3 mm2 and 0.4 mm2 in the SI and AP directions, while it was 0.4 mm2 and 1.3 mm2 when only the diaphragm surrogate was used. We evaluated intra-fraction pancreatic tumor motion and demonstrated prediction models between the tumor and surrogate. The models calculated the pancreatic tumor position from diaphragm, abdominal, or both contours within standard pancreatic cancer target margin, and the process could be applied to other disease sites in the abdominothoracic cavity.

Analyses of canine cancer mutations and treatment outcomes using real-world clinico-genomics data of 2119 dogs.

Spontaneous tumors in canines share significant genetic and histological similarities with human tumors, positioning them as valuable models to guide drug development. However, current translational studies have limited real world evidence as cancer outcomes are dispersed across veterinary clinics and genomic tests are rarely performed on dogs. In this study, we aim to expand the value of canine models by systematically characterizing genetic mutations in tumors and their response to targeted treatments. In total, we collect and analyze survival outcomes for 2119 tumor-bearing dogs and the prognostic effect of genomic alterations in a subset of 1108 dogs. Our analysis identifies prognostic concordance between canines and humans in several key oncogenes, including TP53 and PIK3CA. We also find that several targeted treatments designed for humans are associated with a positive prognosis when used to treat canine tumors with specific genomic alterations, underscoring the value of canine models in advancing drug discovery for personalized oncology.

Assessment of a novel commercial large field of view phantom for comprehensive MR imaging quality assurance of a 0.35T MRgRT system.

Consistent quality assurance (QA) programs are vital to MR-guided radiotherapy (MRgRT), for ensuring treatment is delivered accurately and the onboard MRI system is providing the expected image quality. However, daily imaging QA with a dedicated phantom is not common at many MRgRT centers, especially with large phantoms that cover a field of view (FOV), similar to the human torso. This work presents the first clinical experience with a purpose-built phantom for large FOV daily and periodic comprehensive quality assurance (QUASAR™ MRgRT Insight Phantom (beta)) from Modus Medical Devices Inc. (Modus QA) on an MRgRT system. A monthly American College of Radiology (ACR) QA phantom was also imaged for reference. Both phantoms were imaged on a 0.35T MR-Linac, a 1.5T Philips wide bore MRI, and a 3.0T Siemens MRI, with T1-weighted and T2-weighted acquisitions. The Insight phantom was imaged in axial and sagittal orientations. Image quality tests including geometric accuracy, spatial resolution accuracy, slice thickness accuracy, slice position accuracy, and image intensity uniformity were performed on each phantom, following their respective instruction manuals. The geometric distortion test showed similar distortions of -1.7 mm and -1.9 mm across a 190 mm and a 283 mm lengths for the ACR and MRgRT Insight phantoms, respectively. The MRgRT Insight phantom utilized a modulation transform function (MTF) for spatial resolution evaluation, which showed decreased performance on the lower B0 strength MRIs, as expected, and could provide a good daily indicator of machine performance. Both the Insight and ACR phantoms showed a match with scan parameters for slice thickness analysis. During the imaging and analysis of this novel MRgRT Insight phantom the authors found setup to be straightforward allowing for easy acquisition each day, and useful image analysis parameters for tracking MRI performance.

Visually guided respiratory motion management for Ethos adaptive radiotherapy.

PURPOSE: Ethos adaptive radiotherapy (ART) is emerging with AI-enhanced adaptive planning and high-quality cone-beam computed tomography (CBCT). Although a respiratory motion management solution is critical for reducing motion artifacts on abdominothoracic CBCT and improving tumor motion control during beam delivery, our institutional Ethos system has not incorporated a commercial solution. Here we developed an institutional visually guided respiratory motion management system to coach patients in regular breathing or breath hold during intrafractional CBCT scans and beam delivery with Ethos ART. METHODS: The institutional visual-guidance respiratory motion management system has three components: (1) a respiratory motion detection system, (2) an in-room display system, and (3) a respiratory motion trace management software. Each component has been developed and implemented in the clinical Ethos ART workflow. The applicability of the solution was demonstrated in installation, routine QA, and clinical workflow. RESULTS: An air pressure sensor has been utilized to detect patient respiratory motion in real time. Either a commercial or in-house software handled respiratory motion trace display, collection and visualization for operators, and visual guidance for patients. An extended screen and a projector on an adjustable stand were installed as the in-room visual guidance solution for the closed-bore ring gantry medical linear accelerator utilized by Ethos. Consistent respiratory motion traces and organ positions on intrafractional CBCTs demonstrated the clinical suitability of the proposed solution in Ethos ART. CONCLUSION: The study demonstrated the utilization of an institutional visually guided respiratory motion management system for Ethos ART. The proposed solution can be easily applied for Ethos ART and adapted for use with any closed bore-type system, such as computed tomography and magnetic resonance imaging, through incorporation with appropriate respiratory motion sensors.

