Development of zonal cartilage constructs : effects of chondrocyte subpopulation, compressive stimulation, and culture models

Articular cartilage is a highly resilient tissue located at the ends of long bones. It has a zonal structure, which has functional significance in load-bearing. Cartilage does not spontaneously heal itself when damaged, and untreated cartilage lesions or age-related wear often lead to osteoarthritis (OA). OA is a degenerative condition that is highly prevalent, age-associated, and significantly affects patient mobility and quality of life. There is no cure for OA, and patients usually resort to replacing the biological joint with an artificial prosthesis. An alternative approach is to dynamically regenerate damaged or diseased cartilage through cartilage tissue engineering, where cells, materials, and stimuli are combined to form new cartilage. However, despite extensive research, major limitations remain that have prevented the wide-spread application of tissue-engineered cartilage. Critically, there is a dearth of information on whether autologous chondrocytes obtained from OA patients can be used to successfully generate cartilage tissues with structural hierarchy typically found in normal articular cartilage. I aim to address these limitations in this thesis by showing that chondrocyte subpopulations isolated from macroscopically normal areas of the cartilage can be used to engineer stratified cartilage tissues and that compressive loading plays an important role in zone-dependent biosynthesis of these chondrocytes. I first demonstrate that chondrocyte subpopulations from the superficial (S) and middle/deep (MD) zones of OA cartilage are responsive to compressive stimulation in vitro, and that the effect of compression on construct quality is zone-dependent. I also show that compressive stimulation can influence pericelluar matrix production, matrix metalloproteinase secretion, and cytokine expression in zonal chondrocytes in an alginate hydrogel model. Subsequently, I focus on recreating the zonal structure by forming layered constructs using the alginate-released chondrocyte (ARC) method either with or without polymeric scaffolds. Resulting zonal ARC constructs had hyaline morphology, and expressed cartilage matrix molecules such as proteoglycans and collagen type II in both scaffold-free and scaffold-based approaches. Overall, my findings demonstrate that chondrocyte subpopulations obtained from OA joints respond sensitively to compressive stimulation, and are able to form cartilaginous constructs with stratified organization similar to native cartilage using the scaffold-free and scaffold-based ARC technique. The ultimate goal in tissue engineering is to help provide improved treatment options for patients suffering from debilitating conditions such as OA. Further investigations in developing functional cartilage replacement tissues using autologous chondrocytes will bring us a step closer to improving the quality of life for millions of OA patients worldwide.

Analysis of acoustic emission data for accurate damage assessment for structural health monitoring applications

Structural health monitoring (SHM) refers to the procedure used to assess the condition of structures so that their performance can be monitored and any damage can be detected early. Early detection of damage and appropriate retrofitting will aid in preventing failure of the structure and save money spent on maintenance or replacement and ensure the structure operates safely and efficiently during its whole intended life. Though visual inspection and other techniques such as vibration based ones are available for SHM of structures such as bridges, the use of acoustic emission (AE) technique is an attractive option and is increasing in use. AE waves are high frequency stress waves generated by rapid release of energy from localised sources within a material, such as crack initiation and growth. AE technique involves recording these waves by means of sensors attached on the surface and then analysing the signals to extract information about the nature of the source. High sensitivity to crack growth, ability to locate source, passive nature (no need to supply energy from outside, but energy from damage source itself is utilised) and possibility to perform real time monitoring (detecting crack as it occurs or grows) are some of the attractive features of AE technique. In spite of these advantages, challenges still exist in using AE technique for monitoring applications, especially in the area of analysis of recorded AE data, as large volumes of data are usually generated during monitoring. The need for effective data analysis can be linked with three main aims of monitoring: (a) accurately locating the source of damage; (b) identifying and discriminating signals from different sources of acoustic emission and (c) quantifying the level of damage of AE source for severity assessment. In AE technique, the location of the emission source is usually calculated using the times of arrival and velocities of the AE signals recorded by a number of sensors. But complications arise as AE waves can travel in a structure in a number of different modes that have different velocities and frequencies. Hence, to accurately locate a source it is necessary to identify the modes recorded by the sensors. This study has proposed and tested the use of time-frequency analysis tools such as short time Fourier transform to identify the modes and the use of the velocities of these modes to achieve very accurate results. Further, this study has explored the possibility of reducing the number of sensors needed for data capture by using the velocities of modes captured by a single sensor for source localization. A major problem in practical use of AE technique is the presence of sources of AE other than crack related, such as rubbing and impacts between different components of a structure. These spurious AE signals often mask the signals from the crack activity; hence discrimination of signals to identify the sources is very important. This work developed a model that uses different signal processing tools such as cross-correlation, magnitude squared coherence and energy distribution in different frequency bands as well as modal analysis (comparing amplitudes of identified modes) for accurately differentiating signals from different simulated AE sources. Quantification tools to assess the severity of the damage sources are highly desirable in practical applications. Though different damage quantification methods have been proposed in AE technique, not all have achieved universal approval or have been approved as suitable for all situations. The b-value analysis, which involves the study of distribution of amplitudes of AE signals, and its modified form (known as improved b-value analysis), was investigated for suitability for damage quantification purposes in ductile materials such as steel. This was found to give encouraging results for analysis of data from laboratory, thereby extending the possibility of its use for real life structures. By addressing these primary issues, it is believed that this thesis has helped improve the effectiveness of AE technique for structural health monitoring of civil infrastructures such as bridges.

