Mineralized human primary osteoblast matrices as a model system to analyse interactions of prostate cancer cells with the bone microenvironment

Prostate cancer metastasis is reliant on the reciprocal interactions between cancer cells and the bone niche/micro-environment. The production of suitable matrices to study metastasis, carcinogenesis and in particular prostate cancer/bone micro-environment interaction has been limited to specific protein matrices or matrix secreted by immortalised cell lines that may have undergone transformation processes altering signaling pathways and modifying gene or receptor expression. We hypothesize that matrices produced by primary human osteoblasts are a suitable means to develop an in vitro model system for bone metastasis research mimicking in vivo conditions. We have used a decellularized matrix secreted from primary human osteoblasts as a model for prostate cancer function in the bone micro-environment. We show that this collagen I rich matrix is of fibrillar appearance, highly mineralized, and contains proteins, such as osteocalcin, osteonectin and osteopontin, and growth factors characteristic of bone extracellular matrix (ECM). LNCaP and PC3 cells grown on this matrix, adhere strongly, proliferate, and express markers consistent with a loss of epithelial phenotype. Moreover, growth of these cells on the matrix is accompanied by the induction of genes associated with attachment, migration, increased invasive potential, Ca2+ signaling and osteolysis. In summary, we show that growth of prostate cancer cells on matrices produced by primary human osteoblasts mimics key features of prostate cancer bone metastases and thus is a suitable model system to study the tumor/bone micro-environment interaction in this disease.

Osteogenic induction of human bone marrow-derived mesenchymal progenitor cells in novel synthetic polymer-hydrogel matrices

The aim of this project was to investigate the in vitro osteogenic potential of human mesenchymal progenitor cells in novel matrix architectures built by means of a three-dimensional bioresorbable synthetic framework in combination with a hydrogel. Human mesenchymal progenitor cells (hMPCs) were isolated from a human bone marrow aspirate by gradient centrifugation. Before in vitro engineering of scaffold-hMPC constructs, the adipogenic and osteogenic differentiation potential was demonstrated by staining of neutral lipids and induction of bone-specific proteins, respectively. After expansion in monolayer cultures, the cells were enzymatically detached and then seeded in combination with a hydrogel into polycaprolactone (PCL) and polycaprolactone-hydroxyapatite (PCL-HA) frameworks. This scaffold design concept is characterized by novel matrix architecture, good mechanical properties, and slow degradation kinetics of the framework and a biomimetic milieu for cell delivery and proliferation. To induce osteogenic differentiation, the specimens were cultured in an osteogenic cell culture medium and were maintained in vitro for 6 weeks. Cellular distribution and viability within three-dimensional hMPC bone grafts were documented by scanning electron microscopy, cell metabolism assays, and confocal laser microscopy. Secretion of the osteogenic marker molecules type I procollagen and osteocalcin was analyzed by semiquantitative immunocytochemistry assays. Alkaline phosphatase activity was visualized by p-nitrophenyl phosphate substrate reaction. During osteogenic stimulation, hMPCs proliferated toward and onto the PCL and PCL-HA scaffold surfaces and metabolic activity increased, reaching a plateau by day 15. The temporal pattern of bone-related marker molecules produced by in vitro tissue-engineered scaffold-cell constructs revealed that hMPCs differentiated better within the biomimetic matrix architecture along the osteogenic lineage.

In vitro characterization of natural and synthetic dermal matrices cultured with human dermal fibroblasts

The ideal dermal matrix should be able to provide the right biological and physical environment to ensure homogenous cell and extracellular matrix (ECM) distribution, as well as the right size and morphology of the neo-tissue required. Four natural and synthetic 3D matrices were evaluated in vitro as dermal matrices, namely (1) equine collagen foam, TissuFleece®, (2) acellular dermal replacement, Alloderm®, (3) knitted poly(lactic-co-glycolic acid) (10:90)–poly(-caprolactone) (PLGA–PCL) mesh, (4) chitosan scaffold. Human dermal fibroblasts were cultured on the specimens over 3 weeks. Cell morphology, distribution and viability were assessed by electron microscopy, histology and confocal laser microscopy. Metabolic activity and DNA synthesis were analysed via MTS metabolic assay and [3H]-thymidine uptake, while ECM protein expression was determined by immunohistochemistry. TissuFleece®, Alloderm® and PLGA–PCL mesh supported cell attachment, proliferation and neo-tissue formation. However, TissuFleece® contracted to 10% of the original size while Alloderm® supported cell proliferation predominantly on the surface of the material. PLGA–PCL mesh promoted more homogenous cell distribution and tissue formation. Chitosan scaffolds did not support cell attachment and proliferation. These results demonstrated that physical characteristics including porosity and mechanical stability to withstand cell contraction forces are important in determining the success of a dermal matrix material.

