Multi-spectral fusion for surveillance systems

Surveillance systems such as object tracking and abandoned object detection systems typically rely on a single modality of colour video for their input. These systems work well in controlled conditions but often fail when low lighting, shadowing, smoke, dust or unstable backgrounds are present, or when the objects of interest are a similar colour to the background. Thermal images are not affected by lighting changes or shadowing, and are not overtly affected by smoke, dust or unstable backgrounds. However, thermal images lack colour information which makes distinguishing between different people or objects of interest within the same scene difficult. ----- By using modalities from both the visible and thermal infrared spectra, we are able to obtain more information from a scene and overcome the problems associated with using either modality individually. We evaluate four approaches for fusing visual and thermal images for use in a person tracking system (two early fusion methods, one mid fusion and one late fusion method), in order to determine the most appropriate method for fusing multiple modalities. We also evaluate two of these approaches for use in abandoned object detection, and propose an abandoned object detection routine that utilises multiple modalities. To aid in the tracking and fusion of the modalities we propose a modified condensation filter that can dynamically change the particle count and features used according to the needs of the system. ----- We compare tracking and abandoned object detection performance for the proposed fusion schemes and the visual and thermal domains on their own. Testing is conducted using the OTCBVS database to evaluate object tracking, and data captured in-house to evaluate the abandoned object detection. Our results show that significant improvement can be achieved, and that a middle fusion scheme is most effective.

Investigation of highly regulated promoters for the production of recombinant proteins in plants