First clinical experience of correcting phantom-based image distortion related to gantry position on a 0.35T MR-Linac.

MR-guided radiotherapy requires strong imaging spatial integrity to deliver high quality plans and provide accurate dose calculation. The MRI system, however, can be compromised by the integrated linear accelerator (Linac), resulting in inaccurate imaging isocenter position and geometric distortion. Dependence on gantry position further complicates the correction of distortions. This work presents a new clinical application of a commercial phantom and software system that quantifies isocenter alignment and geometric distortion, as well as providing a deformation vector field (DVF). A large distortion phantom and a smaller grid phantom were imaged at multiple gantry angles from 0 to 330° on a 0.35 T integrated MR-Linac. The software package was used to assess geometric distortion and generate DVFs to correct distortions within the phantom volume. The DVFs were applied to the grid phantom with resampling software then evaluated using structural similarity index measure (SSIM). Scans were also performed with a ferromagnetic clip near the phantom to investigate the correction of more severe artifacts. The mean magnitude isocenter shift was 0.67 mm, ranging from 0.25 to 1.04 mm across all angles. The DVF had a mean component value of 0.27 ± 0.02, 0.24 ± 0.01, and 0.19 ± 0.01 mm in the right-left (RL), anterior-posterior (AP), and superior-inferior (SI) directions. The ferromagnetic clip increased isocenter position error from 1.98 mm to 2.20 mm and increased mean DVF component values in the RL and AP directions. The resampled grid phantom had an increased SSIM for all gantry angles compared to original images, increasing from 0.26 ± 0.001 to 0.70 ± 0.004. Through this clinical assessment, we were able to correct geometric distortion and isocenter shift related to gantry position on a 0.35 T MR-Linac using the distortion phantom and software package. This provides encouragement that it could be used for quality assurance and clinically to correct systematic distortion caused by imaging at different gantry angles.

Evaluation of super-resolution on 50 pancreatic cancer patients with real-time cine MRI from 0.35T MRgRT.

MR-guided radiotherapy (MRgRT) systems provide excellent soft tissue imaging immediately prior to and in real time during radiation delivery for cancer treatment. However, 2D cine MRI often has limited spatial resolution due to high temporal resolution. This work applies a super resolution machine learning framework to 3.5 mm pixel edge length, low resolution (LR), sagittal 2D cine MRI images acquired on a MRgRT system to generate 0.9 mm pixel edge length, super resolution (SR), images originally acquired at 4 frames per second (FPS). LR images were collected from 50 pancreatic cancer patients treated on a ViewRay MR-LINAC. SR images were evaluated using three methods. 1) The first method utilized intrinsic image quality metrics for evaluation. 2) The second used relative metrics including edge detection and structural similarity index (SSIM). 3) Finally, automatically generated tumor contours were created on both low resolution and super resolution images to evaluate target delineation and compared with DICE and SSIM. Intrinsic image quality metrics all had statistically significant improvements for SR images versus LR images, with mean (±1 SD) BRISQUE scores of 29.65 ± 2.98 and 42.48 ± 0.98 for SR and LR, respectively. SR images showed good agreement with LR images in SSIM evaluation, indicating there was not significant distortion of the images. Comparison of LR and SR images with paired high resolution (HR) 3D images showed that SR images had a mean (±1 SD) SSIM value of 0.633 ± 0.063 and LR a value of 0.587 ± 0.067 (p ≪ 0.05). Contours generated on SR images were also more robust to noise addition than those generated on LR images. This study shows that super resolution with a machine learning framework can generate high spatial resolution images from 4fps low spatial resolution cine MRI acquired on the ViewRay MR-LINAC while maintaining tumor contour quality and without significant acquisition or post processing delay.

