The sustained delivery of antisense oligodeoxynucleotides using biodegradable polymer microspheres

Antisense technology is a novel drug discovery method, which provides an essential tool for directly using gene sequence information to rationally design specific inhibitions of mRNA, to treat a wide range of diseases. The efficacy of naked oligodeoxynucleotides (ODNs) is relatively short lived due to rapid degradation in vivo. The entrapment of ODNs within biodegradable sustained-release delivery systems may improve ODN stability and reduce dose required for efficacy. Biodegradable polymer microspheres were evaluated as delivery devices for ODNs and ribozymes. Poly(lactide-co-glycolide) polymers were used due to their biocompatibility and non toxic degradation products. Microspheres were prepared using a double emulsion-deposition method and the formulations characterised. In vitro release profiles were characterised by an initial burst effect during the first 48 hours of release followed by a more sustained release. The release profiles were influenced by microsphere size, copolymer molecular weight, copolymer ratio, ODN loading, ODN length, and ODN chemistry. The serum stability of ODNs was significantly improved when entrapped within polymer microspheres. The cellular association of ODNs entrapped within small spheres (1-2μm) was improved by approximately 20-fold in A431 carcinoma cells compared with free ODNs. Fluorescence microscopy studies showed a more diffuse subcellular distribution when delivered as a microsphere formulation compared with free ODNs, which exhibited the characteristic punctate periplasmic distribution. For in vivo evaluation, polymer microspheres containing fluorescently-labelled ODNs were stereo-taxically administered to the neostriatum of the rat brain. Free ODN resulted in a punctate cellular distribution after 24 hours. In comparison ODN delivered using polymer microspheres were intensely visible in cells 48 hours post administration, and fluorescence appeared to be diffuse covering both cytosolic and nuclear regions. Whole-body autoradiography was also used to evaluate the biodistribution of free tritium labelled ODN and ODN entrapped microspheres, following subcutaneous administration to Balb-C mice. Polymer entrapped ODN gave a similar biodistribution to free ODN. Free ODN was distributed within 24 hours, whereas polymer released ODN was observed still presented in organs and at the site of administration seven days post administration.

Visual performance of the human eye with soft hydrophilic contact lenses

The recording of visual acuity using the Snellen letter chart is only a limited measure of the visual performance of an eye wearing a refractive aid. Qualitative in addition to quantitative information is required to establish such a parameter: spatial, temporal and photometric aspects must all be incorporated into the test procedure. The literature relating to the correction of ametropia by refractive aids was reviewed. Selected aspects of a comparison between the correction provided by spectacles and contact lenses were considered. Special attention was directed to soft hydrophilic contact lenses. Despite technological advances which have produced physiologically acceptable soft lenses, there still remain associated with this recent form of refractive aid unpredictable visual factors. Several techniques for vision assessment were described, and previous studies of visual performance were discussed. To facilitate the investigation of visual performance in a clinical environment, a new semi-automated system was described: this utilized the presentation of broken ring test stimuli on a television screen. The research project comprised two stages. Initial work was concerned with the validation of the television system, including the optimization of its several operational variables. The second phase involved the utilization of the system in an investigation of visual performance aspects of the first month of regular daily soft contact lens wear by experimentally-naive subjects. On the basis of the results of this work an ‘homoeostatic’ model has been proposed to represent the strategy which an observer adopts in order to optimize his visual performance with soft contact lenses.

The chromatic visual evoked response as an indication of visual development

In an endeavour to provide further insight into the maturation of the cortical visual system in human infants, chromatic transient pattern reversal visual evoked potentials to red/green stimuli, were studied in a group of normal full term infants between the ages of 1 and 14 weeks post term in both cross sectional and longitudinal studies. In order to produce stimuli in which luminance cues had been eliminated with an aim to eliciting a chromatic response, preliminary studies of isoluminance determination in adults and infants were undertaken using behavioural and electrophysiological techniques. The results showed close similarity between the isoluminant ratio for adults and infants and all values were close to photometric isoluminance. Pattern reversal VEPs were recorded to stimuli of a range of red/green luminance ratios and an achromatic checkerboard. No transient VEP could be elicited with an isoluminant chromatic pattern reversal stimulus from any infant less than 7 weeks post term and similarly, all infants more than 7 weeks post term showed clear chromatic VEPs. The chromatic response first appeared at that age as a major positive component (P1) of long latency. This was delayed and reduced in comparison to the achromatic response. As the infant grew older, the latency of the P1 component decreased with the appearance of N1 and N by the 10th week post term. This finding was consistent throughout all infants assessed. In a behavioural study, no infant less than 7 weeks post term demonstrated clear discrimination of the chromatic stimulus, while those infants older than 7 weeks could do so. These findings are reviewed with respect to current neural models of visual development.

