Full-scale simulated live load tests were conducted in a controlled laboratory setting using a single-axle frame on 600-mm-inner-diameter reinforced concrete pipe (RCP) and corrugated steel pipe (CSP) when buried in dense, well-graded sand and gravel. Measurements of the RCP at nominal and working forces and beyond are reported for 0.3, 0.6 and 0.9 m of soil cover above the pipe crown. The RCP experienced no cracking when buried at 0.3 m under nominal and working CL-625 and CL-800 single-axle design loads. At these loads, the vertical contraction of the pipe diameter was less than 0.08 and 0.10 mm and the largest tensile strains in the pipe were 75 and 100 με (50-60% of the cracking strain), respectively. A 0.15 (±0.05)-mm-wide axial crack developed at the inner crown in the presence of a 6 kNm/m circumferential bending moment (70% of the theoretical ultimate moment capacity) at the fully factored CL-625 load. This crack did not propagate or widen from 3 series of cyclic load-unload tests. At 1300 kN of applied load the change in pipe diameter was less than 3.5 mm. Increasing soil cover from 0.3 to 0.6 to 0.9 m reduced the circumferential crown bending moment from 6.0 to 3.9 to 2.1 kNm/m, respectively, at 400 kN of axle load. A 1.6- and a 2.8-mm-thick CSP were also subjected to axle loading. No yielding or limit states occurred in the 1.6-mm-thick CSP when buried 0.9-m-deep. However, at 0.6 m of cover a 300 kN axle load caused local yielding at the pipe crown. Increasing soil cover from 0.6 to 0.9 m decreased the vertical diameter change from -3.0 to -1.2 mm and the crown bending moment from 0.7 to 0.2 kNm/m (75% and 20% of the yield moment), respectively, at a 250 kN axle load. Deflections of the thicker CSP were less than the thinner pipe below the CL-625 single-axle load, however further increases in applied load produced a greater response in the thicker pipe, likely due to a haunch support issue. Shallow axle loading produced a greater 3-dimensional response and a larger bending effect in both CSPs. / Thesis (Master, Civil Engineering) -- Queen's University, 2012-05-06 15:10:12.754

No description available.

Combination chemotherapies that introduce multi-agent treatments to target cancer cells are emerging as new paradigms to overcome chemotherapy resistance and side effects involved with conventional monotherapies. In environmental toxicology, characterizing effects of mixtures of toxins rather than simply analyzing the effect of single toxins are of significant interest. In order to determine such combination effects, it is necessary to systematically investigate interactions between different concentration-dependent components of a mixture. Conventional microtiter plate format based assays are efficient and cost-effective, however are not practical as the number of combinations increases drastically. Although robotic pipetting systems can overcome the labor-intensive and time-consuming limitations, they are too costly for general users. Microfluidic live cell screening platforms can allow precise control of cell culture microenvironments by applying accurate doses of biomolecular mixtures with specific mixing ratios generated through integrated on-chip microfluidic gradient generators. This thesis first presents a live cell array platform with integrated microfluidic network-based gradient generator which enables generation and dosing of 64 unique combinations of two cancer drugs at different concentrations to an 8 by 8 cell culture chamber array. We have developed the system into a fully automated microfluidic live cell screening platform with uniform cell seeding capability and pair-wise gradient profile generation. This platform was utilized to investigate the gene expression regulation of colorectal cancer cells in response to combination cancer drug treatment. The resulting cell responses indicate that the two cancer drugs show additive effect when sequential drug treatment scheme is applied, demonstrating the utility of the microfluidic live cell assay platform. However, large reagent consumption and difficulties of repeatedly generating the exact same concentrations and mixture profiles from batch to batch and device to device due to the fact that the generated gradient profiles or mixing ratios of chemicals have to rely on stable flow at optimized flow rate throughout the entire multi-day experiment limit the widespread use of this method. Moreover, producing three or more reagent mixtures require complicated microchannel structures and operating procedures when using traditional microfluidic network-based gradient generators. Therefore, an on-demand geometric metering-based mixture generator which facilitates robust, scalable, and accurate multi-reagent mixing in a high-throughput fashion has been developed and incorporated with a live cell array as a microfluidic screening platform for conducting combination drug or toxin assays. Integrated single cell trapping array allowed single cell resolution analysis of drugs and toxin effects. Reagent mixture generation and precise application of the mixtures to arrays of cell culture chambers repeatedly over time were successfully demonstrated, showing significantly improved repeatability and accuracy than those from conventional microfluidic network-based gradient generators. The influence of this improved repeatability and accuracy in generating concentration specified mixtures on obtaining more reliable and repeatable biological data sets were studied.

