A bioengineered 3D ovarian cancer model for the assessment of peptidase-mediated enhancement of spheroid growth and intraperitoneal spread

Cancer-associated proteases promote peritoneal dissemination and chemoresistance in malignant progression. In this study, kallikrein-related peptidases 4, 5, 6, and 7 (KLK4-7)-cotransfected OV-MZ-6 ovarian cancer cells were embedded in a bioengineered three-dimensional (3D) microenvironment that contains RGD motifs for integrin engagement to analyze their spheroid growth and survival after chemotreatment. KLK4-7-cotransfected cells formed larger spheroids and proliferated more than controls in 3D, particularly within RGD-functionalized matrices, which was reduced upon integrin inhibition. In contrast, KLK4-7-expressing cell monolayers proliferated less than controls, emphasizing the relevance of the 3D microenvironment and integrin engagement. In a spheroid-based animal model, KLK4-7-overexpression induced tumor growth after 4 weeks and intraperitoneal spread after 8 weeks. Upon paclitaxel administration, KLK4-7-expressing tumors declined in size by 91% (controls: 87%) and showed 90% less metastatic outgrowth (controls: 33%, P<0.001). KLK4-7-expressing spheroids showed 53% survival upon paclitaxel treatment (controls: 51%), accompanied by enhanced chemoresistance-related factors, and their survival was further reduced by combination treatment of paclitaxel with KLK4/5/7 (22%, P=0.007) or MAPK (6%, P=0.006) inhibition. The concomitant presence of KLK4-7 in ovarian cancer cells together with integrin activation drives spheroid formation and proliferation. Combinatorial approaches of paclitaxel and KLK/MAPK inhibition may be more efficient for late-stage disease than chemotherapeutics alone as these inhibitory regimens reduced cancer spheroid growth to a greater extent than paclitaxel alone.

Advantages of exploiting projection structure for segmenting dense 3D point clouds

Timely and comprehensive scene segmentation is often a critical step for many high level mobile robotic tasks. This paper examines a projected area based neighbourhood lookup approach with the motivation towards faster unsupervised segmentation of dense 3D point clouds. The proposed algorithm exploits the projection geometry of a depth camera to find nearest neighbours which is time independent of the input data size. Points near depth discontinuations are also detected to reinforce object boundaries in the clustering process. The search method presented is evaluated using both indoor and outdoor dense depth images and demonstrates significant improvements in speed and precision compared to the commonly used Fast library for approximate nearest neighbour (FLANN) [Muja and Lowe, 2009].

Real-time mobile 3D temperature mapping

The ability to measure surface temperature and represent it on a metrically accurate 3D model has proven applications in many areas such as medical imaging, building energy auditing, and search and rescue. A system is proposed that enables this task to be performed with a handheld sensor, and for the first time with results able to be visualized and analyzed in real-time. A device comprising a thermal-infrared camera and range sensor is calibrated geometrically and used for data capture. The device is localized using a combination of ICP and video-based pose estimation from the thermal-infrared video footage which is shown to reduce the occurrence of failure modes. Furthermore, the problem of misregistration which can introduce severe distortions in assigned surface temperatures is avoided through the use of a risk-averse neighborhood weighting mechanism. Results demonstrate that the system is more stable and accurate than previous approaches, and can be used to accurately model complex objects and environments for practical tasks.

Constructing a frame of cube : connecting 3D shapes with direction, location and movement

With the advancement of new technologies, this author has in 2010 started to engineer an online learning environment for investigating the nature and development of spatial abilities, and the teaching and learning of geometry. This paper documents how this new digital learning environment can afford the opportunity to integrate the learning about 3D shapes with direction, location and movement, and how young children can mentally and visually construct virtual 3D shapes using movements in both egocentric and fixed frames of reference (FOR). Findings suggest that year 4 (aged 9) children can develop the capacity to construct a cube using egocentric FOR only, fixed FOR only or a combination of both FOR. However, these young participants were unable to articulate the effect of individual or combined FOR movements. Directions for future research are proposed.

A 3D virtual medical imaging suite : bridging the academic-clinical training boundary

Aims The Medical Imaging Training Immersive Environment (MITIE) system is a recently developed virtual reality (VR) platform that allows students to practice a range of medical imaging techniques. The aim of this pilot study was to harvest user feedback about the educational value of the application and inform future pedagogical development. This presentation explores the use of this technology for skills training and blurring the boundaries between academic learning and clinical skills training. Background MITIE is a 3D VR environment that allows students to manipulate a patient and radiographic equipment in order to produce a VR-generated image for comparison with a gold standard. As with VR initiatives in other health disciplines (1-6) the software mimics clinical practice as much as possible and uses 3D technology to enhance immersion and realism. The software was developed by the Medical Imaging Course Team at a provider University with funding from a Health Workforce Australia “Simulated Learning Environments” grant. Methods Over 80 students undertaking the Bachelor of Medical Imaging Course were randomised to receive practical experience with either MITIE or radiographic equipment in the medical radiation laboratory. Student feedback about the educational value of the software was collected and performance with an assessed setup was measured for both groups for comparison. Ethical approval for the project was provided by the university ethics panel. Results This presentation provides qualitative analysis of student perceptions relating to satisfaction, usability and educational value as well as comparative quantitative performance data. Students reported high levels of satisfaction and both feedback and assessment results confirmed the application’s significance as a pre-clinical training tool. There was a clear emerging theme that MITIE could be a useful learning tool that students could access to consolidate their clinical learning, either during their academic timetables or their clinical placement. Conclusion Student feedback and performance data indicate that MITIE has a valuable role to play in the clinical skills training for medical imaging students both in the academic and the clinical environment. Future work will establish a framework for an appropriate supporting pedagogy that can cross the boundary between the two environments. This project was possible due to funding made available by Health Workforce Australia.