Development of visual evoked responses to tritan,red-green and luminance stimuli in human infants

The principal aim of this work was to investigate the development of the S-cone colour-opponent pathway in human infants aged 4 weeks to 6 months. This was achieved by recording transient visual evoked responses to pattern-onset stimuli along a tritanopic confusion axis (tritan stimuli) at and around the adult isoluminant match. For comparison, visual evoked responses to red-green and luminance-modulated stimuli were recorded from the same infants at the same ages. Evoked responses were also recorded from colour-normal adults for comparison with those of the infants. The transient VEP allowed observation of response morphology as luminance differences were introduced to the chromatic stimuli. In this way, an estimate of isoluminance was possible in infants. Estimated isoluminant points for a group of six infants aged 6 to 10 weeks closely approximated the adult isoluminant match. This finding has implications for the use of photometric isoluminance in infant work, and suggests that photopic spectral sensitivity is similar in infants and adults. Abnormalities of the visual evoked responses to tritan, red-green and luminance-modulated stimuli in an infant with cystic fibrosis are reported. The results suggest abnormal function of the retino-striate visual pathway in this infant, and it is argued that these may be secondary to his illness, although data from more infants with cystic fibrosis are needed to clarify this further. A group of nine healthy infants demonstrated evoked responses to tritan stimuli by 4 to 10 weeks and to red-green stimuli by 6 to 11 weeks post-term age. Responses to luminance-modulated stimuli were present in all nine infants at the earliest age tested, namely 4 weeks post-term. The slightly earlier age of onset of evoked responses to tritan stimuli than for red-green may be explained by the relatively lower cone contrast afforded by red-green stimuli. Latency of the evoked response to both types of chromatic stimuli and to luminance-modulated stimuli decreased with age at a similar rate, suggesting that the visual pathways transmitting luminance and chromatic information mature at similar rates in young infants.

Influence of impact angles on penetration rates of CRAs exposed to a high velocity multiphase flow

A combined flow loop - jet impingement pilot plant has been used to determine mass loss rates in a mixed gas - saltwater - sand multiphase flow at impact velocities up to 70 m/s. Artificial brine with a salt content of 27 g/1 was used as liquid phase. Sand content, with grain size below 150 µ, was 2.7 g/l brine. CO at a pressure of 15 bar was used as gas phase. The impact angle between jet stream (nozzle) and sample surface was varied between 30 and 90°. Rectangular stainless steel disc samples with a size of 20 × 15 × 5 mm were used. They were mechanically ground and polished prior to testing. Damaged surfaces of specimens exposed to the high velocity multiphase flow were investigated by stereo microscopy, scanning electron microscopy (SEM) and an optical device for 3D surface measurements. Furthermore, samples were investigated by applying atomic force microscopy (AFM), magnetic force microscopy (MFM) and nanoindentation. Influence of impact velocity and impact angle on penetration rates (mass loss rates) of two CRAs (UNS S30400 and N08028) are presented. Moreover effects of chemical composition and mechanical properties are critically discussed. © 2008 by NACE International.

Retinal drusen detection using the Nidek F-10 SLO in retro-mode

Presentaton Purpose:We conducted a small study to assess the novel, retro - mode imaging technique of the NIDEK F-10 scanning laser ophthalmoscope, for detecting and quantifying retinal drusen. Methods:Fundus photographs of 4 eyes of 2 patients taken in retro-mode on the Nidek F-10 SLO were graded independently by 6,experienced, masked fundus graders for the presence of retinal drusen , and compared to stereo colour fundus photographs taken with a Topcon TRC-50DX camera. Results:The mean number of retinal drusen detected in retro mode was 142.96+/- 60.8, range 63-265, and on colour fundus photography mean of 66.6+/-32.6, range 26-177. All observers independently detected approximately twice as many drusen on retro-mode than colour fundus photography (p<0.0001, Student’s paired t-test) . The statistical significance of interobserver variation in drusen detection was p=0.07 on colour fundus photography , and p=0.02 on retro mode ( ANOVA) . Conclusions:The retro-mode of the F-10 camera uses infrared laser and an aperture with a modified central stop, with the aperture deviated laterally from the confocal light path. This forms a pseudo -3D image which is a new means of detecting abnomalites in the deeper retinal layers. Retro-mode imaging of retinal drusen using the F-10 Nidek SLO is a highly sensitive technique for detecting and quantifying retinal drusen , and detected twice as many drusen than colour fundus photography. This small pilot study suggests that this novel type of imaging may have a role in the future detection and analysis of retinal drusen, a field that is likely to become increasingly important in future AMD prevention studies.