Structural performance of reinforced concrete beams repairing from spalling

The effectiveness of a repair work for the restoration of spalled reinforced concrete (r.c.) structures depends to a great extent, on their ability to restore the structural integrity of the r.c. element, to restore its serviceability and to protect the reinforcements from further deterioration. This paper presents results of a study concocted to investigate the structural performance of eight spalled r.c. beams repaired using two advanced repair materials in various zones for comparison purposes, namely a free flowing self compacting mortar (FFSCM) and a polymer Modified cementitious mortar (PMCM). The repair technique adopted was that for the repair of spalled concrete in which the bond between the concrete and steel was completely lost due to reinforcement corrosion or the effect of fire or impact. The beams used for the experiment were first cast, then hacked at various zones before they were repaired except for the control beam. The beam specimens were then loaded to failure under four point loadings. The structural response of each beam was evaluated in terms of first crack load, cracking behavior, crack pattern, deflection, variation of strains in the concrete and steel, collapse load and the modes of failure. The results of the test showed that, the repair materials applied on the various zones of the beams were able to restore more than 100% of the beams’ capacity and that FFSCM gave a better overall performance.

Measurement of structural changes to a food material during dehydration

Rates of dehydration/rehydration are important quality parameters for dried products. Theoretically, if there are no adverse effects on the integrity of the tissue structure, it should absorb water to the same moisture content of the initial product before drying.The purpose of this work is to semi-automate the process of detection of cell structure boundaries as a food is dehydrated and rehydrated. This will enable food materials researchers to quantify changes to material’s structure as these processes take place. Images of potato cells as they were dehydrated and rehydrated were taken using an electron microscope. Cell boundaries were detected using an image processing algorithm. Average cell area and perimeter at each stage of dehydration were calculated and plotted versus time. The results show that the algorithm can successfully identify cell boundaries.

A particle-based micromechanics approach to simulate structural changes of plant cells during drying

This paper is concerned with applying a particle-based approach to simulate the micro-level cellular structural changes of plant cells during drying. The objective of the investigation was to relate the micro-level structural properties such as cell area, diameter and perimeter to the change of moisture content of the cell. Model assumes a simplified cell which consists of two basic components, cell wall and cell fluid. The cell fluid is assumed to be a Newtonian fluid with higher viscosity compared to water and cell wall is assumed to be a visco-elastic solid boundary located around the cell fluid. Cell fluid is modelled with Smoothed Particle Hydrodynamics (SPH) technique and for the cell wall; a Discrete Element Method (DEM) is used. The developed model is two-dimensional, but accounts for three-dimensional physical properties of real plant cells. Drying phenomena is simulated as fluid mass reductions and the model is used to predict the above mentioned structural properties as a function of cell fluid mass. Model predictions are found to be in fairly good agreement with experimental data in literature and the particle-based approach is demonstrated to be suitable for numerical studies of drying related structural deformations. Also a sensitivity analysis is included to demonstrate the influence of key model parameters to model predictions.