Selective Detection of Cu+ Ions in Live Cells via Fluorescence Lifetime Imaging Microscopy.

Copper is an essential trace element in living organisms with its levels and localisation being carefully managed by the cellular machinery. However, if misregulated, deficiency or excess of copper ions can lead to several diseases. Therefore, it is important to have reliable methods to detect, monitor and visualise this metal in cells. Herein we report a new optical probe based on BODIPY, which shows a switch-on in its fluorescence intensity upon binding to copper(I), but not in the presence of high concentration of other physiologically relevant metal ions. More interestingly, binding to copper(I) leads to significant changes in the fluorescence lifetime of the new probe, which can be used to visualize copper(I) pools in lysosomes of live cells via fluorescence lifetime imaging microscopy (FLIM).

Effects of B0 eddy currents on imaging isocenter shifts in 0.35-T MRI-guided radiotherapy (MR-IGRT) system.

PURPOSE: The purpose of this study was to measure gantry angle-related eddy currents in a 0.35-T MRI-Linac and determine if B0 (zeroth order) eddy currents are the primary cause of gantry angle-dependent imaging isocenter shifts vs other potential causes like B0 inhomogeneities and gradient (first order) eddy currents. For conventional Cartesian acquisitions, B0 eddy currents can cause imaging isocenter shifts along both phase encode and readout directions. Gradient eddy currents can cause spatial distortion along both the phase encode and readout directions. Center frequency offsets can cause imaging isocenter shifts along the readout direction that vary with readout gradient polarity. METHODS: MRI-related eddy currents and imaging isocenter shifts were measured on a 0.35-T MRI-Linac at gantry angles from 0° to 330° in increments of 30° . All measurements were made after gradient shimming and center frequency tuning at each planned gantry angle. Eddy current and field homogeneity measurements were conducted using a 24-cm diameter spherical phantom. Gradient and B0 eddy currents were calculated from the free induction decays (FIDs) resulting from selective excitation of slices located ±5 cm from isocenter. B0 eddy currents were also calculated from FIDs acquired with nonselective excitation and compared with B0 eddy current values derived using selective excitation. B0 inhomogeneities and center frequency offsets were measured by acquiring FIDs with nonselective excitation. Imaging isocenter shifts were measured using a 33x33x10.5 cm3 uniformity linearity (grid) phantom and a 3D true fast imaging with steady-state precession (TrueFISP) sequence used in MRI-guided radiation therapy. Eddy currents were compared to vendor specifications and correlated with the imaging isocenter shifts. Measurements were conducted before and after the MRI-Linac's waveguide was replaced with an updated design to reduce eddy currents. RESULTS: B0 eddy currents were highly correlated (r = 0.986, P << 0.001) for measurements made with vs without selective excitation. Transverse (X and Y) axis B0 eddy currents before and after the waveguide upgrade were out of specification (specification: ≤0.1 µT m/mT for delays < 10 ms) for most of the measured gantry angles. Gradient eddy currents before and after the upgrade were within specifications for the measured gantry angles (≤0.1% for delays < 10 ms). B0 eddy currents and imaging isocenter shifts were highly correlated (r = 0.965, P << 0.001). After the Linac waveguide upgrade, root mean square (RMS) peak B0 and gradient eddy currents dropped 45% and 11%, respectively, for delays <10 ms, while imaging isocenter shifts dropped 53%. Isocenter shifts were observed in both phase encode and readout directions. Center frequency offsets were <26 Hz while B0 inhomogeneities were <33 Hz full width at half maximum (FWHM). CONCLUSIONS: Imaging isocenter shifts measured in a 0.35-T MRI-Linac were highly correlated with B0 eddy currents. The eddy currents and imaging isocenter shifts decreased after the MRI-Linac's waveguide was replaced.

Evaluation of diffusion-weighted MRI and geometric distortion on a 0.35T MR-LINAC at multiple gantry angles.

