Absorbed dose calculation for a realistic CT-derived mouse phantom irradiated with a standard Cs-137 cell irradiator using a Monte Carlo method.

Computed tomography (CT) derived Monte Carlo (MC) phantoms allow dose determination within small animal models that is not feasible with in-vivo dosimetry. The aim of this study was to develop a CT-derived MC phantom generated from a mouse with a xenograft tumour that could then be used to calculate both the dose heterogeneity in the tumour volume and out of field scattered dose for pre-clinical small animal irradiation experiments. A BEAMnrc Monte-Carlo model has been built of our irradiation system that comprises a lead collimator with a 1 cm diameter aperture fitted to a Cs-137 gamma irradiator. The MC model of the irradiation system was validated by comparing the calculated dose results with dosimetric film measurement in a polymethyl methacrylate (PMMA) phantom using a 1D gamma-index analysis. Dose distributions in the MC mouse phantom were calculated and visualized on the CT-image data. Dose volume histograms (DVHs) were generated for the tumour and organs at risk (OARs). The effect of the xenographic tumour volume on the scattered out of field dose was also investigated. The defined gamma index analysis criteria were met, indicating that our MC simulation is a valid model for MC mouse phantom dose calculations. MC dose calculations showed a maximum out of field dose to the mouse of 7% of Dmax. Absorbed dose to the tumour varies in the range 60%-100% of Dmax. DVH analysis demonstrated that tumour received an inhomogeneous dose of 12 Gy-20 Gy (for 20 Gy prescribed dose) while out of field doses to all OARs were minimized (1.29 Gy-1.38 Gy). Variation of the xenographic tumour volume exhibited no significant effect on the out of field scattered dose to OARs. The CT derived MC mouse model presented here is a useful tool for tumour dose verifications as well as investigating the doses to normal tissue (in out of field) for preclinical radiobiological research.

Aggressive uveal melanoma displays a high degree of centrosome amplification, opening the door to therapeutic intervention.

Uveal melanoma (UM) is the most common intraocular cancer in adults. Whilst treatment of primary UM (PUM) is often successful, around 50% of patients develop metastatic disease with poor outcomes, linked to chromosome 3 loss (monosomy 3, M3). Advances in understanding UM cell biology may indicate new therapeutic options. We report that UM exhibits centrosome abnormalities, which in other cancers are associated with increased invasiveness and worse prognosis, but also represent a potential Achilles' heel for cancer-specific therapeutics. Analysis of 75 PUM patient samples revealed both higher centrosome numbers and an increase in centrosomes with enlarged pericentriolar matrix (PCM) compared to surrounding normal tissue, both indicative of centrosome amplification. The PCM phenotype was significantly associated with M3 (t-test, p < 0.01). Centrosomes naturally enlarge as cells approach mitosis; however, whilst UM with higher mitotic scores had enlarged PCM regardless of genetic status, the PCM phenotype remained significantly associated with M3 in UM with low mitotic scores (ANOVA, p = 0.021) suggesting that this is independent of proliferation. Phenotypic analysis of patient-derived cultures and established UM lines revealed comparable levels of centrosome amplification in PUM cells to archetypal triple-negative breast cancer cell lines, whilst metastatic UM (MUM) cell lines had even higher levels. Importantly, many UM cells also exhibit centrosome clustering, a common strategy employed by other cancer cells with centrosome amplification to survive cell division. As UM samples with M3 display centrosome abnormalities indicative of amplification, this phenotype may contribute to the development of MUM, suggesting that centrosome de-clustering drugs may provide a novel therapeutic approach.

A feasibility study on the development and use of a deep learning model to automate real-time monitoring of tumor position and assessment of interfraction fiducial marker migration in prostate radiotherapy patients.