Automatic object segmentation from calibrated images

This paper addresses the problem of automatically obtaining the object/background segmentation of a rigid 3D object observed in a set of images that have been calibrated for camera pose and intrinsics. Such segmentations can be used to obtain a shape representation of a potentially texture-less object by computing a visual hull. We propose an automatic approach where the object to be segmented is identified by the pose of the cameras instead of user input such as 2D bounding rectangles or brush-strokes. The key behind our method is a pairwise MRF framework that combines (a) foreground/background appearance models, (b) epipolar constraints and (c) weak stereo correspondence into a single segmentation cost function that can be efficiently solved by Graph-cuts. The segmentation thus obtained is further improved using silhouette coherency and then used to update the foreground/background appearance models which are fed into the next Graph-cut computation. These two steps are iterated until segmentation convergences. Our method can automatically provide a 3D surface representation even in texture-less scenes where MVS methods might fail. Furthermore, it confers improved performance in images where the object is not readily separable from the background in colour space, an area that previous segmentation approaches have found challenging. © 2011 IEEE.

Large scale multi-view stereopsis evaluation

The seminal multiple view stereo benchmark evaluations from Middlebury and by Strecha et al. have played a major role in propelling the development of multi-view stereopsis methodology. Although seminal, these benchmark datasets are limited in scope with few reference scenes. Here, we try to take these works a step further by proposing a new multi-view stereo dataset, which is an order of magnitude larger in number of scenes and with a significant increase in diversity. Specifically, we propose a dataset containing 80 scenes of large variability. Each scene consists of 49 or 64 accurate camera positions and reference structured light scans, all acquired by a 6-axis industrial robot. To apply this dataset we propose an extension of the evaluation protocol from the Middlebury evaluation, reflecting the more complex geometry of some of our scenes. The proposed dataset is used to evaluate the state of the art multiview stereo algorithms of Tola et al., Campbell et al. and Furukawa et al. Hereby we demonstrate the usability of the dataset as well as gain insight into the workings and challenges of multi-view stereopsis. Through these experiments we empirically validate some of the central hypotheses of multi-view stereopsis, as well as determining and reaffirming some of the central challenges.

Large-scale data for multiple-view stereopsis

The seminal multiple-view stereo benchmark evaluations from Middlebury and by Strecha et al. have played a major role in propelling the development of multi-view stereopsis (MVS) methodology. The somewhat small size and variability of these data sets, however, limit their scope and the conclusions that can be derived from them. To facilitate further development within MVS, we here present a new and varied data set consisting of 80 scenes, seen from 49 or 64 accurate camera positions. This is accompanied by accurate structured light scans for reference and evaluation. In addition all images are taken under seven different lighting conditions. As a benchmark and to validate the use of our data set for obtaining reasonable and statistically significant findings about MVS, we have applied the three state-of-the-art MVS algorithms by Campbell et al., Furukawa et al., and Tola et al. to the data set. To do this we have extended the evaluation protocol from the Middlebury evaluation, necessitated by the more complex geometry of some of our scenes. The data set and accompanying evaluation framework are made freely available online. Based on this evaluation, we are able to observe several characteristics of state-of-the-art MVS, e.g. that there is a tradeoff between the quality of the reconstructed 3D points (accuracy) and how much of an object’s surface is captured (completeness). Also, several issues that we hypothesized would challenge MVS, such as specularities and changing lighting conditions did not pose serious problems. Our study finds that the two most pressing issues for MVS are lack of texture and meshing (forming 3D points into closed triangulated surfaces).