HeatWave : a handheld 3D thermography system for energy auditing

Energy auditing is an effective but costly approach for reducing the long-term energy consumption of buildings. When well-executed, energy loss can be quickly identified in the building structure and its subsystems. This then presents opportunities for improving energy efficiency. We present a low-cost, portable technology called "HeatWave" which allows non-experts to generate detailed 3D surface temperature models for energy auditing. This handheld 3D thermography system consists of two commercially available imaging sensors and a set of software algorithms which can be run on a laptop. The 3D model can be visualized in real-time by the operator so that they can monitor their degree of coverage as the sensors are used to capture data. In addition, results can be analyzed offline using the proposed "Spectra" multispectral visualization toolbox. The presence of surface temperature data in the generated 3D model enables the operator to easily identify and measure thermal irregularities such as thermal bridges, insulation leaks, moisture build-up and HVAC faults. Moreover, 3D models generated from subsequent audits of the same environment can be automatically compared to detect temporal changes in conditions and energy use over time.

Handheld 3D thermography using range sensing and computer vision

This thesis developed a method for real-time and handheld 3D temperature mapping using a combination of off-the-shelf devices and efficient computer algorithms. It contributes a new sensing and data processing framework to the science of 3D thermography, unlocking its potential for application areas such as building energy auditing and industrial monitoring. New techniques for the precise calibration of multi-sensor configurations were developed, along with several algorithms that ensure both accurate and comprehensive surface temperature estimates can be made for rich 3D models as they are generated by a non-expert user.

Evaluation of MegaVoltage Cone Beam CT image quality with an unmodified Elekta Precise Linac and EPID : a feasibility study

In order to increase the accuracy of patient positioning for complex radiotherapy treatments various 3D imaging techniques have been developed. MegaVoltage Cone Beam CT (MVCBCT) can utilise existing hardware to implement a 3D imaging modality to aid patient positioning. MVCBCT has been investigated using an unmodified Elekta Precise linac and 15 iView amorphous silicon electronic portal imaging device (EPID). Two methods of delivery and acquisition have been investigated for imaging an anthropomorphic head phantom and quality assurance phantom. Phantom projections were successfully acquired and CT datasets reconstructed using both acquisition methods. Bone, tissue and air were 20 clearly resolvable in both phantoms even with low dose (22 MU) scans. The feasibility of MegaVoltage Cone beam CT was investigated using a standard linac, amorphous silicon EPID and a combination of a free open source reconstruction toolkit as well as custom in-house software written in Matlab. The resultant image quality has 25 been assessed and presented. Although bone, tissue and air were resolvable 2 in all scans, artifacts are present and scan doses are increased when compared with standard portal imaging. The feasibility of MVCBCT with unmodified Elekta Precise linac and EPID has been considered as well as the identification of possible areas for future development in artifact correction techniques to 30 further improve image quality.

Retrospective evaluation of dosimetric quality for prostate carcinomas treated with 3D conformal, intensity-modulated and volumetric-modulated arc radiotherapy

Introduction This study examines and compares the dosimetric quality of radiotherapy treatment plans for prostate carcinoma across a cohort of 163 patients treated across 5 centres: 83 treated with three-dimensional conformal radiotherapy (3DCRT), 33 treated with intensity-modulated radiotherapy (IMRT) and 47 treated with volumetric-modulated arc therapy (VMAT). Methods Treatment plan quality was evaluated in terms of target dose homogeneity and organ-at-risk sparing, through the use of a set of dose metrics. These included the mean, maximum and minimum doses; the homogeneity and conformity indices for the target volumes; and a selection of dose coverage values that were relevant to each organ-at-risk. Statistical significance was evaluated using two-tailed Welch’s T-tests. The Monte Carlo DICOM ToolKit software was adapted to permit the evaluation of dose metrics from DICOM data exported from a commercial radiotherapy treatment planning system. Results The 3DCRT treatment plans offered greater planning target volume dose homogeneity than the other two treatment modalities. The IMRT and VMAT plans offered greater dose reduction in the organs-at-risk: with increased compliance with recommended organ-at-risk dose constraints, compared to conventional 3DCRT treatments. When compared to each other, IMRT and VMAT did not provide significantly different treatment plan quality for like-sized tumour volumes. Conclusions This study indicates that IMRT and VMAT have provided similar dosimetric quality, which is superior to the dosimetric quality achieved with 3DCRT.