Surface expression has attracted much recent interest, and it has been suggested for a variety of applications. Two such applications are whole-cell biocatalysis and the creation of live vaccines. For successful implementation of these applications there is a need for flexible surface expression systems that can yield a high level of expression with a variety of recombinant fusion proteins. The aim of this work was thus to create a surface expression system that would fulfil these requirements.   A novel surface expression system based on the AIDA-I autotransporter was created with the key qualities being are good, protein-independent detection of the expression through the presence of two epitope tags flanking the recombinant protein, and full modularity of the different components of the expression cassette. To evaluate the flexibility of this construct, 8 different model proteins with potential use as live-vaccines or biocatalysts were expressed and their surface expression levels were analysed.   Positive signals were detected for all of the studied proteins using antibody labelling followed by flow cytometric analysis, showing the functionality of the expression system. The ratio of the signal from the two epitope tags indicated that several of the studied proteins were present mainly in proteolytically degraded forms, which was confirmed by Western blot analysis of the outer membrane protein fraction. This proteolysis was suggested to be due to protein-dependent stalling of translocation intermediates in the periplasm, with indications that larger size and higher cysteine content had a negative impact on expression levels. Process design with reduced cultivation pH and temperature was used to increase total surface expression yield of one of the model proteins by 400 %, with a simultaneous reduction of proteolysis by a third. While not sufficient to completely remove proteolysis, this shows that process design can be used to greatly increase surface expression. Thus, it is recommended that future work combine this with engineering of the bacterial strain or the expression system in order to overcome the observed proteolysis and maximise the yield of surface expressed protein. / <p>QC 20130516</p>

The intellectual property concept of ‘communication to the public’ in Article 3(1) of the InfoSoc Directive is a current issue within the EU, as the means of communication increase all the time due to the fast development of the information society. Accordingly, the CJEU has more than occasionally been instructed to rule upon this provision, as the Member States of the EU are unconfident as to how it should be interpreted. The poor guidance provided for in the preamble of the InfoSoc Directive in terms of how to construe the provision does probably contribute to this confusion. Article 3(1) of the InfoSoc Directive has been subject to disputes as concerns different means of communication, that is to say broadcasting, live performances, streaming and hyperlinking. Broadcasting and streaming are generally considered as communications within the meaning of the provision whereas live performances and hyperlinking have been deemed as falling outside the scope. More in detail, the most important criteria of a ‘communication to the public’ – laid down by the CJEU – suggest that the concept should be interpreted broadly, covering all transmissions and retransmissions where equipment facilitates for communications of signs, sounds or images, regardless of the means used and the nature of the place for the communication. A public refers to an indeterminate but fairly high number of people – considering the potential number rather than the exact number – not present at the place where the communication originates. The gap between the wording of Article 3(1) of the InfoSoc Directive and the developments carried out by the CJEU is considered troublesome since it decreases the legal certainty of the provision. A future recasting of these matters should thus contemplate the possibility to integrate the criteria laid down by the CJEU into the law.

The present thesis comprises a musical score and a set of specifications for a programmer to employ in the creation of a computer patch required to run the electronics. Scored for symphony orchestra and solo cello, the work also incorporates a computer operator and sound engineer who function as performers, following detailed instructions to store, activate, shape, spatialize, and regulate electronic material. The computer patch manifests two main components: the recording and playback of live sound (with the solo cello acting as the input source) and a virtual instrument feedback module. Electronic superimpositions of solo cello samples, ranging in density between a single layer and many simultaneous layers, intertwine with the live instrumentalist, forming timbrally distinct polyrhythmic contrapuntal lines. The feedback component allows for an interaction between the soloist and electronics, in which the computer operator manipulates faders connected to a series of feedback sub-patches, each employing a range of diverse delay parameters. The musical language of the work, which evolves from a series of expanding and contracting pitch clusters, continually transmutes, shifting back and forth from atonality, tonality, and polytonality. Formally, the work both opposes and espouses traditional paradigms; and it is this dichotomy, the tension created between old and new, which ultimately unifies the structure. Finally, the expansion of the role of the soloist, which entails not only the integration of multiple superimposed electronic layers but of the immersive diffusion of sound in multidimensional space, broadens the concerto form.