Purpose. The aim of this study was to assess the feasibility of the development and training of a deep learning object detection model for automating the assessment of fiducial marker migration and tracking of the prostate in radiotherapy patients.Methods and Materials. A fiducial marker detection model was trained on the YOLO v2 detection framework using approximately 20,000 pelvis kV projection images with fiducial markers labelled. The ability of the trained model to detect marker positions was validated by tracking the motion of markers in a respiratory phantom and comparing detection data with the expected displacement from a reference position. Marker migration was then assessed in 14 prostate radiotherapy patients using the detector for comparison with previously conducted studies. This was done by determining variations in intermarker distance between the first and subsequent fractions in each patient.Results. On completion of training, a detection model was developed that operated at a 96% detection efficacy and with a root mean square error of 0.3 pixels. By determining the displacement from a reference position in a respiratory phantom, experimentally and with the detector it was found that the detector was able to compute displacements with a mean accuracy of 97.8% when compared to the actual values. Interfraction marker migration was measured in 14 patients and the average and maximum±standard deviation marker migration were found to be2.0±0.9mmand2.3±0.9mm,respectively.Conclusion. This study demonstrates the benefits of pairing deep learning object detection, and image-guided radiotherapy and how a workflow to automate the assessment of organ motion and seed migration during prostate radiotherapy can be developed. The high detection efficacy and low error make evident the advantages of using a pre-trained model to automate the assessment of the target volume positional variation and the migration of fiducial markers between fractions.

Comparison and Validation of Multiple Detectors against Monte Carlo Simulation for the Use of Small-Field Dosimetry.

Aim: The aim of this study was to compare the Exradin W2 scintillator, PTW microDiamond, IBA Razor Nano, and IBA Razor chamber detectors for small-field dose measurements and validate the measured data against the EGSnrc user code and observe the variation between daisy-chained and direct measurement methods for the above detectors. Materials and Methods: The W2 scintillator, microDiamond, Razor Nano, and Razor chamber detectors were used to measure the in-plane and cross-plane profiles and the output factors (OFs) at 10 cm depth, and 90 source-to-surface distance for 6MV X-rays (Elekta Versa HD). The field sizes ranged from 0.5 cm × 0.5 cm to 5 cm × 5 cm. The BEAMnrc/DOSXYZnrc user codes (EGSnrc) were used to simulate the reference profiles. Gamma analysis was performed to compare the measured and simulated dose distributions. Results: The OFs measured with the W2 scintillator, microDiamond, Razor Nano chamber, Razor chamber, and the calculated Monte Carlo (MC) showed agreement to within 1% for the 3 cm × 3 cm field size. The uncertainty in the MC simulation was observed to be 0.4%. The percent difference in OFs measured using daisy-chained and direct measurement methods was within 0.15%, 0.4%, 1.4%, and 2.4% for microDiamond, W2 scintillator, Nano, and Razor chamber detectors, respectively. Conclusion: The lateral beam profiles and OFs of W2 scintillator, microDiamond, Razor Nano, and Razor chambers exhibit good agreement with the MC simulation within the nominal field sizes. Our results demonstrate that we can achieve considerable time-saving by directly measuring small-field OFs without daisy-chained methods using microDiamond and W2 scintillator. In terms of ease of use, sensitivity, reproducibility, and from a practical standpoint, we recommend microDiamond for small-field dosimetry.

3D Breast Tumor Models for Radiobiology Applications.

Breast cancer is a leading cause of cancer-associated death in women. The clinical management of breast cancers is normally carried out using a combination of chemotherapy, surgery and radiation therapy. The majority of research investigating breast cancer therapy until now has mainly utilized two-dimensional (2D) in vitro cultures or murine models of disease. However, there has been significant uptake of three-dimensional (3D) in vitro models by cancer researchers over the past decade, highlighting a complimentary model for studies of radiotherapy, especially in conjunction with chemotherapy. In this review, we underline the effects of radiation therapy on normal and malignant breast cells and tissues, and explore the emerging opportunities that pre-clinical 3D models offer in improving our understanding of this treatment modality.

Automatic Detection and Tracking of Marker Seeds Implanted in Prostate Cancer Patients using a Deep Learning Algorithm.