Expanding guitar production techniques: Building the Guitar Application Toolkit (GATK)

The purpose of this thesis was to build the Guitar Application ToolKit (GATK), a series of applications used to expand the sonic capabilities of the acoustic/electric stereo guitar. Furthermore, the goal of the GATK was to extend improvisational capabilities and the compositional techniques generated by this innovative instrument. ^ During the GATK creation process, the current production guitar techniques and overall sonic result were enhanced by planning and implementing a personalized electro-acoustic performance set up, designing custom-made performance interfaces, creating interactive compositional strategies, crafting non-standardized sounds, and controlling various music parameters in real-time using the Max/MSP programming environment. ^ This was the fast thesis project of its kind. It is expected that this thesis will be useful as a reference paper for electronic musicians and music technology students; as a product demonstration for companies that manufacture the relevant software; and as a personal portfolio for future technology related jobs. ^

The development of optimized high performance liquid chromatography systems for the analysis of improvised explosives

Existing instrumental techniques must be adaptable to the analysis of novel explosives if science is to keep up with the practices of terrorists and criminals. The focus of this work has been the development of analytical techniques for the analysis of two types of novel explosives: ascorbic acid-based propellants, and improvised mixtures of concentrated hydrogen peroxide/fuel. In recent years, the use of these explosives in improvised explosive devices (IEDs) has increased. It is therefore important to develop methods which permit the identification of the nature of the original explosive from post-blast residues. Ascorbic acid-based propellants are low explosives which employ an ascorbic acid fuel source with a nitrate/perchlorate oxidizer. A method which utilized ion chromatography with indirect photometric detection was optimized for the analysis of intact propellants. Post-burn and post-blast residues if these propellants were analyzed. It was determined that the ascorbic acid fuel and nitrate oxidizer could be detected in intact propellants, as well as in the post-burn and post-blast residues. Degradation products of the nitrate and perchlorate oxidizers were also detected. With a quadrupole time-of-flight mass spectrometer (QToFMS), exact mass measurements are possible. When an HPLC instrument is coupled to a QToFMS, the combination of retention time with accurate mass measurements, mass spectral fragmentation information, and isotopic abundance patterns allows for the unequivocal identification of a target analyte. An optimized HPLC-ESI-QToFMS method was applied to the analysis of ascorbic acid-based propellants. Exact mass measurements were collected for the fuel and oxidizer anions, and their degradation products. Ascorbic acid was detected in the intact samples and half of the propellants subjected to open burning; the intact fuel molecule was not detected in any of the post-blast residue. Two methods were optimized for the analysis of trace levels of hydrogen peroxide: HPLC with fluorescence detection (HPLC-FD), and HPLC with electrochemical detection (HPLC-ED). Both techniques were extremely selective for hydrogen peroxide. Both methods were applied to the analysis of post-blast debris from improvised mixtures of concentrated hydrogen peroxide/fuel; hydrogen peroxide was detected on variety of substrates. Hydrogen peroxide was detected in the post-blast residues of the improvised explosives TATP and HMTD.

Using sonic enhancement to augment non-visual tabular navigation

More information is now readily available to computer users than at any time in human history; however, much of this information is often inaccessible to people with blindness or low-vision, for whom information must be presented non-visually. Currently, screen readers are able to verbalize on-screen text using text-to-speech (TTS) synthesis; however, much of this vocalization is inadequate for browsing the Internet. An auditory interface that incorporates auditory-spatial orientation was created and tested. For information that can be structured as a two-dimensional table, links can be semantically grouped as cells in a row within an auditory table, which provides a consistent structure for auditory navigation. An auditory display prototype was tested.^ Sixteen legally blind subjects participated in this research study. Results demonstrated that stereo panning was an effective technique for audio-spatially orienting non-visual navigation in a five-row, six-column HTML table as compared to a centered, stationary synthesized voice. These results were based on measuring the time- to-target (TTT), or the amount of time elapsed from the first prompting to the selection of each tabular link. Preliminary analysis of the TTT values recorded during the experiment showed that the populations did not conform to the ANOVA requirements of normality and equality of variances. Therefore, the data were transformed using the natural logarithm. The repeated-measures two-factor ANOVA results show that the logarithmically-transformed TTTs were significantly affected by the tonal variation method, F(1,15) = 6.194, p= 0.025. Similarly, the results show that the logarithmically transformed TTTs were marginally affected by the stereo spatialization method, F(1,15) = 4.240, p=0.057. The results show that the logarithmically transformed TTTs were not significantly affected by the interaction of both methods, F(1,15) = 1.381, p=0.258. These results suggest that some confusion may be caused in the subject when employing both of these methods simultaneously. The significant effect of tonal variation indicates that the effect is actually increasing the average TTT. In other words, the presence of preceding tones increases task completion time on average. The marginally-significant effect of stereo spatialization decreases the average log(TTT) from 2.405 to 2.264.^