Robust multidisciplinary design optimisation using CFD and advanced evolutionary algorithms

Computation Fluid Dynamics (CFD) has become an important tool in optimization and has seen successful in many real world applications. Most important among these is in the optimisation of aerodynamic surfaces which has become Multi-Objective (MO) and Multidisciplinary (MDO) in nature. Most of these have been carried out for a given set of input parameters such as free stream Mach number and angle of attack. One cannot ignore the fact that in aerospace engineering one frequently deals with situations where the design input parameters and flight/flow conditions have some amount of uncertainty attached to them. When the optimisation is carried out for fixed values of design variables and parameters however, one arrives at an optimised solution that results in good performance at design condition but poor drag or lift to drag ratio at slightly off-design conditions. The challenge is still to develop a robust design that accounts for uncertainty in the design in aerospace applications. In this paper this issue is taken up and an attempt is made to prevent the fluctuation of objective performance by using robust design technique or Uncertainty.

An innovative robust reactive surgery assignment model

Online scheduling in the Operating Theatre Department is a dynamic process that deals with both elective and emergency patients. Each business day begins with an elective schedule determined in advance based on a mastery surgery schedule. Throughout the course of the day however, disruptions to this baseline schedule occur due to variations in treatment time, emergency arrivals, equipment failure and resource unavailability. An innovative robust reactive surgery assignment model is developed for the operating theatre department. Following the completion of each surgery, the schedule is re-solved taking into account any disruptions in order to minimise cancellations of pre-planned patients and maximise throughput of emergency cases. The single theatre case is solved and future work on the computationally more complex multiple theatre case under resource constraints is discussed.

A robust structure and motion replacement for bundle adjustment

This paper demonstrates the application of a robust form of pose estimation and scene reconstruction using data from camera images. We demonstrate results that suggest the ability of the algorithm to rival methods of RANSAC based pose estimation polished by bundle adjustment in terms of solution robustness, speed and accuracy, even when given poor initialisations. Our simulated results show the behaviour of the algorithm in a number of novel simulated scenarios reflective of real world cases that show the ability of the algorithm to handle large observation noise and difficult reconstruction scenes. These results have a number of implications for the vision and robotics community, and show that the application of visual motion estimation on robotic platforms in an online fashion is approaching real-world feasibility.

A simple orientation robust transmit polarization signalling scheme

A special transmit polarization signalling scheme is presented to alleviate the power reduction as a result of polarization mismatch from random antenna orientations. This is particularly useful for hand held mobile terminals typically equipped with only a single linearly polarized antenna, since the average signal power is desensitized against receiver orientations. Numerical simulations also show adequate robustness against incorrect channel estimations.

Robust tracking control of autonomous underwater vehicles in the presence of disturbance inputs

An autonomous underwater vehicle (AUV) is expected to operate in an ocean in the presence of poorly known disturbance forces and moments. The uncertainties of the environment makes it difficult to apply open-loop control scheme for the motion planning of the vehicle. The objective of this paper is to develop a robust feedback trajectory tracking control scheme for an AUV that can track a prescribed trajectory amidst such disturbances. We solve a general problem of feedback trajectory tracking of an AUV in SE(3). The feedback control scheme is derived using Lyapunov-type analysis. The results obtained from numerical simulations confirm the asymptotic tracking properties of the feedback control law. We apply the feedback control scheme to different mission scenarios, with the disturbances being initial errors in the state of the AUV.

Engineered silk fibroin protein 3D matrices for in vitro tumor model

3D in vitro model systems that are able to mimic the in vivo microenvironment are now highly sought after in cancer research. Antheraea mylitta silk fibroin protein matrices were investigated as potential biomaterial for in vitro tumor modeling. We compared the characteristics of MDA-MB-231 cells on A. mylitta, Bombyx mori silk matrices, Matrigel, and tissue culture plates. The attachment and morphology of the MDA-MB-231 cell line on A. mylitta silk matrices was found to be better than on B. mori matrices and comparable to Matrigel and tissue culture plates. The cells grown in all 3D cultures showed more MMP-9 activity, indicating a more invasive potential. In comparison to B. mori fibroin, A. mylitta fibroin not only provided better cell adhesion, but also improved cell viability and proliferation. Yield coefficient of glucose consumed to lactate produced by cells on 3D A. mylitta fibroin was found to be similar to that of cancer cells in vivo. LNCaP prostate cancer cells were also cultured on 3D A. mylitta fibroin and they grew as clumps in long term culture. The results indicate that A. mylitta fibroin scaffold can provide an easily manipulated microenvironment system to investigate individual factors such as growth factors and signaling peptides, as well as evaluation of anticancer drugs.