PURPOSE: Fiducial marker seeds are often used as a surrogate to identify and track the positioning of prostate volume in the treatment of prostate cancer. Tracking the movement of prostate seeds aids in minimizing the prescription dose spillage outside the target volume to reduce normal tissue complications. In this study, You Only Look Once (YOLO) v2™ (MathWorks™) convolutional neural network was employed to train ground truth datasets and develop a program in MATLAB that can visualize and detect the seeds on projection images obtained from kilovoltage (kV) X-ray volume imaging (XVI) panel (Elekta™). METHODS: As a proof of concept, a wax phantom containing three gold marker seeds was imaged, and kV XVI seed images were labeled and used as ground truth to train the model. The projection images were corrected for any panel shift using flex map data. Upon successful testing, labeled marker seeds and projection images of three patients were used to train a model to detect fiducial marker seeds. A software program was developed to display the projection images in real-time and predict the seeds using YOLO v2 and determine the centers of the marker seeds on each image. RESULTS: The fiducial marker seeds were successfully detected in 98% of images from all gantry angles; the variation in the position of the seed center was within ± 1 mm. The percentage difference between the ground truth and the detected seeds was within 3%. CONCLUSION: Our study shows that deep learning can be used to detect fiducial marker seeds in kV images in real time. This is an ongoing study, and work is underway to extend it to other sites for tracking moving structures with minimal effort.

A novel add-on collimator for preclinical radiotherapy applications using a standard cell irradiator: design, construction, and validation.

PURPOSE: Preclinical radiotherapy applications require dedicated irradiation systems which are expensive and not widely available. In this work, a clinical dual source 137 Cs cell irradiator was adapted to deliver 1-cm diameter preclinical treatment beams using a lead and stainless steel custom-made collimator to treat one or two mice at a time. METHODS: The dosimetric characteristics of all the different components of the system (including collimator, phantoms, and radiation sources) have been simulated with EGSnrc Monte Carlo methods. The collimator was constructed based on these simulations and the calculated results were verified against dosimetric measurements with MOSKin detectors, GAFchromic films, and dosimetric gels. RESULTS: The comparisons showed an agreement, in terms of full width half maximum values, between the simulated and the measured dose cross profiles at the midline within 4% for both gel dosimetry and GAFchromic films. Out of beam dose, measured in air at the collimator midplane with MOSFET detectors was between 6% and 10% of the beam axis dose. The dimensions of the beam are constant along the vertical axis of the collimator and also the simulated and measured Percentage Depth Dose (PDD) curves show an agreement within 1%. CONCLUSIONS: The collimator design developed in this work allows the creation of a beam with the necessary characteristics for ablative radiotherapy treatments on small animals using a standard clinical cell irradiator. This collimator design will make advanced preclinical studies with ablative beams possible for all those institutions which do not have collimated preclinical irradiators available.

Developing a contextualised blended learning framework to enhance medical physics student learning and engagement.

As a specialist field of study, medical physicists require a broad range of knowledge and skills to operate competently in their workplace. In Australia, these competencies are accredited by the Australasian College of Physical Scientists and Engineers in Medicine (ACPSEM). Education and training for medical physicists therefore consists of an exhaustive range of knowledge areas. This is made even more challenging due to the extremely diverse backgrounds of students in these specialist courses of study. These factors frequently lead to a disengagement by students with learning activities. To address some of these challenges, the Radiotherapy Physics unit in a Masters level Medical Physics course of study was re-designed to increase active learning that included scaffolded in-class and online tasks and supported by virtual reality simulations. These re-design initiatives were informed by a diverse team including academic and clinical medical physicists as well as education experts. A survey, conducted over two consecutive years was used to gain students perceptions about the re-design. The questions were designed to see if the students felt engaged with the various learning activities. Analysis of the survey data indicates that there was an overall improvement in students' engagement with the learning activities and the learning content. The paper further discusses nuanced understanding about the ways in which students engaged with the various learning activities including online, in-class, practicals and industry attachments. The paper discusses the appropriately informed learning activities that can be used to improve student engagement for highly specialised, content heavy areas of study.

Targeting centrosome amplification, an Achilles' heel of cancer.