3D navigation with six degrees-of-freedom using a multi-touch display

With the introduction of new input devices, such as multi-touch surface displays, the Nintendo WiiMote, the Microsoft Kinect, and the Leap Motion sensor, among others, the field of Human-Computer Interaction (HCI) finds itself at an important crossroads that requires solving new challenges. Given the amount of three-dimensional (3D) data available today, 3D navigation plays an important role in 3D User Interfaces (3DUI). This dissertation deals with multi-touch, 3D navigation, and how users can explore 3D virtual worlds using a multi-touch, non-stereo, desktop display. ^ The contributions of this dissertation include a feature-extraction algorithm for multi-touch displays (FETOUCH), a multi-touch and gyroscope interaction technique (GyroTouch), a theoretical model for multi-touch interaction using high-level Petri Nets (PeNTa), an algorithm to resolve ambiguities in the multi-touch gesture classification process (Yield), a proposed technique for navigational experiments (FaNS), a proposed gesture (Hold-and-Roll), and an experiment prototype for 3D navigation (3DNav). The verification experiment for 3DNav was conducted with 30 human-subjects of both genders. The experiment used the 3DNav prototype to present a pseudo-universe, where each user was required to find five objects using the multi-touch display and five objects using a game controller (GamePad). For the multi-touch display, 3DNav used a commercial library called GestureWorks in conjunction with Yield to resolve the ambiguity posed by the multiplicity of gestures reported by the initial classification. The experiment compared both devices. The task completion time with multi-touch was slightly shorter, but the difference was not statistically significant. The design of experiment also included an equation that determined the level of video game console expertise of the subjects, which was used to break down users into two groups: casual users and experienced users. The study found that experienced gamers performed significantly faster with the GamePad than casual users. When looking at the groups separately, casual gamers performed significantly better using the multi-touch display, compared to the GamePad. Additional results are found in this dissertation.^

The Development of High Performance Liquid Chromatography Systems for the Analysis of Improvised Explosives

Existing instrumental techniques must be adaptable to the analysis of novel explosives if science is to keep up with the practices of terrorists and criminals. The focus of this work has been the development of analytical techniques for the analysis of two types of novel explosives: ascorbic acid-based propellants, and improvised mixtures of concentrated hydrogen peroxide/fuel. In recent years, the use of these explosives in improvised explosive devices (IEDs) has increased. It is therefore important to develop methods which permit the identification of the nature of the original explosive from post-blast residues. Ascorbic acid-based propellants are low explosives which employ an ascorbic acid fuel source with a nitrate/perchlorate oxidizer. A method which utilized ion chromatography with indirect photometric detection was optimized for the analysis of intact propellants. Post-burn and post-blast residues if these propellants were analyzed. It was determined that the ascorbic acid fuel and nitrate oxidizer could be detected in intact propellants, as well as in the post-burn and post-blast residues. Degradation products of the nitrate and perchlorate oxidizers were also detected. With a quadrupole time-of-flight mass spectrometer (QToFMS), exact mass measurements are possible. When an HPLC instrument is coupled to a QToFMS, the combination of retention time with accurate mass measurements, mass spectral fragmentation information, and isotopic abundance patterns allows for the unequivocal identification of a target analyte. An optimized HPLC-ESI-QToFMS method was applied to the analysis of ascorbic acid-based propellants. Exact mass measurements were collected for the fuel and oxidizer anions, and their degradation products. Ascorbic acid was detected in the intact samples and half of the propellants subjected to open burning; the intact fuel molecule was not detected in any of the post-blast residue. Two methods were optimized for the analysis of trace levels of hydrogen peroxide: HPLC with fluorescence detection (HPLC-FD), and HPLC with electrochemical detection (HPLC-ED). Both techniques were extremely selective for hydrogen peroxide. Both methods were applied to the analysis of post-blast debris from improvised mixtures of concentrated hydrogen peroxide/fuel; hydrogen peroxide was detected on variety of substrates. Hydrogen peroxide was detected in the post-blast residues of the improvised explosives TATP and HMTD.

Análise do comportamento de técnicas de correspondência binocular para imagens em alta definição

This work presents an analysis of the behavior of some algorithms usually available in stereo correspondence literature, with full HD images (1920x1080 pixels) to establish, within the precision dilemma versus runtime applications which these methods can be better used. The images are obtained by a system composed of a stereo camera coupled to a computer via a capture board. The OpenCV library is used for computer vision operations and processing images involved. The algorithms discussed are an overall method of search for matching blocks with the Sum of the Absolute Value of the difference (Sum of Absolute Differences - SAD), a global technique based on cutting energy graph cuts, and a so-called matching technique semi -global. The criteria for analysis are processing time, the consumption of heap memory and the mean absolute error of disparity maps generated.