Diffusion-weighted imaging (DWI) provides a valuable diagnostic tool for tumor evaluation. Yet, it is difficult to acquire daily MRI data sets in the traditional radiotherapy clinical setting due to patient burden and limited resources. However, integrated MRI radiotherapy treatment systems facilitate daily functional MRI acquisitions like DWI during treatment exams. Before ADC values from MR-RT systems can be used clinically their reproducibility and accuracy must be quantified. This study used a NIST traceable DWI phantom to verify ADC values acquired on a 0.35 T MR-LINAC system at multiple gantry angles. A diffusion-weighted echo planar imaging sequence was used for all image acquisitions, with b-values of 0, 500, 900, 2000 s/mm2 for the 1.5 T and 3.0 T systems and 0, 200, 500, 800 s/mm2 for the 0.35 T system. Images were acquired at multiple gantry angles on the MR-LINAC system from 0° to 330° in 30° increments to assess the impact of gantry angle on geometric distortion and ADC values. CT images, and three fiducial markers were used as ground truth for geometric distortion measurements. The distance between fiducial markers increased by as much as 7.2 mm on the MR-LINAC at gantry angle 60°. ADC values of deionized water vials from the 1.5 T and 3.0 T systems were 8.30 × 10-6  mm2 /s and -0.85 × 10-6  mm2 /s off, respectively, from the expected value of 1127 × 10-6  mm2 /s. The MR-LINAC system provided an ADC value of the pure water vials that was -116.63 × 10-6  mm2 /s off from the expected value of 1127 × 10-6  mm2 /s. The MR-LINAC also showed a variation in ADC across all gantry angles of 33.72 × 10-6  mm2 /s and 20.41 × 10-6  mm2 /s for the vials with expected values of 1127 × 10-6  mm2 /s and 248 × 10-6  mm2 /s, respectively. This study showed that variation of the ADC values and geometric information on the 0.35 T MR-LINAC system was dependent on the gantry angle at acquisition.

Visualising G-quadruplex DNA dynamics in live cells by fluorescence lifetime imaging microscopy.

Guanine rich regions of oligonucleotides fold into quadruple-stranded structures called G-quadruplexes (G4s). Increasing evidence suggests that these G4 structures form in vivo and play a crucial role in cellular processes. However, their direct observation in live cells remains a challenge. Here we demonstrate that a fluorescent probe (DAOTA-M2) in conjunction with fluorescence lifetime imaging microscopy (FLIM) can identify G4s within nuclei of live and fixed cells. We present a FLIM-based cellular assay to study the interaction of non-fluorescent small molecules with G4s and apply it to a wide range of drug candidates. We also demonstrate that DAOTA-M2 can be used to study G4 stability in live cells. Reduction of FancJ and RTEL1 expression in mammalian cells increases the DAOTA-M2 lifetime and therefore suggests an increased number of G4s in these cells, implying that FancJ and RTEL1 play a role in resolving G4 structures in cellulo.

Characterization of radiotherapy component impact on MR imaging quality for an MRgRT system.

Radiotherapy components of an magnetic resonnace-guided radiotherapy (MRgRT) system can alter the magnetic fields, causing spatial distortion and image deformation, altering imaging and radiation isocenter coincidence and the accuracy of dose calculations. This work presents a characterization of radiotherapy component impact on MR imaging quality in terms of imaging isocenter variation and spatial integrity changes on a 0.35T MRgRT system, pre- and postupgrade of the system. The impact of gantry position, MLC field size, and treatment table power state on imaging isocenter and spatial integrity were investigated. A spatial integrity phantom was used for all tests. Images were acquired for gantry angles 0-330° at 30° increments to assess the impact of gantry position. For MLC and table power state tests all images were acquired at the home gantry position (330°). MLC field sizes ranged from 1.66 to 27.4 cm edge length square fields. Imaging isocenter shift caused by gantry position was reduced from 1.7 mm at gantry 150° preupgrade to 0.9 mm at gantry 120° postupgrade. Maximum spatial integrity errors were 0.5 mm or less pre- and postupgrade for all gantry angles, MLC field sizes, and treatment table power states. However, when the treatment table was powered on, there was significant reduction in SNR. This study showed that gantry position can impact imaging isocenter, but spatial integrity errors were not dependent on gantry position, MLC field size, or treatment table power state. Significant isocenter variation, while reduced postupgrade, is cause for further investigation.