Factors affecting the extraction of absorbed dose information in a 3D polymer gel dosimeter by X ray computed tomography (IAEA-CN-96/122P)

Two sources of uncertainty in the X ray computed tomography imaging of polymer gel dosimeters are investigated in the paper.The first cause is a change in postirradiation density, which is proportional to the computed tomography signal and is associated with a volume change. The second cause of uncertainty is reconstruction noise.A simple technique that increases the residual signal to noise ratio by almost two orders of magnitude is examined.

Numerical estimation of 3D mechanical forces exerted by cells on non-linear materials

The exchange of physical forces in both cell-cell and cell-matrix interactions play a significant role in a variety of physiological and pathological processes, such as cell migration, cancer metastasis, inflammation and wound healing. Therefore, great interest exists in accurately quantifying the forces that cells exert on their substrate during migration. Traction Force Microscopy (TFM) is the most widely used method for measuring cell traction forces. Several mathematical techniques have been developed to estimate forces from TFM experiments. However, certain simplifications are commonly assumed, such as linear elasticity of the materials and/or free geometries, which in some cases may lead to inaccurate results. Here, cellular forces are numerically estimated by solving a minimization problem that combines multiple non-linear FEM solutions. Our simulations, free from constraints on the geometrical and the mechanical conditions, show that forces are predicted with higher accuracy than when using the standard approaches.

A derivation of high-frequency asymptotic values of 3D added mass and damping based on properties of the Cummins' equation

This brief paper provides a novel derivation of the known asymptotic values of three-dimensional (3D) added mass and damping of marine structures in waves. The derivation is based on the properties of the convolution terms in the Cummins's Equation as derived by Ogilvie. The new derivation is simple and no approximations or series expansions are made. The results follow directly from the relative degree and low-frequency asymptotic properties of the rational representation of the convolution terms in the frequency domain. As an application, the extrapolation of damping values at high frequencies for the computation of retardation functions is also discussed.

Joint 2D and 3D cues for image segmentation towards robotic applications

This thesis investigates the fusion of 3D visual information with 2D image cues to provide 3D semantic maps of large-scale environments in which a robot traverses for robotic applications. A major theme of this thesis was to exploit the availability of 3D information acquired from robot sensors to improve upon 2D object classification alone. The proposed methods have been evaluated on several indoor and outdoor datasets collected from mobile robotic platforms including a quadcopter and ground vehicle covering several kilometres of urban roads.

Paracrine interactions between LNCaP prostate cancer cells and bioengineered bone in 3D in vitro culture reflect molecular changes during bone metastasis

As microenvironmental factors such as three-dimensionality and cell–matrix interactions are increasingly being acknowledged by cancer biologists, more complex 3D in vitro models are being developed to study tumorigenesis and cancer progression. To better understand the pathophysiology of bone metastasis, we have established and validated a 3D indirect co-culture model to investigate the paracrine interactions between prostate cancer (PCa) cells and human osteoblasts. Co-culture of the human PCa, LNCaP cells embedded within polyethylene glycol hydrogels with human osteoblasts in the form of a tissue engineered bone construct (TEB), resulted in reduced proliferation of LNCaP cells. LNCaP cells in both monoculture and co-culture were responsive to the androgen analog, R1881, as indicated by an increase in the expression (mRNA and/or protein induction) of androgen-regulated genes including prostate specific antigen and fatty acid synthase. Microarray gene expression analysis further revealed an up-regulation of bone markers and other genes associated with skeletal and vasculature development and a significant activation of transforming growth factor β1 downstream genes in LNCaP cells after co-culture with TEB. LNCaP cells co-cultured with TEB also unexpectedly showed similar changes in classical androgen-responsive genes under androgen-deprived conditions not seen in LNCaP monocultures. The molecular changes of LNCaP cells after co-culturing with TEBs suggest that osteoblasts exert a paracrine effect that may promote osteomimicry and modulate the expression of androgen-responsive genes in LNCaP cells. Taken together, we have presented a novel 3D in vitro model that allows the study of cellular and molecular changes occurring in PCa cells and osteoblasts that are relevant to metastatic colonization of bone. This unique in vitro model could also facilitate cancer biologists to dissect specific biological hypotheses via extensive genomic or proteomic assessments to further our understanding of the PCa-bone crosstalk.

Atmospheric microplasma-functionalized 3D microfluidic strips within dense carbon nanotube arrays confine Au nanodots for SERS sensing

An atmospheric microplasma jet produces three-dimensional (3D) microfluidic channels on dense arrays of vertically aligned carbon nanotubes, which confines Au nanodot aqueous solution. The resulting hybrid 3D nanostructure is exploited as an effective microscopic area-selective sensing platform based on surface-enhanced Raman scattering.