Due to cell-cycle dysregulation, many cancer cells contain more than the normal compliment of centrosomes, a state referred to as centrosome amplification (CA). CA can drive oncogenic phenotypes and indeed can cause cancer in flies and mammals. However, cells have to actively manage CA, often by centrosome clustering, in order to divide. Thus, CA is also an Achilles' Heel of cancer cells. In recent years, there have been many important studies identifying proteins required for the management of CA and it has been demonstrated that disruption of some of these proteins can cause cancer-specific inhibition of cell growth. For certain targets therapeutically relevant interventions are being investigated, for example, small molecule inhibitors, although none are yet in clinical trials. As the field is now poised to move towards clinically relevant interventions, it is opportune to summarise the key work in targeting CA thus far, with particular emphasis on recent developments where small molecule or other strategies have been proposed. We also highlight the relatively unexplored paradigm of reversing CA, and thus its oncogenic effects, for therapeutic gain.

The effect of respiratory motion on electronic portal imaging device dosimetry.

There is an increasing need to develop methods for in vivo verification of the delivery of radiotherapy treatments. Electronic portal imaging devices (EPID's) have been demonstrated to be of use for this application. The basic principle is relatively straightforward, the EPID is used to measure a two-dimensional (2D) planar exit or portal dose map behind the patient during the treatment delivery that can provide information on any errors in linear accelerator output or changes in the patient anatomy. In this paper we focused on the effect of intra-fraction motion, particularly respiratory motion, on the measured 2D EPID dose-response. Measurements were made with a breast phantom undergoing one-dimensional (1D) sinusoidal motion with a range of amplitudes (0.5, 1.0, and 1.5 cm) and frequencies (12, 15, and 20  cycles/min). Further measurements were made with the phantom undergoing breathing sequences measured during patient planning computed tomography simulation. We made use of the quadratic calibration method that converts the EPID images to a surrogate for dose, equivalent thickness of Plastic Water® . Comparisons were made of the 2D thickness maps derived for the different motions compared to the static phantom case and the resulting dose difference analyzed over the "breast" region of interest. A 2D gamma analysis within the same region of interest was performed of the motion images compared to static reference image. Comparisons were made of 1D thickness profiles for the moving and static phantom. The 1D and 2D analyses show the method to be sensitive to the smallest motion amplitude of 0.5 cm tested in the phantom measurements. The results using the phantom demonstrate the method to be a potentially useful tool for monitoring intra-fraction motion during the delivery of patient radiotherapy treatments as well as more generally providing information on the effects of motion on EPID based in vivo dosimetric verification.

Utilising the Virtual Environment for Radiotherapy Training System to Support Undergraduate Teaching of IMRT, VMAT, DCAT Treatment Planning, and QA Concepts.

The use of three-dimensional virtual reality technologies in education has been widely reported in the literature. The goal of this article is to review how the virtual environment for radiotherapy training (VERT) can be utilised to support the teaching of intensity-modulated radiotherapy (IMRT), volumetric-modulated arc therapy, and dynamic conformal arc therapy techniques. Utilising Pinnacle v14 (Philips Radiation Oncology Systems, Madison, WI) and Monaco v 5.10 (Elekta CMS, Maryland Heights, MO), we exported IMRT, volumetric-modulated arc therapy, and dynamic conformal arc therapy treatment plans to VERT. Quality assurance (QA) plans were also exported from the Monaco planning software to demonstrate theoretical concepts and virtual plan delivery using the ArcCHECK phantom (Sun Nuclear Corp, Melbourne, FL) and solid water IMRT phantom. Several VERT features were utilised to critically evaluate dose coverage and organ at risk sparing on inverse treatment plans. The display of gantry and multileaf collimator motion and fluence maps could be a useful feature in the education of treatment planning concepts. QA could be delivered on VERT to demonstrate patient-specific QA concepts in a virtual environment. Anecdotal evidence shows that the use of VERT for collaborative plan evaluation sessions is likely to engage the students and improve their ability to evaluate treatment plans. VERT can be utilised to reinforce understanding of treatment plan evaluation skills, QA, and treatment delivery of inverse plans in educational environments.

The deubiquitylase USP15 regulates topoisomerase II alpha to maintain genome integrity.