Plants have been identified as promising expression systems for the commercial production of recombinant proteins. Plant-based protein production or “biofarming” offers a number of advantages over traditional expression systems in terms of scale of production, the capacity for post-translation processing, providing a product free of contaminants and cost effectiveness. A number of pharmaceutically important and commercially valuable proteins, such as antibodies, biopharmaceuticals and industrial enzymes are currently being produced in plant expression systems. However, several challenges still remain to improve recombinant protein yield with no ill effect on the host plant. The ability for transgenic plants to produce foreign proteins at commercially viable levels can be directly related to the level and cell specificity of the selected promoter driving the transgene. The accumulation of recombinant proteins may be controlled by a tissue-specific, developmentally-regulated or chemically-inducible promoter such that expression of recombinant proteins can be spatially- or temporally- controlled. The strict control of gene expression is particularly useful for proteins that are considered toxic and whose expression is likely to have a detrimental effect on plant growth. To date, the most commonly used promoter in plant biotechnology is the cauliflower mosaic virus (CaMV) 35S promoter which is used to drive strong, constitutive transgene expression in most organs of transgenic plants. Of particular interest to researchers in the Centre for Tropical Crops and Biocommodities at QUT are tissue-specific promoters for the accumulation of foreign proteins in the roots, seeds and fruit of various plant species, including tobacco, banana and sugarcane. Therefore this Masters project aimed to isolate and characterise root- and seed-specific promoters for the control of genes encoding recombinant proteins in plant-based expression systems. Additionally, the effects of matching cognate terminators with their respective gene promoters were assessed. The Arabidopsis root promoters ARSK1 and EIR1 were selected from the literature based on their reported limited root expression profiles. Both promoters were analysed using the PlantCARE database to identify putative motifs or cis-acting elements that may be associated with this activity. A number of motifs were identified in the ARSK1 promoter region including, WUN (wound-inducible), MBS (MYB binding site), Skn-1, and a RY core element (seed-specific) and in the EIR1 promoter region including, Skn-1 (seed-specific), Box-W1 (fungal elicitor), Aux-RR core (auxin response) and ABRE (ABA response). However, no previously reported root-specific cis-acting elements were observed in either promoter region. To confirm root specificity, both promoters, and truncated versions, were fused to the GUS reporter gene and the expression cassette introduced into Arabidopsis via Agrobacterium-mediated transformation. Despite the reported tissue-specific nature of these promoters, both upstream regulatory regions directed constitutive GUS expression in all transgenic plants. Further, similar levels of GUS expression from the ARSK1 promoter were directed by the control CaMV 35S promoter. The truncated version of the EIR1 promoter (1.2 Kb) showed some differences in the level of GUS expression compared to the 2.2 Kb promoter. Therefore, this suggests an enhancer element is contained in the 2.2 Kb upstream region that increases transgene expression. The Arabidopsis seed-specific genes ATS1 and ATS3 were selected from the literature based on their seed-specific expression profiles and gene expression confirmed in this study as seed-specific by RT-PCR analysis. The selected promoter regions were analysed using the PlantCARE database in order to identify any putative cis elements. The seed-specific motifs GCN4 and Skn-1 were identified in both promoter regions that are associated with elevated expression levels in the endosperm. Additionaly, the seed-specific RY element and the ABRE were located in the ATS1 promoter. Both promoters were fused to the GUS reporter gene and used to transform Arabidopsis plants. GUS expression from the putative promoters was consitutive in all transgenic Arabidopsis tissue tested. Importantly, the positive control FAE1 seed-specific promoter also directed constitutive GUS expression throughout transgenic Arabidopsis plants. The constitutive nature seen in all of the promoters used in this study was not anticipated. While variations in promoter activity can be caused by a number of influencing factors, the variation in promoter activity observed here would imply a major contributing factor common to all plant expression cassettes tested. All promoter constructs generated in this study were based on the binary vector pCAMBIA2300. This vector contains the plant selection gene (NPTII) under the transcriptional control of the duplicated CaMV 35S promoter. This CaMV 35S promoter contains two enhancer domains that confer strong, constitutive expression of the selection gene and is located immediately upstream of the promoter-GUS fusion. During the course of this project, Yoo et al. (2005) reported that transgene expression is significantly affected when the expression cassette is located on the same T-DNA as the 35S enhancer. It was concluded, the trans-acting effects of the enhancer activate and control transgene expression causing irregular expression patterns. This phenomenon seems the most plausible reason for the constitutive expression profiles observed with the root- and seed-specific promoters assessed in this study. The expression from some promoters can be influenced by their cognate terminator sequences. Therefore, the Arabidopsis ARSK1, EIR1, ATS1 and ATS3 terminator sequences were isolated and incorporated into expression cassettes containing the GUS reporter gene under the control of their cognate promoters. Again, unrestricted GUS activity was displayed throughout transgenic plants transformed with these reporter gene fusions. As previously discussed constitutive GUS expression was most likely due to the trans-acting effect of the upstream CaMV 35S promoter in the selection cassette located on the same T-DNA. The results obtained in this study make it impossible to assess the influence matching terminators with their cognate promoters have on transgene expression profiles. The obvious future direction of research continuing from this study would be to transform pBIN-based promoter-GUS fusions (ie. constructs containing no CaMV 35S promoter driving the plant selection gene) into Arabidopsis in order to determine the true tissue specificity of these promoters and evaluate the effects of their cognate 3’ terminator sequences. Further, promoter truncations based around the cis-elements identified here may assist in determining whether these motifs are in fact involved in the overall activity of the promoter.

Bioactive mesopore-glass microspheres with controllable protein-delivery properties by biomimetic surface modification

Microsphere systems with the ideal properties for bone regeneration need to be bioactive, and at the same time possess the capacity for controlled protein/drug-delivery; however, the current crop of microsphere system fails to fulfill these properties. The aim of this study was to develop a novel protein-delivery system of bioactive mesoporous glass (MBG) microspheres by a biomimetic method through controlling the density of apatite on the surface of microspheres, for potential bone tissue regeneration. MBG microspheres were prepared by using the method of alginate cross-linking with Ca2+ ions. The cellular bioactivity of MBG microspheres was evaluated by investigating the proliferation and attachment of bone marrow stromal cell (BMSC). The loading efficiency and release kinetics of bovine serum albumin (BSA) on MBG microspheres were investigated after coprecipitating with biomimetic apatite in simulated body fluids (SBF). The results showed that MBG microspheres supported BMSC attachment and the Si containing ionic products from MBG microspheres stimulated BMSCs proliferation. The density of apatite on MBG microspheres increased with the length of soaking time in SBF. BSA-loading efficiency of MBG was significantly enhanced by co-precipitating with apatite. Furthermore, the loading efficiency and release kinetics of BSA could be controlled by controlling the density of apatite formed on MBG microspheres. Our results suggest that MBG microspheres are a promising protein-delivery system as a filling material for bone defect healing and regeneration.