During inflammatory responses, macrophages make and secrete cytokines, including the proinflammatory cytokine TNF-alpha (TNF). TNF is a highly potent activator of immune responses with pleiomorphic effects throughout the body. TNF is a key causative agent of chronic inflammatory diseases and is of an intense clinical interest as a therapeutic target. At the outset of this thesis, little was known about how macrophages secrete TNF. Notably, the pathways, carriers and molecules that regulate TNF secretion had not been characterised. A main goal of this work was to identify compartment and molecules involved in the intracellular trafficking of TNF. Live cell imaging of GFP-TNF was established and this provided novel and important new insights into trafficking. Both endogenous and GFP-tagged TNF were followed in macrophages using fluorescence microscopy. The trafficking of other molecules in macrophages was also studied. The major findings of this work include the identification of a new two-step secretory pathway for TNF and other proteins from the trans-Golgi Network (TGN) to the cell surface. This pathway goes via the recycling endosome as an intermediate station. Pleiotrophic tubular-vesicular carriers containing TNF bud off the TGN for the post-Golgi trafficking of TNF and their characterization both in live cell imaging and in biochemical analysis of isolated vesicles constituted the main parts of this work. Functional studies, including endosome inactivation and overexpression of Rab11 mutants (proteins functioning at the level of the recycling endosome) revealed that recycling endosomes have indeed an essential role in the exocytic trafficking of TNF in macrophages. This thesis also provides further insight into recycling endosomes as a possible intermediate step in the exocytic trafficking of several other proteins including the adhesion protein E-Cadherin, that function at the cell surface. Finally, the last chapter of this thesis examines endocytic pathways in activated macrophages. Assays for fluid phase endocytosis and receptor-mediated endocytosis were established and the regulation of both pathways was compared. The results show that LPS has opposite effects on fluid phase and receptor mediated endocytosis, decreasing and increasing their activity respectively. Recycling of transferrin through the recycling endosome was also measured, providing a link with studies on TNF exocytosis. Overall, the work in this thesis has made a major contribution to our understanding of TNF trafficking in macrophages, of macrophage pathways more generally and of trafficking at a fundamental level. The findings herein set the stage for more in depth analysis at a single molecular level to explore TNF regulation in normal and disease cells.

The live export of cattle is an important industry for Australia. Concerns have arisen about animal welfare and in particular heat stress which may cause production losses and death. Cattle shipped live to the Middle East from a southern Australian winter can face continuous and prolonged periods of high heat and humidity as they cross the equator and arrive into a northern hemisphere summer, leading to heat stress and excessive heat load. Some live animal exporters treat heat stressed cattle with electrolyte supplements, but no scientific data exists as to whether this is beneficial to cattle in these unique environmental conditions. In response to industry’s concerns, the experiments described here monitored the physiological responses of Bos taurus and Bos indicus to conditions similar to those experienced by cattle being shipped from southern Australian to the Middle East. Initial experiments were conducted in climate controlled rooms at Murdoch University where intensive monitoring was possible. In the Bos taurus, increases in core body temperature, reductions in feed intake, and increased water intake were measured. There were also changes in blood gas variables consistent with the observed panting causing a compensated respiratory alkalosis. Following the heating period, there were decreases in blood and urinary pH. Bos indicus showed similar responses to the heat, but the changes were less pronounced at the temperatures tested. A pair feeding experiment was conducted to separate the effects of heat from the reductions in feed intake, and this indicated that the major measured effects were due to the responses to heat. On the basis of the measured responses, an electrolyte supplement was formulated and tested on Bos taurus, in the climate controlled rooms, and then on a commercial live export vessel. Results from these experiments indicated improved buffering capacity and a weight advantage for supplemented cattle, even in the absence of extreme heat stress. A final experiment investigated the effects of amount and quality of roughage in a pelleted feed on core and rumen temperature and feed intakes in Bos taurus subjected to hot environmental conditions in climate controlled rooms at Murdoch University. Both pelleted feeds had approximately the same metabolisable energy and crude protein but differed in content and type of roughage. There were no differences in feed intake, core temperature or rumen temperature between diets. This work has led to a greater understanding of the physiological responses of cattle to prolonged and continuous high heat and humidity, the requirements and effects of supplemental electrolytes in these conditions, and the effect of manipulating export diets. The demonstration of advantages in weight and buffering capacity with the electrolyte supplement highlights future areas of research to investigate electrolyte doses, route and types of supplementation, and dietary manipulation.