Ubiquitin-specific protease 15 (USP15) is a widely expressed deubiquitylase that has been implicated in diverse cellular processes in cancer. Here we identify topoisomerase II (TOP2A) as a novel protein that is regulated by USP15. TOP2A accumulates during G2 and functions to decatenate intertwined sister chromatids at prophase, ensuring the replicated genome can be accurately divided into daughter cells at anaphase. We show that USP15 is required for TOP2A accumulation, and that USP15 depletion leads to the formation of anaphase chromosome bridges. These bridges fail to decatenate, and at mitotic exit form micronuclei that are indicative of genome instability. We also describe the cell cycle-dependent behaviour for two major isoforms of USP15, which differ by a short serine-rich insertion that is retained in isoform-1 but not in isoform-2. Although USP15 is predominantly cytoplasmic in interphase, we show that both isoforms move into the nucleus at prophase, but that isoform-1 is phosphorylated on its unique S229 residue at mitotic entry. The micronuclei phenotype we observe on USP15 depletion can be rescued by either USP15 isoform and requires USP15 catalytic activity. Importantly, however, an S229D phospho-mimetic mutant of USP15 isoform-1 cannot rescue either the micronuclei phenotype, or accumulation of TOP2A. Thus, S229 phosphorylation selectively abrogates this role of USP15 in maintaining genome integrity in an isoform-specific manner. Finally, we show that USP15 isoform-1 is preferentially upregulated in a panel of non-small cell lung cancer cell lines, and propose that isoform imbalance may contribute to genome instability in cancer. Our data provide the first example of isoform-specific deubiquitylase phospho-regulation and reveal a novel role for USP15 in guarding genome integrity.

Obesity does not influence prostate intrafractional motion.

INTRODUCTION: Motion of the prostate is problematic in the accurate delivery of external beam radiation therapy (EBRT) for prostate cancer. This study investigated the relationship between body mass index (BMI), an easily measured indicator of obesity, and prostate motion. METHODS: Prostate motion during EBRT was assessed by measuring the displacement of fiducial markers implanted within the prostate in 130 prostate cancer patients. Interfractional motion was corrected on daily imaging through pre-treatment cone-beam-computed tomography (CBCT) and intrafractional motion measured using movie sequences captured using an electronic portal imaging device (EPID) during treatment delivery. RESULTS: There was no statistically significant relationship between the mean intrafractional motion and BMI, except in the left-right (LR) translation (P = 0.049) over the study population. For each BMI category, there was no statistical significance (P > 0.05) between any of the translations/rotations except LR (P = 0.003). CONCLUSION: While intrafractional motion is an important consideration, prostate motion cannot be reliably predicted through measurement of patient's BMI.

Probing Ligand Exchange in the P450 Enzyme CYP121 from Mycobacterium tuberculosis: Dynamic Equilibrium of the Distal Heme Ligand as a Function of pH and Temperature.

CYP121 is a cytochrome P450 enzyme from Mycobacterium tuberculosis that catalyzes the formation of a C-C bond between the aromatic groups of its cyclodityrosine substrate (cYY). The crystal structure of CYP121 in complex with cYY reveals that the solvent-derived ligand remains bound to the ferric ion in the enzyme-substrate complex. Whereas in the generally accepted P450 mechanism, binding of the primary substrate in the active-site triggers the release of the solvent-derived ligand, priming the metal center for reduction and subsequent O2 binding. Here we employed sodium cyanide to probe the metal-ligand exchange of the enzyme and the enzyme-substrate complex. The cyano adducts were characterized by UV-vis, EPR, and ENDOR spectroscopies and X-ray crystallography. A 100-fold increase in the affinity of cyanide binding to the enzyme-substrate complex over the ligand-free enzyme was observed. The crystal structure of the [CYP121(cYY)CN] ternary complex showed a rearrangement of the substrate in the active-site, when compared to the structure of the binary [CYP121(cYY)] complex. Transient kinetic studies showed that cYY binding resulted in a lower second-order rate constant (kon (CN)) but a much more stable cyanide adduct with 3 orders of magnitude slower koff (CN) rate. A dynamic equilibrium between multiple high- and low-spin species for both the enzyme and enzyme-substrate complex was also observed, which is sensitive to changes in both pH and temperature. Our data reveal the chemical and physical properties of the solvent-derived ligand of the enzyme, which will help to understand the initial steps of the catalytic mechanism.