Direct tumor visual feedback during free breathing in 0.35T MRgRT.

To present a tumor motion control system during free breathing using direct tumor visual feedback to patients in 0.35 T magnetic resonance-guided radiotherapy (MRgRT). We present direct tumor visualization to patients by projecting real-time cine MR images on an MR-compatible display system inside a 0.35 T MRgRT bore. The direct tumor visualization included anatomical images with a target contour and an auto-segmented gating contour. In addition, a beam-status sign was added for patient guidance. The feasibility was investigated with a six-patient clinical evaluation of the system in terms of tumor motion range and beam-on time. Seven patients without visual guidance were used for comparison. Positions of the tumor and the auto-segmented gating contour from the cine MR images were used in probability analysis to evaluate tumor motion control. In addition, beam-on time was recorded to assess the efficacy of the visual feedback system. The direct tumor visualization system was developed and implemented in our clinic. The target contour extended 3 mm outside of the gating contour for 33.6 ± 24.9% of the time without visual guidance, and 37.2 ± 26.4% of the time with visual guidance. The average maximum motion outside of the gating contour was 14.4 ± 11.1 mm without and 13.0 ± 7.9 mm with visual guidance. Beam-on time as a percentage was 43.9 ± 15.3% without visual guidance, and 48.0 ± 21.2% with visual guidance, but was not significantly different (P = 0.34). We demonstrated the clinical feasibility and potential benefits of presenting direct tumor visual feedback to patients in MRgRT. The visual feedback allows patients to visualize and attempt to minimize tumor motion in free breathing. The proposed system and associated clinical workflow can be easily adapted for any type of MRgRT.

Characterizing MR Imaging isocenter variation in MRgRT.

We characterized MRI isocenter variation at various gantry positions in two 0.35 T MRgRT systems using two independent methods. First, image center-based quantification was employed on 3D volumetric and 2D cine images of a 24 cm diameter spherical phantom at various gantry positions in the MRI QA mode. The center of the phantom images was identified to quantify the variation of the imaging center at each gantry position. Second, image registration-based quantification was used in radiotherapy mode. 3D volumetric MRIs of a cylindrical phantom were acquired and corresponding image registration from MRI to planning CT was performed. The shifts of the couch were identified to quantify the variation of the imaging center. For verification of noticeable MRI isocenter variation, star-shot pattern measurements with five beams were delivered on the radio-chromic film inserted into the phantom after the couch was shifted. The center of the star-shot pattern was identified to quantify the variation of the imaging center. The proposed methods for measuring MRI isocenter variation were demonstrated with MR-LINAC and MR-60Co systems. Both of the MRgRT systems had field inhomogeneities <5 ppm over a 24 cm diameter spherical volume (DSV) and spatial integrity distortion: <1 mm within 100 mm radius and <2 mm within 175 mm radius. The MRI isocenter of the MR-LINAC system showed noticeable 3D variation (max magnitude: 1.8 mm) compared to that of MR-60Co system (max magnitude: 0.9 mm) relative to the reference gantry positions. In addition, 2D variations (max magnitude) of the MRI isocenter from sagittal cine images were 0.9 mm for the MR-LINAC system and 0.5 mm for the MR-60Co system. Two proposed methods quantified the MRI isocenter variation for various gantry positions in two 0.35 T MRgRT systems. The results of significant isocenter variation in the MR-LINAC system requires further investigation to determine the cause.

X-ray crystallographic structure of BshB, the zinc-dependent deacetylase involved in bacillithiol biosynthesis.

Many gram-positive bacteria produce bacillithiol to aid in the maintenance of redox homeostasis and degradation of toxic compounds, including the antibiotic fosfomycin. Bacillithiol is produced via a three-enzyme pathway that includes the action of the zinc-dependent deacetylase BshB. Previous studies identified conserved aspartate and histidine residues within the active site that are involved in metal binding and catalysis, but the enzymatic mechanism is not fully understood. Here we report two X-ray crystallographic structures of BshB from Bacillus subtilis that provide insight into the BshB catalytic mechanism.

A retrospective 4D-MRI based on 2D diaphragm profiles for lung cancer patients.