Structural characterisation of kaolinite : NaCl intercalate and its derivatives

Kaolinite:NaCl intercalates with basal layer dimensions of 0.95 and 1.25 nm have been prepared by direct reaction of saturated aqueous NaCl solution with well-crystallized source clay KGa-1. The intercalates and their thermal decomposition products have been studied by XRD, solid-state 23Na, 27Al, and 29Si MAS NMR, and FTIR. Intercalate yield is enhanced by dry grinding of kaolinite with NaCl prior to intercalation. The layered structure survives dehydroxylation of the kaolinite at 500°–600°C and persists to above 800°C with a resultant tetrahedral aluminosilicate framework. Excess NaCl can be readily removed by rinsing with water, producing an XRD ‘amorphous’ material. Upon heating at 900°C this material converts to a well-crystallized framework aluminosilicate closely related to low-camegieite, NaAlSiO4, some 350°C below its stability field. Reaction mechanisms are discussed and structural models proposed for each of these novel materials.

Protein monoubiquitination and polyubiquitination generate structural diversity to control distinct biological processes

Ubiquitination involves the attachment of ubiquitin (Ub) to lysine residues on substrate proteins or itself, which can result in protein monoubiquitination or polyubiquitination. Polyubiquitination through different lysines (seven) or the N-terminus of Ub can generate different protein-Ub structures. These include monoubiquitinated proteins, polyubiqutinated proteins with homotypic chains through a particular lysine on Ub or mixed polyubiquitin chains generated by polymerization through different Ub lysines. The ability of the ubiquitination pathway to generate different protein-Ub structures provides versatility of this pathway to target proteins to different fates. Protein ubiquitination is catalyzed by Ub-conjugating and Ub-ligase enzymes, with different combinations of these enzymes specifying the type of Ub modification on protein substrates. How Ub-conjugating and Ub-ligase enzymes generate this structural diversity is not clearly understood. In the current review, we discuss mechanisms utilized by the Ub-conjugating and Ub-ligase enzymes to generate structural diversity during protein ubiquitination, with a focus on recent mechanistic insights into protein monoubiquitination and polyubiquitination.

Contribution of DEAF1 structural domains to the interaction with the breast cancer oncogene LMO4

The proteins LMO4 and DEAF1 contribute to the proliferation of mammary epithelial cells. During breast cancer LMO4 is upregulated, affecting its interaction with other protein partners. This may set cells on a path to tumour formation. LMO4 and DEAF1 interact, but it is unknown how they cooperate to regulate cell proliferation. In this study, we identify a specific LMO4-binding domain in DEAF1. This domain contains an unstructured region that directly contacts LMO4, and a coiled coil that contains the DEAF1 nuclear export signal (NES). The coiled coil region can form tetramers and has the typical properties of a coiled coil domain. Using a simple cell-based assay, we show that LMO4 modulates the activity of the DEAF NES, causing nuclear accumulation of a construct containing the LMO4-interaction region of DEAF1.

Socio-structural influences on the work participation of refugees : an exploratory systematic mixed studies review

This systematic mixed studies review aimed at synthesizing evidence from studies related to the influences on the work participation of people with refugee status (PWRS). The review focused on the role of proximal socio-structural barriers on work participation by PWRS while foregrounding related distal, intermediate, proximal, and meta-systemic influences. For the systematic search of the literature, we focused on databases that addressed work, well-being, and social policy in refugee populations, including, Medline, CINAHL, PsycInfo, Web of Science, Scopus, and Sociological Abstracts. Of the studies reviewed, 16 of 39 met the inclusion criteria and were retained for the final analysis. We performed a narrative synthesis of the evidence on barriers to work participation by PWRS, interlinking clusters of barriers potent to their effects on work participation. Findings from the narrative synthesis suggest that proximal factors, those at point of entry to the labor market, influence work participation more directly than distal or intermediate factors. Distal and intermediate factors achieve their effects on work participation by PWRS primarily through meta-systemic interlinkages, including host-country documentation and refugee administration provisions.

Reliable and valid survey design for structural equation modelling

My quantitative study asks how Chinese Australians’ “Chineseness” and their various resources influence their Chinese language proficiency, using online survey and snowball sampling. ‘Operationalization’ is a challenging process which ensures that the survey design talks back to the informing theory and forwards to the analysis model. It requires the attention to two core methodological concerns, namely ‘validity’ and ‘reliability’. Construction of a high-quality questionnaire is critical to the achievement of valid and reliable operationalization. A series of strategies were chosen to ensure the quality of the questions, and thus the eventual data. These strategies enable the use of structural equation modelling to examine how well the data fits the theoretical framework, which was constructed in light of Bourdieu’s theory of habitus, capital and field.