Regulation of the cell cycle and centrosome biology by deubiquitylases.

Post-translational modification of proteins by ubiquitylation is increasingly recognised as a highly complex code that contributes to the regulation of diverse cellular processes. In humans, a family of almost 100 deubiquitylase enzymes (DUBs) are assigned to six subfamilies and many of these DUBs can remove ubiquitin from proteins to reverse signals. Roles for individual DUBs have been delineated within specific cellular processes, including many that are dysregulated in diseases, particularly cancer. As potentially druggable enzymes, disease-associated DUBs are of increasing interest as pharmaceutical targets. The biology, structure and regulation of DUBs have been extensively reviewed elsewhere, so here we focus specifically on roles of DUBs in regulating cell cycle processes in mammalian cells. Over a quarter of all DUBs, representing four different families, have been shown to play roles either in the unidirectional progression of the cell cycle through specific checkpoints, or in the DNA damage response and repair pathways. We catalogue these roles and discuss specific examples. Centrosomes are the major microtubule nucleating centres within a cell and play a key role in forming the bipolar mitotic spindle required to accurately divide genetic material between daughter cells during cell division. To enable this mitotic role, centrosomes undergo a complex replication cycle that is intimately linked to the cell division cycle. Here, we also catalogue and discuss DUBs that have been linked to centrosome replication or function, including centrosome clustering, a mitotic survival strategy unique to cancer cells with supernumerary centrosomes.

Cross-linking of dicyclotyrosine by the cytochrome P450 enzyme CYP121 from Mycobacterium tuberculosis proceeds through a catalytic shunt pathway.

CYP121, the cytochrome P450 enzyme in Mycobacterium tuberculosis that catalyzes a single intramolecular C-C cross-linking reaction in the biosynthesis of mycocyclosin, is crucial for the viability of this pathogen. This C-C coupling reaction represents an expansion of the activities carried out by P450 enzymes distinct from oxygen insertion. Although the traditional mechanism for P450 enzymes has been well studied, it is unclear whether CYP121 follows the general P450 mechanism or uses a different catalytic strategy for generating an iron-bound oxidant. To gain mechanistic insight into the CYP121-catalyzed reaction, we tested the peroxide shunt pathway by using rapid kinetic techniques to monitor the enzyme activity with its substrate dicyclotyrosine (cYY) and observed the formation of the cross-linked product mycocyclosin by LC-MS. In stopped-flow experiments, we observed that cYY binding to CYP121 proceeds in a two-step process, and EPR spectroscopy indicates that the binding induces active site reorganization and uniformity. Using rapid freeze-quenching EPR, we observed the formation of a high-spin intermediate upon the addition of peracetic acid to the enzyme-substrate complex. This intermediate exhibits a high-spin (S = 5/2) signal with g values of 2.00, 5.77, and 6.87. Likewise, iodosylbenzene could also produce mycocyclosin, implicating compound I as the initial oxidizing species. Moreover, we also demonstrated that CYP121 performs a standard peroxidase type of reaction by observing substrate-based radicals. On the basis of these results, we propose plausible free radical-based mechanisms for the C-C bond coupling reaction.

Qualitative Evaluation of a Novel 3D Volumetric Radiotherapy Segmentation Tool.