Ο σκοπός της διπλωματικής εργασίας ήταν η βελτιστοποίηση ενός αλγορίθμου αναγνώρισης ανθρώπινων προσώπων και η εφαρμογή του σε μια έξυπνη κάμερα. Για την αναγνώριση των προσώπων χρησιμοποιήσαμε τον αλγόριθμο PCA ( Αλγόριθμος ανάλυσης κύριων συνιστωσών ). Η εφαρμογή του αλγορίθμου έγινε σε μια εργαστηριακή έξυπνη κάμερα εξοπλισμένη με τον επεξεργαστή LEON 2 όπως επίσης και με ενσωματωμένη μνήμη Sdram μεγέθους 16Mbytes. Η βελτιστοποίηση του αλγορίθμου έγινε με γνώμονα την δυνατότητα εφαρμογής του στην έξυπνη κάμερα. Έτσι η υλοποίηση έγινε αρχικά στο περιβάλλον Matlab στην συνέχεια υλοποιήθηκε σε C γλώσσα προγραμματισμού ενώ τέλος εφαρμόστηκε, μετά από κατάλληλες παραμετροποιήσεις, στην έξυπνη κάμερα. Η έξυπνη κάμερα είναι δυνατόν να καταγράφει και να αναγνωρίζει πρόσωπα με ικανοποιητική ακρίβεια, σε χρόνο μικρότερο του ενός δευτερολέπτου. Τα αποτελέσματα ήταν αρκετά ικανοποιητικά καθώς η κάμερα μπορεί και αναγνωρίζει συγκεκριμένα πρόσωπα, μέσα από ένα σύνολο ανθρώπων που παρακολουθεί. Κύριο πλεονέκτημα της υλοποίησης είναι η μεταφερσιμότητά της που την καθιστά εύχρηστη σε πολλές εφαρμογές που απαιτούν αναγνώριση προσώπων, καθώς επίσης θα μπορούσε να αποτελέσει την βάση για επιπλέον εφαρμογές που θα αποσκοπούσαν στην αναγνώριση διαφορετικών ειδώλων. / The propose of this thesis was the optimizing of an algorithm for Face Recognition, and the implementation of this algorithm to a smart camera. In order to identify the Faces we use the PCA algorithm (Principal Component Analysis) . The implementation of the algorithm was done in a laboratory smart camera, equipped with a LEON2 processor as well as embedded 16Mbytes Sdram memory. The optimization of the algorithm was based on the applicability in the smart camera. This implementation was done originally in Matlab environment, then implemented in C programming code and after the appropriate configurations applied to the smart camera. The smart camera can record and recognize faceswith sufficient accuracy, in less than one second. The results were quite good as the camera can also recognize individual Faces within a group of observed people. Main advantage of the implementation is the portability, which makes it useful in many applications that require identification of persons. Also could be the basis for further applications aimed at identifying different kind of images.

The convergence of inexpensive video-enabled mobile phones, high-speed mobile data networks and ubiquitous sensing devices opens up a new design space called “live mobile video interaction”. It gives rise to a new genre of applications concerning live mobile video production, which can be seen as an instance of the said space. In this work we are particularly interested to explore potential technical challenges and opportunities presented by “live mobile video interaction”. We started our investigation by studying two existing prototypes from the said genre i.e. the Instant Broadcasting System (IBS) and the Mobile Vision Mixer (MVM). We studied their applicability for amateur users of collaborative mobile video production tools and the problems caused by inherent communication delays in the Internet. We acquired initial user feedback and conducted technical tests on Instant Broadcasting System (IBS) and Mobile Vision Mixer (MVM). Our results indicate that lack of synchronisation among video streams causes problems for directors in such systems that were not present in professional systems. We also identified two distinct video production modes depending on visual access of the director to the event that is being filmed. Based on our study we proposed technical design suggestions and indications on how to solve the synchronisation problems in respective mixing modes. We also proposed an algorithm for frame-rate exclusive synchronisation management of live streams in a collaborative mobile production environment. We further probed the design space using the research through design method, which resulted in a fully functional prototype system called “Livenature” that would incite an emotional connection that exists between people and the places they cherish. Further investigation of Livenature allowed us to produce detailed studies about experiential and technical aspects of the system, thus revealing phenomenological and technical dimensions of the design space.