Porous PLGA microspheres effectively loaded with BSA protein by electrospraying combined with phase separation in liquid nitrogen

Polymer microspheres loaded with bioactive particles, biomolecules, proteins, and/or growth factors play important roles in tissue engineering, drug delivery, and cell therapy. The conventional double emulsion method and a new method of electrospraying into liquid nitrogen were used to prepare bovine serum albumin (BAS)-loaded poly(lactic-co-glycolic acid) (PLGA) porous microspheres. The particle size, the surface morphology and the internal porous structure of the microspheres were observed using scanning electron microscopy (SEM). The loading efficiency, the encapsulation efficiency, and the release profile of the BSA-loaded PLGA microspheres were measured and studied. It was shown that the microspheres from double emulsion had smaller particle sizes (3-50 m), a less porous structure, a poor loading efficiency (5.2 %), and a poor encapsulation efficiency (43.5%). However, the microspheres from the electrospraying into liquid nitrogen had larger particle sizes (400-600 m), a highly porous structure, a high loading efficiency (12.2%), and a high encapsulation efficiency (93.8%). Thus the combination of electrospraying with freezing in liquid nitrogen and subsequent freeze drying represented a suitable way to produce polymer microspheres for effective loading and sustained release of proteins.

Sequential decision fusion for controlled detection errors

Information fusion in biometrics has received considerable attention. The architecture proposed here is based on the sequential integration of multi-instance and multi-sample fusion schemes. This method is analytically shown to improve the performance and allow a controlled trade-off between false alarms and false rejects when the classifier decisions are statistically independent. Equations developed for detection error rates are experimentally evaluated by considering the proposed architecture for text dependent speaker verification using HMM based digit dependent speaker models. The tuning of parameters, n classifiers and m attempts/samples, is investigated and the resultant detection error trade-off performance is evaluated on individual digits. Results show that performance improvement can be achieved even for weaker classifiers (FRR-19.6%, FAR-16.7%). The architectures investigated apply to speaker verification from spoken digit strings such as credit card numbers in telephone or VOIP or internet based applications.

Secondary curve behaviour in Lenke IC class adolescent idiopathic scoliosis following video assisted thoracoscopic spinal fusion

Introduction. Ideally after selective thoracic fusion for Lenke Class IC (i.e. major thoracic / secondary lumbar) curves, the lumbar spine will spontaneously accommodate to the corrected position of the thoracic curve, thereby achieving a balanced spine, avoiding the need for fusion of lumbar spinal segments1. The purpose of this study was to evaluate the behaviour of the lumbar curve in Lenke IC class adolescent idiopathic scoliosis (AIS) following video-assisted thoracoscopic spinal fusion and instrumentation (VATS) of the major thoracic curve. Methods. A retrospective review of 22 consecutive patients with AIS who underwent VATS by a single surgeon was conducted. The results were compared to published literature examining the behaviour of the secondary lumbar curve where other surgical approaches were employed. Results. Twenty-two patients (all female) with AIS underwent VATS. All major thoracic curves were right convex. The average age at surgery was 14 years (range 10 to 22 years). On average 6.7 levels (6 to 8) were instrumented. The mean follow-up was 25.1 months (6 to 36). The pre-operative major thoracic Cobb angle mean was 53.8° (40° to 75°). The pre-operative secondary lumbar Cobb angle mean was 43.9° (34° to 55°). On bending radiographs, the secondary curve corrected to 11.3° (0° to 35°). The rib hump mean measurement was 15.0° (7° to 21°). At latest follow-up the major thoracic Cobb angle measured on average 27.2° (20° to 41°) (p<0.001 – univariate ANOVA) and the mean secondary lumbar curve was 27.3° (15° to 42°) (p<0.001). This represented an uninstrumented secondary curve correction factor of 37.8%. The mean rib hump measured was 6.5° (2° to 15°) (p<0.001). The results above were comparable to published series when open surgery was performed. Discussion. VATS is an effective method of correcting major thoracic curves with secondary lumbar curves. The behaviour of the secondary lumbar curve is consistent with published series when open surgery, both anterior and posterior, is performed.