Molecular and structural requirements of the ß1L-adrenoceptor

Noradrenaline which occurs naturally in the body binds to beta-adrenoceptors on the heart, causing the heart to beat faster and with greater force in response to increased demand. This enables the heart to provide oxygenated blood to vital organs. Prolonged overstimulation by noradrenaline can be harmful to the heart and lead to the progression of heart disease. In these circumstances beta-adrenoceptors are blocked with drugs called beta-blockers. Beta-blockers block the effects of noradrenaline by binding to the same site on the beta-adrenoceptor. Some beta-blockers such as CGP12177 can also cause increases in heart rate. Therefore it was proposed that CGP12177 could bind in a different place to noradrenaline. The aim of this study was to determine where CGP12177 binds to on the beta-adrenoceptor. The results have revealed a separate binding site named beta-1-low. These results may lead to the development of improved -blockers for the management of heart conditions.

Towards a typology of structural arrangements for shared services : evidence from the higher education sector

Shared services are increasingly prevalent in practice, their introduction potentially entailing substantive and highly consequential organizational redesign. Yet, attention to the structural arrangements of shared services has been limited. This study explores types of structural arrangements for shared services that are observed in practice, and the salient dimensions along which those types can be usefully differentiated. Through inductive attention to the shared services literature, and content analysis of 36 secondary case studies of shared services in the higher education sector, three salient dimensions emerged: (1) the existence or not of a separate organizational entity, (2) an intra- or inter-organizational sharing boundary, and (3) involvement or not of a third party. Each dimension being dichotomous yields 23 combinations, or eight shared services structural arrangement types. Each of the eight structural arrangement types is defined and demonstrated through case examples. The typology offers clarity around shared services structural arrangements. It can serve as a useful analytical tool for researchers investigating the phenomenon further, and for practitioners considering the introduction or further development of shared services arrangements. Important follow on research is suggested too.

Structural dissection of Ebola virus and its assembly determinants using cryo-electron tomography

Ebola virus is a highly pathogenic filovirus causing severe hemorrhagic fever with high mortality rates. It assembles heterogenous, filamentous, enveloped virus particles containing a negative-sense, single-stranded RNA genome packaged within a helical nucleocapsid (NC). We have used cryo-electron microscopy and tomography to visualize Ebola virus particles, as well as Ebola virus-like particles, in three dimensions in a near-native state. The NC within the virion forms a left-handed helix with an inner nucleoprotein layer decorated with protruding arms composed of VP24 and VP35. A comparison with the closely related Marburg virus shows that the N-terminal region of nucleoprotein defines the inner diameter of the Ebola virus NC, whereas the RNA genome defines its length. Binding of the nucleoprotein to RNA can assemble a loosely coiled NC-like structure; the loose coil can be condensed by binding of the viral matrix protein VP40 to the C terminus of the nucleoprotein, and rigidified by binding of VP24 and VP35 to alternate copies of the nucleoprotein. Four proteins (NP, VP24, VP35, and VP40) are necessary and sufficient to mediate assembly of an NC with structure, symmetry, variability, and flexibility indistinguishable from that in Ebola virus particles released from infected cells. Together these data provide a structural and architectural description of Ebola virus and define the roles of viral proteins in its structure and assembly

Extending and validating the is-impact model in Saudi Arabia : accounting for computer network quality