INTRODUCTION: 4D-MRI, compared to 4D-CT, provides better soft-tissue contrast for target delineation. However, motion artefacts are often observed due to residual breathing variations. This study is to present a retrospective 4D-MRI reconstruction method based on 2D diaphragm profiles to improve the quality of 4D-MR images in the presence of significant breathing variations. METHODS: The proposed 4D-MRI reconstruction method utilized diaphragm profiles (2D cine images on a single sagittal plan at the peak diaphragm) in conjunction with 4D-MR scans (2D-cine images on multiple pre-determined coronal planes along the anterior-posterior direction over a volume of interest). The diaphragm profile images were exploited to sort the 4D-MR scans by matching respiratory amplitude of diaphragm on the 4D-MR scans to the diaphragm profiles. To evaluate reconstructed 4D-MR images (ten 3D-MR images), sagittal images on ten 3D-MR images under free breathing (FB) and respiratory guidance (GB) were compared with diaphragm profile images (reference) from 13 healthy volunteers. RESULTS: Forty-four 4D-MR scan datasets were successfully reconstructed without distinct respiratory-related motion artefacts even with the presence of breathing variation. The differences in diaphragm profiles between the reference and corresponding reconstructed images in the mean of root mean square were similar between FB (3.5 mm) and GB (3.0 mm), confirming that the 4D-MRI reconstruction method was effective even with significant breathing variation. CONCLUSIONS: The diaphragm profiles were utilized to reconstruct 4D-MR images with spatial reliability and a fixed scan time under FB and GB. Our method can provide reliable 4D information of thoracic and abdominal regions for MRI-guided radiotherapy.

Autologous and Heterologous Cell Therapy for Hemophilia B toward Functional Restoration of Factor IX.

Hemophilia B is an ideal target for gene- and cell-based therapies because of its monogenic nature and broad therapeutic index. Here, we demonstrate the use of cell therapy as a potential long-term cure for hemophilia B in our FIX-deficient mouse model. We show that transplanted, cryopreserved, cadaveric human hepatocytes remain functional for more than a year and secrete FIX at therapeutic levels. Hepatocytes from different sources (companies and donors) perform comparably in curing the bleeding defect. We also generated induced pluripotent stem cells (iPSCs) from two hemophilia B patients and corrected the disease-causing mutations in them by two different approaches (mutation specific and universal). These corrected iPSCs were differentiated into hepatocyte-like cells (HLCs) and transplanted into hemophilic mice. We demonstrate these iPSC-HLCs to be viable and functional in mouse models for 9-12 months. This study aims to establish the use of cells from autologous and heterologous sources to treat hemophilia B.

Monitoring frequency of intra-fraction patient motion using the ExacTrac system for LINAC-based SRS treatments.

PURPOSE: The aim of this study was to investigate the intra-fractional patient motion using the ExacTrac system in LINAC-based stereotactic radiosurgery (SRS). METHOD: A retrospective analysis of 104 SRS patients with kilovoltage image-guided setup (Brainlab ExacTrac) data was performed. Each patient was imaged pre-treatment, and at two time points during treatment (1st and 2nd mid-treatment), and bony anatomy of the skull was used to establish setup error at each time point. The datasets included the translational and rotational setup error, as well as the time period between image acquisitions. After each image acquisition, the patient was repositioned using the calculated shift to correct the setup error. Only translational errors were corrected due to the absence of a 6D treatment table. Setup time and directional shift values were analyzed to determine correlation between shift magnitudes as well as time between acquisitions. RESULTS: The average magnitude translation was 0.64 ± 0.59 mm, 0.79 ± 0.45 mm, and 0.65 ± 0.35 mm for the pre-treatment, 1st mid-treatment, and 2nd mid-treatment imaging time points. The average time from pre-treatment image acquisition to 1st mid-treatment image acquisition was 7.98 ± 0.45 min, from 1st to 2nd mid-treatment image was 4.87 ± 1.96 min. The greatest translation was 3.64 mm, occurring in the pre-treatment image. No patient had a 1st or 2nd mid-treatment image with greater than 2 mm magnitude shifts. CONCLUSION: There was no correlation between patient motion over time, in direction or magnitude, and duration of treatment. The imaging frequency could be reduced to decrease imaging dose and treatment time without significant changes in patient position.