Computed tomography analysis of sagittal plane parameters in adolescent idiopathic scoliosis treated by video assisted thoracoscopic spinal fusion and instrumentation

One of the primary treatment goals of adolescent idiopathic scoliosis (AIS) surgery is to achieve maximum coronal plane correction while maintaining coronal balance. However maintaining or restoring sagittal plane spinal curvature has become increasingly important in maintaining the long-term health of the spine. Patients with AIS are characterised by pre-operative thoracic hypokyphosis, and it is generally agreed that operative treatment of thoracic idiopathic scoliosis should aim to restore thoracic kyphosis to normal values while maintaining lumbar lordosis and good overall sagittal balance. The aim of this study was to evaluate CT sagittal plane parameters, with particular emphasis on thoracolumbar junctional alignment, in patients with AIS who underwent Video Assisted Thoracoscopic Spinal Fusion and Instrumentation (VATS). This study concluded that video-assisted thoracoscopic spinal fusion and instrumentation reliably increases thoracic kyphosis while preserving junctional alignment and lumbar lordosis in thoracic AIS.

Image acquisition and manipulation protocols for CT-PET fusion to improve the accuracy of gross tumour volume localisation for 3D conformal radiotherapy for lung cancer

Aims: To develop clinical protocols for acquiring PET images, performing CT-PET registration and tumour volume definition based on the PET image data, for radiotherapy for lung cancer patients and then to test these protocols with respect to levels of accuracy and reproducibility. Method: A phantom-based quality assurance study of the processes associated with using registered CT and PET scans for tumour volume definition was conducted to: (1) investigate image acquisition and manipulation techniques for registering and contouring CT and PET images in a radiotherapy treatment planning system, and (2) determine technology-based errors in the registration and contouring processes. The outcomes of the phantom image based quality assurance study were used to determine clinical protocols. Protocols were developed for (1) acquiring patient PET image data for incorporation into the 3DCRT process, particularly for ensuring that the patient is positioned in their treatment position; (2) CT-PET image registration techniques and (3) GTV definition using the PET image data. The developed clinical protocols were tested using retrospective clinical trials to assess levels of inter-user variability which may be attributed to the use of these protocols. A Siemens Somatom Open Sensation 20 slice CT scanner and a Philips Allegro stand-alone PET scanner were used to acquire the images for this research. The Philips Pinnacle3 treatment planning system was used to perform the image registration and contouring of the CT and PET images. Results: Both the attenuation-corrected and transmission images obtained from standard whole-body PET staging clinical scanning protocols were acquired and imported into the treatment planning system for the phantom-based quality assurance study. Protocols for manipulating the PET images in the treatment planning system, particularly for quantifying uptake in volumes of interest and window levels for accurate geometric visualisation were determined. The automatic registration algorithms were found to have sub-voxel levels of accuracy, with transmission scan-based CT-PET registration more accurate than emission scan-based registration of the phantom images. Respiration induced image artifacts were not found to influence registration accuracy while inadequate pre-registration over-lap of the CT and PET images was found to result in large registration errors. A threshold value based on a percentage of the maximum uptake within a volume of interest was found to accurately contour the different features of the phantom despite the lower spatial resolution of the PET images. Appropriate selection of the threshold value is dependant on target-to-background ratios and the presence of respiratory motion. The results from the phantom-based study were used to design, implement and test clinical CT-PET fusion protocols. The patient PET image acquisition protocols enabled patients to be successfully identified and positioned in their radiotherapy treatment position during the acquisition of their whole-body PET staging scan. While automatic registration techniques were found to reduce inter-user variation compared to manual techniques, there was no significant difference in the registration outcomes for transmission or emission scan-based registration of the patient images, using the protocol. Tumour volumes contoured on registered patient CT-PET images using the tested threshold values and viewing windows determined from the phantom study, demonstrated less inter-user variation for the primary tumour volume contours than those contoured using only the patient’s planning CT scans. Conclusions: The developed clinical protocols allow a patient’s whole-body PET staging scan to be incorporated, manipulated and quantified in the treatment planning process to improve the accuracy of gross tumour volume localisation in 3D conformal radiotherapy for lung cancer. Image registration protocols which factor in potential software-based errors combined with adequate user training are recommended to increase the accuracy and reproducibility of registration outcomes. A semi-automated adaptive threshold contouring technique incorporating a PET windowing protocol, accurately defines the geometric edge of a tumour volume using PET image data from a stand alone PET scanner, including 4D target volumes.