Organizations from every industry sector seek to enhance their business performance and competitiveness through the deployment of contemporary information systems (IS), such as Enterprise Systems (ERP). Investments in ERP are complex and costly, attracting scrutiny and pressure to justify their cost. Thus, IS researchers highlight the need for systematic evaluation of information system success, or impact, which has resulted in the introduction of varied models for evaluating information systems. One of these systematic measurement approaches is the IS-Impact Model introduced by a team of researchers at Queensland University of technology (QUT) (Gable, Sedera, & Chan, 2008). The IS-Impact Model is conceptualized as a formative, multidimensional index that consists of four dimensions. Gable et al. (2008) define IS-Impact as "a measure at a point in time, of the stream of net benefits from the IS, to date and anticipated, as perceived by all key-user-groups" (p.381). The IT Evaluation Research Program (ITE-Program) at QUT has grown the IS-Impact Research Track with the central goal of conducting further studies to enhance and extend the IS-Impact Model. The overall goal of the IS-Impact research track at QUT is "to develop the most widely employed model for benchmarking information systems in organizations for the joint benefit of both research and practice" (Gable, 2009). In order to achieve that, the IS-Impact research track advocates programmatic research having the principles of tenacity, holism, and generalizability through extension research strategies. This study was conducted within the IS-Impact Research Track, to further generalize the IS-Impact Model by extending it to the Saudi Arabian context. According to Hofsted (2012), the national culture of Saudi Arabia is significantly different from the Australian national culture making the Saudi Arabian culture an interesting context for testing the external validity of the IS-Impact Model. The study re-visits the IS-Impact Model from the ground up. Rather than assume the existing instrument is valid in the new context, or simply assess its validity through quantitative data collection, the study takes a qualitative, inductive approach to re-assessing the necessity and completeness of existing dimensions and measures. This is done in two phases: Exploratory Phase and Confirmatory Phase. The exploratory phase addresses the first research question of the study "Is the IS-Impact Model complete and able to capture the impact of information systems in Saudi Arabian Organization?". The content analysis, used to analyze the Identification Survey data, indicated that 2 of the 37 measures of the IS-Impact Model are not applicable for the Saudi Arabian Context. Moreover, no new measures or dimensions were identified, evidencing the completeness and content validity of the IS-Impact Model. In addition, the Identification Survey data suggested several concepts related to IS-Impact, the most prominent of which was "Computer Network Quality" (CNQ). The literature supported the existence of a theoretical link between IS-Impact and CNQ (CNQ is viewed as an antecedent of IS-Impact). With the primary goal of validating the IS-Impact model within its extended nomological network, CNQ was introduced to the research model. The Confirmatory Phase addresses the second research question of the study "Is the Extended IS-Impact Model Valid as a Hierarchical Multidimensional Formative Measurement Model?". The objective of the Confirmatory Phase was to test the validity of IS-Impact Model and CNQ Model. To achieve that, IS-Impact, CNQ, and IS-Satisfaction were operationalized in a survey instrument, and then the research model was assessed by employing the Partial Least Squares (PLS) approach. The CNQ model was validated as a formative model. Similarly, the IS-Impact Model was validated as a hierarchical multidimensional formative construct. However, the analysis indicated that one of the IS-Impact Model indicators was insignificant and can be removed from the model. Thus, the resulting Extended IS-Impact Model consists of 4 dimensions and 34 measures. Finally, the structural model was also assessed against two aspects: explanatory and predictive power. The analysis revealed that the path coefficient between CNQ and IS-Impact is significant with t-value= (4.826) and relatively strong with â = (0.426) with CNQ explaining 18% of the variance in IS-Impact. These results supported the hypothesis that CNQ is antecedent of IS-Impact. The study demonstrates that the quality of Computer Network affects the quality of the Enterprise System (ERP) and consequently the impacts of the system. Therefore, practitioners should pay attention to the Computer Network quality. Similarly, the path coefficient between IS-Impact and IS-Satisfaction was significant t-value = (17.79) and strong â = (0.744), with IS-Impact alone explaining 55% of the variance in Satisfaction, consistent with results of the original IS-Impact study (Gable et al., 2008). The research contributions include: (a) supporting the completeness and validity of IS-Impact Model as a Hierarchical Multi-dimensional Formative Measurement Model in the Saudi Arabian context, (b) operationalizing Computer Network Quality as conceptualized in the ITU-T Recommendation E.800 (ITU-T, 1993), (c) validating CNQ as a formative measurement model and as an antecedent of IS Impact, and (d) conceptualizing and validating IS-Satisfaction as a reflective measurement model and as an immediate consequence of IS Impact. The CNQ model provides a framework to perceptually measure Computer Network Quality from multiple perspectives. The CNQ model features an easy-to-understand, easy-to-use, and economical survey instrument.

In pursuit of tourism destination brand loyalty : visitor relationship management orientation of destination marketing organisations

The proposition underpinning this study is engaging in meaningful dialogue with previous visitors represents an efficient and effective use of resources for a destination marketing organization (DMO), compared to above the line advertising in broadcast media. However there has been a lack of attention in the tourism literature relating to destination switching, loyalty and customer relationship management (CRM) to test such a proposition. This paper reports an investigation of visitor relationship marketing (VRM) orientation among DMOs. A model of CRM orientation, which was developed from the wider marketing literature and a prior qualitative study, was used to develop a scale to operationalise DMO visitor relationship orientation. Due to a small sample, the Partial Least Squares (PLS) method of structural equation modelling was used to analyse the data. Although the sample limits the ability to generalise, the results indicated the DMOs’ visitor orientation is generally responsive and reactive rather than proactive.