INTRODUCTION: A novel 3D volumetric segmentation tool allows the user to outline using a small number of points on a range of planes. Unique 3D volumetric "sculpting" tools enable editing of the resulting structures across multiple slices concurrently. This article reports the results of radiation oncologists' preclinical evaluation of the tool. METHODS: Three clinicians outlined prostate and seminal vesicles on 14 data sets using the traditional slice-by-slice method and the new 3D tool. The project gathered focus-group feedback to gather rich data relating to clinician perceptions of the new 3D outlining paradigm. Emergent themes were identified and categorised for discussion. RESULTS: Radiation oncologists reported high levels of satisfaction with the outlines arising from both paradigms. The volumetric sculpting was a challenge, but participants enjoyed using points in orthogonal planes and felt that the paradigm had potential value in terms of speed and smooth volume creation. CONCLUSION: This study has demonstrated that a 3D volumetric outlining system is felt to have potential value by radiation oncologists for accelerating clinician-directed prostate and seminal vesicle segmentation. The new tool was well-received and reported to be capable of producing very rapid and smooth volumes. The novelty of the approach required significant training input and a radically different approach of minimal point placement. Further testing of this software with a less time-poor cohort may be indicated to gain reliable quantitative data relating to the impact on segmentation time.

A Comparison of Non-coplanar Three-dimensional Conformal Radiation Therapy, Intensity Modulated Radiation Therapy, and Volumetric Modulated Radiation Therapy for the Delivery of Stereotactic Ablative Radiation Therapy to Peripheral Lung Cancer.

AIM: The objective of the study was to compare three noncoplanar delivery techniques (three-dimensional conformal radiation therapy [3DCRT], intensity-modulated radiation therapy [IMRT], and volumetric-modulated arc therapy [VMAT]) for the delivery of lung stereotactic ablative radiation therapy to peripheral lung tumours. METHODS AND MATERIALS: The plans were compared by assessing the planning target volume coverage, doses to organs at risk, high and intermediate dose constraints (D2cm and R50%) and delivery times using analysis of variance for repeated measurements or Friedman's test when appropriate. RESULTS: Mean PTV54 Gy coverage was found to be 95.6%, 95.7%, and 95.6% for the 3DCRT, IMRT, and VMAT techniques, respectively. No deviations to the intermediate dose constraints were found in 65%, 65%, and 85% of the patients for the 3DCRT, IMRT, and VMAT plans, respectively. Mean treatment times (excluding setup and imaging) were 20.0 minutes (±1.67), 25.2 minutes (±2.15), and 11.7 (±2.0) minutes respectively for 3DCRT, IMRT, and VMAT. CONCLUSION: A noncoplanar VMAT technique was found to provide superior intermediate dose sparing with comparable prescription dose coverage when compared with noncoplanar 3DCRT or IMRT. In addition, VMAT was found to reduce the treatment times of stereotactic ablative radiation therapy delivery for peripheral lung tumours.

Exploring Electron/Proton Transfer and Conformational Changes in the Nitrogenase MoFe Protein and FeMo-cofactor Through Cryoreduction/EPR Measurements.

We combine cryoreduction/annealing/EPR measurements of nitrogenase MoFe protein with results of earlier investigations to provide a detailed view of the electron/proton transfer events and conformational changes that occur during early stages of [e-/H+] accumulation by the MoFe protein. This includes reduction of (i) the non-catalytic state of the iron-molybdenum cofactor (FeMo-co) active site that is generated by chemical oxidation of the resting-state cofactor (S = 3/2)) within resting MoFe (E0), and (ii) the catalytic state that has accumulated n =1 [e-/H+] above the resting-state level, denoted E1(1H) (S ≥ 1) in the Lowe-Thorneley kinetic scheme. FeMo-co does not undergo a major change of conformation during reduction of oxidized FeMo-co. In contrast, FeMo-co undergoes substantial conformational changes during the reduction of E0 to E1(1H), and of E1(1H) to E2(2H) (n = 2; S = 3/2). The experimental results further suggest that the E1(1H) → E2(2H) step involves coupled delivery of a proton and electron (PCET) to FeMo-co of E1(H) to generate a non-equilibrium S = ½ form E2(2H)*. This subsequently undergoes conformational relaxation and attendant change in FeMo-co spin state, to generate the equilibrium E2(2H) (S = 3/2) state. Unexpectedly, these experiments also reveal conformational coupling between FeMo-co and P-cluster, and between Fe protein binding and FeMo-co, which might play a role in gated ET from reduced Fe protein to FeMo-co.