Noise robust voice activity detection using normal probability testing and time-domain histogram analysis

This paper presents a method of voice activity detection (VAD) suitable for high noise scenarios, based on the fusion of two complementary systems. The first system uses a proposed non-Gaussianity score (NGS) feature based on normal probability testing. The second system employs a histogram distance score (HDS) feature that detects changes in the signal through conducting a template-based similarity measure between adjacent frames. The decision outputs by the two systems are then merged using an open-by-reconstruction fusion stage. Accuracy of the proposed method was compared to several baseline VAD methods on a database created using real recordings of a variety of high-noise environments.

PLGA-based microparticles for the sustained release of BMP-2

The development of growth factor delivery strategies to circumvent the burst release phenomenon prevalent in most current systems has driven research towards encapsulating molecules in resorbable polymer matrices. For these polymer release techniques to be efficacious in a clinical setting, several key points need to be addressed. This present study has investigated the encapsulation of the growth factor, BMP-2 within PLGA/PLGA-PEG-PLGA microparticles. Morphology, size distribution, encapsulation efficiency and release kinetics were investigated and we have demonstrated a sustained release of bioactive BMP-2. Furthermore, biocompatibility of the PLGA microparticles was established and released BMP-2 was shown to promote the differentiation of MC3T3-E1 cells towards the osteogenic lineage to a greater extent than osteogenic supplements (as early as day 10 in culture), as determined using alkaline phosphatase and alizarin red assays. This study showcases a potential BMP-2 delivery system which may now be translated into more complex delivery systems, such as 3D, mechanically robust scaffolds for bone tissue regeneration applications.

Noise robust voice activity detection using features extracted from the time-domain autocorrelation function

This paper presents a method of voice activity detection (VAD) for high noise scenarios, using a noise robust voiced speech detection feature. The developed method is based on the fusion of two systems. The first system utilises the maximum peak of the normalised time-domain autocorrelation function (MaxPeak). The second zone system uses a novel combination of cross-correlation and zero-crossing rate of the normalised autocorrelation to approximate a measure of signal pitch and periodicity (CrossCorr) that is hypothesised to be noise robust. The score outputs by the two systems are then merged using weighted sum fusion to create the proposed autocorrelation zero-crossing rate (AZR) VAD. Accuracy of AZR was compared to state of the art and standardised VAD methods and was shown to outperform the best performing system with an average relative improvement of 24.8% in half-total error rate (HTER) on the QUT-NOISE-TIMIT database created using real recordings from high-noise environments.

Toward a more robust facial expression recognition in occluded images using randomly sampled Gabor based templates

Occlusion is a big challenge for facial expression recognition (FER) in real-world situations. Previous FER efforts to address occlusion suffer from loss of appearance features and are largely limited to a few occlusion types and single testing strategy. This paper presents a robust approach for FER in occluded images and addresses these issues. A set of Gabor based templates is extracted from images in the gallery using a Monte Carlo algorithm. These templates are converted into distance features using template matching. The resulting feature vectors are robust to occlusion. Occluded eyes and mouth regions and randomly places occlusion patches are used for testing. Two testing strategies analyze the effects of these occlusions on the overall recognition performance as well as each facial expression. Experimental results on the Cohn-Kanade database confirm the high robustness of our approach and provide useful insights about the effects of occlusion on FER. Performance is also compared with previous approaches.

Cell sourcing for bone tissue engineering : amniotic fluid stem cells have a delayed, robust differentiation compared to mesenchymal stem cells

Cell based therapies for bone regeneration are an exciting emerging technology, but the availability of osteogenic cells is limited and an ideal cell source has not been identified. Amniotic fluid-derived stem (AFS) cells and bone-marrow derived mesenchymal stem cells (MSCs) were compared to determine their osteogenic differentiation capacity in both 2D and 3D environments. In 2D culture, the AFS cells produced more mineralized matrix but delayed peaks in osteogenic markers. Cells were also cultured on 3D scaffolds constructed of poly-e-caprolactone for 15 weeks. MSCs differentiated more quickly than AFS cells on 3D scaffolds, but mineralized matrix production slowed considerably after 5 weeks. In contrast, the rate of AFS cell mineralization continued to increase out to 15 weeks, at which time AFS constructs contained 5-fold more mineralized matrix than MSC constructs. Therefore, cell source should be taken into consideration when used for cell therapy, as the MSCs would be a good choice for immediate matrix production, but the AFS cells would continue robust mineralization for an extended period of time. This study demonstrates that stem cell source can dramatically influence the magnitude and rate of osteogenic differentiation in vitro.

Robust mean super-resolution for less cooperative NIR iris recognition at a distance and on the move

Less cooperative iris identification systems at a distance and on the move often suffers from poor resolution. The lack of pixel resolution significantly degrades the iris recognition performance. Super-resolution has been considered to enhance resolution of iris images. This paper proposes a pixelwise super-resolution technique to reconstruct a high resolution iris image from a video sequence of an eye. A novel fusion approach is proposed to incorporate information details from multiple frames using robust mean. Experiments on the MBGC NIR portal database show the validity of the proposed approach in comparison with other resolution enhancement techniques.